TY - CHAP AB - The analysis of dynamic cellular processes such as plant cytokinesis stands and falls with live-cell time-lapse confocal imaging. Conventional approaches to time-lapse imaging of cell division in Arabidopsis root tips are tedious and have low throughput. Here, we describe a protocol for long-term time-lapse simultaneous imaging of multiple root tips on a vertical-stage confocal microscope with automated root tracking. We also provide modifications of the basic protocol to implement this imaging method in the analysis of genetic, pharmacological or laser ablation wounding-mediated experimental manipulations. Our method dramatically improves the efficiency of cell division time-lapse imaging by increasing the throughput, while reducing the person-hour requirements of such experiments. AU - Hörmayer, Lukas AU - Friml, Jiří AU - Glanc, Matous ID - 10268 SN - 1064-3745 T2 - Plant Cell Division TI - Automated time-lapse imaging and manipulation of cell divisions in Arabidopsis roots by vertical-stage confocal microscopy VL - 2382 ER - TY - JOUR AB - Cell and tissue polarization is fundamental for plant growth and morphogenesis. The polar, cellular localization of Arabidopsis PIN‐FORMED (PIN) proteins is crucial for their function in directional auxin transport. The clustering of PIN polar cargoes within the plasma membrane has been proposed to be important for the maintenance of their polar distribution. However, the more detailed features of PIN clusters and the cellular requirements of cargo clustering remain unclear. Here, we characterized PIN clusters in detail by means of multiple advanced microscopy and quantification methods, such as 3D quantitative imaging or freeze‐fracture replica labeling. The size and aggregation types of PIN clusters were determined by electron microscopy at the nanometer level at different polar domains and at different developmental stages, revealing a strong preference for clustering at the polar domains. Pharmacological and genetic studies revealed that PIN clusters depend on phosphoinositol pathways, cytoskeletal structures and specific cell‐wall components as well as connections between the cell wall and the plasma membrane. This study identifies the role of different cellular processes and structures in polar cargo clustering and provides initial mechanistic insight into the maintenance of polarity in plants and other systems. AU - Li, Hongjiang AU - von Wangenheim, Daniel AU - Zhang, Xixi AU - Tan, Shutang AU - Darwish-Miranda, Nasser AU - Naramoto, Satoshi AU - Wabnik, Krzysztof T AU - de Rycke, Riet AU - Kaufmann, Walter AU - Gütl, Daniel J AU - Tejos, Ricardo AU - Grones, Peter AU - Ke, Meiyu AU - Chen, Xu AU - Dettmer, Jan AU - Friml, Jiří ID - 8582 IS - 1 JF - New Phytologist SN - 0028646X TI - Cellular requirements for PIN polar cargo clustering in Arabidopsis thaliana VL - 229 ER - TY - JOUR AB - The leaf is a crucial organ evolved with remarkable morphological diversity to maximize plant photosynthesis. The leaf shape is a key trait that affects photosynthesis, flowering rates, disease resistance, and yield. Although many genes regulating leaf development have been identified in the past years, the precise regulatory architecture underlying the generation of diverse leaf shapes remains to be elucidated. We used cotton as a reference model to probe the genetic framework underlying divergent leaf forms. Comparative transcriptome analysis revealed that the GhARF16‐1 and GhKNOX2‐1 genes might be potential regulators of leaf shape. We functionally characterized the auxin‐responsive factor ARF16‐1 acting upstream of GhKNOX2‐1 to determine leaf morphology in cotton. The transcription of GhARF16‐1 was significantly higher in lobed‐leaved cotton than in smooth‐leaved cotton. Furthermore, the overexpression of GhARF16‐1 led to the upregulation of GhKNOX2‐1 and resulted in more and deeper serrations in cotton leaves, similar to the leaf shape of cotton plants overexpressing GhKNOX2‐1. We found that GhARF16‐1 specifically bound to the promoter of GhKNOX2‐1 to induce its expression. The heterologous expression of GhARF16‐1 and GhKNOX2‐1 in Arabidopsis led to lobed and curly leaves, and a genetic analysis revealed that GhKNOX2‐1 is epistatic to GhARF16‐1 in Arabidopsis, suggesting that the GhARF16‐1 and GhKNOX2‐1 interaction paradigm also functions to regulate leaf shape in Arabidopsis. To our knowledge, our results uncover a novel mechanism by which auxin, through the key component ARF16‐1 and its downstream‐activated gene KNOX2‐1, determines leaf morphology in eudicots. AU - He, P AU - Zhang, Yuzhou AU - Li, H AU - Fu, X AU - Shang, H AU - Zou, C AU - Friml, Jiří AU - Xiao, G ID - 8606 IS - 3 JF - Plant Biotechnology Journal SN - 1467-7644 TI - GhARF16-1 modulates leaf development by transcriptionally regulating the GhKNOX2-1 gene in cotton VL - 19 ER - TY - JOUR AB - The phytohormone auxin plays a central role in shaping plant growth and development. With decades of genetic and biochemical studies, numerous core molecular components and their networks, underlying auxin biosynthesis, transport, and signaling, have been identified. Notably, protein phosphorylation, catalyzed by kinases and oppositely hydrolyzed by phosphatases, has been emerging to be a crucial type of post-translational modification, regulating physiological and developmental auxin output at all levels. In this review, we comprehensively discuss earlier and recent advances in our understanding of genetics, biochemistry, and cell biology of the kinases and phosphatases participating in auxin action. We provide insights into the mechanisms by which reversible protein phosphorylation defines developmental auxin responses, discuss current challenges, and provide our perspectives on future directions involving the integration of the control of protein phosphorylation into the molecular auxin network. AU - Tan, Shutang AU - Luschnig, Christian AU - Friml, Jiří ID - 8992 IS - 1 JF - Molecular Plant SN - 16742052 TI - Pho-view of auxin: Reversible protein phosphorylation in auxin biosynthesis, transport and signaling VL - 14 ER - TY - JOUR AB - N-1-naphthylphthalamic acid (NPA) is a key inhibitor of directional (polar) transport of the hormone auxin in plants. For decades, it has been a pivotal tool in elucidating the unique polar auxin transport-based processes underlying plant growth and development. Its exact mode of action has long been sought after and is still being debated, with prevailing mechanistic schemes describing only indirect connections between NPA and the main transporters responsible for directional transport, namely PIN auxin exporters. Here we present data supporting a model in which NPA associates with PINs in a more direct manner than hitherto postulated. We show that NPA inhibits PIN activity in a heterologous oocyte system and that expression of NPA-sensitive PINs in plant, yeast, and oocyte membranes leads to specific saturable NPA binding. We thus propose that PINs are a bona fide NPA target. This offers a straightforward molecular basis for NPA inhibition of PIN-dependent auxin transport and a logical parsimonious explanation for the known physiological effects of NPA on plant growth, as well as an alternative hypothesis to interpret past and future results. We also introduce PIN dimerization and describe an effect of NPA on this, suggesting that NPA binding could be exploited to gain insights into structural aspects of PINs related to their transport mechanism. AU - Abas, Lindy AU - Kolb, Martina AU - Stadlmann, Johannes AU - Janacek, Dorina P. AU - Lukic, Kristina AU - Schwechheimer, Claus AU - Sazanov, Leonid A AU - Mach, Lukas AU - Friml, Jiří AU - Hammes, Ulrich Z. ID - 8993 IS - 1 JF - PNAS SN - 00278424 TI - Naphthylphthalamic acid associates with and inhibits PIN auxin transporters VL - 118 ER - TY - JOUR AB - Auxin is a key regulator of plant growth and development. Local auxin biosynthesis and intercellular transport generates regional gradients in the root that are instructive for processes such as specification of developmental zones that maintain root growth and tropic responses. Here we present a toolbox to study auxin-mediated root development that features: (i) the ability to control auxin synthesis with high spatio-temporal resolution and (ii) single-cell nucleus tracking and morphokinetic analysis infrastructure. Integration of these two features enables cutting-edge analysis of root development at single-cell resolution based on morphokinetic parameters under normal growth conditions and during cell-type-specific induction of auxin biosynthesis. We show directional auxin flow in the root and refine the contributions of key players in this process. In addition, we determine the quantitative kinetics of Arabidopsis root meristem skewing, which depends on local auxin gradients but does not require PIN2 and AUX1 auxin transporter activities. Beyond the mechanistic insights into root development, the tools developed here will enable biologists to study kinetics and morphology of various critical processes at the single cell-level in whole organisms. AU - Hu, Yangjie AU - Omary, Moutasem AU - Hu, Yun AU - Doron, Ohad AU - Hörmayer, Lukas AU - Chen, Qingguo AU - Megides, Or AU - Chekli, Ori AU - Ding, Zhaojun AU - Friml, Jiří AU - Zhao, Yunde AU - Tsarfaty, Ilan AU - Shani, Eilon ID - 9254 JF - Nature Communications TI - Cell kinetics of auxin transport and activity in Arabidopsis root growth and skewing VL - 12 ER - TY - JOUR AB - Endoplasmic reticulum–plasma membrane contact sites (ER–PM CS) play fundamental roles in all eukaryotic cells. Arabidopsis thaliana mutants lacking the ER–PM protein tether synaptotagmin1 (SYT1) exhibit decreased PM integrity under multiple abiotic stresses, such as freezing, high salt, osmotic stress, and mechanical damage. Here, we show that, together with SYT1, the stress-induced SYT3 is an ER–PM tether that also functions in maintaining PM integrity. The ER–PM CS localization of SYT1 and SYT3 is dependent on PM phosphatidylinositol-4-phosphate and is regulated by abiotic stress. Lipidomic analysis revealed that cold stress increased the accumulation of diacylglycerol at the PM in a syt1/3 double mutant relative to wild-type while the levels of most glycerolipid species remain unchanged. In addition, the SYT1-green fluorescent protein fusion preferentially binds diacylglycerol in vivo with little affinity for polar glycerolipids. Our work uncovers a SYT-dependent mechanism of stress adaptation counteracting the detrimental accumulation of diacylglycerol at the PM produced during episodes of abiotic stress. AU - Ruiz-Lopez, N AU - Pérez-Sancho, J AU - Esteban Del Valle, A AU - Haslam, RP AU - Vanneste, S AU - Catalá, R AU - Perea-Resa, C AU - Van Damme, D AU - García-Hernández, S AU - Albert, A AU - Vallarino, J AU - Lin, J AU - Friml, Jiří AU - Macho, AP AU - Salinas, J AU - Rosado, A AU - Napier, JA AU - Amorim-Silva, V AU - Botella, MA ID - 9443 IS - 7 JF - Plant Cell SN - 1040-4651 TI - Synaptotagmins at the endoplasmic reticulum-plasma membrane contact sites maintain diacylglycerol homeostasis during abiotic stress VL - 33 ER - TY - JOUR AB - To overcome nitrogen deficiency, legume roots establish symbiotic interactions with nitrogen-fixing rhizobia that is fostered in specialized organs (nodules). Similar to other organs, nodule formation is determined by a local maximum of the phytohormone auxin at the primordium site. However, how auxin regulates nodule development remains poorly understood. Here, we found that in soybean, (Glycine max), dynamic auxin transport driven by PIN-FORMED (PIN) transporter GmPIN1 is involved in nodule primordium formation. GmPIN1 was specifically expressed in nodule primordium cells and GmPIN1 was polarly localized in these cells. Two nodulation regulators, (iso)flavonoids trigger expanded distribution of GmPIN1b to root cortical cells, and cytokinin rearranges GmPIN1b polarity. Gmpin1abc triple mutants generated with CRISPR-Cas9 showed impaired establishment of auxin maxima in nodule meristems and aberrant divisions in the nodule primordium cells. Moreover, overexpression of GmPIN1 suppressed nodule primordium initiation. GmPIN9d, an ortholog of Arabidopsis thaliana PIN2, acts together with GmPIN1 later in nodule development to acropetally transport auxin in vascular bundles, fine-tuning the auxin supply for nodule enlargement. Our findings reveal how PIN-dependent auxin transport modulates different aspects of soybean nodule development and suggest that establishment of auxin gradient is a prerequisite for the proper interaction between legumes and rhizobia. AU - Gao, Z AU - Chen, Z AU - Cui, Y AU - Ke, M AU - Xu, H AU - Xu, Q AU - Chen, J AU - Li, Y AU - Huang, L AU - Zhao, H AU - Huang, D AU - Mai, S AU - Xu, T AU - Liu, X AU - Li, S AU - Guan, Y AU - Yang, W AU - Friml, Jiří AU - Petrášek, J AU - Zhang, J AU - Chen, X ID - 9657 IS - 9 JF - Plant Cell SN - 1040-4651 TI - GmPIN-dependent polar auxin transport is involved in soybean nodule development VL - 33 ER - TY - JOUR AB - Tropisms, growth responses to environmental stimuli such as light or gravity, are spectacular examples of adaptive plant development. The plant hormone auxin serves as a major coordinative signal. The PIN auxin exporters, through their dynamic polar subcellular localizations, redirect auxin fluxes in response to environmental stimuli and the resulting auxin gradients across organs underly differential cell elongation and bending. In this review, we discuss recent advances concerning regulations of PIN polarity during tropisms, focusing on PIN phosphorylation and trafficking. We also cover how environmental cues regulate PIN actions during tropisms, and a crucial role of auxin feedback on PIN polarity during bending termination. Finally, the interactions between different tropisms are reviewed to understand plant adaptive growth in the natural environment. AU - Han, Huibin AU - Adamowski, Maciek AU - Qi, Linlin AU - Alotaibi, SS AU - Friml, Jiří ID - 9656 IS - 2 JF - New Phytologist SN - 0028-646x TI - PIN-mediated polar auxin transport regulations in plant tropic responses VL - 232 ER - TY - JOUR AB - Roots are composed of different root types and, in the dicotyledonous Arabidopsis, typically consist of a primary root that branches into lateral roots. Adventitious roots emerge from non-root tissue and are formed upon wounding or other types of abiotic stress. Here, we investigated adventitious root (AR) formation in Arabidopsis hypocotyls under conditions of altered abscisic acid (ABA) signaling. Exogenously applied ABA suppressed AR formation at 0.25 µM or higher doses. AR formation was less sensitive to the synthetic ABA analog pyrabactin (PB). However, PB was a more potent inhibitor at concentrations above 1 µM, suggesting that it was more selective in triggering a root inhibition response. Analysis of a series of phosphonamide and phosphonate pyrabactin analogs suggested that adventitious root formation and lateral root branching are differentially regulated by ABA signaling. ABA biosynthesis and signaling mutants affirmed a general inhibitory role of ABA and point to PYL1 and PYL2 as candidate ABA receptors that regulate AR inhibition. AU - Zeng, Yinwei AU - Verstraeten, Inge AU - Trinh, Hoang Khai AU - Heugebaert, Thomas AU - Stevens, Christian V. AU - Garcia-Maquilon, Irene AU - Rodriguez, Pedro L. AU - Vanneste, Steffen AU - Geelen, Danny ID - 9909 IS - 8 JF - Genes TI - Arabidopsis hypocotyl adventitious root formation is suppressed by ABA signaling VL - 12 ER - TY - JOUR AB - Advanced transcriptome sequencing has revealed that the majority of eukaryotic genes undergo alternative splicing (AS). Nonetheless, little effort has been dedicated to investigating the functional relevance of particular splicing events, even those in the key developmental and hormonal regulators. Combining approaches of genetics, biochemistry and advanced confocal microscopy, we describe the impact of alternative splicing on the PIN7 gene in the model plant Arabidopsis thaliana. PIN7 encodes a polarly localized transporter for the phytohormone auxin and produces two evolutionarily conserved transcripts, PIN7a and PIN7b. PIN7a and PIN7b, differing in a four amino acid stretch, exhibit almost identical expression patterns and subcellular localization. We reveal that they are closely associated and mutually influence each other's mobility within the plasma membrane. Phenotypic complementation tests indicate that the functional contribution of PIN7b per se is minor, but it markedly reduces the prominent PIN7a activity, which is required for correct seedling apical hook formation and auxin-mediated tropic responses. Our results establish alternative splicing of the PIN family as a conserved, functionally relevant mechanism, revealing an additional regulatory level of auxin-mediated plant development. AU - Kashkan, Ivan AU - Hrtyan, Mónika AU - Retzer, Katarzyna AU - Humpolíčková, Jana AU - Jayasree, Aswathy AU - Filepová, Roberta AU - Vondráková, Zuzana AU - Simon, Sibu AU - Rombaut, Debbie AU - Jacobs, Thomas B. AU - Frilander, Mikko J. AU - Hejátko, Jan AU - Friml, Jiří AU - Petrášek, Jan AU - Růžička, Kamil ID - 10282 JF - New Phytologist SN - 0028-646X TI - Mutually opposing activity of PIN7 splicing isoforms is required for auxin-mediated tropic responses in Arabidopsis thaliana VL - 233 ER - TY - JOUR AB - Strigolactones (SLs) are carotenoid-derived plant hormones that control shoot branching and communications between host plants and symbiotic fungi or root parasitic plants. Extensive studies have identified the key components participating in SL biosynthesis and signalling, whereas the catabolism or deactivation of endogenous SLs in planta remains largely unknown. Here, we report that the Arabidopsis carboxylesterase 15 (AtCXE15) and its orthologues function as efficient hydrolases of SLs. We show that overexpression of AtCXE15 promotes shoot branching by dampening SL-inhibited axillary bud outgrowth. We further demonstrate that AtCXE15 could bind and efficiently hydrolyse SLs both in vitro and in planta. We also provide evidence that AtCXE15 is capable of catalysing hydrolysis of diverse SL analogues and that such CXE15-dependent catabolism of SLs is evolutionarily conserved in seed plants. These results disclose a catalytic mechanism underlying homoeostatic regulation of SLs in plants, which also provides a rational approach to spatial-temporally manipulate the endogenous SLs and thus architecture of crops and ornamental plants. AU - Xu, Enjun AU - Chai, Liang AU - Zhang, Shiqi AU - Yu, Ruixue AU - Zhang, Xixi AU - Xu, Chongyi AU - Hu, Yuxin ID - 10326 JF - Nature Plants TI - Catabolism of strigolactones by a carboxylesterase VL - 7 ER - TY - JOUR AB - The quality control system for messenger RNA (mRNA) is fundamental for cellular activities in eukaryotes. To elucidate the molecular mechanism of 3'-Phosphoinositide-Dependent Protein Kinase1 (PDK1), a master regulator that is essential throughout eukaryotic growth and development, we employed a forward genetic approach to screen for suppressors of the loss-of-function T-DNA insertion double mutant pdk1.1 pdk1.2 in Arabidopsis thaliana. Notably, the severe growth attenuation of pdk1.1 pdk1.2 was rescued by sop21 (suppressor of pdk1.1 pdk1.2), which harbours a loss-of-function mutation in PELOTA1 (PEL1). PEL1 is a homologue of mammalian PELOTA and yeast (Saccharomyces cerevisiae) DOM34p, which each form a heterodimeric complex with the GTPase HBS1 (HSP70 SUBFAMILY B SUPPRESSOR1, also called SUPERKILLER PROTEIN7, SKI7), a protein that is responsible for ribosomal rescue and thereby assures the quality and fidelity of mRNA molecules during translation. Genetic analysis further revealed that a dysfunctional PEL1-HBS1 complex failed to degrade the T-DNA-disrupted PDK1 transcripts, which were truncated but functional, and thus rescued the growth and developmental defects of pdk1.1 pdk1.2. Our studies demonstrated the functionality of a homologous PELOTA-HBS1 complex and identified its essential regulatory role in plants, providing insights into the mechanism of mRNA quality control. AU - Kong, W AU - Tan, Shutang AU - Zhao, Q AU - Lin, DL AU - Xu, ZH AU - Friml, Jiří AU - Xue, HW ID - 9368 IS - 4 JF - Plant Physiology SN - 0032-0889 TI - mRNA surveillance complex PELOTA-HBS1 eegulates phosphoinositide-sependent protein kinase1 and plant growth VL - 186 ER - TY - JOUR AB - Polar subcellular localization of the PIN exporters of the phytohormone auxin is a key determinant of directional, intercellular auxin transport and thus a central topic of both plant cell and developmental biology. Arabidopsis mutants lacking PID, a kinase that phosphorylates PINs, or the MAB4/MEL proteins of unknown molecular function display PIN polarity defects and phenocopy pin mutants, but mechanistic insights into how these factors convey PIN polarity are missing. Here, by combining protein biochemistry with quantitative live-cell imaging, we demonstrate that PINs, MAB4/MELs, and AGC kinases interact in the same complex at the plasma membrane. MAB4/MELs are recruited to the plasma membrane by the PINs and in concert with the AGC kinases maintain PIN polarity through limiting lateral diffusion-based escape of PINs from the polar domain. The PIN-MAB4/MEL-PID protein complex has self-reinforcing properties thanks to positive feedback between AGC kinase-mediated PIN phosphorylation and MAB4/MEL recruitment. We thus uncover the molecular mechanism by which AGC kinases and MAB4/MEL proteins regulate PIN localization and plant development. AU - Glanc, Matous AU - Van Gelderen, K AU - Hörmayer, Lukas AU - Tan, Shutang AU - Naramoto, S AU - Zhang, Xixi AU - Domjan, David AU - Vcelarova, L AU - Hauschild, Robert AU - Johnson, Alexander J AU - de Koning, E AU - van Dop, M AU - Rademacher, E AU - Janson, S AU - Wei, X AU - Molnar, Gergely AU - Fendrych, Matyas AU - De Rybel, B AU - Offringa, R AU - Friml, Jiří ID - 9290 IS - 9 JF - Current Biology SN - 0960-9822 TI - AGC kinases and MAB4/MEL proteins maintain PIN polarity by limiting lateral diffusion in plant cells VL - 31 ER - TY - JOUR AB - Plants are able to orient their growth according to gravity, which ultimately controls both shoot and root architecture.1 Gravitropism is a dynamic process whereby gravistimulation induces the asymmetric distribution of the plant hormone auxin, leading to asymmetric growth, organ bending, and subsequent reset of auxin distribution back to the original pre-gravistimulation situation.1, 2, 3 Differential auxin accumulation during the gravitropic response depends on the activity of polarly localized PIN-FORMED (PIN) auxin-efflux carriers.1, 2, 3, 4 In particular, the timing of this dynamic response is regulated by PIN2,5,6 but the underlying molecular mechanisms are poorly understood. Here, we show that MEMBRANE ASSOCIATED KINASE REGULATOR2 (MAKR2) controls the pace of the root gravitropic response. We found that MAKR2 is required for the PIN2 asymmetry during gravitropism by acting as a negative regulator of the cell-surface signaling mediated by the receptor-like kinase TRANSMEMBRANE KINASE1 (TMK1).2,7, 8, 9, 10 Furthermore, we show that the MAKR2 inhibitory effect on TMK1 signaling is antagonized by auxin itself, which triggers rapid MAKR2 membrane dissociation in a TMK1-dependent manner. Our findings suggest that the timing of the root gravitropic response is orchestrated by the reversible inhibition of the TMK1 signaling pathway at the cell surface. AU - Marquès-Bueno, MM AU - Armengot, L AU - Noack, LC AU - Bareille, J AU - Rodriguez Solovey, Lesia AU - Platre, MP AU - Bayle, V AU - Liu, M AU - Opdenacker, D AU - Vanneste, S AU - Möller, BK AU - Nimchuk, ZL AU - Beeckman, T AU - Caño-Delgado, AI AU - Friml, Jiří AU - Jaillais, Y ID - 8824 IS - 1 JF - Current Biology SN - 0960-9822 TI - Auxin-regulated reversible inhibition of TMK1 signaling by MAKR2 modulates the dynamics of root gravitropism VL - 31 ER - TY - JOUR AB - • The phenylpropanoid pathway serves a central role in plant metabolism, providing numerous compounds involved in diverse physiological processes. Most carbon entering the pathway is incorporated into lignin. Although several phenylpropanoid pathway mutants show seedling growth arrest, the role for lignin in seedling growth and development is unexplored. • We use complementary pharmacological and genetic approaches to block CINNAMATE‐4‐HYDROXYLASE (C4H) functionality in Arabidopsis seedlings and a set of molecular and biochemical techniques to investigate the underlying phenotypes. • Blocking C4H resulted in reduced lateral rooting and increased adventitious rooting apically in the hypocotyl. These phenotypes coincided with an inhibition in auxin transport. The upstream accumulation in cis‐cinnamic acid was found to likely cause polar auxin transport inhibition. Conversely, a downstream depletion in lignin perturbed phloem‐mediated auxin transport. Restoring lignin deposition effectively reestablished phloem transport and, accordingly, auxin homeostasis. • Our results show that the accumulation of bioactive intermediates and depletion in lignin jointly cause the aberrant phenotypes upon blocking C4H, and demonstrate that proper deposition of lignin is essential for the establishment of auxin distribution in seedlings. Our data position the phenylpropanoid pathway and lignin in a new physiological framework, consolidating their importance in plant growth and development. AU - El Houari, I AU - Van Beirs, C AU - Arents, HE AU - Han, Huibin AU - Chanoca, A AU - Opdenacker, D AU - Pollier, J AU - Storme, V AU - Steenackers, W AU - Quareshy, M AU - Napier, R AU - Beeckman, T AU - Friml, Jiří AU - De Rybel, B AU - Boerjan, W AU - Vanholme, B ID - 9288 IS - 6 JF - New Phytologist SN - 0028-646x TI - Seedling developmental defects upon blocking CINNAMATE-4-HYDROXYLASE are caused by perturbations in auxin transport VL - 230 ER - TY - JOUR AB - To adapt to the diverse array of biotic and abiotic cues, plants have evolved sophisticated mechanisms to sense changes in environmental conditions and modulate their growth. Growth-promoting hormones and defence signalling fine tune plant development antagonistically. During host-pathogen interactions, this defence-growth trade-off is mediated by the counteractive effects of the defence hormone salicylic acid (SA) and the growth hormone auxin. Here we revealed an underlying mechanism of SA regulating auxin signalling by constraining the plasma membrane dynamics of PIN2 auxin efflux transporter in Arabidopsis thaliana roots. The lateral diffusion of PIN2 proteins is constrained by SA signalling, during which PIN2 proteins are condensed into hyperclusters depending on REM1.2-mediated nanodomain compartmentalisation. Furthermore, membrane nanodomain compartmentalisation by SA or Remorin (REM) assembly significantly suppressed clathrin-mediated endocytosis. Consequently, SA-induced heterogeneous surface condensation disrupted asymmetric auxin distribution and the resultant gravitropic response. Our results demonstrated a defence-growth trade-off mechanism by which SA signalling crosstalked with auxin transport by concentrating membrane-resident PIN2 into heterogeneous compartments. AU - Ke, M AU - Ma, Z AU - Wang, D AU - Sun, Y AU - Wen, C AU - Huang, D AU - Chen, Z AU - Yang, L AU - Tan, Shutang AU - Li, R AU - Friml, Jiří AU - Miao, Y AU - Chen, X ID - 8608 IS - 2 JF - New Phytologist SN - 0028-646x TI - Salicylic acid regulates PIN2 auxin transporter hyper-clustering and root gravitropic growth via Remorin-dependent lipid nanodomain organization in Arabidopsis thaliana VL - 229 ER - TY - JOUR AB - In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field. AU - Klionsky, Daniel J. AU - Abdel-Aziz, Amal Kamal AU - Abdelfatah, Sara AU - Abdellatif, Mahmoud AU - Abdoli, Asghar AU - Abel, Steffen AU - Abeliovich, Hagai AU - Abildgaard, Marie H. AU - Abudu, Yakubu Princely AU - Acevedo-Arozena, Abraham AU - Adamopoulos, Iannis E. AU - Adeli, Khosrow AU - Adolph, Timon E. AU - Adornetto, Annagrazia AU - Aflaki, Elma AU - Agam, Galila AU - Agarwal, Anupam AU - Aggarwal, Bharat B. AU - Agnello, Maria AU - Agostinis, Patrizia AU - Agrewala, Javed N. AU - Agrotis, Alexander AU - Aguilar, Patricia V. AU - Ahmad, S. Tariq AU - Ahmed, Zubair M. AU - Ahumada-Castro, Ulises AU - Aits, Sonja AU - Aizawa, Shu AU - Akkoc, Yunus AU - Akoumianaki, Tonia AU - Akpinar, Hafize Aysin AU - Al-Abd, Ahmed M. AU - Al-Akra, Lina AU - Al-Gharaibeh, Abeer AU - Alaoui-Jamali, Moulay A. AU - Alberti, Simon AU - Alcocer-Gómez, Elísabet AU - Alessandri, Cristiano AU - Ali, Muhammad AU - Alim Al-Bari, M. Abdul AU - Aliwaini, Saeb AU - Alizadeh, Javad AU - Almacellas, Eugènia AU - Almasan, Alexandru AU - Alonso, Alicia AU - Alonso, Guillermo D. AU - Altan-Bonnet, Nihal AU - Altieri, Dario C. AU - Álvarez, Élida M.C. AU - Alves, Sara AU - Alves Da Costa, Cristine AU - Alzaharna, Mazen M. AU - Amadio, Marialaura AU - Amantini, Consuelo AU - Amaral, Cristina AU - Ambrosio, Susanna AU - Amer, Amal O. AU - Ammanathan, Veena AU - An, Zhenyi AU - Andersen, Stig U. AU - Andrabi, Shaida A. AU - Andrade-Silva, Magaiver AU - Andres, Allen M. AU - Angelini, Sabrina AU - Ann, David AU - Anozie, Uche C. AU - Ansari, Mohammad Y. AU - Antas, Pedro AU - Antebi, Adam AU - Antón, Zuriñe AU - Anwar, Tahira AU - Apetoh, Lionel AU - Apostolova, Nadezda AU - Araki, Toshiyuki AU - Araki, Yasuhiro AU - Arasaki, Kohei AU - Araújo, Wagner L. AU - Araya, Jun AU - Arden, Catherine AU - Arévalo, Maria Angeles AU - Arguelles, Sandro AU - Arias, Esperanza AU - Arikkath, Jyothi AU - Arimoto, Hirokazu AU - Ariosa, Aileen R. AU - Armstrong-James, Darius AU - Arnauné-Pelloquin, Laetitia AU - Aroca, Angeles AU - Arroyo, Daniela S. AU - Arsov, Ivica AU - Artero, Rubén AU - Asaro, Dalia Maria Lucia AU - Aschner, Michael AU - Ashrafizadeh, Milad AU - Ashur-Fabian, Osnat AU - Atanasov, Atanas G. AU - Au, Alicia K. AU - Auberger, Patrick AU - Auner, Holger W. AU - Aurelian, Laure AU - Autelli, Riccardo AU - Avagliano, Laura AU - Ávalos, Yenniffer AU - Aveic, Sanja AU - Aveleira, Célia Alexandra AU - Avin-Wittenberg, Tamar AU - Aydin, Yucel AU - Ayton, Scott AU - Ayyadevara, Srinivas AU - Azzopardi, Maria AU - Baba, Misuzu AU - Backer, Jonathan M. AU - Backues, Steven K. AU - Bae, Dong Hun AU - Bae, Ok Nam AU - Bae, Soo Han AU - Baehrecke, Eric H. AU - Baek, Ahruem AU - Baek, Seung Hoon AU - Baek, Sung Hee AU - Bagetta, Giacinto AU - Bagniewska-Zadworna, Agnieszka AU - Bai, Hua AU - Bai, Jie AU - Bai, Xiyuan AU - Bai, Yidong AU - Bairagi, Nandadulal AU - Baksi, Shounak AU - Balbi, Teresa AU - Baldari, Cosima T. AU - Balduini, Walter AU - Ballabio, Andrea AU - Ballester, Maria AU - Balazadeh, Salma AU - Balzan, Rena AU - Bandopadhyay, Rina AU - Banerjee, Sreeparna AU - Banerjee, Sulagna AU - Bánréti, Ágnes AU - Bao, Yan AU - Baptista, Mauricio S. AU - Baracca, Alessandra AU - Barbati, Cristiana AU - Bargiela, Ariadna AU - Barilà, Daniela AU - Barlow, Peter G. AU - Barmada, Sami J. AU - Barreiro, Esther AU - Barreto, George E. AU - Bartek, Jiri AU - Bartel, Bonnie AU - Bartolome, Alberto AU - Barve, Gaurav R. AU - Basagoudanavar, Suresh H. AU - Bassham, Diane C. AU - Bast, Robert C. AU - Basu, Alakananda AU - Batoko, Henri AU - Batten, Isabella AU - Baulieu, Etienne E. AU - Baumgarner, Bradley L. AU - Bayry, Jagadeesh AU - Beale, Rupert AU - Beau, Isabelle AU - Beaumatin, Florian AU - Bechara, Luiz R.G. AU - Beck, George R. AU - Beers, Michael F. AU - Begun, Jakob AU - Behrends, Christian AU - Behrens, Georg M.N. AU - Bei, Roberto AU - Bejarano, Eloy AU - Bel, Shai AU - Behl, Christian AU - Belaid, Amine AU - Belgareh-Touzé, Naïma AU - Bellarosa, Cristina AU - Belleudi, Francesca AU - Belló Pérez, Melissa AU - Bello-Morales, Raquel AU - Beltran, Jackeline Soares De Oliveira AU - Beltran, Sebastián AU - Benbrook, Doris Mangiaracina AU - Bendorius, Mykolas AU - Benitez, Bruno A. AU - Benito-Cuesta, Irene AU - Bensalem, Julien AU - Berchtold, Martin W. AU - Berezowska, Sabina AU - Bergamaschi, Daniele AU - Bergami, Matteo AU - Bergmann, Andreas AU - Berliocchi, Laura AU - Berlioz-Torrent, Clarisse AU - Bernard, Amélie AU - Berthoux, Lionel AU - Besirli, Cagri G. AU - Besteiro, Sebastien AU - Betin, Virginie M. AU - Beyaert, Rudi AU - Bezbradica, Jelena S. AU - Bhaskar, Kiran AU - Bhatia-Kissova, Ingrid AU - Bhattacharya, Resham AU - Bhattacharya, Sujoy AU - Bhattacharyya, Shalmoli AU - Bhuiyan, Md Shenuarin AU - Bhutia, Sujit Kumar AU - Bi, Lanrong AU - Bi, Xiaolin AU - Biden, Trevor J. AU - Bijian, Krikor AU - Billes, Viktor A. AU - Binart, Nadine AU - Bincoletto, Claudia AU - Birgisdottir, Asa B. AU - Bjorkoy, Geir AU - Blanco, Gonzalo AU - Blas-Garcia, Ana AU - Blasiak, Janusz AU - Blomgran, Robert AU - Blomgren, Klas AU - Blum, Janice S. AU - Boada-Romero, Emilio AU - Boban, Mirta AU - Boesze-Battaglia, Kathleen AU - Boeuf, Philippe AU - Boland, Barry AU - Bomont, Pascale AU - Bonaldo, Paolo AU - Bonam, Srinivasa Reddy AU - Bonfili, Laura AU - Bonifacino, Juan S. AU - Boone, Brian A. AU - Bootman, Martin D. AU - Bordi, Matteo AU - Borner, Christoph AU - Bornhauser, Beat C. AU - Borthakur, Gautam AU - Bosch, Jürgen AU - Bose, Santanu AU - Botana, Luis M. AU - Botas, Juan AU - Boulanger, Chantal M. AU - Boulton, Michael E. AU - Bourdenx, Mathieu AU - Bourgeois, Benjamin AU - Bourke, Nollaig M. AU - Bousquet, Guilhem AU - Boya, Patricia AU - Bozhkov, Peter V. AU - Bozi, Luiz H.M. AU - Bozkurt, Tolga O. AU - Brackney, Doug E. AU - Brandts, Christian H. AU - Braun, Ralf J. AU - Braus, Gerhard H. AU - Bravo-Sagua, Roberto AU - Bravo-San Pedro, José M. AU - Brest, Patrick AU - Bringer, Marie Agnès AU - Briones-Herrera, Alfredo AU - Broaddus, V. Courtney AU - Brodersen, Peter AU - Brodsky, Jeffrey L. AU - Brody, Steven L. AU - Bronson, Paola G. AU - Bronstein, Jeff M. AU - Brown, Carolyn N. AU - Brown, Rhoderick E. AU - Brum, Patricia C. AU - Brumell, John H. AU - Brunetti-Pierri, Nicola AU - Bruno, Daniele AU - Bryson-Richardson, Robert J. AU - Bucci, Cecilia AU - Buchrieser, Carmen AU - Bueno, Marta AU - Buitrago-Molina, Laura Elisa AU - Buraschi, Simone AU - Buch, Shilpa AU - Buchan, J. Ross AU - Buckingham, Erin M. AU - Budak, Hikmet AU - Budini, Mauricio AU - Bultynck, Geert AU - Burada, Florin AU - Burgoyne, Joseph R. AU - Burón, M. Isabel AU - Bustos, Victor AU - Büttner, Sabrina AU - Butturini, Elena AU - Byrd, Aaron AU - Cabas, Isabel AU - Cabrera-Benitez, Sandra AU - Cadwell, Ken AU - Cai, Jingjing AU - Cai, Lu AU - Cai, Qian AU - Cairó, Montserrat AU - Calbet, Jose A. AU - Caldwell, Guy A. AU - Caldwell, Kim A. AU - Call, Jarrod A. AU - Calvani, Riccardo AU - Calvo, Ana C. AU - Calvo-Rubio Barrera, Miguel AU - Camara, Niels O.S. AU - Camonis, Jacques H. AU - Camougrand, Nadine AU - Campanella, Michelangelo AU - Campbell, Edward M. AU - Campbell-Valois, François Xavier AU - Campello, Silvia AU - Campesi, Ilaria AU - Campos, Juliane C. AU - Camuzard, Olivier AU - Cancino, Jorge AU - Candido De Almeida, Danilo AU - Canesi, Laura AU - Caniggia, Isabella AU - Canonico, Barbara AU - Cantí, Carles AU - Cao, Bin AU - Caraglia, Michele AU - Caramés, Beatriz AU - Carchman, Evie H. AU - Cardenal-Muñoz, Elena AU - Cardenas, Cesar AU - Cardenas, Luis AU - Cardoso, Sandra M. AU - Carew, Jennifer S. AU - Carle, Georges F. AU - Carleton, Gillian AU - Carloni, Silvia AU - Carmona-Gutierrez, Didac AU - Carneiro, Leticia A. AU - Carnevali, Oliana AU - Carosi, Julian M. AU - Carra, Serena AU - Carrier, Alice AU - Carrier, Lucie AU - Carroll, Bernadette AU - Carter, A. Brent AU - Carvalho, Andreia Neves AU - Casanova, Magali AU - Casas, Caty AU - Casas, Josefina AU - Cassioli, Chiara AU - Castillo, Eliseo F. AU - Castillo, Karen AU - Castillo-Lluva, Sonia AU - Castoldi, Francesca AU - Castori, Marco AU - Castro, Ariel F. AU - Castro-Caldas, Margarida AU - Castro-Hernandez, Javier AU - Castro-Obregon, Susana AU - Catz, Sergio D. AU - Cavadas, Claudia AU - Cavaliere, Federica AU - Cavallini, Gabriella AU - Cavinato, Maria AU - Cayuela, Maria L. AU - Cebollada Rica, Paula AU - Cecarini, Valentina AU - Cecconi, Francesco AU - Cechowska-Pasko, Marzanna AU - Cenci, Simone AU - Ceperuelo-Mallafré, Victòria AU - Cerqueira, João J. AU - Cerutti, Janete M. AU - Cervia, Davide AU - Cetintas, Vildan Bozok AU - Cetrullo, Silvia AU - Chae, Han Jung AU - Chagin, Andrei S. AU - Chai, Chee Yin AU - Chakrabarti, Gopal AU - Chakrabarti, Oishee AU - Chakraborty, Tapas AU - Chakraborty, Trinad AU - Chami, Mounia AU - Chamilos, Georgios AU - Chan, David W. AU - Chan, Edmond Y.W. AU - Chan, Edward D. AU - Chan, H. Y.Edwin AU - Chan, Helen H. AU - Chan, Hung AU - Chan, Matthew T.V. AU - Chan, Yau Sang AU - Chandra, Partha K. AU - Chang, Chih Peng AU - Chang, Chunmei AU - Chang, Hao Chun AU - Chang, Kai AU - Chao, Jie AU - Chapman, Tracey AU - Charlet-Berguerand, Nicolas AU - Chatterjee, Samrat AU - Chaube, Shail K. AU - Chaudhary, Anu AU - Chauhan, Santosh AU - Chaum, Edward AU - Checler, Frédéric AU - Cheetham, Michael E. AU - Chen, Chang Shi AU - Chen, Guang Chao AU - Chen, Jian Fu AU - Chen, Liam L. AU - Chen, Leilei AU - Chen, Lin AU - Chen, Mingliang AU - Chen, Mu Kuan AU - Chen, Ning AU - Chen, Quan AU - Chen, Ruey Hwa AU - Chen, Shi AU - Chen, Wei AU - Chen, Weiqiang AU - Chen, Xin Ming AU - Chen, Xiong Wen AU - Chen, Xu AU - Chen, Yan AU - Chen, Ye Guang AU - Chen, Yingyu AU - Chen, Yongqiang AU - Chen, Yu Jen AU - Chen, Yue Qin AU - Chen, Zhefan Stephen AU - Chen, Zhi AU - Chen, Zhi Hua AU - Chen, Zhijian J. AU - Chen, Zhixiang AU - Cheng, Hanhua AU - Cheng, Jun AU - Cheng, Shi Yuan AU - Cheng, Wei AU - Cheng, Xiaodong AU - Cheng, Xiu Tang AU - Cheng, Yiyun AU - Cheng, Zhiyong AU - Chen, Zhong AU - Cheong, Heesun AU - Cheong, Jit Kong AU - Chernyak, Boris V. AU - Cherry, Sara AU - Cheung, Chi Fai Randy AU - Cheung, Chun Hei Antonio AU - Cheung, King Ho AU - Chevet, Eric AU - Chi, Richard J. AU - Chiang, Alan Kwok Shing AU - Chiaradonna, Ferdinando AU - Chiarelli, Roberto AU - Chiariello, Mario AU - Chica, Nathalia AU - Chiocca, Susanna AU - Chiong, Mario AU - Chiou, Shih Hwa AU - Chiramel, Abhilash I. AU - Chiurchiù, Valerio AU - Cho, Dong Hyung AU - Choe, Seong Kyu AU - Choi, Augustine M.K. AU - Choi, Mary E. AU - Choudhury, Kamalika Roy AU - Chow, Norman S. AU - Chu, Charleen T. AU - Chua, Jason P. AU - Chua, John Jia En AU - Chung, Hyewon AU - Chung, Kin Pan AU - Chung, Seockhoon AU - Chung, So Hyang AU - Chung, Yuen Li AU - Cianfanelli, Valentina AU - Ciechomska, Iwona A. AU - Cifuentes, Mariana AU - Cinque, Laura AU - Cirak, Sebahattin AU - Cirone, Mara AU - Clague, Michael J. AU - Clarke, Robert AU - Clementi, Emilio AU - Coccia, Eliana M. AU - Codogno, Patrice AU - Cohen, Ehud AU - Cohen, Mickael M. AU - Colasanti, Tania AU - Colasuonno, Fiorella AU - Colbert, Robert A. AU - Colell, Anna AU - Čolić, Miodrag AU - Coll, Nuria S. AU - Collins, Mark O. AU - Colombo, María I. AU - Colón-Ramos, Daniel A. AU - Combaret, Lydie AU - Comincini, Sergio AU - Cominetti, Márcia R. AU - Consiglio, Antonella AU - Conte, Andrea AU - Conti, Fabrizio AU - Contu, Viorica Raluca AU - Cookson, Mark R. AU - Coombs, Kevin M. AU - Coppens, Isabelle AU - Corasaniti, Maria Tiziana AU - Corkery, Dale P. AU - Cordes, Nils AU - Cortese, Katia AU - Costa, Maria Do Carmo AU - Costantino, Sarah AU - Costelli, Paola AU - Coto-Montes, Ana AU - Crack, Peter J. AU - Crespo, Jose L. AU - Criollo, Alfredo AU - Crippa, Valeria AU - Cristofani, Riccardo AU - Csizmadia, Tamas AU - Cuadrado, Antonio AU - Cui, Bing AU - Cui, Jun AU - Cui, Yixian AU - Cui, Yong AU - Culetto, Emmanuel AU - Cumino, Andrea C. AU - Cybulsky, Andrey V. AU - Czaja, Mark J. AU - Czuczwar, Stanislaw J. AU - D’Adamo, Stefania AU - D’Amelio, Marcello AU - D’Arcangelo, Daniela AU - D’Lugos, Andrew C. AU - D’Orazi, Gabriella AU - Da Silva, James A. AU - Dafsari, Hormos Salimi AU - Dagda, Ruben K. AU - Dagdas, Yasin AU - Daglia, Maria AU - Dai, Xiaoxia AU - Dai, Yun AU - Dai, Yuyuan AU - Dal Col, Jessica AU - Dalhaimer, Paul AU - Dalla Valle, Luisa AU - Dallenga, Tobias AU - Dalmasso, Guillaume AU - Damme, Markus AU - Dando, Ilaria AU - Dantuma, Nico P. AU - Darling, April L. AU - Das, Hiranmoy AU - Dasarathy, Srinivasan AU - Dasari, Santosh K. AU - Dash, Srikanta AU - Daumke, Oliver AU - Dauphinee, Adrian N. AU - Davies, Jeffrey S. AU - Dávila, Valeria A. AU - Davis, Roger J. AU - Davis, Tanja AU - Dayalan Naidu, Sharadha AU - De Amicis, Francesca AU - De Bosscher, Karolien AU - De Felice, Francesca AU - De Franceschi, Lucia AU - De Leonibus, Chiara AU - De Mattos Barbosa, Mayara G. AU - De Meyer, Guido R.Y. AU - De Milito, Angelo AU - De Nunzio, Cosimo AU - De Palma, Clara AU - De Santi, Mauro AU - De Virgilio, Claudio AU - De Zio, Daniela AU - Debnath, Jayanta AU - Debosch, Brian J. AU - Decuypere, Jean Paul AU - Deehan, Mark A. AU - Deflorian, Gianluca AU - Degregori, James AU - Dehay, Benjamin AU - Del Rio, Gabriel AU - Delaney, Joe R. AU - Delbridge, Lea M.D. AU - Delorme-Axford, Elizabeth AU - Delpino, M. Victoria AU - Demarchi, Francesca AU - Dembitz, Vilma AU - Demers, Nicholas D. AU - Deng, Hongbin AU - Deng, Zhiqiang AU - Dengjel, Joern AU - Dent, Paul AU - Denton, Donna AU - Depamphilis, Melvin L. AU - Der, Channing J. AU - Deretic, Vojo AU - Descoteaux, Albert AU - Devis, Laura AU - Devkota, Sushil AU - Devuyst, Olivier AU - Dewson, Grant AU - Dharmasivam, Mahendiran AU - Dhiman, Rohan AU - Di Bernardo, Diego AU - Di Cristina, Manlio AU - Di Domenico, Fabio AU - Di Fazio, Pietro AU - Di Fonzo, Alessio AU - Di Guardo, Giovanni AU - Di Guglielmo, Gianni M. AU - Di Leo, Luca AU - Di Malta, Chiara AU - Di Nardo, Alessia AU - Di Rienzo, Martina AU - Di Sano, Federica AU - Diallinas, George AU - Diao, Jiajie AU - Diaz-Araya, Guillermo AU - Díaz-Laviada, Inés AU - Dickinson, Jared M. AU - Diederich, Marc AU - Dieudé, Mélanie AU - Dikic, Ivan AU - Ding, Shiping AU - Ding, Wen Xing AU - Dini, Luciana AU - Dinić, Jelena AU - Dinic, Miroslav AU - Dinkova-Kostova, Albena T. AU - Dionne, Marc S. AU - Distler, Jörg H.W. AU - Diwan, Abhinav AU - Dixon, Ian M.C. AU - Djavaheri-Mergny, Mojgan AU - Dobrinski, Ina AU - Dobrovinskaya, Oxana AU - Dobrowolski, Radek AU - Dobson, Renwick C.J. AU - Đokić, Jelena AU - Dokmeci Emre, Serap AU - Donadelli, Massimo AU - Dong, Bo AU - Dong, Xiaonan AU - Dong, Zhiwu AU - Dorn, Gerald W. AU - Dotsch, Volker AU - Dou, Huan AU - Dou, Juan AU - Dowaidar, Moataz AU - Dridi, Sami AU - Drucker, Liat AU - Du, Ailian AU - Du, Caigan AU - Du, Guangwei AU - Du, Hai Ning AU - Du, Li Lin AU - Du Toit, André AU - Duan, Shao Bin AU - Duan, Xiaoqiong AU - Duarte, Sónia P. AU - Dubrovska, Anna AU - Dunlop, Elaine A. AU - Dupont, Nicolas AU - Durán, Raúl V. AU - Dwarakanath, Bilikere S. AU - Dyshlovoy, Sergey A. AU - Ebrahimi-Fakhari, Darius AU - Eckhart, Leopold AU - Edelstein, Charles L. AU - Efferth, Thomas AU - Eftekharpour, Eftekhar AU - Eichinger, Ludwig AU - Eid, Nabil AU - Eisenberg, Tobias AU - Eissa, N. Tony AU - Eissa, Sanaa AU - Ejarque, Miriam AU - El Andaloussi, Abdeljabar AU - El-Hage, Nazira AU - El-Naggar, Shahenda AU - Eleuteri, Anna Maria AU - El-Shafey, Eman S. AU - Elgendy, Mohamed AU - Eliopoulos, Aristides G. AU - Elizalde, María M. AU - Elks, Philip M. AU - Elsasser, Hans Peter AU - Elsherbiny, Eslam S. AU - Emerling, Brooke M. AU - Emre, N. C.Tolga AU - Eng, Christina H. AU - Engedal, Nikolai AU - Engelbrecht, Anna Mart AU - Engelsen, Agnete S.T. AU - Enserink, Jorrit M. AU - Escalante, Ricardo AU - Esclatine, Audrey AU - Escobar-Henriques, Mafalda AU - Eskelinen, Eeva Liisa AU - Espert, Lucile AU - Eusebio, Makandjou Ola AU - Fabrias, Gemma AU - Fabrizi, Cinzia AU - Facchiano, Antonio AU - Facchiano, Francesco AU - Fadeel, Bengt AU - Fader, Claudio AU - Faesen, Alex C. AU - Fairlie, W. Douglas AU - Falcó, Alberto AU - Falkenburger, Bjorn H. AU - Fan, Daping AU - Fan, Jie AU - Fan, Yanbo AU - Fang, Evandro F. AU - Fang, Yanshan AU - Fang, Yognqi AU - Fanto, Manolis AU - Farfel-Becker, Tamar AU - Faure, Mathias AU - Fazeli, Gholamreza AU - Fedele, Anthony O. AU - Feldman, Arthur M. AU - Feng, Du AU - Feng, Jiachun AU - Feng, Lifeng AU - Feng, Yibin AU - Feng, Yuchen AU - Feng, Wei AU - Fenz Araujo, Thais AU - Ferguson, Thomas A. AU - Fernández, Álvaro F. AU - Fernandez-Checa, Jose C. AU - Fernández-Veledo, Sonia AU - Fernie, Alisdair R. AU - Ferrante, Anthony W. AU - Ferraresi, Alessandra AU - Ferrari, Merari F. AU - Ferreira, Julio C.B. AU - Ferro-Novick, Susan AU - Figueras, Antonio AU - Filadi, Riccardo AU - Filigheddu, Nicoletta AU - Filippi-Chiela, Eduardo AU - Filomeni, Giuseppe AU - Fimia, Gian Maria AU - Fineschi, Vittorio AU - Finetti, Francesca AU - Finkbeiner, Steven AU - Fisher, Edward A. AU - Fisher, Paul B. AU - Flamigni, Flavio AU - Fliesler, Steven J. AU - Flo, Trude H. AU - Florance, Ida AU - Florey, Oliver AU - Florio, Tullio AU - Fodor, Erika AU - Follo, Carlo AU - Fon, Edward A. AU - Forlino, Antonella AU - Fornai, Francesco AU - Fortini, Paola AU - Fracassi, Anna AU - Fraldi, Alessandro AU - Franco, Brunella AU - Franco, Rodrigo AU - Franconi, Flavia AU - Frankel, Lisa B. AU - Friedman, Scott L. AU - Fröhlich, Leopold F. AU - Frühbeck, Gema AU - Fuentes, Jose M. AU - Fujiki, Yukio AU - Fujita, Naonobu AU - Fujiwara, Yuuki AU - Fukuda, Mitsunori AU - Fulda, Simone AU - Furic, Luc AU - Furuya, Norihiko AU - Fusco, Carmela AU - Gack, Michaela U. AU - Gaffke, Lidia AU - Galadari, Sehamuddin AU - Galasso, Alessia AU - Galindo, Maria F. AU - Gallolu Kankanamalage, Sachith AU - Galluzzi, Lorenzo AU - Galy, Vincent AU - Gammoh, Noor AU - Gan, Boyi AU - Ganley, Ian G. AU - Gao, Feng AU - Gao, Hui AU - Gao, Minghui AU - Gao, Ping AU - Gao, Shou Jiang AU - Gao, Wentao AU - Gao, Xiaobo AU - Garcera, Ana AU - Garcia, Maria Noé AU - Garcia, Verónica E. AU - García-Del Portillo, Francisco AU - Garcia-Escudero, Vega AU - Garcia-Garcia, Aracely AU - Garcia-Macia, Marina AU - García-Moreno, Diana AU - Garcia-Ruiz, Carmen AU - García-Sanz, Patricia AU - Garg, Abhishek D. AU - Gargini, Ricardo AU - Garofalo, Tina AU - Garry, Robert F. AU - Gassen, Nils C. AU - Gatica, Damian AU - Ge, Liang AU - Ge, Wanzhong AU - Geiss-Friedlander, Ruth AU - Gelfi, Cecilia AU - Genschik, Pascal AU - Gentle, Ian E. AU - Gerbino, Valeria AU - Gerhardt, Christoph AU - Germain, Kyla AU - Germain, Marc AU - Gewirtz, David A. AU - Ghasemipour Afshar, Elham AU - Ghavami, Saeid AU - Ghigo, Alessandra AU - Ghosh, Manosij AU - Giamas, Georgios AU - Giampietri, Claudia AU - Giatromanolaki, Alexandra AU - Gibson, Gary E. AU - Gibson, Spencer B. AU - Ginet, Vanessa AU - Giniger, Edward AU - Giorgi, Carlotta AU - Girao, Henrique AU - Girardin, Stephen E. AU - Giridharan, Mridhula AU - Giuliano, Sandy AU - Giulivi, Cecilia AU - Giuriato, Sylvie AU - Giustiniani, Julien AU - Gluschko, Alexander AU - Goder, Veit AU - Goginashvili, Alexander AU - Golab, Jakub AU - Goldstone, David C. AU - Golebiewska, Anna AU - Gomes, Luciana R. AU - Gomez, Rodrigo AU - Gómez-Sánchez, Rubén AU - Gomez-Puerto, Maria Catalina AU - Gomez-Sintes, Raquel AU - Gong, Qingqiu AU - Goni, Felix M. AU - González-Gallego, Javier AU - Gonzalez-Hernandez, Tomas AU - Gonzalez-Polo, Rosa A. AU - Gonzalez-Reyes, Jose A. AU - González-Rodríguez, Patricia AU - Goping, Ing Swie AU - Gorbatyuk, Marina S. AU - Gorbunov, Nikolai V. AU - Görgülü, Kıvanç AU - Gorojod, Roxana M. AU - Gorski, Sharon M. AU - Goruppi, Sandro AU - Gotor, Cecilia AU - Gottlieb, Roberta A. AU - Gozes, Illana AU - Gozuacik, Devrim AU - Graef, Martin AU - Gräler, Markus H. AU - Granatiero, Veronica AU - Grasso, Daniel AU - Gray, Joshua P. AU - Green, Douglas R. AU - Greenhough, Alexander AU - Gregory, Stephen L. AU - Griffin, Edward F. AU - Grinstaff, Mark W. AU - Gros, Frederic AU - Grose, Charles AU - Gross, Angelina S. AU - Gruber, Florian AU - Grumati, Paolo AU - Grune, Tilman AU - Gu, Xueyan AU - Guan, Jun Lin AU - Guardia, Carlos M. AU - Guda, Kishore AU - Guerra, Flora AU - Guerri, Consuelo AU - Guha, Prasun AU - Guillén, Carlos AU - Gujar, Shashi AU - Gukovskaya, Anna AU - Gukovsky, Ilya AU - Gunst, Jan AU - Günther, Andreas AU - Guntur, Anyonya R. AU - Guo, Chuanyong AU - Guo, Chun AU - Guo, Hongqing AU - Guo, Lian Wang AU - Guo, Ming AU - Gupta, Pawan AU - Gupta, Shashi Kumar AU - Gupta, Swapnil AU - Gupta, Veer Bala AU - Gupta, Vivek AU - Gustafsson, Asa B. AU - Gutterman, David D. AU - H.B, Ranjitha AU - Haapasalo, Annakaisa AU - Haber, James E. AU - Hać, Aleksandra AU - Hadano, Shinji AU - Hafrén, Anders J. AU - Haidar, Mansour AU - Hall, Belinda S. AU - Halldén, Gunnel AU - Hamacher-Brady, Anne AU - Hamann, Andrea AU - Hamasaki, Maho AU - Han, Weidong AU - Hansen, Malene AU - Hanson, Phyllis I. . AU - Hao, Zijian AU - Harada, Masaru AU - Harhaji-Trajkovic, Ljubica AU - Hariharan, Nirmala AU - Haroon, Nigil AU - Harris, James AU - Hasegawa, Takafumi AU - Hasima Nagoor, Noor AU - Haspel, Jeffrey A. AU - Haucke, Volker AU - Hawkins, Wayne D. AU - Hay, Bruce A. AU - Haynes, Cole M. AU - Hayrabedyan, Soren B. AU - Hays, Thomas S. AU - He, Congcong AU - He, Qin AU - He, Rong Rong AU - He, You Wen AU - He, Yu Ying AU - Heakal, Yasser AU - Heberle, Alexander M. AU - Hejtmancik, J. Fielding AU - Helgason, Gudmundur Vignir AU - Henkel, Vanessa AU - Herb, Marc AU - Hergovich, Alexander AU - Herman-Antosiewicz, Anna AU - Hernández, Agustín AU - Hernandez, Carlos AU - Hernandez-Diaz, Sergio AU - Hernandez-Gea, Virginia AU - Herpin, Amaury AU - Herreros, Judit AU - Hervás, Javier H. AU - Hesselson, Daniel AU - Hetz, Claudio AU - Heussler, Volker T. AU - Higuchi, Yujiro AU - Hilfiker, Sabine AU - Hill, Joseph A. AU - Hlavacek, William S. AU - Ho, Emmanuel A. AU - Ho, Idy H.T. AU - Ho, Philip Wing Lok AU - Ho, Shu Leong AU - Ho, Wan Yun AU - Hobbs, G. Aaron AU - Hochstrasser, Mark AU - Hoet, Peter H.M. AU - Hofius, Daniel AU - Hofman, Paul AU - Höhn, Annika AU - Holmberg, Carina I. AU - Hombrebueno, Jose R. AU - Yi-Ren Hong, Chang Won Hong AU - Hooper, Lora V. AU - Hoppe, Thorsten AU - Horos, Rastislav AU - Hoshida, Yujin AU - Hsin, I. Lun AU - Hsu, Hsin Yun AU - Hu, Bing AU - Hu, Dong AU - Hu, Li Fang AU - Hu, Ming Chang AU - Hu, Ronggui AU - Hu, Wei AU - Hu, Yu Chen AU - Hu, Zhuo Wei AU - Hua, Fang AU - Hua, Jinlian AU - Hua, Yingqi AU - Huan, Chongmin AU - Huang, Canhua AU - Huang, Chuanshu AU - Huang, Chuanxin AU - Huang, Chunling AU - Huang, Haishan AU - Huang, Kun AU - Huang, Michael L.H. AU - Huang, Rui AU - Huang, Shan AU - Huang, Tianzhi AU - Huang, Xing AU - Huang, Yuxiang Jack AU - Huber, Tobias B. AU - Hubert, Virginie AU - Hubner, Christian A. AU - Hughes, Stephanie M. AU - Hughes, William E. AU - Humbert, Magali AU - Hummer, Gerhard AU - Hurley, James H. AU - Hussain, Sabah AU - Hussain, Salik AU - Hussey, Patrick J. AU - Hutabarat, Martina AU - Hwang, Hui Yun AU - Hwang, Seungmin AU - Ieni, Antonio AU - Ikeda, Fumiyo AU - Imagawa, Yusuke AU - Imai, Yuzuru AU - Imbriano, Carol AU - Imoto, Masaya AU - Inman, Denise M. AU - Inoki, Ken AU - Iovanna, Juan AU - Iozzo, Renato V. AU - Ippolito, Giuseppe AU - Irazoqui, Javier E. AU - Iribarren, Pablo AU - Ishaq, Mohd AU - Ishikawa, Makoto AU - Ishimwe, Nestor AU - Isidoro, Ciro AU - Ismail, Nahed AU - Issazadeh-Navikas, Shohreh AU - Itakura, Eisuke AU - Ito, Daisuke AU - Ivankovic, Davor AU - Ivanova, Saška AU - Iyer, Anand Krishnan V. AU - Izquierdo, José M. AU - Izumi, Masanori AU - Jäättelä, Marja AU - Jabir, Majid Sakhi AU - Jackson, William T. AU - Jacobo-Herrera, Nadia AU - Jacomin, Anne Claire AU - Jacquin, Elise AU - Jadiya, Pooja AU - Jaeschke, Hartmut AU - Jagannath, Chinnaswamy AU - Jakobi, Arjen J. AU - Jakobsson, Johan AU - Janji, Bassam AU - Jansen-Dürr, Pidder AU - Jansson, Patric J. AU - Jantsch, Jonathan AU - Januszewski, Sławomir AU - Jassey, Alagie AU - Jean, Steve AU - Jeltsch-David, Hélène AU - Jendelova, Pavla AU - Jenny, Andreas AU - Jensen, Thomas E. AU - Jessen, Niels AU - Jewell, Jenna L. AU - Ji, Jing AU - Jia, Lijun AU - Jia, Rui AU - Jiang, Liwen AU - Jiang, Qing AU - Jiang, Richeng AU - Jiang, Teng AU - Jiang, Xuejun AU - Jiang, Yu AU - Jimenez-Sanchez, Maria AU - Jin, Eun Jung AU - Jin, Fengyan AU - Jin, Hongchuan AU - Jin, Li AU - Jin, Luqi AU - Jin, Meiyan AU - Jin, Si AU - Jo, Eun Kyeong AU - Joffre, Carine AU - Johansen, Terje AU - Johnson, Gail V.W. AU - Johnston, Simon A. AU - Jokitalo, Eija AU - Jolly, Mohit Kumar AU - Joosten, Leo A.B. AU - Jordan, Joaquin AU - Joseph, Bertrand AU - Ju, Dianwen AU - Ju, Jeong Sun AU - Ju, Jingfang AU - Juárez, Esmeralda AU - Judith, Delphine AU - Juhász, Gábor AU - Jun, Youngsoo AU - Jung, Chang Hwa AU - Jung, Sung Chul AU - Jung, Yong Keun AU - Jungbluth, Heinz AU - Jungverdorben, Johannes AU - Just, Steffen AU - Kaarniranta, Kai AU - Kaasik, Allen AU - Kabuta, Tomohiro AU - Kaganovich, Daniel AU - Kahana, Alon AU - Kain, Renate AU - Kajimura, Shinjo AU - Kalamvoki, Maria AU - Kalia, Manjula AU - Kalinowski, Danuta S. AU - Kaludercic, Nina AU - Kalvari, Ioanna AU - Kaminska, Joanna AU - Kaminskyy, Vitaliy O. AU - Kanamori, Hiromitsu AU - Kanasaki, Keizo AU - Kang, Chanhee AU - Kang, Rui AU - Kang, Sang Sun AU - Kaniyappan, Senthilvelrajan AU - Kanki, Tomotake AU - Kanneganti, Thirumala Devi AU - Kanthasamy, Anumantha G. AU - Kanthasamy, Arthi AU - Kantorow, Marc AU - Kapuy, Orsolya AU - Karamouzis, Michalis V. AU - Karim, Md Razaul AU - Karmakar, Parimal AU - Katare, Rajesh G. AU - Kato, Masaru AU - Kaufmann, Stefan H.E. AU - Kauppinen, Anu AU - Kaushal, Gur P. AU - Kaushik, Susmita AU - Kawasaki, Kiyoshi AU - Kazan, Kemal AU - Ke, Po Yuan AU - Keating, Damien J. AU - Keber, Ursula AU - Kehrl, John H. AU - Keller, Kate E. AU - Keller, Christian W. AU - Kemper, Jongsook Kim AU - Kenific, Candia M. AU - Kepp, Oliver AU - Kermorgant, Stephanie AU - Kern, Andreas AU - Ketteler, Robin AU - Keulers, Tom G. AU - Khalfin, Boris AU - Khalil, Hany AU - Khambu, Bilon AU - Khan, Shahid Y. AU - Khandelwal, Vinoth Kumar Megraj AU - Khandia, Rekha AU - Kho, Widuri AU - Khobrekar, Noopur V. AU - Khuansuwan, Sataree AU - Khundadze, Mukhran AU - Killackey, Samuel A. AU - Kim, Dasol AU - Kim, Deok Ryong AU - Kim, Do Hyung AU - Kim, Dong Eun AU - Kim, Eun Young AU - Kim, Eun Kyoung AU - Kim, Hak Rim AU - Kim, Hee Sik AU - Hyung-Ryong Kim, Unknown AU - Kim, Jeong Hun AU - Kim, Jin Kyung AU - Kim, Jin Hoi AU - Kim, Joungmok AU - Kim, Ju Hwan AU - Kim, Keun Il AU - Kim, Peter K. AU - Kim, Seong Jun AU - Kimball, Scot R. AU - Kimchi, Adi AU - Kimmelman, Alec C. AU - Kimura, Tomonori AU - King, Matthew A. AU - Kinghorn, Kerri J. AU - Kinsey, Conan G. AU - Kirkin, Vladimir AU - Kirshenbaum, Lorrie A. AU - Kiselev, Sergey L. AU - Kishi, Shuji AU - Kitamoto, Katsuhiko AU - Kitaoka, Yasushi AU - Kitazato, Kaio AU - Kitsis, Richard N. AU - Kittler, Josef T. AU - Kjaerulff, Ole AU - Klein, Peter S. AU - Klopstock, Thomas AU - Klucken, Jochen AU - Knævelsrud, Helene AU - Knorr, Roland L. AU - Ko, Ben C.B. AU - Ko, Fred AU - Ko, Jiunn Liang AU - Kobayashi, Hotaka AU - Kobayashi, Satoru AU - Koch, Ina AU - Koch, Jan C. AU - Koenig, Ulrich AU - Kögel, Donat AU - Koh, Young Ho AU - Koike, Masato AU - Kohlwein, Sepp D. AU - Kocaturk, Nur M. AU - Komatsu, Masaaki AU - König, Jeannette AU - Kono, Toru AU - Kopp, Benjamin T. AU - Korcsmaros, Tamas AU - Korkmaz, Gözde AU - Korolchuk, Viktor I. AU - Korsnes, Mónica Suárez AU - Koskela, Ali AU - Kota, Janaiah AU - Kotake, Yaichiro AU - Kotler, Monica L. AU - Kou, Yanjun AU - Koukourakis, Michael I. AU - Koustas, Evangelos AU - Kovacs, Attila L. AU - Kovács, Tibor AU - Koya, Daisuke AU - Kozako, Tomohiro AU - Kraft, Claudine AU - Krainc, Dimitri AU - Krämer, Helmut AU - Krasnodembskaya, Anna D. AU - Kretz-Remy, Carole AU - Kroemer, Guido AU - Ktistakis, Nicholas T. AU - Kuchitsu, Kazuyuki AU - Kuenen, Sabine AU - Kuerschner, Lars AU - Kukar, Thomas AU - Kumar, Ajay AU - Kumar, Ashok AU - Kumar, Deepak AU - Kumar, Dhiraj AU - Kumar, Sharad AU - Kume, Shinji AU - Kumsta, Caroline AU - Kundu, Chanakya N. AU - Kundu, Mondira AU - Kunnumakkara, Ajaikumar B. AU - Kurgan, Lukasz AU - Kutateladze, Tatiana G. AU - Kutlu, Ozlem AU - Kwak, Seong Ae AU - Kwon, Ho Jeong AU - Kwon, Taeg Kyu AU - Kwon, Yong Tae AU - Kyrmizi, Irene AU - La Spada, Albert AU - Labonté, Patrick AU - Ladoire, Sylvain AU - Laface, Ilaria AU - Lafont, Frank AU - Lagace, Diane C. AU - Lahiri, Vikramjit AU - Lai, Zhibing AU - Laird, Angela S. AU - Lakkaraju, Aparna AU - Lamark, Trond AU - Lan, Sheng Hui AU - Landajuela, Ane AU - Lane, Darius J.R. AU - Lane, Jon D. AU - Lang, Charles H. AU - Lange, Carsten AU - Langel, Ülo AU - Langer, Rupert AU - Lapaquette, Pierre AU - Laporte, Jocelyn AU - Larusso, Nicholas F. AU - Lastres-Becker, Isabel AU - Lau, Wilson Chun Yu AU - Laurie, Gordon W. AU - Lavandero, Sergio AU - Law, Betty Yuen Kwan AU - Law, Helen Ka Wai AU - Layfield, Rob AU - Le, Weidong AU - Le Stunff, Herve AU - Leary, Alexandre Y. AU - Lebrun, Jean Jacques AU - Leck, Lionel Y.W. AU - Leduc-Gaudet, Jean Philippe AU - Lee, Changwook AU - Lee, Chung Pei AU - Lee, Da Hye AU - Lee, Edward B. AU - Lee, Erinna F. AU - Lee, Gyun Min AU - Lee, He Jin AU - Lee, Heung Kyu AU - Lee, Jae Man AU - Lee, Jason S. AU - Lee, Jin A. AU - Lee, Joo Yong AU - Lee, Jun Hee AU - Lee, Michael AU - Lee, Min Goo AU - Lee, Min Jae AU - Lee, Myung Shik AU - Lee, Sang Yoon AU - Lee, Seung Jae AU - Lee, Stella Y. AU - Lee, Sung Bae AU - Lee, Won Hee AU - Lee, Ying Ray AU - Lee, Yong Ho AU - Lee, Youngil AU - Lefebvre, Christophe AU - Legouis, Renaud AU - Lei, Yu L. AU - Lei, Yuchen AU - Leikin, Sergey AU - Leitinger, Gerd AU - Lemus, Leticia AU - Leng, Shuilong AU - Lenoir, Olivia AU - Lenz, Guido AU - Lenz, Heinz Josef AU - Lenzi, Paola AU - León, Yolanda AU - Leopoldino, Andréia M. AU - Leschczyk, Christoph AU - Leskelä, Stina AU - Letellier, Elisabeth AU - Leung, Chi Ting AU - Leung, Po Sing AU - Leventhal, Jeremy S. AU - Levine, Beth AU - Lewis, Patrick A. AU - Ley, Klaus AU - Li, Bin AU - Li, Da Qiang AU - Li, Jianming AU - Li, Jing AU - Li, Jiong AU - Li, Ke AU - Li, Liwu AU - Li, Mei AU - Li, Min AU - Li, Min AU - Li, Ming AU - Li, Mingchuan AU - Li, Pin Lan AU - Li, Ming Qing AU - Li, Qing AU - Li, Sheng AU - Li, Tiangang AU - Li, Wei AU - Li, Wenming AU - Li, Xue AU - Li, Yi Ping AU - Li, Yuan AU - Li, Zhiqiang AU - Li, Zhiyong AU - Li, Zhiyuan AU - Lian, Jiqin AU - Liang, Chengyu AU - Liang, Qiangrong AU - Liang, Weicheng AU - Liang, Yongheng AU - Liang, Yong Tian AU - Liao, Guanghong AU - Liao, Lujian AU - Liao, Mingzhi AU - Liao, Yung Feng AU - Librizzi, Mariangela AU - Lie, Pearl P.Y. AU - Lilly, Mary A. AU - Lim, Hyunjung J. AU - Lima, Thania R.R. AU - Limana, Federica AU - Lin, Chao AU - Lin, Chih Wen AU - Lin, Dar Shong AU - Lin, Fu Cheng AU - Lin, Jiandie D. AU - Lin, Kurt M. AU - Lin, Kwang Huei AU - Lin, Liang Tzung AU - Lin, Pei Hui AU - Lin, Qiong AU - Lin, Shaofeng AU - Lin, Su Ju AU - Lin, Wenyu AU - Lin, Xueying AU - Lin, Yao Xin AU - Lin, Yee Shin AU - Linden, Rafael AU - Lindner, Paula AU - Ling, Shuo Chien AU - Lingor, Paul AU - Linnemann, Amelia K. AU - Liou, Yih Cherng AU - Lipinski, Marta M. AU - Lipovšek, Saška AU - Lira, Vitor A. AU - Lisiak, Natalia AU - Liton, Paloma B. AU - Liu, Chao AU - Liu, Ching Hsuan AU - Liu, Chun Feng AU - Liu, Cui Hua AU - Liu, Fang AU - Liu, Hao AU - Liu, Hsiao Sheng AU - Liu, Hua Feng AU - Liu, Huifang AU - Liu, Jia AU - Liu, Jing AU - Liu, Julia AU - Liu, Leyuan AU - Liu, Longhua AU - Liu, Meilian AU - Liu, Qin AU - Liu, Wei AU - Liu, Wende AU - Liu, Xiao Hong AU - Liu, Xiaodong AU - Liu, Xingguo AU - Liu, Xu AU - Liu, Xuedong AU - Liu, Yanfen AU - Liu, Yang AU - Liu, Yang AU - Liu, Yueyang AU - Liu, Yule AU - Livingston, J. Andrew AU - Lizard, Gerard AU - Lizcano, Jose M. AU - Ljubojevic-Holzer, Senka AU - Lleonart, Matilde E. AU - Llobet-Navàs, David AU - Llorente, Alicia AU - Lo, Chih Hung AU - Lobato-Márquez, Damián AU - Long, Qi AU - Long, Yun Chau AU - Loos, Ben AU - Loos, Julia A. AU - López, Manuela G. AU - López-Doménech, Guillermo AU - López-Guerrero, José Antonio AU - López-Jiménez, Ana T. AU - López-Pérez, Óscar AU - López-Valero, Israel AU - Lorenowicz, Magdalena J. AU - Lorente, Mar AU - Lorincz, Peter AU - Lossi, Laura AU - Lotersztajn, Sophie AU - Lovat, Penny E. AU - Lovell, Jonathan F. AU - Lovy, Alenka AU - Lőw, Péter AU - Lu, Guang AU - Lu, Haocheng AU - Lu, Jia Hong AU - Lu, Jin Jian AU - Lu, Mengji AU - Lu, Shuyan AU - Luciani, Alessandro AU - Lucocq, John M. AU - Ludovico, Paula AU - Luftig, Micah A. AU - Luhr, Morten AU - Luis-Ravelo, Diego AU - Lum, Julian J. AU - Luna-Dulcey, Liany AU - Lund, Anders H. AU - Lund, Viktor K. AU - Lünemann, Jan D. AU - Lüningschrör, Patrick AU - Luo, Honglin AU - Luo, Rongcan AU - Luo, Shouqing AU - Luo, Zhi AU - Luparello, Claudio AU - Lüscher, Bernhard AU - Luu, Luan AU - Lyakhovich, Alex AU - Lyamzaev, Konstantin G. AU - Lystad, Alf Håkon AU - Lytvynchuk, Lyubomyr AU - Ma, Alvin C. AU - Ma, Changle AU - Ma, Mengxiao AU - Ma, Ning Fang AU - Ma, Quan Hong AU - Ma, Xinliang AU - Ma, Yueyun AU - Ma, Zhenyi AU - Macdougald, Ormond A. AU - Macian, Fernando AU - Macintosh, Gustavo C. AU - Mackeigan, Jeffrey P. AU - Macleod, Kay F. AU - Maday, Sandra AU - Madeo, Frank AU - Madesh, Muniswamy AU - Madl, Tobias AU - Madrigal-Matute, Julio AU - Maeda, Akiko AU - Maejima, Yasuhiro AU - Magarinos, Marta AU - Mahavadi, Poornima AU - Maiani, Emiliano AU - Maiese, Kenneth AU - Maiti, Panchanan AU - Maiuri, Maria Chiara AU - Majello, Barbara AU - Major, Michael B. AU - Makareeva, Elena AU - Malik, Fayaz AU - Mallilankaraman, Karthik AU - Malorni, Walter AU - Maloyan, Alina AU - Mammadova, Najiba AU - Man, Gene Chi Wai AU - Manai, Federico AU - Mancias, Joseph D. AU - Mandelkow, Eva Maria AU - Mandell, Michael A. AU - Manfredi, Angelo A. AU - Manjili, Masoud H. AU - Manjithaya, Ravi AU - Manque, Patricio AU - Manshian, Bella B. AU - Manzano, Raquel AU - Manzoni, Claudia AU - Mao, Kai AU - Marchese, Cinzia AU - Marchetti, Sandrine AU - Marconi, Anna Maria AU - Marcucci, Fabrizio AU - Mardente, Stefania AU - Mareninova, Olga A. AU - Margeta, Marta AU - Mari, Muriel AU - Marinelli, Sara AU - Marinelli, Oliviero AU - Mariño, Guillermo AU - Mariotto, Sofia AU - Marshall, Richard S. AU - Marten, Mark R. AU - Martens, Sascha AU - Martin, Alexandre P.J. AU - Martin, Katie R. AU - Martin, Sara AU - Martin, Shaun AU - Martín-Segura, Adrián AU - Martín-Acebes, Miguel A. AU - Martin-Burriel, Inmaculada AU - Martin-Rincon, Marcos AU - Martin-Sanz, Paloma AU - Martina, José A. AU - Martinet, Wim AU - Martinez, Aitor AU - Martinez, Ana AU - Martinez, Jennifer AU - Martinez Velazquez, Moises AU - Martinez-Lopez, Nuria AU - Martinez-Vicente, Marta AU - Martins, Daniel O. AU - Martins, Joilson O. AU - Martins, Waleska K. AU - Martins-Marques, Tania AU - Marzetti, Emanuele AU - Masaldan, Shashank AU - Masclaux-Daubresse, Celine AU - Mashek, Douglas G. AU - Massa, Valentina AU - Massieu, Lourdes AU - Masson, Glenn R. AU - Masuelli, Laura AU - Masyuk, Anatoliy I. AU - Masyuk, Tetyana V. AU - Matarrese, Paola AU - Matheu, Ander AU - Matoba, Satoaki AU - Matsuzaki, Sachiko AU - Mattar, Pamela AU - Matte, Alessandro AU - Mattoscio, Domenico AU - Mauriz, José L. AU - Mauthe, Mario AU - Mauvezin, Caroline AU - Maverakis, Emanual AU - Maycotte, Paola AU - Mayer, Johanna AU - Mazzoccoli, Gianluigi AU - Mazzoni, Cristina AU - Mazzulli, Joseph R. AU - Mccarty, Nami AU - Mcdonald, Christine AU - Mcgill, Mitchell R. AU - Mckenna, Sharon L. AU - Mclaughlin, Beth Ann AU - Mcloughlin, Fionn AU - Mcniven, Mark A. AU - Mcwilliams, Thomas G. AU - Mechta-Grigoriou, Fatima AU - Medeiros, Tania Catarina AU - Medina, Diego L. AU - Megeney, Lynn A. AU - Megyeri, Klara AU - Mehrpour, Maryam AU - Mehta, Jawahar L. AU - Meijer, Alfred J. AU - Meijer, Annemarie H. AU - Mejlvang, Jakob AU - Meléndez, Alicia AU - Melk, Annette AU - Memisoglu, Gonen AU - Mendes, Alexandrina F. AU - Meng, Delong AU - Meng, Fei AU - Meng, Tian AU - Menna-Barreto, Rubem AU - Menon, Manoj B. AU - Mercer, Carol AU - Mercier, Anne E. AU - Mergny, Jean Louis AU - Merighi, Adalberto AU - Merkley, Seth D. AU - Merla, Giuseppe AU - Meske, Volker AU - Mestre, Ana Cecilia AU - Metur, Shree Padma AU - Meyer, Christian AU - Meyer, Hemmo AU - Mi, Wenyi AU - Mialet-Perez, Jeanne AU - Miao, Junying AU - Micale, Lucia AU - Miki, Yasuo AU - Milan, Enrico AU - Milczarek, Małgorzata AU - Miller, Dana L. AU - Miller, Samuel I. AU - Miller, Silke AU - Millward, Steven W. AU - Milosevic, Ira AU - Minina, Elena A. AU - Mirzaei, Hamed AU - Mirzaei, Hamid Reza AU - Mirzaei, Mehdi AU - Mishra, Amit AU - Mishra, Nandita AU - Mishra, Paras Kumar AU - Misirkic Marjanovic, Maja AU - Misasi, Roberta AU - Misra, Amit AU - Misso, Gabriella AU - Mitchell, Claire AU - Mitou, Geraldine AU - Miura, Tetsuji AU - Miyamoto, Shigeki AU - Miyazaki, Makoto AU - Miyazaki, Mitsunori AU - Miyazaki, Taiga AU - Miyazawa, Keisuke AU - Mizushima, Noboru AU - Mogensen, Trine H. AU - Mograbi, Baharia AU - Mohammadinejad, Reza AU - Mohamud, Yasir AU - Mohanty, Abhishek AU - Mohapatra, Sipra AU - Möhlmann, Torsten AU - Mohmmed, Asif AU - Moles, Anna AU - Moley, Kelle H. AU - Molinari, Maurizio AU - Mollace, Vincenzo AU - Møller, Andreas Buch AU - Mollereau, Bertrand AU - Mollinedo, Faustino AU - Montagna, Costanza AU - Monteiro, Mervyn J. AU - Montella, Andrea AU - Montes, L. Ruth AU - Montico, Barbara AU - Mony, Vinod K. AU - Monzio Compagnoni, Giacomo AU - Moore, Michael N. AU - Moosavi, Mohammad A. AU - Mora, Ana L. AU - Mora, Marina AU - Morales-Alamo, David AU - Moratalla, Rosario AU - Moreira, Paula I. AU - Morelli, Elena AU - Moreno, Sandra AU - Moreno-Blas, Daniel AU - Moresi, Viviana AU - Morga, Benjamin AU - Morgan, Alwena H. AU - Morin, Fabrice AU - Morishita, Hideaki AU - Moritz, Orson L. AU - Moriyama, Mariko AU - Moriyasu, Yuji AU - Morleo, Manuela AU - Morselli, Eugenia AU - Moruno-Manchon, Jose F. AU - Moscat, Jorge AU - Mostowy, Serge AU - Motori, Elisa AU - Moura, Andrea Felinto AU - Moustaid-Moussa, Naima AU - Mrakovcic, Maria AU - Muciño-Hernández, Gabriel AU - Mukherjee, Anupam AU - Mukhopadhyay, Subhadip AU - Mulcahy Levy, Jean M. AU - Mulero, Victoriano AU - Muller, Sylviane AU - Münch, Christian AU - Munjal, Ashok AU - Munoz-Canoves, Pura AU - Muñoz-Galdeano, Teresa AU - Münz, Christian AU - Murakawa, Tomokazu AU - Muratori, Claudia AU - Murphy, Brona M. AU - Murphy, J. Patrick AU - Murthy, Aditya AU - Myöhänen, Timo T. AU - Mysorekar, Indira U. AU - Mytych, Jennifer AU - Nabavi, Seyed Mohammad AU - Nabissi, Massimo AU - Nagy, Péter AU - Nah, Jihoon AU - Nahimana, Aimable AU - Nakagawa, Ichiro AU - Nakamura, Ken AU - Nakatogawa, Hitoshi AU - Nandi, Shyam S. AU - Nanjundan, Meera AU - Nanni, Monica AU - Napolitano, Gennaro AU - Nardacci, Roberta AU - Narita, Masashi AU - Nassif, Melissa AU - Nathan, Ilana AU - Natsumeda, Manabu AU - Naude, Ryno J. AU - Naumann, Christin AU - Naveiras, Olaia AU - Navid, Fatemeh AU - Nawrocki, Steffan T. AU - Nazarko, Taras Y. AU - Nazio, Francesca AU - Negoita, Florentina AU - Neill, Thomas AU - Neisch, Amanda L. AU - Neri, Luca M. AU - Netea, Mihai G. AU - Neubert, Patrick AU - Neufeld, Thomas P. AU - Neumann, Dietbert AU - Neutzner, Albert AU - Newton, Phillip T. AU - Ney, Paul A. AU - Nezis, Ioannis P. AU - Ng, Charlene C.W. AU - Ng, Tzi Bun AU - Nguyen, Hang T.T. AU - Nguyen, Long T. AU - Ni, Hong Min AU - Ní Cheallaigh, Clíona AU - Ni, Zhenhong AU - Nicolao, M. Celeste AU - Nicoli, Francesco AU - Nieto-Diaz, Manuel AU - Nilsson, Per AU - Ning, Shunbin AU - Niranjan, Rituraj AU - Nishimune, Hiroshi AU - Niso-Santano, Mireia AU - Nixon, Ralph A. AU - Nobili, Annalisa AU - Nobrega, Clevio AU - Noda, Takeshi AU - Nogueira-Recalde, Uxía AU - Nolan, Trevor M. AU - Nombela, Ivan AU - Novak, Ivana AU - Novoa, Beatriz AU - Nozawa, Takashi AU - Nukina, Nobuyuki AU - Nussbaum-Krammer, Carmen AU - Nylandsted, Jesper AU - O’Donovan, Tracey R. AU - O’Leary, Seónadh M. AU - O’Rourke, Eyleen J. AU - O’Sullivan, Mary P. AU - O’Sullivan, Timothy E. AU - Oddo, Salvatore AU - Oehme, Ina AU - Ogawa, Michinaga AU - Ogier-Denis, Eric AU - Ogmundsdottir, Margret H. AU - Ogretmen, Besim AU - Oh, Goo Taeg AU - Oh, Seon Hee AU - Oh, Young J. AU - Ohama, Takashi AU - Ohashi, Yohei AU - Ohmuraya, Masaki AU - Oikonomou, Vasileios AU - Ojha, Rani AU - Okamoto, Koji AU - Okazawa, Hitoshi AU - Oku, Masahide AU - Oliván, Sara AU - Oliveira, Jorge M.A. AU - Ollmann, Michael AU - Olzmann, James A. AU - Omari, Shakib AU - Omary, M. Bishr AU - Önal, Gizem AU - Ondrej, Martin AU - Ong, Sang Bing AU - Ong, Sang Ging AU - Onnis, Anna AU - Orellana, Juan A. AU - Orellana-Muñoz, Sara AU - Ortega-Villaizan, Maria Del Mar AU - Ortiz-Gonzalez, Xilma R. AU - Ortona, Elena AU - Osiewacz, Heinz D. AU - Osman, Abdel Hamid K. AU - Osta, Rosario AU - Otegui, Marisa S. AU - Otsu, Kinya AU - Ott, Christiane AU - Ottobrini, Luisa AU - Ou, Jing Hsiung James AU - Outeiro, Tiago F. AU - Oynebraten, Inger AU - Ozturk, Melek AU - Pagès, Gilles AU - Pahari, Susanta AU - Pajares, Marta AU - Pajvani, Utpal B. AU - Pal, Rituraj AU - Paladino, Simona AU - Pallet, Nicolas AU - Palmieri, Michela AU - Palmisano, Giuseppe AU - Palumbo, Camilla AU - Pampaloni, Francesco AU - Pan, Lifeng AU - Pan, Qingjun AU - Pan, Wenliang AU - Pan, Xin AU - Panasyuk, Ganna AU - Pandey, Rahul AU - Pandey, Udai B. AU - Pandya, Vrajesh AU - Paneni, Francesco AU - Pang, Shirley Y. AU - Panzarini, Elisa AU - Papademetrio, Daniela L. AU - Papaleo, Elena AU - Papinski, Daniel AU - Papp, Diana AU - Park, Eun Chan AU - Park, Hwan Tae AU - Park, Ji Man AU - Park, Jong In AU - Park, Joon Tae AU - Park, Junsoo AU - Park, Sang Chul AU - Park, Sang Youel AU - Parola, Abraham H. AU - Parys, Jan B. AU - Pasquier, Adrien AU - Pasquier, Benoit AU - Passos, João F. AU - Pastore, Nunzia AU - Patel, Hemal H. AU - Patschan, Daniel AU - Pattingre, Sophie AU - Pedraza-Alva, Gustavo AU - Pedraza-Chaverri, Jose AU - Pedrozo, Zully AU - Pei, Gang AU - Pei, Jianming AU - Peled-Zehavi, Hadas AU - Pellegrini, Joaquín M. AU - Pelletier, Joffrey AU - Peñalva, Miguel A. AU - Peng, Di AU - Peng, Ying AU - Penna, Fabio AU - Pennuto, Maria AU - Pentimalli, Francesca AU - Pereira, Cláudia M.F. AU - Pereira, Gustavo J.S. AU - Pereira, Lilian C. AU - Pereira De Almeida, Luis AU - Perera, Nirma D. AU - Pérez-Lara, Ángel AU - Perez-Oliva, Ana B. AU - Pérez-Pérez, María Esther AU - Periyasamy, Palsamy AU - Perl, Andras AU - Perrotta, Cristiana AU - Perrotta, Ida AU - Pestell, Richard G. AU - Petersen, Morten AU - Petrache, Irina AU - Petrovski, Goran AU - Pfirrmann, Thorsten AU - Pfister, Astrid S. AU - Philips, Jennifer A. AU - Pi, Huifeng AU - Picca, Anna AU - Pickrell, Alicia M. AU - Picot, Sandy AU - Pierantoni, Giovanna M. AU - Pierdominici, Marina AU - Pierre, Philippe AU - Pierrefite-Carle, Valérie AU - Pierzynowska, Karolina AU - Pietrocola, Federico AU - Pietruczuk, Miroslawa AU - Pignata, Claudio AU - Pimentel-Muiños, Felipe X. AU - Pinar, Mario AU - Pinheiro, Roberta O. AU - Pinkas-Kramarski, Ronit AU - Pinton, Paolo AU - Pircs, Karolina AU - Piya, Sujan AU - Pizzo, Paola AU - Plantinga, Theo S. AU - Platta, Harald W. AU - Plaza-Zabala, Ainhoa AU - Plomann, Markus AU - Plotnikov, Egor Y. AU - Plun-Favreau, Helene AU - Pluta, Ryszard AU - Pocock, Roger AU - Pöggeler, Stefanie AU - Pohl, Christian AU - Poirot, Marc AU - Poletti, Angelo AU - Ponpuak, Marisa AU - Popelka, Hana AU - Popova, Blagovesta AU - Porta, Helena AU - Porte Alcon, Soledad AU - Portilla-Fernandez, Eliana AU - Post, Martin AU - Potts, Malia B. AU - Poulton, Joanna AU - Powers, Ted AU - Prahlad, Veena AU - Prajsnar, Tomasz K. AU - Praticò, Domenico AU - Prencipe, Rosaria AU - Priault, Muriel AU - Proikas-Cezanne, Tassula AU - Promponas, Vasilis J. AU - Proud, Christopher G. AU - Puertollano, Rosa AU - Puglielli, Luigi AU - Pulinilkunnil, Thomas AU - Puri, Deepika AU - Puri, Rajat AU - Puyal, Julien AU - Qi, Xiaopeng AU - Qi, Yongmei AU - Qian, Wenbin AU - Qiang, Lei AU - Qiu, Yu AU - Quadrilatero, Joe AU - Quarleri, Jorge AU - Raben, Nina AU - Rabinowich, Hannah AU - Ragona, Debora AU - Ragusa, Michael J. AU - Rahimi, Nader AU - Rahmati, Marveh AU - Raia, Valeria AU - Raimundo, Nuno AU - Rajasekaran, Namakkal Soorappan AU - Ramachandra Rao, Sriganesh AU - Rami, Abdelhaq AU - Ramírez-Pardo, Ignacio AU - Ramsden, David B. AU - Randow, Felix AU - Rangarajan, Pundi N. AU - Ranieri, Danilo AU - Rao, Hai AU - Rao, Lang AU - Rao, Rekha AU - Rathore, Sumit AU - Ratnayaka, J. Arjuna AU - Ratovitski, Edward A. AU - Ravanan, Palaniyandi AU - Ravegnini, Gloria AU - Ray, Swapan K. AU - Razani, Babak AU - Rebecca, Vito AU - Reggiori, Fulvio AU - Régnier-Vigouroux, Anne AU - Reichert, Andreas S. AU - Reigada, David AU - Reiling, Jan H. AU - Rein, Theo AU - Reipert, Siegfried AU - Rekha, Rokeya Sultana AU - Ren, Hongmei AU - Ren, Jun AU - Ren, Weichao AU - Renault, Tristan AU - Renga, Giorgia AU - Reue, Karen AU - Rewitz, Kim AU - Ribeiro De Andrade Ramos, Bruna AU - Riazuddin, S. Amer AU - Ribeiro-Rodrigues, Teresa M. AU - Ricci, Jean Ehrland AU - Ricci, Romeo AU - Riccio, Victoria AU - Richardson, Des R. AU - Rikihisa, Yasuko AU - Risbud, Makarand V. AU - Risueño, Ruth M. AU - Ritis, Konstantinos AU - Rizza, Salvatore AU - Rizzuto, Rosario AU - Roberts, Helen C. AU - Roberts, Luke D. AU - Robinson, Katherine J. AU - Roccheri, Maria Carmela AU - Rocchi, Stephane AU - Rodney, George G. AU - Rodrigues, Tiago AU - Rodrigues Silva, Vagner Ramon AU - Rodriguez, Amaia AU - Rodriguez-Barrueco, Ruth AU - Rodriguez-Henche, Nieves AU - Rodriguez-Rocha, Humberto AU - Roelofs, Jeroen AU - Rogers, Robert S. AU - Rogov, Vladimir V. AU - Rojo, Ana I. AU - Rolka, Krzysztof AU - Romanello, Vanina AU - Romani, Luigina AU - Romano, Alessandra AU - Romano, Patricia S. AU - Romeo-Guitart, David AU - Romero, Luis C. AU - Romero, Montserrat AU - Roney, Joseph C. AU - Rongo, Christopher AU - Roperto, Sante AU - Rosenfeldt, Mathias T. AU - Rosenstiel, Philip AU - Rosenwald, Anne G. AU - Roth, Kevin A. AU - Roth, Lynn AU - Roth, Steven AU - Rouschop, Kasper M.A. AU - Roussel, Benoit D. AU - Roux, Sophie AU - Rovere-Querini, Patrizia AU - Roy, Ajit AU - Rozieres, Aurore AU - Ruano, Diego AU - Rubinsztein, David C. AU - Rubtsova, Maria P. AU - Ruckdeschel, Klaus AU - Ruckenstuhl, Christoph AU - Rudolf, Emil AU - Rudolf, Rüdiger AU - Ruggieri, Alessandra AU - Ruparelia, Avnika Ashok AU - Rusmini, Paola AU - Russell, Ryan R. AU - Russo, Gian Luigi AU - Russo, Maria AU - Russo, Rossella AU - Ryabaya, Oxana O. AU - Ryan, Kevin M. AU - Ryu, Kwon Yul AU - Sabater-Arcis, Maria AU - Sachdev, Ulka AU - Sacher, Michael AU - Sachse, Carsten AU - Sadhu, Abhishek AU - Sadoshima, Junichi AU - Safren, Nathaniel AU - Saftig, Paul AU - Sagona, Antonia P. AU - Sahay, Gaurav AU - Sahebkar, Amirhossein AU - Sahin, Mustafa AU - Sahin, Ozgur AU - Sahni, Sumit AU - Saito, Nayuta AU - Saito, Shigeru AU - Saito, Tsunenori AU - Sakai, Ryohei AU - Sakai, Yasuyoshi AU - Sakamaki, Jun Ichi AU - Saksela, Kalle AU - Salazar, Gloria AU - Salazar-Degracia, Anna AU - Salekdeh, Ghasem H. AU - Saluja, Ashok K. AU - Sampaio-Marques, Belém AU - Sanchez, Maria Cecilia AU - Sanchez-Alcazar, Jose A. AU - Sanchez-Vera, Victoria AU - Sancho-Shimizu, Vanessa AU - Sanderson, J. Thomas AU - Sandri, Marco AU - Santaguida, Stefano AU - Santambrogio, Laura AU - Santana, Magda M. AU - Santoni, Giorgio AU - Sanz, Alberto AU - Sanz, Pascual AU - Saran, Shweta AU - Sardiello, Marco AU - Sargeant, Timothy J. AU - Sarin, Apurva AU - Sarkar, Chinmoy AU - Sarkar, Sovan AU - Sarrias, Maria Rosa AU - Sarkar, Surajit AU - Sarmah, Dipanka Tanu AU - Sarparanta, Jaakko AU - Sathyanarayan, Aishwarya AU - Sathyanarayanan, Ranganayaki AU - Scaglione, K. Matthew AU - Scatozza, Francesca AU - Schaefer, Liliana AU - Schafer, Zachary T. AU - Schaible, Ulrich E. AU - Schapira, Anthony H.V. AU - Scharl, Michael AU - Schatzl, Hermann M. AU - Schein, Catherine H. AU - Scheper, Wiep AU - Scheuring, David AU - Schiaffino, Maria Vittoria AU - Schiappacassi, Monica AU - Schindl, Rainer AU - Schlattner, Uwe AU - Schmidt, Oliver AU - Schmitt, Roland AU - Schmidt, Stephen D. AU - Schmitz, Ingo AU - Schmukler, Eran AU - Schneider, Anja AU - Schneider, Bianca E. AU - Schober, Romana AU - Schoijet, Alejandra C. AU - Schott, Micah B. AU - Schramm, Michael AU - Schröder, Bernd AU - Schuh, Kai AU - Schüller, Christoph AU - Schulze, Ryan J. AU - Schürmanns, Lea AU - Schwamborn, Jens C. AU - Schwarten, Melanie AU - Scialo, Filippo AU - Sciarretta, Sebastiano AU - Scott, Melanie J. AU - Scotto, Kathleen W. AU - Scovassi, A. Ivana AU - Scrima, Andrea AU - Scrivo, Aurora AU - Sebastian, David AU - Sebti, Salwa AU - Sedej, Simon AU - Segatori, Laura AU - Segev, Nava AU - Seglen, Per O. AU - Seiliez, Iban AU - Seki, Ekihiro AU - Selleck, Scott B. AU - Sellke, Frank W. AU - Selsby, Joshua T. AU - Sendtner, Michael AU - Senturk, Serif AU - Seranova, Elena AU - Sergi, Consolato AU - Serra-Moreno, Ruth AU - Sesaki, Hiromi AU - Settembre, Carmine AU - Setty, Subba Rao Gangi AU - Sgarbi, Gianluca AU - Sha, Ou AU - Shacka, John J. AU - Shah, Javeed A. AU - Shang, Dantong AU - Shao, Changshun AU - Shao, Feng AU - Sharbati, Soroush AU - Sharkey, Lisa M. AU - Sharma, Dipali AU - Sharma, Gaurav AU - Sharma, Kulbhushan AU - Sharma, Pawan AU - Sharma, Surendra AU - Shen, Han Ming AU - Shen, Hongtao AU - Shen, Jiangang AU - Shen, Ming AU - Shen, Weili AU - Shen, Zheni AU - Sheng, Rui AU - Sheng, Zhi AU - Sheng, Zu Hang AU - Shi, Jianjian AU - Shi, Xiaobing AU - Shi, Ying Hong AU - Shiba-Fukushima, Kahori AU - Shieh, Jeng Jer AU - Shimada, Yohta AU - Shimizu, Shigeomi AU - Shimozawa, Makoto AU - Shintani, Takahiro AU - Shoemaker, Christopher J. AU - Shojaei, Shahla AU - Shoji, Ikuo AU - Shravage, Bhupendra V. AU - Shridhar, Viji AU - Shu, Chih Wen AU - Shu, Hong Bing AU - Shui, Ke AU - Shukla, Arvind K. AU - Shutt, Timothy E. AU - Sica, Valentina AU - Siddiqui, Aleem AU - Sierra, Amanda AU - Sierra-Torre, Virginia AU - Signorelli, Santiago AU - Sil, Payel AU - Silva, Bruno J.De Andrade AU - Silva, Johnatas D. AU - Silva-Pavez, Eduardo AU - Silvente-Poirot, Sandrine AU - Simmonds, Rachel E. AU - Simon, Anna Katharina AU - Simon, Hans Uwe AU - Simons, Matias AU - Singh, Anurag AU - Singh, Lalit P. AU - Singh, Rajat AU - Singh, Shivendra V. AU - Singh, Shrawan K. AU - Singh, Sudha B. AU - Singh, Sunaina AU - Singh, Surinder Pal AU - Sinha, Debasish AU - Sinha, Rohit Anthony AU - Sinha, Sangita AU - Sirko, Agnieszka AU - Sirohi, Kapil AU - Sivridis, Efthimios L. AU - Skendros, Panagiotis AU - Skirycz, Aleksandra AU - Slaninová, Iva AU - Smaili, Soraya S. AU - Smertenko, Andrei AU - Smith, Matthew D. AU - Soenen, Stefaan J. AU - Sohn, Eun Jung AU - Sok, Sophia P.M. AU - Solaini, Giancarlo AU - Soldati, Thierry AU - Soleimanpour, Scott A. AU - Soler, Rosa M. AU - Solovchenko, Alexei AU - Somarelli, Jason A. AU - Sonawane, Avinash AU - Song, Fuyong AU - Song, Hyun Kyu AU - Song, Ju Xian AU - Song, Kunhua AU - Song, Zhiyin AU - Soria, Leandro R. AU - Sorice, Maurizio AU - Soukas, Alexander A. AU - Soukup, Sandra Fausia AU - Sousa, Diana AU - Sousa, Nadia AU - Spagnuolo, Paul A. AU - Spector, Stephen A. AU - Srinivas Bharath, M. M. AU - St. Clair, Daret AU - Stagni, Venturina AU - Staiano, Leopoldo AU - Stalnecker, Clint A. AU - Stankov, Metodi V. AU - Stathopulos, Peter B. AU - Stefan, Katja AU - Stefan, Sven Marcel AU - Stefanis, Leonidas AU - Steffan, Joan S. AU - Steinkasserer, Alexander AU - Stenmark, Harald AU - Sterneckert, Jared AU - Stevens, Craig AU - Stoka, Veronika AU - Storch, Stephan AU - Stork, Björn AU - Strappazzon, Flavie AU - Strohecker, Anne Marie AU - Stupack, Dwayne G. AU - Su, Huanxing AU - Su, Ling Yan AU - Su, Longxiang AU - Suarez-Fontes, Ana M. AU - Subauste, Carlos S. AU - Subbian, Selvakumar AU - Subirada, Paula V. AU - Sudhandiran, Ganapasam AU - Sue, Carolyn M. AU - Sui, Xinbing AU - Summers, Corey AU - Sun, Guangchao AU - Sun, Jun AU - Sun, Kang AU - Sun, Meng Xiang AU - Sun, Qiming AU - Sun, Yi AU - Sun, Zhongjie AU - Sunahara, Karen K.S. AU - Sundberg, Eva AU - Susztak, Katalin AU - Sutovsky, Peter AU - Suzuki, Hidekazu AU - Sweeney, Gary AU - Symons, J. David AU - Sze, Stephen Cho Wing AU - Szewczyk, Nathaniel J. AU - Tabęcka-Łonczynska, Anna AU - Tabolacci, Claudio AU - Tacke, Frank AU - Taegtmeyer, Heinrich AU - Tafani, Marco AU - Tagaya, Mitsuo AU - Tai, Haoran AU - Tait, Stephen W.G. AU - Takahashi, Yoshinori AU - Takats, Szabolcs AU - Talwar, Priti AU - Tam, Chit AU - Tam, Shing Yau AU - Tampellini, Davide AU - Tamura, Atsushi AU - Tan, Chong Teik AU - Tan, Eng King AU - Tan, Ya Qin AU - Tanaka, Masaki AU - Tanaka, Motomasa AU - Tang, Daolin AU - Tang, Jingfeng AU - Tang, Tie Shan AU - Tanida, Isei AU - Tao, Zhipeng AU - Taouis, Mohammed AU - Tatenhorst, Lars AU - Tavernarakis, Nektarios AU - Taylor, Allen AU - Taylor, Gregory A. AU - Taylor, Joan M. AU - Tchetina, Elena AU - Tee, Andrew R. AU - Tegeder, Irmgard AU - Teis, David AU - Teixeira, Natercia AU - Teixeira-Clerc, Fatima AU - Tekirdag, Kumsal A. AU - Tencomnao, Tewin AU - Tenreiro, Sandra AU - Tepikin, Alexei V. AU - Testillano, Pilar S. AU - Tettamanti, Gianluca AU - Tharaux, Pierre Louis AU - Thedieck, Kathrin AU - Thekkinghat, Arvind A. AU - Thellung, Stefano AU - Thinwa, Josephine W. AU - Thirumalaikumar, V. P. AU - Thomas, Sufi Mary AU - Thomes, Paul G. AU - Thorburn, Andrew AU - Thukral, Lipi AU - Thum, Thomas AU - Thumm, Michael AU - Tian, Ling AU - Tichy, Ales AU - Till, Andreas AU - Timmerman, Vincent AU - Titorenko, Vladimir I. AU - Todi, Sokol V. AU - Todorova, Krassimira AU - Toivonen, Janne M. AU - Tomaipitinca, Luana AU - Tomar, Dhanendra AU - Tomas-Zapico, Cristina AU - Tomić, Sergej AU - Tong, Benjamin Chun Kit AU - Tong, Chao AU - Tong, Xin AU - Tooze, Sharon A. AU - Torgersen, Maria L. AU - Torii, Satoru AU - Torres-López, Liliana AU - Torriglia, Alicia AU - Towers, Christina G. AU - Towns, Roberto AU - Toyokuni, Shinya AU - Trajkovic, Vladimir AU - Tramontano, Donatella AU - Tran, Quynh Giao AU - Travassos, Leonardo H. AU - Trelford, Charles B. AU - Tremel, Shirley AU - Trougakos, Ioannis P. AU - Tsao, Betty P. AU - Tschan, Mario P. AU - Tse, Hung Fat AU - Tse, Tak Fu AU - Tsugawa, Hitoshi AU - Tsvetkov, Andrey S. AU - Tumbarello, David A. AU - Tumtas, Yasin AU - Tuñón, María J. AU - Turcotte, Sandra AU - Turk, Boris AU - Turk, Vito AU - Turner, Bradley J. AU - Tuxworth, Richard I. AU - Tyler, Jessica K. AU - Tyutereva, Elena V. AU - Uchiyama, Yasuo AU - Ugun-Klusek, Aslihan AU - Uhlig, Holm H. AU - Ułamek-Kozioł, Marzena AU - Ulasov, Ilya V. AU - Umekawa, Midori AU - Ungermann, Christian AU - Unno, Rei AU - Urbe, Sylvie AU - Uribe-Carretero, Elisabet AU - Üstün, Suayib AU - Uversky, Vladimir N. AU - Vaccari, Thomas AU - Vaccaro, Maria I. AU - Vahsen, Björn F. AU - Vakifahmetoglu-Norberg, Helin AU - Valdor, Rut AU - Valente, Maria J. AU - Valko, Ayelén AU - Vallee, Richard B. AU - Valverde, Angela M. AU - Van Den Berghe, Greet AU - Van Der Veen, Stijn AU - Van Kaer, Luc AU - Van Loosdregt, Jorg AU - Van Wijk, Sjoerd J.L. AU - Vandenberghe, Wim AU - Vanhorebeek, Ilse AU - Vannier-Santos, Marcos A. AU - Vannini, Nicola AU - Vanrell, M. Cristina AU - Vantaggiato, Chiara AU - Varano, Gabriele AU - Varela-Nieto, Isabel AU - Varga, Máté AU - Vasconcelos, M. Helena AU - Vats, Somya AU - Vavvas, Demetrios G. AU - Vega-Naredo, Ignacio AU - Vega-Rubin-De-Celis, Silvia AU - Velasco, Guillermo AU - Velázquez, Ariadna P. AU - Vellai, Tibor AU - Vellenga, Edo AU - Velotti, Francesca AU - Verdier, Mireille AU - Verginis, Panayotis AU - Vergne, Isabelle AU - Verkade, Paul AU - Verma, Manish AU - Verstreken, Patrik AU - Vervliet, Tim AU - Vervoorts, Jörg AU - Vessoni, Alexandre T. AU - Victor, Victor M. AU - Vidal, Michel AU - Vidoni, Chiara AU - Vieira, Otilia V. AU - Vierstra, Richard D. AU - Viganó, Sonia AU - Vihinen, Helena AU - Vijayan, Vinoy AU - Vila, Miquel AU - Vilar, Marçal AU - Villalba, José M. AU - Villalobo, Antonio AU - Villarejo-Zori, Beatriz AU - Villarroya, Francesc AU - Villarroya, Joan AU - Vincent, Olivier AU - Vindis, Cecile AU - Viret, Christophe AU - Viscomi, Maria Teresa AU - Visnjic, Dora AU - Vitale, Ilio AU - Vocadlo, David J. AU - Voitsekhovskaja, Olga V. AU - Volonté, Cinzia AU - Volta, Mattia AU - Vomero, Marta AU - Von Haefen, Clarissa AU - Vooijs, Marc A. AU - Voos, Wolfgang AU - Vucicevic, Ljubica AU - Wade-Martins, Richard AU - Waguri, Satoshi AU - Waite, Kenrick A. AU - Wakatsuki, Shuji AU - Walker, David W. AU - Walker, Mark J. AU - Walker, Simon A. AU - Walter, Jochen AU - Wandosell, Francisco G. AU - Wang, Bo AU - Wang, Chao Yung AU - Wang, Chen AU - Wang, Chenran AU - Wang, Chenwei AU - Wang, Cun Yu AU - Wang, Dong AU - Wang, Fangyang AU - Wang, Feng AU - Wang, Fengming AU - Wang, Guansong AU - Wang, Han AU - Wang, Hao AU - Wang, Hexiang AU - Wang, Hong Gang AU - Wang, Jianrong AU - Wang, Jigang AU - Wang, Jiou AU - Wang, Jundong AU - Wang, Kui AU - Wang, Lianrong AU - Wang, Liming AU - Wang, Maggie Haitian AU - Wang, Meiqing AU - Wang, Nanbu AU - Wang, Pengwei AU - Wang, Peipei AU - Wang, Ping AU - Wang, Ping AU - Wang, Qing Jun AU - Wang, Qing AU - Wang, Qing Kenneth AU - Wang, Qiong A. AU - Wang, Wen Tao AU - Wang, Wuyang AU - Wang, Xinnan AU - Wang, Xuejun AU - Wang, Yan AU - Wang, Yanchang AU - Wang, Yanzhuang AU - Wang, Yen Yun AU - Wang, Yihua AU - Wang, Yipeng AU - Wang, Yu AU - Wang, Yuqi AU - Wang, Zhe AU - Wang, Zhenyu AU - Wang, Zhouguang AU - Warnes, Gary AU - Warnsmann, Verena AU - Watada, Hirotaka AU - Watanabe, Eizo AU - Watchon, Maxinne AU - Wawrzyńska, Anna AU - Weaver, Timothy E. AU - Wegrzyn, Grzegorz AU - Wehman, Ann M. AU - Wei, Huafeng AU - Wei, Lei AU - Wei, Taotao AU - Wei, Yongjie AU - Weiergräber, Oliver H. AU - Weihl, Conrad C. AU - Weindl, Günther AU - Weiskirchen, Ralf AU - Wells, Alan AU - Wen, Runxia H. AU - Wen, Xin AU - Werner, Antonia AU - Weykopf, Beatrice AU - Wheatley, Sally P. AU - Whitton, J. Lindsay AU - Whitworth, Alexander J. AU - Wiktorska, Katarzyna AU - Wildenberg, Manon E. AU - Wileman, Tom AU - Wilkinson, Simon AU - Willbold, Dieter AU - Williams, Brett AU - Williams, Robin S.B. AU - Williams, Roger L. AU - Williamson, Peter R. AU - Wilson, Richard A. AU - Winner, Beate AU - Winsor, Nathaniel J. AU - Witkin, Steven S. AU - Wodrich, Harald AU - Woehlbier, Ute AU - Wollert, Thomas AU - Wong, Esther AU - Wong, Jack Ho AU - Wong, Richard W. AU - Wong, Vincent Kam Wai AU - Wong, W. Wei Lynn AU - Wu, An Guo AU - Wu, Chengbiao AU - Wu, Jian AU - Wu, Junfang AU - Wu, Kenneth K. AU - Wu, Min AU - Wu, Shan Ying AU - Wu, Shengzhou AU - Wu, Shu Yan AU - Wu, Shufang AU - Wu, William K.K. AU - Wu, Xiaohong AU - Wu, Xiaoqing AU - Wu, Yao Wen AU - Wu, Yihua AU - Xavier, Ramnik J. AU - Xia, Hongguang AU - Xia, Lixin AU - Xia, Zhengyuan AU - Xiang, Ge AU - Xiang, Jin AU - Xiang, Mingliang AU - Xiang, Wei AU - Xiao, Bin AU - Xiao, Guozhi AU - Xiao, Hengyi AU - Xiao, Hong Tao AU - Xiao, Jian AU - Xiao, Lan AU - Xiao, Shi AU - Xiao, Yin AU - Xie, Baoming AU - Xie, Chuan Ming AU - Xie, Min AU - Xie, Yuxiang AU - Xie, Zhiping AU - Xie, Zhonglin AU - Xilouri, Maria AU - Xu, Congfeng AU - Xu, En AU - Xu, Haoxing AU - Xu, Jing AU - Xu, Jin Rong AU - Xu, Liang AU - Xu, Wen Wen AU - Xu, Xiulong AU - Xue, Yu AU - Yakhine-Diop, Sokhna M.S. AU - Yamaguchi, Masamitsu AU - Yamaguchi, Osamu AU - Yamamoto, Ai AU - Yamashina, Shunhei AU - Yan, Shengmin AU - Yan, Shian Jang AU - Yan, Zhen AU - Yanagi, Yasuo AU - Yang, Chuanbin AU - Yang, Dun Sheng AU - Yang, Huan AU - Yang, Huang Tian AU - Yang, Hui AU - Yang, Jin Ming AU - Yang, Jing AU - Yang, Jingyu AU - Yang, Ling AU - Yang, Liu AU - Yang, Ming AU - Yang, Pei Ming AU - Yang, Qian AU - Yang, Seungwon AU - Yang, Shu AU - Yang, Shun Fa AU - Yang, Wannian AU - Yang, Wei Yuan AU - Yang, Xiaoyong AU - Yang, Xuesong AU - Yang, Yi AU - Yang, Ying AU - Yao, Honghong AU - Yao, Shenggen AU - Yao, Xiaoqiang AU - Yao, Yong Gang AU - Yao, Yong Ming AU - Yasui, Takahiro AU - Yazdankhah, Meysam AU - Yen, Paul M. AU - Yi, Cong AU - Yin, Xiao Ming AU - Yin, Yanhai AU - Yin, Zhangyuan AU - Yin, Ziyi AU - Ying, Meidan AU - Ying, Zheng AU - Yip, Calvin K. AU - Yiu, Stephanie Pei Tung AU - Yoo, Young H. AU - Yoshida, Kiyotsugu AU - Yoshii, Saori R. AU - Yoshimori, Tamotsu AU - Yousefi, Bahman AU - Yu, Boxuan AU - Yu, Haiyang AU - Yu, Jun AU - Yu, Jun AU - Yu, Li AU - Yu, Ming Lung AU - Yu, Seong Woon AU - Yu, Victor C. AU - Yu, W. Haung AU - Yu, Zhengping AU - Yu, Zhou AU - Yuan, Junying AU - Yuan, Ling Qing AU - Yuan, Shilin AU - Yuan, Shyng Shiou F. AU - Yuan, Yanggang AU - Yuan, Zengqiang AU - Yue, Jianbo AU - Yue, Zhenyu AU - Yun, Jeanho AU - Yung, Raymond L. AU - Zacks, David N. AU - Zaffagnini, Gabriele AU - Zambelli, Vanessa O. AU - Zanella, Isabella AU - Zang, Qun S. AU - Zanivan, Sara AU - Zappavigna, Silvia AU - Zaragoza, Pilar AU - Zarbalis, Konstantinos S. AU - Zarebkohan, Amir AU - Zarrouk, Amira AU - Zeitlin, Scott O. AU - Zeng, Jialiu AU - Zeng, Ju Deng AU - Žerovnik, Eva AU - Zhan, Lixuan AU - Zhang, Bin AU - Zhang, Donna D. AU - Zhang, Hanlin AU - Zhang, Hong AU - Zhang, Hong AU - Zhang, Honghe AU - Zhang, Huafeng AU - Zhang, Huaye AU - Zhang, Hui AU - Zhang, Hui Ling AU - Zhang, Jianbin AU - Zhang, Jianhua AU - Zhang, Jing Pu AU - Zhang, Kalin Y.B. AU - Zhang, Leshuai W. AU - Zhang, Lin AU - Zhang, Lisheng AU - Zhang, Lu AU - Zhang, Luoying AU - Zhang, Menghuan AU - Zhang, Peng AU - Zhang, Sheng AU - Zhang, Wei AU - Zhang, Xiangnan AU - Zhang, Xiao Wei AU - Zhang, Xiaolei AU - Zhang, Xiaoyan AU - Zhang, Xin AU - Zhang, Xinxin AU - Zhang, Xu Dong AU - Zhang, Yang AU - Zhang, Yanjin AU - Zhang, Yi AU - Zhang, Ying Dong AU - Zhang, Yingmei AU - Zhang, Yuan Yuan AU - Zhang, Yuchen AU - Zhang, Zhe AU - Zhang, Zhengguang AU - Zhang, Zhibing AU - Zhang, Zhihai AU - Zhang, Zhiyong AU - Zhang, Zili AU - Zhao, Haobin AU - Zhao, Lei AU - Zhao, Shuang AU - Zhao, Tongbiao AU - Zhao, Xiao Fan AU - Zhao, Ying AU - Zhao, Yongchao AU - Zhao, Yongliang AU - Zhao, Yuting AU - Zheng, Guoping AU - Zheng, Kai AU - Zheng, Ling AU - Zheng, Shizhong AU - Zheng, Xi Long AU - Zheng, Yi AU - Zheng, Zu Guo AU - Zhivotovsky, Boris AU - Zhong, Qing AU - Zhou, Ao AU - Zhou, Ben AU - Zhou, Cefan AU - Zhou, Gang AU - Zhou, Hao AU - Zhou, Hong AU - Zhou, Hongbo AU - Zhou, Jie AU - Zhou, Jing AU - Zhou, Jing AU - Zhou, Jiyong AU - Zhou, Kailiang AU - Zhou, Rongjia AU - Zhou, Xu Jie AU - Zhou, Yanshuang AU - Zhou, Yinghong AU - Zhou, Yubin AU - Zhou, Zheng Yu AU - Zhou, Zhou AU - Zhu, Binglin AU - Zhu, Changlian AU - Zhu, Guo Qing AU - Zhu, Haining AU - Zhu, Hongxin AU - Zhu, Hua AU - Zhu, Wei Guo AU - Zhu, Yanping AU - Zhu, Yushan AU - Zhuang, Haixia AU - Zhuang, Xiaohong AU - Zientara-Rytter, Katarzyna AU - Zimmermann, Christine M. AU - Ziviani, Elena AU - Zoladek, Teresa AU - Zong, Wei Xing AU - Zorov, Dmitry B. AU - Zorzano, Antonio AU - Zou, Weiping AU - Zou, Zhen AU - Zou, Zhengzhi AU - Zuryn, Steven AU - Zwerschke, Werner AU - Brand-Saberi, Beate AU - Dong, X. Charlie AU - Kenchappa, Chandra Shekar AU - Li, Zuguo AU - Lin, Yong AU - Oshima, Shigeru AU - Rong, Yueguang AU - Sluimer, Judith C. AU - Stallings, Christina L. AU - Tong, Chun Kit ID - 9298 IS - 1 JF - Autophagy SN - 1554-8627 TI - Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition) VL - 17 ER - TY - JOUR AB - Growth regulation tailors development in plants to their environment. A prominent example of this is the response to gravity, in which shoots bend up and roots bend down1. This paradox is based on opposite effects of the phytohormone auxin, which promotes cell expansion in shoots while inhibiting it in roots via a yet unknown cellular mechanism2. Here, by combining microfluidics, live imaging, genetic engineering and phosphoproteomics in Arabidopsis thaliana, we advance understanding of how auxin inhibits root growth. We show that auxin activates two distinct, antagonistically acting signalling pathways that converge on rapid regulation of apoplastic pH, a causative determinant of growth. Cell surface-based TRANSMEMBRANE KINASE1 (TMK1) interacts with and mediates phosphorylation and activation of plasma membrane H+-ATPases for apoplast acidification, while intracellular canonical auxin signalling promotes net cellular H+ influx, causing apoplast alkalinization. Simultaneous activation of these two counteracting mechanisms poises roots for rapid, fine-tuned growth modulation in navigating complex soil environments. AU - Li, Lanxin AU - Verstraeten, Inge AU - Roosjen, Mark AU - Takahashi, Koji AU - Rodriguez Solovey, Lesia AU - Merrin, Jack AU - Chen, Jian AU - Shabala, Lana AU - Smet, Wouter AU - Ren, Hong AU - Vanneste, Steffen AU - Shabala, Sergey AU - De Rybel, Bert AU - Weijers, Dolf AU - Kinoshita, Toshinori AU - Gray, William M. AU - Friml, Jiří ID - 10223 IS - 7884 JF - Nature KW - Multidisciplinary SN - 00280836 TI - Cell surface and intracellular auxin signalling for H+ fluxes in root growth VL - 599 ER - TY - JOUR AB - Transposable elements exist widely throughout plant genomes and play important roles in plant evolution. Auxin is an important regulator that is traditionally associated with root development and drought stress adaptation. The DEEPER ROOTING 1 (DRO1) gene is a key component of rice drought avoidance. Here, we identified a transposon that acts as an autonomous auxin‐responsive promoter and its presence at specific genome positions conveys physiological adaptations related to drought avoidance. Rice varieties with high and auxin‐mediated transcription of DRO1 in the root tip show deeper and longer root phenotypes and are thus better adapted to drought. The INDITTO2 transposon contains an auxin response element and displays auxin‐responsive promoter activity; it is thus able to convey auxin regulation of transcription to genes in its proximity. In the rice Acuce, which displays DRO1‐mediated drought adaptation, the INDITTO2 transposon was found to be inserted at the promoter region of the DRO1 locus. Transgenesis‐based insertion of the INDITTO2 transposon into the DRO1 promoter of the non‐adapted rice variety Nipponbare was sufficient to promote its drought avoidance. Our data identify an example of how transposons can act as promoters and convey hormonal regulation to nearby loci, improving plant fitness in response to different abiotic stresses. AU - Zhao, Y AU - Wu, L AU - Fu, Q AU - Wang, D AU - Li, J AU - Yao, B AU - Yu, S AU - Jiang, L AU - Qian, J AU - Zhou, X AU - Han, L AU - Zhao, S AU - Ma, C AU - Zhang, Y AU - Luo, C AU - Dong, Q AU - Li, S AU - Zhang, L AU - Jiang, X AU - Li, Y AU - Luo, H AU - Li, K AU - Yang, J AU - Luo, Q AU - Li, L AU - Peng, S AU - Huang, H AU - Zuo, Z AU - Liu, C AU - Wang, L AU - Li, C AU - He, X AU - Friml, Jiří AU - Du, Y ID - 9189 IS - 6 JF - Plant, Cell & Environment SN - 0140-7791 TI - INDITTO2 transposon conveys auxin-mediated DRO1 transcription for rice drought avoidance VL - 44 ER - TY - JOUR AB - Clathrin-mediated endocytosis is the major route of entry of cargos into cells and thus underpins many physiological processes. During endocytosis, an area of flat membrane is remodeled by proteins to create a spherical vesicle against intracellular forces. The protein machinery which mediates this membrane bending in plants is unknown. However, it is known that plant endocytosis is actin independent, thus indicating that plants utilize a unique mechanism to mediate membrane bending against high-turgor pressure compared to other model systems. Here, we investigate the TPLATE complex, a plant-specific endocytosis protein complex. It has been thought to function as a classical adaptor functioning underneath the clathrin coat. However, by using biochemical and advanced live microscopy approaches, we found that TPLATE is peripherally associated with clathrin-coated vesicles and localizes at the rim of endocytosis events. As this localization is more fitting to the protein machinery involved in membrane bending during endocytosis, we examined cells in which the TPLATE complex was disrupted and found that the clathrin structures present as flat patches. This suggests a requirement of the TPLATE complex for membrane bending during plant clathrin–mediated endocytosis. Next, we used in vitro biophysical assays to confirm that the TPLATE complex possesses protein domains with intrinsic membrane remodeling activity. These results redefine the role of the TPLATE complex and implicate it as a key component of the evolutionarily distinct plant endocytosis mechanism, which mediates endocytic membrane bending against the high-turgor pressure in plant cells. AU - Johnson, Alexander J AU - Dahhan, Dana A AU - Gnyliukh, Nataliia AU - Kaufmann, Walter AU - Zheden, Vanessa AU - Costanzo, Tommaso AU - Mahou, Pierre AU - Hrtyan, Mónika AU - Wang, Jie AU - Aguilera Servin, Juan L AU - van Damme, Daniël AU - Beaurepaire, Emmanuel AU - Loose, Martin AU - Bednarek, Sebastian Y AU - Friml, Jiří ID - 9887 IS - 51 JF - Proceedings of the National Academy of Sciences TI - The TPLATE complex mediates membrane bending during plant clathrin-mediated endocytosis VL - 118 ER - TY - GEN AB - Raw data generated from the publication - The TPLATE complex mediates membrane bending during plant clathrin-mediated endocytosis by Johnson et al., 2021 In PNAS AU - Johnson, Alexander J ID - 14988 TI - Raw data from Johnson et al, PNAS, 2021 ER - TY - THES AB - Blood – this is what animals use to heal wounds fast and efficient. Plants do not have blood circulation and their cells cannot move. However, plants have evolved remarkable capacities to regenerate tissues and organs preventing further damage. In my PhD research, I studied the wound healing in the Arabidopsis root. I used a UV laser to ablate single cells in the root tip and observed the consequent wound healing. Interestingly, the inner adjacent cells induced a division plane switch and subsequently adopted the cell type of the killed cell to replace it. We termed this form of wound healing “restorative divisions”. This initial observation triggered the questions of my PhD studies: How and why do cells orient their division planes, how do they feel the wound and why does this happen only in inner adjacent cells. For answering these questions, I used a quite simple experimental setup: 5 day - old seedlings were stained with propidium iodide to visualize cell walls and dead cells; ablation was carried out using a special laser cutter and a confocal microscope. Adaptation of the novel vertical microscope system made it possible to observe wounds in real time. This revealed that restorative divisions occur at increased frequency compared to normal divisions. Additionally, the major plant hormone auxin accumulates in wound adjacent cells and drives the expression of the wound-stress responsive transcription factor ERF115. Using this as a marker gene for wound responses, we found that an important part of wound signalling is the sensing of the collapse of the ablated cell. The collapse causes a radical pressure drop, which results in strong tissue deformations. These deformations manifest in an invasion of the now free spot specifically by the inner adjacent cells within seconds, probably because of higher pressure of the inner tissues. Long-term imaging revealed that those deformed cells continuously expand towards the wound hole and that this is crucial for the restorative division. These wound-expanding cells exhibit an abnormal, biphasic polarity of microtubule arrays before the division. Experiments inhibiting cell expansion suggest that it is the biphasic stretching that induces those MT arrays. Adapting the micromanipulator aspiration system from animal scientists at our institute confirmed the hypothesis that stretching influences microtubule stability. In conclusion, this shows that microtubules react to tissue deformation and this facilitates the observed division plane switch. This puts mechanical cues and tensions at the most prominent position for explaining the growth and wound healing properties of plants. Hence, it shines light onto the importance of understanding mechanical signal transduction. AU - Hörmayer, Lukas ID - 9992 SN - 2663-337X TI - Wound healing in the Arabidopsis root meristem ER - TY - JOUR AB - Availability of the essential macronutrient nitrogen in soil plays a critical role in plant growth, development, and impacts agricultural productivity. Plants have evolved different strategies for sensing and responding to heterogeneous nitrogen distribution. Modulation of root system architecture, including primary root growth and branching, is among the most essential plant adaptions to ensure adequate nitrogen acquisition. However, the immediate molecular pathways coordinating the adjustment of root growth in response to distinct nitrogen sources, such as nitrate or ammonium, are poorly understood. Here, we show that growth as manifested by cell division and elongation is synchronized by coordinated auxin flux between two adjacent outer tissue layers of the root. This coordination is achieved by nitrate‐dependent dephosphorylation of the PIN2 auxin efflux carrier at a previously uncharacterized phosphorylation site, leading to subsequent PIN2 lateralization and thereby regulating auxin flow between adjacent tissues. A dynamic computer model based on our experimental data successfully recapitulates experimental observations. Our study provides mechanistic insights broadening our understanding of root growth mechanisms in dynamic environments. AU - Ötvös, Krisztina AU - Marconi, Marco AU - Vega, Andrea AU - O’Brien, Jose AU - Johnson, Alexander J AU - Abualia, Rashed AU - Antonielli, Livio AU - Montesinos López, Juan C AU - Zhang, Yuzhou AU - Tan, Shutang AU - Cuesta, Candela AU - Artner, Christina AU - Bouguyon, Eleonore AU - Gojon, Alain AU - Friml, Jiří AU - Gutiérrez, Rodrigo A. AU - Wabnik, Krzysztof T AU - Benková, Eva ID - 9010 IS - 3 JF - EMBO Journal SN - 02614189 TI - Modulation of plant root growth by nitrogen source-defined regulation of polar auxin transport VL - 40 ER - TY - JOUR AB - Auxin is a major plant growth regulator, but current models on auxin perception and signaling cannot explain the whole plethora of auxin effects, in particular those associated with rapid responses. A possible candidate for a component of additional auxin perception mechanisms is the AUXIN BINDING PROTEIN 1 (ABP1), whose function in planta remains unclear. Here we combined expression analysis with gain- and loss-of-function approaches to analyze the role of ABP1 in plant development. ABP1 shows a broad expression largely overlapping with, but not regulated by, transcriptional auxin response activity. Furthermore, ABP1 activity is not essential for the transcriptional auxin signaling. Genetic in planta analysis revealed that abp1 loss-of-function mutants show largely normal development with minor defects in bolting. On the other hand, ABP1 gain-of-function alleles show a broad range of growth and developmental defects, including root and hypocotyl growth and bending, lateral root and leaf development, bolting, as well as response to heat stress. At the cellular level, ABP1 gain-of-function leads to impaired auxin effect on PIN polar distribution and affects BFA-sensitive PIN intracellular aggregation. The gain-of-function analysis suggests a broad, but still mechanistically unclear involvement of ABP1 in plant development, possibly masked in abp1 loss-of-function mutants by a functional redundancy. AU - Gelová, Zuzana AU - Gallei, Michelle C AU - Pernisová, Markéta AU - Brunoud, Géraldine AU - Zhang, Xixi AU - Glanc, Matous AU - Li, Lanxin AU - Michalko, Jaroslav AU - Pavlovicova, Zlata AU - Verstraeten, Inge AU - Han, Huibin AU - Hajny, Jakub AU - Hauschild, Robert AU - Čovanová, Milada AU - Zwiewka, Marta AU - Hörmayer, Lukas AU - Fendrych, Matyas AU - Xu, Tongda AU - Vernoux, Teva AU - Friml, Jiří ID - 8931 JF - Plant Science KW - Agronomy and Crop Science KW - Plant Science KW - Genetics KW - General Medicine SN - 0168-9452 TI - Developmental roles of auxin binding protein 1 in Arabidopsis thaliana VL - 303 ER - TY - JOUR AB - The phytohormone auxin and its directional transport through tissues are intensively studied. However, a mechanistic understanding of auxin-mediated feedback on endocytosis and polar distribution of PIN auxin transporters remains limited due to contradictory observations and interpretations. Here, we used state-of-the-art methods to reexamine the auxin effects on PIN endocytic trafficking. We used high auxin concentrations or longer treatments versus lower concentrations and shorter treatments of natural (IAA) and synthetic (NAA) auxins to distinguish between specific and nonspecific effects. Longer treatments of both auxins interfere with Brefeldin A-mediated intracellular PIN2 accumulation and also with general aggregation of endomembrane compartments. NAA treatment decreased the internalization of the endocytic tracer dye, FM4-64; however, NAA treatment also affected the number, distribution, and compartment identity of the early endosome/trans-Golgi network (EE/TGN), rendering the FM4-64 endocytic assays at high NAA concentrations unreliable. To circumvent these nonspecific effects of NAA and IAA affecting the endomembrane system, we opted for alternative approaches visualizing the endocytic events directly at the plasma membrane (PM). Using Total Internal Reflection Fluorescence (TIRF) microscopy, we saw no significant effects of IAA or NAA treatments on the incidence and dynamics of clathrin foci, implying that these treatments do not affect the overall endocytosis rate. However, both NAA and IAA at low concentrations rapidly and specifically promoted endocytosis of photo-converted PIN2 from the PM. These analyses identify a specific effect of NAA and IAA on PIN2 endocytosis, thus contributing to its polarity maintenance and furthermore illustrate that high auxin levels have nonspecific effects on trafficking and endomembrane compartments. AU - Narasimhan, Madhumitha AU - Gallei, Michelle C AU - Tan, Shutang AU - Johnson, Alexander J AU - Verstraeten, Inge AU - Li, Lanxin AU - Rodriguez Solovey, Lesia AU - Han, Huibin AU - Himschoot, E AU - Wang, R AU - Vanneste, S AU - Sánchez-Simarro, J AU - Aniento, F AU - Adamowski, Maciek AU - Friml, Jiří ID - 9287 IS - 2 JF - Plant Physiology SN - 0032-0889 TI - Systematic analysis of specific and nonspecific auxin effects on endocytosis and trafficking VL - 186 ER - TY - THES AB - Plant motions occur across a wide spectrum of timescales, ranging from seed dispersal through bursting (milliseconds) and stomatal opening (minutes) to long-term adaptation of gross architecture. Relatively fast motions include water-driven growth as exemplified by root cell expansion under abiotic/biotic stresses or during gravitropism. A showcase is a root growth inhibition in 30 seconds triggered by the phytohormone auxin. However, the cellular and molecular mechanisms are still largely unknown. This thesis covers the studies about this topic as follows. By taking advantage of microfluidics combined with live imaging, pharmaceutical tools, and transgenic lines, we examined the kinetics of and causal relationship among various auxininduced rapid cellular changes in root growth, apoplastic pH, cytosolic Ca2+, cortical microtubule (CMT) orientation, and vacuolar morphology. We revealed that CMT reorientation and vacuolar constriction are the consequence of growth itself instead of responding directly to auxin. In contrast, auxin induces apoplast alkalinization to rapidly inhibit root growth in 30 seconds. This auxin-triggered apoplast alkalinization results from rapid H+- influx that is contributed by Ca2+ inward channel CYCLIC NUCLEOTIDE-GATED CHANNEL 14 (CNGC14)-dependent Ca2+ signaling. To dissect which auxin signaling mediates the rapid apoplast alkalinization, we combined microfluidics and genetic engineering to verify that TIR1/AFB receptors conduct a non-transcriptional regulation on Ca2+ and H+ -influx. This non-canonical pathway is mostly mediated by the cytosolic portion of TIR1/AFB. On the other hand, we uncovered, using biochemical and phospho-proteomic analysis, that auxin cell surface signaling component TRANSMEMBRANE KINASE 1 (TMK1) plays a negative role during auxin-trigger apoplast alkalinization and root growth inhibition through directly activating PM H+ -ATPases. Therefore, we discovered that PM H+ -ATPases counteract instead of mediate the auxintriggered rapid H+ -influx, and that TIR1/AFB and TMK1 regulate root growth antagonistically. This opposite effect of TIR1/AFB and TMK1 is consistent during auxin-induced hypocotyl elongation, leading us to explore the relation of two signaling pathways. Assisted with biochemistry and fluorescent imaging, we verified for the first time that TIR1/AFB and TMK1 can interact with each other. The ability of TIR1/AFB binding to membrane lipid provides a basis for the interaction of plasma membrane- and cytosol-localized proteins. Besides, transgenic analysis combined with genetic engineering and biochemistry showed that vi they do function in the same pathway. Particularly, auxin-induced TMK1 increase is TIR1/AFB dependent, suggesting TIR1/AFB regulation on TMK1. Conversely, TMK1 also regulates TIR1/AFB protein levels and thus auxin canonical signaling. To follow the study of rapid growth regulation, we analyzed another rapid growth regulator, signaling peptide RALF1. We showed that RALF1 also triggers a rapid and reversible growth inhibition caused by H + influx, highly resembling but not dependent on auxin. Besides, RALF1 promotes auxin biosynthesis by increasing expression of auxin biosynthesis enzyme YUCCAs and thus induces auxin signaling in ca. 1 hour, contributing to the sustained RALF1-triggered growth inhibition. These studies collectively contribute to understanding rapid regulation on plant cell growth, novel auxin signaling pathway as well as auxin-peptide crosstalk. AU - Li, Lanxin ID - 10083 SN - 2663-337X TI - Rapid cell growth regulation in Arabidopsis ER - TY - JOUR AB - Auxin plays a dual role in growth regulation and, depending on the tissue and concentration of the hormone, it can either promote or inhibit division and expansion processes in plants. Recent studies have revealed that, beyond transcriptional reprogramming, alternative auxincontrolled mechanisms regulate root growth. Here, we explored the impact of different concentrations of the synthetic auxin NAA that establish growth-promoting and -repressing conditions on the root tip proteome and phosphoproteome, generating a unique resource. From the phosphoproteome data, we pinpointed (novel) growth regulators, such as the RALF34-THE1 module. Our results, together with previously published studies, suggest that auxin, H+-ATPases, cell wall modifications and cell wall sensing receptor-like kinases are tightly embedded in a pathway regulating cell elongation. Furthermore, our study assigned a novel role to MKK2 as a regulator of primary root growth and a (potential) regulator of auxin biosynthesis and signalling, and suggests the importance of the MKK2 Thr31 phosphorylation site for growth regulation in the Arabidopsis root tip. AU - Nikonorova, N AU - Murphy, E AU - Fonseca de Lima, CF AU - Zhu, S AU - van de Cotte, B AU - Vu, LD AU - Balcerowicz, D AU - Li, Lanxin AU - Kong, X AU - De Rop, G AU - Beeckman, T AU - Friml, Jiří AU - Vissenberg, K AU - Morris, PC AU - Ding, Z AU - De Smet, I ID - 10015 JF - Cells KW - primary root KW - (phospho)proteomics KW - auxin KW - (receptor) kinase SN - 2073-4409 TI - The Arabidopsis root tip (phospho)proteomes at growth-promoting versus growth-repressing conditions reveal novel root growth regulators VL - 10 ER - TY - GEN AB - Growth regulation tailors plant development to its environment. A showcase is response to gravity, where shoots bend up and roots down1. This paradox is based on opposite effects of the phytohormone auxin, which promotes cell expansion in shoots, while inhibiting it in roots via a yet unknown cellular mechanism2. Here, by combining microfluidics, live imaging, genetic engineering and phospho-proteomics in Arabidopsis thaliana, we advance our understanding how auxin inhibits root growth. We show that auxin activates two distinct, antagonistically acting signalling pathways that converge on the rapid regulation of the apoplastic pH, a causative growth determinant. Cell surface-based TRANSMEMBRANE KINASE1 (TMK1) interacts with and mediates phosphorylation and activation of plasma membrane H+-ATPases for apoplast acidification, while intracellular canonical auxin signalling promotes net cellular H+-influx, causing apoplast alkalinisation. The simultaneous activation of these two counteracting mechanisms poises the root for a rapid, fine-tuned growth modulation while navigating complex soil environment. AU - Li, Lanxin AU - Verstraeten, Inge AU - Roosjen, Mark AU - Takahashi, Koji AU - Rodriguez Solovey, Lesia AU - Merrin, Jack AU - Chen, Jian AU - Shabala, Lana AU - Smet, Wouter AU - Ren, Hong AU - Vanneste, Steffen AU - Shabala, Sergey AU - De Rybel, Bert AU - Weijers, Dolf AU - Kinoshita, Toshinori AU - Gray, William M. AU - Friml, Jiří ID - 10095 SN - 2693-5015 T2 - Research Square TI - Cell surface and intracellular auxin signalling for H+-fluxes in root growth ER - TY - GEN AB - Plasmodesmata (PD) are crucial structures for intercellular communication in multicellular plants with remorins being their crucial plant-specific structural and functional constituents. The PD biogenesis is an intriguing but poorly understood process. By expressing an Arabidopsis remorin protein in mammalian cells, we have reconstituted a PD-like filamentous structure, termed remorin filament (RF), connecting neighboring cells physically and physiologically. Notably, RFs are capable of transporting macromolecules intercellularly, in a way similar to plant PD. With further super-resolution microscopic analysis and biochemical characterization, we found that RFs are also composed of actin filaments, forming the core skeleton structure, aligned with the remorin protein. This unique heterologous filamentous structure might explain the molecular mechanism for remorin function as well as PD construction. Furthermore, remorin protein exhibits a specific distribution manner in the plasma membrane in mammalian cells, representing a lipid nanodomain, depending on its lipid modification status. Our studies not only provide crucial insights into the mechanism of PD biogenesis, but also uncovers unsuspected fundamental mechanistic and evolutionary links between intercellular communication systems of plants and animals. AU - Wei, Zhuang AU - Tan, Shutang AU - Liu, Tao AU - Wu, Yuan AU - Lei, Ji-Gang AU - Chen, ZhengJun AU - Friml, Jiří AU - Xue, Hong-Wei AU - Liao, Kan ID - 7601 T2 - bioRxiv TI - Plasmodesmata-like intercellular connections by plant remorin in animal cells ER - TY - JOUR AU - Zhang, Yuzhou AU - Friml, Jiří ID - 6997 IS - 3 JF - New Phytologist SN - 0028-646x TI - Auxin guides roots to avoid obstacles during gravitropic growth VL - 225 ER - TY - JOUR AB - Plant root architecture dynamically adapts to various environmental conditions, such as salt‐containing soil. The phytohormone abscisic acid (ABA) is involved among others also in these developmental adaptations, but the underlying molecular mechanism remains elusive. Here, a novel branch of the ABA signaling pathway in Arabidopsis involving PYR/PYL/RCAR (abbreviated as PYLs) receptor‐protein phosphatase 2A (PP2A) complex that acts in parallel to the canonical PYLs‐protein phosphatase 2C (PP2C) mechanism is identified. The PYLs‐PP2A signaling modulates root gravitropism and lateral root formation through regulating phytohormone auxin transport. In optimal conditions, PYLs ABA receptor interacts with the catalytic subunits of PP2A, increasing their phosphatase activity and thus counteracting PINOID (PID) kinase‐mediated phosphorylation of PIN‐FORMED (PIN) auxin transporters. By contrast, in salt and osmotic stress conditions, ABA binds to PYLs, inhibiting the PP2A activity, which leads to increased PIN phosphorylation and consequently modulated directional auxin transport leading to adapted root architecture. This work reveals an adaptive mechanism that may flexibly adjust plant root growth to withstand saline and osmotic stresses. It occurs via the cross‐talk between the stress hormone ABA and the versatile developmental regulator auxin. AU - Li, Yang AU - Wang, Yaping AU - Tan, Shutang AU - Li, Zhen AU - Yuan, Zhi AU - Glanc, Matous AU - Domjan, David AU - Wang, Kai AU - Xuan, Wei AU - Guo, Yan AU - Gong, Zhizhong AU - Friml, Jiří AU - Zhang, Jing ID - 7204 IS - 3 JF - Advanced Science TI - Root growth adaptation is mediated by PYLs ABA receptor-PP2A protein phosphatase complex VL - 7 ER - TY - JOUR AB - The phytohormone auxin acts as an amazingly versatile coordinator of plant growth and development. With its morphogen-like properties, auxin controls sites and timing of differentiation and/or growth responses both, in quantitative and qualitative terms. Specificity in the auxin response depends largely on distinct modes of signal transmission, by which individual cells perceive and convert auxin signals into a remarkable diversity of responses. The best understood, or so-called canonical mechanism of auxin perception ultimately results in variable adjustments of the cellular transcriptome, via a short, nuclear signal transduction pathway. Additional findings that accumulated over decades implied that an additional, presumably, cell surface-based auxin perception mechanism mediates very rapid cellular responses and decisively contributes to the cell's overall hormonal response. Recent investigations into both, nuclear and cell surface auxin signalling challenged this assumed partition of roles for different auxin signalling pathways and revealed an unexpected complexity in transcriptional and non-transcriptional cellular responses mediated by auxin. AU - Gallei, Michelle C AU - Luschnig, Christian AU - Friml, Jiří ID - 7142 IS - 2 JF - Current Opinion in Plant Biology SN - 1369-5266 TI - Auxin signalling in growth: Schrödinger's cat out of the bag VL - 53 ER - TY - JOUR AB - Root system architecture (RSA), governed by the phytohormone auxin, endows plants with an adaptive advantage in particular environments. Using geographically representative arabidopsis (Arabidopsis thaliana) accessions as a resource for GWA mapping, Waidmann et al. and Ogura et al. recently identified two novel components involved in modulating auxin-mediated RSA and conferring plant fitness in particular habitats. AU - Xiao, Guanghui AU - Zhang, Yuzhou ID - 7219 IS - 2 JF - Trends in Plant Science SN - 13601385 TI - Adaptive growth: Shaping auxin-mediated root system architecture VL - 25 ER - TY - JOUR AB - The flexible development of plants is characterized by a high capacity for post-embryonic organ formation and tissue regeneration, processes, which require tightly regulated intercellular communication and coordinated tissue (re-)polarization. The phytohormone auxin, the main driver for these processes, is able to establish polarized auxin transport channels, which are characterized by the expression and polar, subcellular localization of the PIN1 auxin transport proteins. These channels are demarcating the position of future vascular strands necessary for organ formation and tissue regeneration. Major progress has been made in the last years to understand how PINs can change their polarity in different contexts and thus guide auxin flow through the plant. However, it still remains elusive how auxin mediates the establishment of auxin conducting channels and the formation of vascular tissue and which cellular processes are involved. By the means of sophisticated regeneration experiments combined with local auxin applications in Arabidopsis thaliana inflorescence stems we show that (i) PIN subcellular dynamics, (ii) PIN internalization by clathrin-mediated trafficking and (iii) an intact actin cytoskeleton required for post-endocytic trafficking are indispensable for auxin channel formation, de novo vascular formation and vascular regeneration after wounding. These observations provide novel insights into cellular mechanism of coordinated tissue polarization during auxin canalization. AU - Mazur, Ewa AU - Gallei, Michelle C AU - Adamowski, Maciek AU - Han, Huibin AU - Robert, Hélène S. AU - Friml, Jiří ID - 7465 IS - 4 JF - Plant Science SN - 01689452 TI - Clathrin-mediated trafficking and PIN trafficking are required for auxin canalization and vascular tissue formation in Arabidopsis VL - 293 ER - TY - JOUR AB - In plants, clathrin mediated endocytosis (CME) represents the major route for cargo internalisation from the cell surface. It has been assumed to operate in an evolutionary conserved manner as in yeast and animals. Here we report characterisation of ultrastructure, dynamics and mechanisms of plant CME as allowed by our advancement in electron microscopy and quantitative live imaging techniques. Arabidopsis CME appears to follow the constant curvature model and the bona fide CME population generates vesicles of a predominantly hexagonal-basket type; larger and with faster kinetics than in other models. Contrary to the existing paradigm, actin is dispensable for CME events at the plasma membrane but plays a unique role in collecting endocytic vesicles, sorting of internalised cargos and directional endosome movement that itself actively promote CME events. Internalized vesicles display a strongly delayed and sequential uncoating. These unique features highlight the independent evolution of the plant CME mechanism during the autonomous rise of multicellularity in eukaryotes. AU - Narasimhan, Madhumitha AU - Johnson, Alexander J AU - Prizak, Roshan AU - Kaufmann, Walter AU - Tan, Shutang AU - Casillas Perez, Barbara E AU - Friml, Jiří ID - 7490 JF - eLife TI - Evolutionarily unique mechanistic framework of clathrin-mediated endocytosis in plants VL - 9 ER - TY - JOUR AB - Endophytic fungi can be beneficial to plant growth. However, the molecular mechanisms underlying colonization of Acremonium spp. remain unclear. In this study, a novel endophytic Acremonium strain was isolated from the buds of Panax notoginseng and named Acremonium sp. D212. The Acremonium sp. D212 could colonize the roots of P. notoginseng, enhance the resistance of P. notoginseng to root rot disease, and promote root growth and saponin biosynthesis in P. notoginseng. Acremonium sp. D212 could secrete indole‐3‐acetic acid (IAA) and jasmonic acid (JA), and inoculation with the fungus increased the endogenous levels of IAA and JA in P. notoginseng. Colonization of the Acremonium sp. D212 in the roots of the rice line Nipponbare was dependent on the concentration of methyl jasmonate (MeJA) (2 to 15 μM) and 1‐naphthalenacetic acid (NAA) (10 to 20 μM). Moreover, the roots of the JA signalling‐defective coi1‐18 mutant were colonized by Acremonium sp. D212 to a lesser degree than those of the wild‐type Nipponbare and miR393b‐overexpressing lines, and the colonization was rescued by MeJA but not by NAA. It suggests that the cross‐talk between JA signalling and the auxin biosynthetic pathway plays a crucial role in the colonization of Acremonium sp. D212 in host plants. AU - Han, L AU - Zhou, X AU - Zhao, Y AU - Zhu, S AU - Wu, L AU - He, Y AU - Ping, X AU - Lu, X AU - Huang, W AU - Qian, J AU - Zhang, L AU - Jiang, X AU - Zhu, D AU - Luo, C AU - Li, S AU - Dong, Q AU - Fu, Q AU - Deng, K AU - Wang, X AU - Wang, L AU - Peng, S AU - Wu, J AU - Li, W AU - Friml, Jiří AU - Zhu, Y AU - He, X AU - Du, Y ID - 7497 IS - 9 JF - Journal of Integrative Plant Biology SN - 1672-9072 TI - Colonization of endophyte Acremonium sp. D212 in Panax notoginseng and rice mediated by auxin and jasmonic acid VL - 62 ER - TY - JOUR AB - In vitro propagation of the ornamentally interesting species Wikstroemia gemmata is limited by the recalcitrance to form adventitious roots. In this article, two strategies to improve the rooting capacity of in vitro microcuttings are presented. Firstly, the effect of exogenous auxin was evaluated in both light and dark cultivated stem segments and also the sucrose-content of the medium was varied in order to determine better rooting conditions. Secondly, different spectral lights were evaluated and the effect on shoot growth and root induction demonstrated that the exact spectral composition of light is important for successful in vitro growth and development of Wikstroemia gemmata. We show that exogenous auxin cannot compensate for the poor rooting under unfavorable light conditions. Adapting the culture conditions is therefore paramount for successful industrial propagation of Wikstroemia gemmata. AU - Verstraeten, Inge AU - Buyle, H. AU - Werbrouck, S. AU - Van Labeke, M.C. AU - Geelen, D. ID - 7540 IS - 1-2 JF - Israel Journal of Plant Sciences SN - 0792-9978 TI - In vitro shoot growth and adventitious rooting of Wikstroemia gemmata depends on light quality VL - 67 ER - TY - JOUR AB - Small RNAs (smRNA, 19–25 nucleotides long), which are transcribed by RNA polymerase II, regulate the expression of genes involved in a multitude of processes in eukaryotes. miRNA biogenesis and the proteins involved in the biogenesis pathway differ across plant and animal lineages. The major proteins constituting the biogenesis pathway, namely, the Dicers (DCL/DCR) and Argonautes (AGOs), have been extensively studied. However, the accessory proteins (DAWDLE (DDL), SERRATE (SE), and TOUGH (TGH)) of the pathway that differs across the two lineages remain largely uncharacterized. We present the first detailed report on the molecular evolution and divergence of these proteins across eukaryotes. Although DDL is present in eukaryotes and prokaryotes, SE and TGH appear to be specific to eukaryotes. The addition/deletion of specific domains and/or domain-specific sequence divergence in the three proteins points to the observed functional divergence of these proteins across the two lineages, which correlates with the differences in miRNA length across the two lineages. Our data enhance the current understanding of the structure–function relationship of these proteins and reveals previous unexplored crucial residues in the three proteins that can be used as a basis for further functional characterization. The data presented here on the number of miRNAs in crown eukaryotic lineages are consistent with the notion of the expansion of the number of miRNA-coding genes in animal and plant lineages correlating with organismal complexity. Whether this difference in functionally correlates with the diversification (or presence/absence) of the three proteins studied here or the miRNA signaling in the plant and animal lineages is unclear. Based on our results of the three proteins studied here and previously available data concerning the evolution of miRNA genes in the plant and animal lineages, we believe that miRNAs probably evolved once in the ancestor to crown eukaryotes and have diversified independently in the eukaryotes. AU - Moturu, Taraka Ramji AU - Sinha, Sansrity AU - Salava, Hymavathi AU - Thula, Sravankumar AU - Nodzyński, Tomasz AU - Vařeková, Radka Svobodová AU - Friml, Jiří AU - Simon, Sibu ID - 7582 IS - 3 JF - Plants TI - Molecular evolution and diversification of proteins involved in miRNA maturation pathway VL - 9 ER - TY - JOUR AB - Directional intercellular transport of the phytohormone auxin mediated by PIN FORMED (PIN) efflux carriers plays essential roles in both coordinating patterning processes and integrating multiple external cues by rapidly redirecting auxin fluxes. Multilevel regulations of PIN activity under internal and external cues are complicated; however, the underlying molecular mechanism remains elusive. Here we demonstrate that 3’-Phosphoinositide-Dependent Protein Kinase1 (PDK1), which is conserved in plants and mammals, functions as a molecular hub integrating the upstream lipid signalling and the downstream substrate activity through phosphorylation. Genetic analysis uncovers that loss-of-function Arabidopsis mutant pdk1.1 pdk1.2 exhibits a plethora of abnormalities in organogenesis and growth, due to the defective PIN-dependent auxin transport. Further cellular and biochemical analyses reveal that PDK1 phosphorylates D6 Protein Kinase to facilitate its activity towards PIN proteins. Our studies establish a lipid-dependent phosphorylation cascade connecting membrane composition-based cellular signalling with plant growth and patterning by regulating morphogenetic auxin fluxes. AU - Tan, Shutang AU - Zhang, Xixi AU - Kong, Wei AU - Yang, Xiao-Li AU - Molnar, Gergely AU - Vondráková, Zuzana AU - Filepová, Roberta AU - Petrášek, Jan AU - Friml, Jiří AU - Xue, Hong-Wei ID - 7600 JF - Nature Plants TI - The lipid code-dependent phosphoswitch PDK1–D6PK activates PIN-mediated auxin efflux in Arabidopsis VL - 6 ER - TY - JOUR AB - In plant cells, environmental stressors promote changes in connectivity between the cortical ER and the PM. Although this process is tightly regulated in space and time, the molecular signals and structural components mediating these changes in inter-organelle communication are only starting to be characterized. In this report, we confirm the presence of a putative tethering complex containing the synaptotagmins 1 and 5 (SYT1 and SYT5) and the Ca2+ and lipid binding protein 1 (CLB1/SYT7). This complex is enriched at ER-PM contact sites (EPCS), have slow responses to changes in extracellular Ca2+, and display severe cytoskeleton-dependent rearrangements in response to the trivalent lanthanum (La3+) and gadolinium (Gd3+) rare earth elements (REEs). Although REEs are generally used as non-selective cation channel blockers at the PM, here we show that the slow internalization of REEs into the cytosol underlies the activation of the Ca2+/Calmodulin intracellular signaling, the accumulation of phosphatidylinositol-4-phosphate (PI4P) at the PM, and the cytoskeleton-dependent rearrangement of the SYT1/SYT5 EPCS complexes. We propose that the observed EPCS rearrangements act as a slow adaptive response to sustained stress conditions, and that this process involves the accumulation of stress-specific phosphoinositides species at the PM. AU - Lee, E AU - Vila Nova Santana, B AU - Samuels, E AU - Benitez-Fuente, F AU - Corsi, E AU - Botella, MA AU - Perez-Sancho, J AU - Vanneste, S AU - Friml, Jiří AU - Macho, A AU - Alves Azevedo, A AU - Rosado, A ID - 7646 IS - 14 JF - Journal of Experimental Botany SN - 0022-0957 TI - Rare earth elements induce cytoskeleton-dependent and PI4P-associated rearrangement of SYT1/SYT5 ER-PM contact site complexes in Arabidopsis VL - 71 ER - TY - JOUR AB - The agricultural green revolution spectacularly enhanced crop yield and lodging resistance with modified DELLA-mediated gibberellin signaling. However, this was achieved at the expense of reduced nitrogen-use efficiency (NUE). Recently, Wu et al. revealed novel gibberellin signaling that provides a blueprint for improving tillering and NUE in Green Revolution varieties (GRVs). AU - Xue, Huidan AU - Zhang, Yuzhou AU - Xiao, Guanghui ID - 7686 IS - 6 JF - Trends in Plant Science SN - 1360-1385 TI - Neo-gibberellin signaling: Guiding the next generation of the green revolution VL - 25 ER - TY - JOUR AB - Hormonal signalling in animals often involves direct transcription factor-hormone interactions that modulate gene expression. In contrast, plant hormone signalling is most commonly based on de-repression via the degradation of transcriptional repressors. Recently, we uncovered a non-canonical signalling mechanism for the plant hormone auxin whereby auxin directly affects the activity of the atypical auxin response factor (ARF), ETTIN towards target genes without the requirement for protein degradation. Here we show that ETTIN directly binds auxin, leading to dissociation from co-repressor proteins of the TOPLESS/TOPLESS-RELATED family followed by histone acetylation and induction of gene expression. This mechanism is reminiscent of animal hormone signalling as it affects the activity towards regulation of target genes and provides the first example of a DNA-bound hormone receptor in plants. Whilst auxin affects canonical ARFs indirectly by facilitating degradation of Aux/IAA repressors, direct ETTIN-auxin interactions allow switching between repressive and de-repressive chromatin states in an instantly-reversible manner. AU - Kuhn, André AU - Ramans Harborough, Sigurd AU - McLaughlin, Heather M AU - Natarajan, Bhavani AU - Verstraeten, Inge AU - Friml, Jiří AU - Kepinski, Stefan AU - Østergaard, Lars ID - 7793 JF - eLife SN - 2050-084X TI - Direct ETTIN-auxin interaction controls chromatin states in gynoecium development VL - 9 ER - TY - JOUR AB - Directional transport of the phytohormone auxin is a versatile, plant-specific mechanism regulating many aspects of plant development. The recently identified plant hormones, strigolactones (SLs), are implicated in many plant traits; among others, they modify the phenotypic output of PIN-FORMED (PIN) auxin transporters for fine-tuning of growth and developmental responses. Here, we show in pea and Arabidopsis that SLs target processes dependent on the canalization of auxin flow, which involves auxin feedback on PIN subcellular distribution. D14 receptor- and MAX2 F-box-mediated SL signaling inhibits the formation of auxin-conducting channels after wounding or from artificial auxin sources, during vasculature de novo formation and regeneration. At the cellular level, SLs interfere with auxin effects on PIN polar targeting, constitutive PIN trafficking as well as clathrin-mediated endocytosis. Our results identify a non-transcriptional mechanism of SL action, uncoupling auxin feedback on PIN polarity and trafficking, thereby regulating vascular tissue formation and regeneration. AU - Zhang, J AU - Mazur, E AU - Balla, J AU - Gallei, Michelle C AU - Kalousek, P AU - Medveďová, Z AU - Li, Y AU - Wang, Y AU - Prat, Tomas AU - Vasileva, Mina K AU - Reinöhl, V AU - Procházka, S AU - Halouzka, R AU - Tarkowski, P AU - Luschnig, C AU - Brewer, PB AU - Friml, Jiří ID - 8138 IS - 1 JF - Nature Communications SN - 2041-1723 TI - Strigolactones inhibit auxin feedback on PIN-dependent auxin transport canalization VL - 11 ER - TY - JOUR AU - He, Peng AU - Zhang, Yuzhou AU - Xiao, Guanghui ID - 8271 IS - 9 JF - Molecular Plant SN - 16742052 TI - Origin of a subgenome and genome evolution of allotetraploid cotton species VL - 13 ER - TY - JOUR AB - Cytokinins are mobile multifunctional plant hormones with roles in development and stress resilience. Although their Histidine Kinase receptors are substantially localised to the endoplasmic reticulum, cellular sites of cytokinin perception and importance of spatially heterogeneous cytokinin distribution continue to be debated. Here we show that cytokinin perception by plasma membrane receptors is an effective additional path for cytokinin response. Readout from a Two Component Signalling cytokinin-specific reporter (TCSn::GFP) closely matches intracellular cytokinin content in roots, yet we also find cytokinins in extracellular fluid, potentially enabling action at the cell surface. Cytokinins covalently linked to beads that could not pass the plasma membrane increased expression of both TCSn::GFP and Cytokinin Response Factors. Super-resolution microscopy of GFP-labelled receptors and diminished TCSn::GFP response to immobilised cytokinins in cytokinin receptor mutants, further indicate that receptors can function at the cell surface. We argue that dual intracellular and surface locations may augment flexibility of cytokinin responses. AU - Antoniadi, Ioanna AU - Novák, Ondřej AU - Gelová, Zuzana AU - Johnson, Alexander J AU - Plíhal, Ondřej AU - Simerský, Radim AU - Mik, Václav AU - Vain, Thomas AU - Mateo-Bonmatí, Eduardo AU - Karady, Michal AU - Pernisová, Markéta AU - Plačková, Lenka AU - Opassathian, Korawit AU - Hejátko, Jan AU - Robert, Stéphanie AU - Friml, Jiří AU - Doležal, Karel AU - Ljung, Karin AU - Turnbull, Colin ID - 8337 JF - Nature Communications TI - Cell-surface receptors enable perception of extracellular cytokinins VL - 11 ER - TY - JOUR AB - Spontaneously arising channels that transport the phytohormone auxin provide positional cues for self-organizing aspects of plant development such as flexible vasculature regeneration or its patterning during leaf venation. The auxin canalization hypothesis proposes a feedback between auxin signaling and transport as the underlying mechanism, but molecular players await discovery. We identified part of the machinery that routes auxin transport. The auxin-regulated receptor CAMEL (Canalization-related Auxin-regulated Malectin-type RLK) together with CANAR (Canalization-related Receptor-like kinase) interact with and phosphorylate PIN auxin transporters. camel and canar mutants are impaired in PIN1 subcellular trafficking and auxin-mediated PIN polarization, which macroscopically manifests as defects in leaf venation and vasculature regeneration after wounding. The CAMEL-CANAR receptor complex is part of the auxin feedback that coordinates polarization of individual cells during auxin canalization. AU - Hajny, Jakub AU - Prat, Tomas AU - Rydza, N AU - Rodriguez Solovey, Lesia AU - Tan, Shutang AU - Verstraeten, Inge AU - Domjan, David AU - Mazur, E AU - Smakowska-Luzan, E AU - Smet, W AU - Mor, E AU - Nolf, J AU - Yang, B AU - Grunewald, W AU - Molnar, Gergely AU - Belkhadir, Y AU - De Rybel, B AU - Friml, Jiří ID - 8721 IS - 6516 JF - Science SN - 0036-8075 TI - Receptor kinase module targets PIN-dependent auxin transport during canalization VL - 370 ER - TY - JOUR AB - Peptides derived from non-functional precursors play important roles in various developmental processes, but also in (a)biotic stress signaling. Our (phospho)proteome-wide analyses of C-terminally encoded peptide 5 (CEP5)-mediated changes revealed an impact on abiotic stress-related processes. Drought has a dramatic impact on plant growth, development and reproduction, and the plant hormone auxin plays a role in drought responses. Our genetic, physiological, biochemical and pharmacological results demonstrated that CEP5-mediated signaling is relevant for osmotic and drought stress tolerance in Arabidopsis, and that CEP5 specifically counteracts auxin effects. Specifically, we found that CEP5 signaling stabilizes AUX/IAA transcriptional repressors, suggesting the existence of a novel peptide-dependent control mechanism that tunes auxin signaling. These observations align with the recently described role of AUX/IAAs in stress tolerance and provide a novel role for CEP5 in osmotic and drought stress tolerance. AU - Smith, S AU - Zhu, S AU - Joos, L AU - Roberts, I AU - Nikonorova, N AU - Vu, LD AU - Stes, E AU - Cho, H AU - Larrieu, A AU - Xuan, W AU - Goodall, B AU - van de Cotte, B AU - Waite, JM AU - Rigal, A AU - R Harborough, SR AU - Persiau, G AU - Vanneste, S AU - Kirschner, GK AU - Vandermarliere, E AU - Martens, L AU - Stahl, Y AU - Audenaert, D AU - Friml, Jiří AU - Felix, G AU - Simon, R AU - Bennett, M AU - Bishopp, A AU - De Jaeger, G AU - Ljung, K AU - Kepinski, S AU - Robert, S AU - Nemhauser, J AU - Hwang, I AU - Gevaert, K AU - Beeckman, T AU - De Smet, I ID - 7949 IS - 8 JF - Molecular & Cellular Proteomics TI - The CEP5 peptide promotes abiotic stress tolerance, as revealed by quantitative proteomics, and attenuates the AUX/IAA equilibrium in Arabidopsis VL - 19 ER - TY - JOUR AB - Cell polarity is a fundamental feature of all multicellular organisms. In plants, prominent cell polarity markers are PIN auxin transporters crucial for plant development. To identify novel components involved in cell polarity establishment and maintenance, we carried out a forward genetic screening with PIN2:PIN1-HA;pin2 Arabidopsis plants, which ectopically express predominantly basally localized PIN1 in the root epidermal cells leading to agravitropic root growth. From the screen, we identified the regulator of PIN polarity 12 (repp12) mutation, which restored gravitropic root growth and caused PIN1-HA polarity switch from basal to apical side of root epidermal cells. Complementation experiments established the repp12 causative mutation as an amino acid substitution in Aminophospholipid ATPase3 (ALA3), a phospholipid flippase with predicted function in vesicle formation. ala3 T-DNA mutants show defects in many auxin-regulated processes, in asymmetric auxin distribution and in PIN trafficking. Analysis of quintuple and sextuple mutants confirmed a crucial role of ALA proteins in regulating plant development and in PIN trafficking and polarity. Genetic and physical interaction studies revealed that ALA3 functions together with GNOM and BIG3 ARF GEFs. Taken together, our results identified ALA3 flippase as an important interactor and regulator of ARF GEF functioning in PIN polarity, trafficking and auxin-mediated development. AU - Zhang, Xixi AU - Adamowski, Maciek AU - Marhavá, Petra AU - Tan, Shutang AU - Zhang, Yuzhou AU - Rodriguez Solovey, Lesia AU - Zwiewka, Marta AU - Pukyšová, Vendula AU - Sánchez, Adrià Sans AU - Raxwal, Vivek Kumar AU - Hardtke, Christian S. AU - Nodzynski, Tomasz AU - Friml, Jiří ID - 7619 IS - 5 JF - The Plant Cell SN - 1040-4651 TI - Arabidopsis flippases cooperate with ARF GTPase exchange factors to regulate the trafficking and polarity of PIN auxin transporters VL - 32 ER - TY - JOUR AB - Clathrin-mediated endocytosis (CME) and its core endocytic machinery are evolutionarily conserved across all eukaryotes. In mammals, the heterotetrameric adaptor protein complex-2 (AP-2) sorts plasma membrane (PM) cargoes into vesicles through the recognition of motifs based on tyrosine or di-leucine in their cytoplasmic tails. However, in plants, very little is known on how PM proteins are sorted for CME and whether similar motifs are required. In Arabidopsis thaliana, the brassinosteroid (BR) receptor, BR INSENSITIVE1 (BRI1), undergoes endocytosis that depends on clathrin and AP-2. Here we demonstrate that BRI1 binds directly to the medium AP-2 subunit, AP2M. The cytoplasmic domain of BRI1 contains five putative canonical surface-exposed tyrosine-based endocytic motifs. The tyrosine-to-phenylalanine substitution in Y898KAI reduced BRI1 internalization without affecting its kinase activity. Consistently, plants carrying the BRI1Y898F mutation were hypersensitive to BRs. Our study demonstrates that AP-2-dependent internalization of PM proteins via the recognition of functional tyrosine motifs also operates in plants. AU - Liu, D AU - Kumar, R AU - LAN, Claus AU - Johnson, Alexander J AU - Siao, W AU - Vanhoutte, I AU - Wang, P AU - Bender, KW AU - Yperman, K AU - Martins, S AU - Zhao, X AU - Vert, G AU - Van Damme, D AU - Friml, Jiří AU - Russinova, E ID - 8607 IS - 11 JF - Plant Cell SN - 1040-4651 TI - Endocytosis of BRASSINOSTEROID INSENSITIVE1 is partly driven by a canonical tyrosine-based Motif VL - 32 ER - TY - JOUR AB - The TPLATE complex (TPC) is a key endocytic adaptor protein complex in plants. TPC in Arabidopsis (Arabidopsis thaliana) contains six evolutionarily conserved subunits and two plant-specific subunits, AtEH1/Pan1 and AtEH2/Pan1, although cytoplasmic proteins are not associated with the hexameric subcomplex in the cytoplasm. To investigate the dynamic assembly of the octameric TPC at the plasma membrane (PM), we performed state-of-the-art dual-color live cell imaging at physiological and lowered temperatures. Lowering the temperature slowed down endocytosis, thereby enhancing the temporal resolution of the differential recruitment of endocytic components. Under both normal and lowered temperature conditions, the core TPC subunit TPLATE and the AtEH/Pan1 proteins exhibited simultaneous recruitment at the PM. These results, together with co-localization analysis of different TPC subunits, allow us to conclude that TPC in plant cells is not recruited to the PM sequentially but as an octameric complex. AU - Wang, J AU - Mylle, E AU - Johnson, Alexander J AU - Besbrugge, N AU - De Jaeger, G AU - Friml, Jiří AU - Pleskot, R AU - van Damme, D ID - 7695 IS - 3 JF - Plant Physiology SN - 0032-0889 TI - High temporal resolution reveals simultaneous plasma membrane recruitment of TPLATE complex subunits VL - 183 ER - TY - JOUR AB - * Morphogenesis and adaptive tropic growth in plants depend on gradients of the phytohormone auxin, mediated by the membrane‐based PIN‐FORMED (PIN) auxin transporters. PINs localize to a particular side of the plasma membrane (PM) or to the endoplasmic reticulum (ER) to directionally transport auxin and maintain intercellular and intracellular auxin homeostasis, respectively. However, the molecular cues that confer their diverse cellular localizations remain largely unknown. * In this study, we systematically swapped the domains between ER‐ and PM‐localized PIN proteins, as well as between apical and basal PM‐localized PINs from Arabidopsis thaliana , to shed light on why PIN family members with similar topological structures reside at different membrane compartments within cells. * Our results show that not only do the N‐ and C‐terminal transmembrane domains (TMDs) and central hydrophilic loop contribute to their differential subcellular localizations and cellular polarity, but that the pairwise‐matched N‐ and C‐terminal TMDs resulting from intramolecular domain–domain coevolution are also crucial for their divergent patterns of localization. * These findings illustrate the complexity of the evolutionary path of PIN proteins in acquiring their plethora of developmental functions and adaptive growth in plants. AU - Zhang, Yuzhou AU - Hartinger, Corinna AU - Wang, Xiaojuan AU - Friml, Jiří ID - 7697 IS - 5 JF - New Phytologist SN - 0028-646X TI - Directional auxin fluxes in plants by intramolecular domain‐domain co‐evolution of PIN auxin transporters VL - 227 ER - TY - JOUR AB - Previously, we reported that the allelic de-etiolated by zinc (dez) and trichome birefringence (tbr) mutants exhibit photomorphogenic development in the dark, which is enhanced by high Zn. TRICHOME BIREFRINGENCE-LIKE proteins had been implicated in transferring acetyl groups to various hemicelluloses. Pectin O-acetylation levels were lower in dark-grown dez seedlings than in the wild type. We observed Zn-enhanced photomorphogenesis in the dark also in the reduced wall acetylation 2 (rwa2-3) mutant, which exhibits lowered O-acetylation levels of cell wall macromolecules including pectins and xyloglucans, supporting a role for cell wall macromolecule O-acetylation in the photomorphogenic phenotypes of rwa2-3 and dez. Application of very short oligogalacturonides (vsOGs) restored skotomorphogenesis in dark-grown dez and rwa2-3. Here we demonstrate that in dez, O-acetylation of non-pectin cell wall components, notably of xyloglucan, is enhanced. Our results highlight the complexity of cell wall homeostasis and indicate against an influence of xyloglucan O-acetylation on light-dependent seedling development. AU - Sinclair, Scott A AU - Gille, S. AU - Pauly, M. AU - Krämer, U. ID - 7417 IS - 1 JF - Plant Signaling & Behavior SN - 1559-2324 TI - Regulation of acetylation of plant cell wall components is complex and responds to external stimuli VL - 15 ER - TY - THES AB - The plant hormone auxin plays indispensable roles in plant growth and development. An essential level of regulation in auxin action is the directional auxin transport within cells. The establishment of auxin gradient in plant tissue has been attributed to local auxin biosynthesis and directional intercellular auxin transport, which both are controlled by various environmental and developmental signals. It is well established that asymmetric auxin distribution in cells is achieved by polarly localized PIN-FORMED (PIN) auxin efflux transporters. Despite the initial insights into cellular mechanisms of PIN polarization obtained from the last decades, the molecular mechanism and specific regulators mediating PIN polarization remains elusive. In this thesis, we aim to find novel players in PIN subcellular polarity regulation during Arabidopsis development. We first characterize the physiological effect of piperonylic acid (PA) on Arabidopsis hypocotyl gravitropic bending and PIN polarization. Secondly, we reveal the importance of SCFTIR1/AFB auxin signaling pathway in shoot gravitropism bending termination. In addition, we also explore the role of myosin XI complex, and actin cytoskeleton in auxin feedback regulation on PIN polarity. In Chapter 1, we give an overview of the current knowledge about PIN-mediated auxin fluxes in various plant tropic responses. In Chapter 2, we study the physiological effect of PA on shoot gravitropic bending. Our results show that PA treatment inhibits auxin-mediated PIN3 repolarization by interfering with PINOID and PIN3 phosphorylation status, ultimately leading to hyperbending hypocotyls. In Chapter 3, we provide evidence to show that the SCFTIR1/AFB nuclear auxin signaling pathway is crucial and required for auxin-mediated PIN3 repolarization and shoot gravitropic bending termination. In Chapter 4, we perform a phosphoproteomics approach and identify the motor protein Myosin XI and its binding protein, the MadB2 family, as an essential regulator of PIN polarity for auxin-canalization related developmental processes. In Chapter 5, we demonstrate the vital role of actin cytoskeleton in auxin feedback on PIN polarity by regulating PIN subcellular trafficking. Overall, the data presented in this PhD thesis brings novel insights into the PIN polar localization regulation that resulted in the (re)establishment of the polar auxin flow and gradient in response to environmental stimuli during plant development. AU - Han, Huibin ID - 8589 SN - 2663-337X TI - Novel insights into PIN polarity regulation during Arabidopsis development ER - TY - JOUR AU - Han, Huibin AU - Rakusova, Hana AU - Verstraeten, Inge AU - Zhang, Yuzhou AU - Friml, Jiří ID - 7643 IS - 5 JF - Plant Physiology SN - 0032-0889 TI - SCF TIR1/AFB auxin signaling for bending termination during shoot gravitropism VL - 183 ER - TY - JOUR AB - Earlier, we demonstrated that transcript levels of METAL TOLERANCE PROTEIN2 (MTP2) and of HEAVY METAL ATPase2 (HMA2) increase strongly in roots of Arabidopsis upon prolonged zinc (Zn) deficiency and respond to shoot physiological Zn status, and not to the local Zn status in roots. This provided evidence for shoot-to-root communication in the acclimation of plants to Zn deficiency. Zn-deficient soils limit both the yield and quality of agricultural crops and can result in clinically relevant nutritional Zn deficiency in human populations. Implementing Zn deficiency during cultivation of the model plant Arabidopsis thaliana on agar-solidified media is difficult because trace element contaminations are present in almost all commercially available agars. Here, we demonstrate root morphological acclimations to Zn deficiency on agar-solidified medium following the effective removal of contaminants. These advancements allow reproducible phenotyping toward understanding fundamental plant responses to deficiencies of Zn and other essential trace elements. AU - Sinclair, Scott A AU - Krämer, U. ID - 7416 IS - 1 JF - Plant Signaling & Behavior SN - 1559-2324 TI - Generation of effective zinc-deficient agar-solidified media allows identification of root morphology changes in response to zinc limitation VL - 15 ER - TY - JOUR AB - The widely used non-steroidal anti-inflammatory drugs (NSAIDs) are derivatives of the phytohormone salicylic acid (SA). SA is well known to regulate plant immunity and development, whereas there have been few reports focusing on the effects of NSAIDs in plants. Our studies here reveal that NSAIDs exhibit largely overlapping physiological activities to SA in the model plant Arabidopsis. NSAID treatments lead to shorter and agravitropic primary roots and inhibited lateral root organogenesis. Notably, in addition to the SA-like action, which in roots involves binding to the protein phosphatase 2A (PP2A), NSAIDs also exhibit PP2A-independent effects. Cell biological and biochemical analyses reveal that many NSAIDs bind directly to and inhibit the chaperone activity of TWISTED DWARF1, thereby regulating actin cytoskeleton dynamics and subsequent endosomal trafficking. Our findings uncover an unexpected bioactivity of human pharmaceuticals in plants and provide insights into the molecular mechanism underlying the cellular action of this class of anti-inflammatory compounds. AU - Tan, Shutang AU - Di Donato, Martin AU - Glanc, Matous AU - Zhang, Xixi AU - Klíma, Petr AU - Liu, Jie AU - Bailly, Aurélien AU - Ferro, Noel AU - Petrášek, Jan AU - Geisler, Markus AU - Friml, Jiří ID - 8943 IS - 9 JF - Cell Reports TI - Non-steroidal anti-inflammatory drugs target TWISTED DWARF1-regulated actin dynamics and auxin transport-mediated plant development VL - 33 ER - TY - JOUR AB - Wound healing in plant tissues, consisting of rigid cell wall-encapsulated cells, represents a considerable challenge and occurs through largely unknown mechanisms distinct from those in animals. Owing to their inability to migrate, plant cells rely on targeted cell division and expansion to regenerate wounds. Strict coordination of these wound-induced responses is essential to ensure efficient, spatially restricted wound healing. Single-cell tracking by live imaging allowed us to gain mechanistic insight into the wound perception and coordination of wound responses after laser-based wounding in Arabidopsis root. We revealed a crucial contribution of the collapse of damaged cells in wound perception and detected an auxin increase specific to cells immediately adjacent to the wound. This localized auxin increase balances wound-induced cell expansion and restorative division rates in a dose-dependent manner, leading to tumorous overproliferation when the canonical TIR1 auxin signaling is disrupted. Auxin and wound-induced turgor pressure changes together also spatially define the activation of key components of regeneration, such as the transcription regulator ERF115. Our observations suggest that the wound signaling involves the sensing of collapse of damaged cells and a local auxin signaling activation to coordinate the downstream transcriptional responses in the immediate wound vicinity. AU - Hörmayer, Lukas AU - Montesinos López, Juan C AU - Marhavá, Petra AU - Benková, Eva AU - Yoshida, Saiko AU - Friml, Jiří ID - 8002 IS - 26 JF - Proceedings of the National Academy of Sciences SN - 0027-8424 TI - Wounding-induced changes in cellular pressure and localized auxin signalling spatially coordinate restorative divisions in roots VL - 117 ER - TY - JOUR AB - Plants, like other multicellular organisms, survive through a delicate balance between growth and defense against pathogens. Salicylic acid (SA) is a major defense signal in plants, and the perception mechanism as well as downstream signaling activating the immune response are known. Here, we identify a parallel SA signaling that mediates growth attenuation. SA directly binds to A subunits of protein phosphatase 2A (PP2A), inhibiting activity of this complex. Among PP2A targets, the PIN2 auxin transporter is hyperphosphorylated in response to SA, leading to changed activity of this important growth regulator. Accordingly, auxin transport and auxin-mediated root development, including growth, gravitropic response, and lateral root organogenesis, are inhibited. This study reveals how SA, besides activating immunity, concomitantly attenuates growth through crosstalk with the auxin distribution network. Further analysis of this dual role of SA and characterization of additional SA-regulated PP2A targets will provide further insights into mechanisms maintaining a balance between growth and defense. AU - Tan, Shutang AU - Abas, Melinda F AU - Verstraeten, Inge AU - Glanc, Matous AU - Molnar, Gergely AU - Hajny, Jakub AU - Lasák, Pavel AU - Petřík, Ivan AU - Russinova, Eugenia AU - Petrášek, Jan AU - Novák, Ondřej AU - Pospíšil, Jiří AU - Friml, Jiří ID - 7427 IS - 3 JF - Current Biology SN - 09609822 TI - Salicylic acid targets protein phosphatase 2A to attenuate growth in plants VL - 30 ER - TY - JOUR AB - Plant survival depends on vascular tissues, which originate in a self‐organizing manner as strands of cells co‐directionally transporting the plant hormone auxin. The latter phenomenon (also known as auxin canalization) is classically hypothesized to be regulated by auxin itself via the effect of this hormone on the polarity of its own intercellular transport. Correlative observations supported this concept, but molecular insights remain limited. In the current study, we established an experimental system based on the model Arabidopsis thaliana, which exhibits auxin transport channels and formation of vasculature strands in response to local auxin application. Our methodology permits the genetic analysis of auxin canalization under controllable experimental conditions. By utilizing this opportunity, we confirmed the dependence of auxin canalization on a PIN‐dependent auxin transport and nuclear, TIR1/AFB‐mediated auxin signaling. We also show that leaf venation and auxin‐mediated PIN repolarization in the root require TIR1/AFB signaling. Further studies based on this experimental system are likely to yield better understanding of the mechanisms underlying auxin transport polarization in other developmental contexts. AU - Mazur, E AU - Kulik, Ivan AU - Hajny, Jakub AU - Friml, Jiří ID - 7500 IS - 5 JF - New Phytologist SN - 0028-646x TI - Auxin canalization and vascular tissue formation by TIR1/AFB-mediated auxin signaling in arabidopsis VL - 226 ER - TY - THES AB - Self-organization is a hallmark of plant development manifested e.g. by intricate leaf vein patterns, flexible formation of vasculature during organogenesis or its regeneration following wounding. Spontaneously arising channels transporting the phytohormone auxin, created by coordinated polar localizations of PIN-FORMED 1 (PIN1) auxin exporter, provide positional cues for these as well as other plant patterning processes. To find regulators acting downstream of auxin and the TIR1/AFB auxin signaling pathway essential for PIN1 coordinated polarization during auxin canalization, we performed microarray experiments. Besides the known components of general PIN polarity maintenance, such as PID and PIP5K kinases, we identified and characterized a new regulator of auxin canalization, the transcription factor WRKY DNA-BINDING PROTEIN 23 (WRKY23). Next, we designed a subsequent microarray experiment to further uncover other molecular players, downstream of auxin-TIR1/AFB-WRKY23 involved in the regulation of auxin-mediated PIN repolarization. We identified a novel and crucial part of the molecular machinery underlying auxin canalization. The auxin-regulated malectin-type receptor-like kinase CAMEL and the associated leucine-rich repeat receptor-like kinase CANAR target and directly phosphorylate PIN auxin transporters. camel and canar mutants are impaired in PIN1 subcellular trafficking and auxin-mediated repolarization leading to defects in auxin transport, ultimately to leaf venation and vasculature regeneration defects. Our results describe the CAMEL-CANAR receptor complex, which is required for auxin feed-back on its own transport and thus for coordinated tissue polarization during auxin canalization. AU - Hajny, Jakub ID - 8822 SN - 2663-337X TI - Identification and characterization of the molecular machinery of auxin-dependent canalization during vasculature formation and regeneration ER - TY - JOUR AB - Flowering plants display the highest diversity among plant species and have notably shaped terrestrial landscapes. Nonetheless, the evolutionary origin of their unprecedented morphological complexity remains largely an enigma. Here, we show that the coevolution of cis-regulatory and coding regions of PIN-FORMED (PIN) auxin transporters confined their expression to certain cell types and directed their subcellular localization to particular cell sides, which together enabled dynamic auxin gradients across tissues critical to the complex architecture of flowering plants. Extensive intraspecies and interspecies genetic complementation experiments with PINs from green alga up to flowering plant lineages showed that PIN genes underwent three subsequent, critical evolutionary innovations and thus acquired a triple function to regulate the development of three essential components of the flowering plant Arabidopsis: shoot/root, inflorescence, and floral organ. Our work highlights the critical role of functional innovations within the PIN gene family as essential prerequisites for the origin of flowering plants. AU - Zhang, Yuzhou AU - Rodriguez Solovey, Lesia AU - Li, Lanxin AU - Zhang, Xixi AU - Friml, Jiří ID - 8986 IS - 50 JF - Science Advances TI - Functional innovations of PIN auxin transporters mark crucial evolutionary transitions during rise of flowering plants VL - 6 ER - TY - JOUR AB - Drought and salt stress are the main environmental cues affecting the survival, development, distribution, and yield of crops worldwide. MYB transcription factors play a crucial role in plants’ biological processes, but the function of pineapple MYB genes is still obscure. In this study, one of the pineapple MYB transcription factors, AcoMYB4, was isolated and characterized. The results showed that AcoMYB4 is localized in the cell nucleus, and its expression is induced by low temperature, drought, salt stress, and hormonal stimulation, especially by abscisic acid (ABA). Overexpression of AcoMYB4 in rice and Arabidopsis enhanced plant sensitivity to osmotic stress; it led to an increase in the number stomata on leaf surfaces and lower germination rate under salt and drought stress. Furthermore, in AcoMYB4 OE lines, the membrane oxidation index, free proline, and soluble sugar contents were decreased. In contrast, electrolyte leakage and malondialdehyde (MDA) content increased significantly due to membrane injury, indicating higher sensitivity to drought and salinity stresses. Besides the above, both the expression level and activities of several antioxidant enzymes were decreased, indicating lower antioxidant activity in AcoMYB4 transgenic plants. Moreover, under osmotic stress, overexpression of AcoMYB4 inhibited ABA biosynthesis through a decrease in the transcription of genes responsible for ABA synthesis (ABA1 and ABA2) and ABA signal transduction factor ABI5. These results suggest that AcoMYB4 negatively regulates osmotic stress by attenuating cellular ABA biosynthesis and signal transduction pathways. AU - Chen, Huihuang AU - Lai, Linyi AU - Li, Lanxin AU - Liu, Liping AU - Jakada, Bello Hassan AU - Huang, Youmei AU - He, Qing AU - Chai, Mengnan AU - Niu, Xiaoping AU - Qin, Yuan ID - 8283 IS - 16 JF - International Journal of Molecular Sciences SN - 16616596 TI - AcoMYB4, an Ananas comosus L. MYB transcription factor, functions in osmotic stress through negative regulation of ABA signaling VL - 21 ER - TY - JOUR AB - Clathrin-mediated endocytosis (CME) is a crucial cellular process implicated in many aspects of plant growth, development, intra- and inter-cellular signaling, nutrient uptake and pathogen defense. Despite these significant roles, little is known about the precise molecular details of how it functions in planta. In order to facilitate the direct quantitative study of plant CME, here we review current routinely used methods and present refined, standardized quantitative imaging protocols which allow the detailed characterization of CME at multiple scales in plant tissues. These include: (i) an efficient electron microscopy protocol for the imaging of Arabidopsis CME vesicles in situ, thus providing a method for the detailed characterization of the ultra-structure of clathrin-coated vesicles; (ii) a detailed protocol and analysis for quantitative live-cell fluorescence microscopy to precisely examine the temporal interplay of endocytosis components during single CME events; (iii) a semi-automated analysis to allow the quantitative characterization of global internalization of cargos in whole plant tissues; and (iv) an overview and validation of useful genetic and pharmacological tools to interrogate the molecular mechanisms and function of CME in intact plant samples. AU - Johnson, Alexander J AU - Gnyliukh, Nataliia AU - Kaufmann, Walter AU - Narasimhan, Madhumitha AU - Vert, G AU - Bednarek, SY AU - Friml, Jiří ID - 8139 IS - 15 JF - Journal of Cell Science SN - 0021-9533 TI - Experimental toolbox for quantitative evaluation of clathrin-mediated endocytosis in the plant model Arabidopsis VL - 133 ER - TY - JOUR AB - The interorganelle communication mediated by membrane contact sites (MCSs) is an evolutionary hallmark of eukaryotic cells. MCS connections enable the nonvesicular exchange of information between organelles and allow them to coordinate responses to changing cellular environments. In plants, the importance of MCS components in the responses to environmental stress has been widely established, but the molecular mechanisms regulating interorganelle connectivity during stress still remain opaque. In this report, we use the model plant Arabidopsis thaliana to show that ionic stress increases endoplasmic reticulum (ER)–plasma membrane (PM) connectivity by promoting the cortical expansion of synaptotagmin 1 (SYT1)-enriched ER–PM contact sites (S-EPCSs). We define differential roles for the cortical cytoskeleton in the regulation of S-EPCS dynamics and ER–PM connectivity, and we identify the accumulation of phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] at the PM as a molecular signal associated with the ER–PM connectivity changes. Our study highlights the functional conservation of EPCS components and PM phosphoinositides as modulators of ER–PM connectivity in eukaryotes, and uncovers unique aspects of the spatiotemporal regulation of ER–PM connectivity in plants. AU - Lee, Eunkyoung AU - Vanneste, Steffen AU - Pérez-Sancho, Jessica AU - Benitez-Fuente, Francisco AU - Strelau, Matthew AU - Macho, Alberto P. AU - Botella, Miguel A. AU - Friml, Jiří AU - Rosado, Abel ID - 5908 IS - 4 JF - Proceedings of the National Academy of Sciences of the United States of America TI - Ionic stress enhances ER–PM connectivity via phosphoinositide-associated SYT1 contact site expansion in Arabidopsis VL - 116 ER - TY - JOUR AB - Multicellular development requires coordinated cell polarization relative to body axes, and translation to oriented cell division 1–3 . In plants, it is unknown how cell polarities are connected to organismal axes and translated to division. Here, we identify Arabidopsis SOSEKI proteins that integrate apical–basal and radial organismal axes to localize to polar cell edges. Localization does not depend on tissue context, requires cell wall integrity and is defined by a transferrable, protein-specific motif. A Domain of Unknown Function in SOSEKI proteins resembles the DIX oligomerization domain in the animal Dishevelled polarity regulator. The DIX-like domain self-interacts and is required for edge localization and for influencing division orientation, together with a second domain that defines the polar membrane domain. Our work shows that SOSEKI proteins locally interpret global polarity cues and can influence cell division orientation. Furthermore, this work reveals that, despite fundamental differences, cell polarity mechanisms in plants and animals converge on a similar protein domain. AU - Yoshida, Saiko AU - Van Der Schuren, Alja AU - Van Dop, Maritza AU - Van Galen, Luc AU - Saiga, Shunsuke AU - Adibi, Milad AU - Möller, Barbara AU - Ten Hove, Colette A. AU - Marhavy, Peter AU - Smith, Richard AU - Friml, Jiří AU - Weijers, Dolf ID - 6023 IS - 2 JF - Nature Plants TI - A SOSEKI-based coordinate system interprets global polarity cues in arabidopsis VL - 5 ER - TY - JOUR AB - Abiotic stress poses constant challenges for plant survival and is a serious problem for global agricultural productivity. On a molecular level, stress conditions result in elevation of reactive oxygen species (ROS) production causing oxidative stress associated with oxidation of proteins and nucleic acids as well as impairment of membrane functions. Adaptation of root growth to ROS accumulation is facilitated through modification of auxin and cytokinin hormone homeostasis. Here, we report that in Arabidopsis root meristem, ROS-induced changes of auxin levels correspond to decreased abundance of PIN auxin efflux carriers at the plasma membrane (PM). Specifically, increase in H2O2 levels affects PIN2 endocytic recycling. We show that the PIN2 intracellular trafficking during adaptation to oxidative stress requires the function of the ADP-ribosylation factor (ARF)-guanine-nucleotide exchange factor (GEF) BEN1, an actin-associated regulator of the trafficking from the PM to early endosomes and, presumably, indirectly, trafficking to the vacuoles. We propose that H2O2 levels affect the actin dynamics thus modulating ARF-GEF-dependent trafficking of PIN2. This mechanism provides a way how root growth acclimates to stress and adapts to a changing environment. AU - Zwiewka, Marta AU - Bielach, Agnieszka AU - Tamizhselvan, Prashanth AU - Madhavan, Sharmila AU - Ryad, Eman Elrefaay AU - Tan, Shutang AU - Hrtyan, Mónika AU - Dobrev, Petre AU - Vanková, Radomira AU - Friml, Jiří AU - Tognetti, Vanesa B. ID - 6104 IS - 2 JF - Plant and Cell Physiology SN - 0032-0781 TI - Root adaptation to H2O2-induced oxidative stress by ARF-GEF BEN1- and cytoskeleton-mediated PIN2 trafficking VL - 60 ER - TY - JOUR AB - Gravitropism is an adaptive response that orients plant growth parallel to the gravity vector. Asymmetric distribution of the phytohormone auxin is a necessary prerequisite to the tropic bending both in roots and shoots. During hypocotyl gravitropic response, the PIN3 auxin transporter polarizes within gravity-sensing cells to redirect intercellular auxin fluxes. First gravity-induced PIN3 polarization to the bottom cell mem- branes leads to the auxin accumulation at the lower side of the organ, initiating bending and, later, auxin feedback-mediated repolarization restores symmetric auxin distribution to terminate bending. Here, we per- formed a forward genetic screen to identify regulators of both PIN3 polarization events during gravitropic response. We searched for mutants with defective PIN3 polarizations based on easy-to-score morphological outputs of decreased or increased gravity-induced hypocotyl bending. We identified the number of hypocotyl reduced bending (hrb) and hypocotyl hyperbending (hhb) mutants, revealing that reduced bending corre- lated typically with defective gravity-induced PIN3 relocation whereas all analyzed hhb mutants showed defects in the second, auxin-mediated PIN3 relocation. Next-generation sequencing-aided mutation map- ping identified several candidate genes, including SCARECROW and ACTIN2, revealing roles of endodermis specification and actin cytoskeleton in the respective gravity- and auxin-induced PIN polarization events. The hypocotyl gravitropism screen thus promises to provide novel insights into mechanisms underlying cell polarity and plant adaptive development. AU - Rakusová, Hana AU - Han, Huibin AU - Valošek, Petr AU - Friml, Jiří ID - 6262 IS - 6 JF - The Plant Journal SN - 0960-7412 TI - Genetic screen for factors mediating PIN polarization in gravistimulated Arabidopsis thaliana hypocotyls VL - 98 ER - TY - JOUR AB - Nitrate regulation of root stem cell activity is auxin-dependent. AU - Wang, Y AU - Gong, Z AU - Friml, Jiří AU - Zhang, J ID - 6261 IS - 1 JF - Plant Physiology SN - 0032-0889 TI - Nitrate modulates the differentiation of root distal stem cells VL - 180 ER - TY - JOUR AB - Root gravitropism is one of the most important processes allowing plant adaptation to the land environment. Auxin plays a central role in mediating root gravitropism, but how auxin contributes to gravitational perception and the subsequent response is still unclear. Here, we showed that the local auxin maximum/gradient within the root apex, which is generated by the PIN directional auxin transporters, regulates the expression of three key starch granule synthesis genes, SS4, PGM and ADG1, which in turn influence the accumulation of starch granules that serve as a statolith perceiving gravity. Moreover, using the cvxIAA‐ccvTIR1 system, we also showed that TIR1‐mediated auxin signaling is required for starch granule formation and gravitropic response within root tips. In addition, axr3 mutants showed reduced auxin‐mediated starch granule accumulation and disruption of gravitropism within the root apex. Our results indicate that auxin‐mediated statolith production relies on the TIR1/AFB‐AXR3‐mediated auxin signaling pathway. In summary, we propose a dual role for auxin in gravitropism: the regulation of both gravity perception and response. AU - Zhang, Yuzhou AU - He, P AU - Ma, X AU - Yang, Z AU - Pang, C AU - Yu, J AU - Wang, G AU - Friml, Jiří AU - Xiao, G ID - 6504 IS - 2 JF - New Phytologist SN - 0028-646x TI - Auxin-mediated statolith production for root gravitropism VL - 224 ER - TY - JOUR AB - Cell polarity is crucial for the coordinated development of all multicellular organisms. In plants, this is exemplified by the PIN-FORMED (PIN) efflux carriers of the phytohormone auxin: The polar subcellular localization of the PINs is instructive to the directional intercellular auxin transport, and thus to a plethora of auxin-regulated growth and developmental processes. Despite its importance, the regulation of PIN polar subcellular localization remains poorly understood. Here, we have employed advanced live-cell imaging techniques to study the roles of microtubules and actin microfilaments in the establishment of apical polar localization of PIN2 in the epidermis of the Arabidopsis root meristem. We report that apical PIN2 polarity requires neither intact actin microfilaments nor microtubules, suggesting that the primary spatial cue for polar PIN distribution is likely independent of cytoskeleton-guided endomembrane trafficking. AU - Glanc, Matous AU - Fendrych, Matyas AU - Friml, Jiří ID - 6611 IS - 6 JF - Biomolecules TI - PIN2 polarity establishment in arabidopsis in the absence of an intact cytoskeleton VL - 9 ER - TY - JOUR AB - An important adaptation during colonization of land by plants is gravitropic growth of roots, which enabled roots to reach water and nutrients, and firmly anchor plants in the ground. Here we provide insights into the evolution of an efficient root gravitropic mechanism in the seed plants. Architectural innovation, with gravity perception constrained in the root tips along with a shootward transport route for the phytohormone auxin, appeared only upon the emergence of seed plants. Interspecies complementation and protein domain swapping revealed functional innovations within the PIN family of auxin transporters leading to the evolution of gravitropism-specific PINs. The unique apical/shootward subcellular localization of PIN proteins is the major evolutionary innovation that connected the anatomically separated sites of gravity perception and growth response via the mobile auxin signal. We conclude that the crucial anatomical and functional components emerged hand-in-hand to facilitate the evolution of fast gravitropic response, which is one of the major adaptations of seed plants to dry land. AU - Zhang, Yuzhou AU - Xiao, G AU - Wang, X AU - Zhang, Xixi AU - Friml, Jiří ID - 6778 JF - Nature Communications SN - 2041-1723 TI - Evolution of fast root gravitropism in seed plants VL - 10 ER - TY - JOUR AB - Plants have a remarkable capacity to adjust their growth and development to elevated ambient temperatures. Increased elongation growth of roots, hypocotyls and petioles in warm temperatures are hallmarks of seedling thermomorphogenesis. In the last decade, significant progress has been made to identify the molecular signaling components regulating these growth responses. Increased ambient temperature utilizes diverse components of the light sensing and signal transduction network to trigger growth adjustments. However, it remains unknown whether temperature sensing and responses are universal processes that occur uniformly in all plant organs. Alternatively, temperature sensing may be confined to specific tissues or organs, which would require a systemic signal that mediates responses in distal parts of the plant. Here we show that Arabidopsis (Arabidopsis thaliana) seedlings show organ-specific transcriptome responses to elevated temperatures, and that thermomorphogenesis involves both autonomous and organ-interdependent temperature sensing and signaling. Seedling roots can sense and respond to temperature in a shoot-independent manner, whereas shoot temperature responses require both local and systemic processes. The induction of cell elongation in hypocotyls requires temperature sensing in cotyledons, followed by generation of a mobile auxin signal. Subsequently, auxin travels to the hypocotyl where it triggers local brassinosteroid-induced cell elongation in seedling stems, which depends upon a distinct, permissive temperature sensor in the hypocotyl. AU - Bellstaedt, Julia AU - Trenner, Jana AU - Lippmann, Rebecca AU - Poeschl, Yvonne AU - Zhang, Xixi AU - Friml, Jiří AU - Quint, Marcel AU - Delker, Carolin ID - 6366 IS - 2 JF - Plant Physiology SN - 0032-0889 TI - A mobile auxin signal connects temperature sensing in cotyledons with growth responses in hypocotyls VL - 180 ER - TY - JOUR AB - The plant hormone auxin has crucial roles in almost all aspects of plant growth and development. Concentrations of auxin vary across different tissues, mediating distinct developmental outcomes and contributing to the functional diversity of auxin. However, the mechanisms that underlie these activities are poorly understood. Here we identify an auxin signalling mechanism, which acts in parallel to the canonical auxin pathway based on the transport inhibitor response1 (TIR1) and other auxin receptor F-box (AFB) family proteins (TIR1/AFB receptors)1,2, that translates levels of cellular auxin to mediate differential growth during apical-hook development. This signalling mechanism operates at the concave side of the apical hook, and involves auxin-mediated C-terminal cleavage of transmembrane kinase 1 (TMK1). The cytosolic and nucleus-translocated C terminus of TMK1 specifically interacts with and phosphorylates two non-canonical transcriptional repressors of the auxin or indole-3-acetic acid (Aux/IAA) family (IAA32 and IAA34), thereby regulating ARF transcription factors. In contrast to the degradation of Aux/IAA transcriptional repressors in the canonical pathway, the newly identified mechanism stabilizes the non-canonical IAA32 and IAA34 transcriptional repressors to regulate gene expression and ultimately inhibit growth. The auxin–TMK1 signalling pathway originates at the cell surface, is triggered by high levels of auxin and shares a partially overlapping set of transcription factors with the TIR1/AFB signalling pathway. This allows distinct interpretations of different concentrations of cellular auxin, and thus enables this versatile signalling molecule to mediate complex developmental outcomes. AU - Cao, Min AU - Chen, Rong AU - Li, Pan AU - Yu, Yongqiang AU - Zheng, Rui AU - Ge, Danfeng AU - Zheng, Wei AU - Wang, Xuhui AU - Gu, Yangtao AU - Gelová, Zuzana AU - Friml, Jiří AU - Zhang, Heng AU - Liu, Renyi AU - He, Jun AU - Xu, Tongda ID - 6259 JF - Nature SN - 0028-0836 TI - TMK1-mediated auxin signalling regulates differential growth of the apical hook VL - 568 ER - TY - JOUR AB - PIN-FORMED (PIN) transporters mediate directional, intercellular movement of the phytohormone auxin in land plants. To elucidate the evolutionary origins of this developmentally crucial mechanism, we analysed the single PIN homologue of a simple green alga Klebsormidium flaccidum. KfPIN functions as a plasma membrane-localized auxin exporter in land plants and heterologous models. While its role in algae remains unclear, PIN-driven auxin export is probably an ancient and conserved trait within streptophytes. AU - Skokan, Roman AU - Medvecká, Eva AU - Viaene, Tom AU - Vosolsobě, Stanislav AU - Zwiewka, Marta AU - Müller, Karel AU - Skůpa, Petr AU - Karady, Michal AU - Zhang, Yuzhou AU - Janacek, Dorina P. AU - Hammes, Ulrich Z. AU - Ljung, Karin AU - Nodzyński, Tomasz AU - Petrášek, Jan AU - Friml, Jiří ID - 7106 IS - 11 JF - Nature Plants SN - 2055-0278 TI - PIN-driven auxin transport emerged early in streptophyte evolution VL - 5 ER - TY - JOUR AB - Roots grow downwards parallel to the gravity vector, to anchor a plant in soil and acquire water and nutrients, using a gravitropic mechanism dependent on the asymmetric distribution of the phytohormone auxin. Recently, Chang et al. demonstrate that asymmetric distribution of another phytohormone, cytokinin, directs root growth towards higher water content. AU - Sinclair, Scott A AU - Friml, Jiří ID - 7143 JF - Cell Research SN - 1001-0602 TI - Defying gravity: a plant's quest for moisture VL - 29 ER - TY - JOUR AB - During infection pathogens secrete small molecules, termed effectors, to manipulate and control the interaction with their specific hosts. Both the pathogen and the plant are under high selective pressure to rapidly adapt and co-evolve in what is usually referred to as molecular arms race. Components of the host’s immune system form a network that processes information about molecules with a foreign origin and damage-associated signals, integrating them with developmental and abiotic cues to adapt the plant’s responses. Both in the case of nucleotide-binding leucine-rich repeat receptors and leucine-rich repeat receptor kinases interaction networks have been extensively characterized. However, little is known on whether pathogenic effectors form complexes to overcome plant immunity and promote disease. Ustilago maydis, a biotrophic fungal pathogen that infects maize plants, produces effectors that target hubs in the immune network of the host cell. Here we assess the capability of U. maydis effector candidates to interact with each other, which may play a crucial role during the infection process. Using a systematic yeast-two-hybrid approach and based on a preliminary pooled screen, we selected 63 putative effectors for one-on-one matings with a library of nearly 300 effector candidates. We found that 126 of these effector candidates interacted either with themselves or other predicted effectors. Although the functional relevance of the observed interactions remains elusive, we propose that the observed abundance in complex formation between effectors adds an additional level of complexity to effector research and should be taken into consideration when studying effector evolution and function. Based on this fundamental finding, we suggest various scenarios which could evolutionarily drive the formation and stabilization of an effector interactome. AU - Alcântara, André AU - Bosch, Jason AU - Nazari, Fahimeh AU - Hoffmann, Gesa AU - Gallei, Michelle C AU - Uhse, Simon AU - Darino, Martin A. AU - Olukayode, Toluwase AU - Reumann, Daniel AU - Baggaley, Laura AU - Djamei, Armin ID - 7182 IS - 11 JF - Frontiers in Plant Science TI - Systematic Y2H screening reveals extensive effector-complex formation VL - 10 ER - TY - JOUR AB - Clathrin-mediated endocytosis (CME) is a highly conserved and essential cellular process in eukaryotic cells, but its dynamic and vital nature makes it challenging to study using classical genetics tools. In contrast, although small molecules can acutely and reversibly perturb CME, the few chemical CME inhibitors that have been applied to plants are either ineffective or show undesirable side effects. Here, we identify the previously described endosidin9 (ES9) as an inhibitor of clathrin heavy chain (CHC) function in both Arabidopsis and human cells through affinity-based target isolation, in vitro binding studies and X-ray crystallography. Moreover, we present a chemically improved ES9 analog, ES9-17, which lacks the undesirable side effects of ES9 while retaining the ability to target CHC. ES9 and ES9-17 have expanded the chemical toolbox used to probe CHC function, and present chemical scaffolds for further design of more specific and potent CHC inhibitors across different systems. AU - Dejonghe, Wim AU - Sharma, Isha AU - Denoo, Bram AU - De Munck, Steven AU - Lu, Qing AU - Mishev, Kiril AU - Bulut, Haydar AU - Mylle, Evelien AU - De Rycke, Riet AU - Vasileva, Mina K AU - Savatin, Daniel V. AU - Nerinckx, Wim AU - Staes, An AU - Drozdzecki, Andrzej AU - Audenaert, Dominique AU - Yperman, Klaas AU - Madder, Annemieke AU - Friml, Jiří AU - Van Damme, Daniël AU - Gevaert, Kris AU - Haucke, Volker AU - Savvides, Savvas N. AU - Winne, Johan AU - Russinova, Eugenia ID - 6377 IS - 6 JF - Nature Chemical Biology SN - 15524450 TI - Disruption of endocytosis through chemical inhibition of clathrin heavy chain function VL - 15 ER - TY - THES AB - The development and growth of Arabidopsis thaliana is regulated by a combination of genetic programing and also by the environmental influences. An important role in these processes play the phytohormones and among them, auxin is crucial as it controls many important functions. It is transported through the whole plant body by creating local and temporal concentration maxima and minima, which have an impact on the cell status, tissue and organ identity. Auxin has the property to undergo a directional and finely regulated cell-to-cell transport, which is enabled by the transport proteins, localized on the plasma membrane. An important role in this process have the PIN auxin efflux proteins, which have an asymmetric/polar subcellular localization and determine the directionality of the auxin transport. During the last years, there were significant advances in understanding how the trafficking molecular machineries function, including studies on molecular interactions, function, subcellular localization and intracellular distribution. However, there is still a lack of detailed characterization on the steps of endocytosis, exocytosis, endocytic recycling and degradation. Due to this fact, I focused on the identification of novel trafficking factors and better characterization of the intracellular trafficking pathways. My PhD thesis consists of an introductory chapter, three experimental chapters, a chapter containing general discussion, conclusions and perspectives and also an appendix chapter with published collaborative papers. The first chapter is separated in two different parts: I start by a general introduction to auxin biology and then I introduce the trafficking pathways in the model plant Arabidopsis thaliana. Then, I explain also the phosphorylation-signals for polar targeting and also the roles of the phytohormone strigolactone. The second chapter includes the characterization of bar1/sacsin mutant, which was identified in a forward genetic screen for novel trafficking components in Arabidopsis thaliana, where by the implementation of an EMS-treated pPIN1::PIN1-GFP marker line and by using the established inhibitor of ARF-GEFs, Brefeldin A (BFA) as a tool to study trafficking processes, we identified a novel factor, which is mediating the adaptation of the plant cell to ARF-GEF inhibition. The mutation is in a previously uncharacterized gene, encoding a very big protein that we, based on its homologies, called SACSIN with domains suggesting roles as a molecular chaperon or as a component of the ubiquitin-proteasome system. Our physiology and imaging studies revealed that SACSIN is a crucial plant cell component of the adaptation to the ARF-GEF inhibition. The third chapter includes six subchapters, where I focus on the role of the phytohormone strigolactone, which interferes with auxin feedback on PIN internalization. Strigolactone moderates the polar auxin transport by increasing the internalization of the PIN auxin efflux carriers, which reduces the canalization related growth responses. In addition, I also studied the role of phosphorylation in the strigolactone regulation of auxin feedback on PIN internalization. In this chapter I also present my results on the MAX2-dependence of strigolactone-mediated root growth inhibition and I also share my results on the auxin metabolomics profiling after application of GR24. In the fourth chapter I studied the effect of two small molecules ES-9 and ES9-17, which were identified from a collection of small molecules with the property to impair the clathrin-mediated endocytosis. In the fifth chapter, I discuss all my observations and experimental findings and suggest alternative hypothesis to interpret my results. In the appendix there are three collaborative published projects. In the first, I participated in the characterization of the role of ES9 as a small molecule, which is inhibitor of clathrin- mediated endocytosis in different model organisms. In the second paper, I contributed to the characterization of another small molecule ES9-17, which is a non-protonophoric analog of ES9 and also impairs the clathrin-mediated endocytosis not only in plant cells, but also in mammalian HeLa cells. Last but not least, I also attach another paper, where I tried to establish the grafting method as a technique in our lab to study canalization related processes. AU - Vasileva, Mina K ID - 7172 TI - Molecular mechanisms of endomembrane trafficking in Arabidopsis thaliana ER - TY - JOUR AB - Plasmodesmata (PD) are plant-specific membrane-lined channels that create cytoplasmic and membrane continuities between adjacent cells, thereby facilitating cell–cell communication and virus movement. Plant cells have evolved diverse mechanisms to regulate PD plasticity in response to numerous environmental stimuli. In particular, during defense against plant pathogens, the defense hormone, salicylic acid (SA), plays a crucial role in the regulation of PD permeability in a callose-dependent manner. Here, we uncover a mechanism by which plants restrict the spreading of virus and PD cargoes using SA signaling by increasing lipid order and closure of PD. We showed that exogenous SA application triggered the compartmentalization of lipid raft nanodomains through a modulation of the lipid raft-regulatory protein, Remorin (REM). Genetic studies, superresolution imaging, and transmission electron microscopy observation together demonstrated that Arabidopsis REM1.2 and REM1.3 are crucial for plasma membrane nanodomain assembly to control PD aperture and functionality. In addition, we also found that a 14-3-3 epsilon protein modulates REM clustering and membrane nanodomain compartmentalization through its direct interaction with REM proteins. This study unveils a molecular mechanism by which the key plant defense hormone, SA, triggers membrane lipid nanodomain reorganization, thereby regulating PD closure to impede virus spreading. AU - Huang, D AU - Sun, Y AU - Ma, Z AU - Ke, M AU - Cui, Y AU - Chen, Z AU - Chen, C AU - Ji, C AU - Tran, TM AU - Yang, L AU - Lam, SM AU - Han, Y AU - Shu, G AU - Friml, Jiří AU - Miao, Y AU - Jiang, L AU - Chen, X ID - 6999 IS - 42 JF - Proceedings of the National Academy of Sciences of the United States of America SN - 0027-8424 TI - Salicylic acid-mediated plasmodesmal closure via Remorin-dependent lipid organization VL - 116 ER - TY - THES AB - Clathrin-Mediated Endocytosis (CME) is an aspect of cellular trafficking that is constantly regulated for mediating developmental and physiological responses. The main aim of my thesis is to decipher the basic mechanisms of CME and post-endocytic trafficking in the whole multicellular organ systems of Arabidopsis. The first chapter of my thesis describes the search for new components involved in CME. Tandem affinity purification was conducted using CLC and its interacting partners were identified. Amongst the identified proteins were the Auxilin-likes1 and 2 (Axl1/2), putative uncoating factors, for which we made a full functional analysis. Over-expression of Axl1/2 causes extreme modifications in the dynamics of the machinery proteins and inhibition of endocytosis altogether. However the loss of function of the axl1/2 did not present any cellular or physiological phenotype, meaning Auxilin-likes do not form the major uncoating machinery. The second chapter of my thesis describes the establishment/utilisation of techniques to capture the dynamicity and the complexity of CME and post-endocytic trafficking. We have studied the development of endocytic pits at the PM – specifically, the mode of membrane remodeling during pit development and the role of actin in it, given plant cells possess high turgor pressure. Utilizing the improved z-resolution of TIRF and VAEM techniques, we captured the time-lapse of the endocytic events at the plasma membrane; and using particle detection software, we quantitatively analysed all the endocytic trajectories in an unbiased way to obtain the endocytic rate of the system. This together with the direct analysis of cargo internalisation from the PM provided an estimate on the endocytic potential of the cell. We also developed a methodology for ultrastructural analysis of different populations of Clathrin-Coated Structures (CCSs) in both PM and endomembranes in unroofed protoplasts. Structural analysis, together with the intensity profile of CCSs at the PM show that the mode of CCP development at the PM follows ‘Constant curvature model’; meaning that clathrin polymerisation energy is a major contributing factor of membrane remodeling. In addition, other analyses clearly show that actin is not required for membrane remodeling during invagination or any other step of CCP development, despite the prevalent high turgor pressure. However, actin is essential in orchestrating the post-endocytic trafficking of CCVs facilitating the EE formation. We also observed that the uncoating process post-endocytosis is not immediate; an alternative mechanism of uncoating – Sequential multi-step process – functions in the cell. Finally we also looked at one of the important physiological stimuli modulating the process – hormone, auxin. auxin has been known to influence CME before. We have made a detailed study on the concentration-time based effect of auxin on the machinery proteins, CCP development, and the specificity of cargoes endocytosed. To this end, we saw no general effect of auxin on CME at earlier time points. However, very low concentration of IAA, such as 50nM, accelerates endocytosis of specifically PIN2 through CME. Such a tight regulatory control with high specificity to PIN2 could be essential in modulating its polarity. AU - Narasimhan, Madhumitha ID - 6269 SN - 2663-337X TI - Clathrin-Mediated endocytosis, post-endocytic trafficking and their regulatory controls in plants ER - TY - JOUR AB - A process of restorative patterning in plant roots correctly replaces eliminated cells to heal local injuries despite the absence of cell migration, which underpins wound healing in animals. Patterning in plants relies on oriented cell divisions and acquisition of specific cell identities. Plants regularly endure wounds caused by abiotic or biotic environmental stimuli and have developed extraordinary abilities to restore their tissues after injuries. Here, we provide insight into a mechanism of restorative patterning that repairs tissues after wounding. Laser-assisted elimination of different cells in Arabidopsis root combined with live-imaging tracking during vertical growth allowed analysis of the regeneration processes in vivo. Specifically, the cells adjacent to the inner side of the injury re-activated their stem cell transcriptional programs. They accelerated their progression through cell cycle, coordinately changed the cell division orientation, and ultimately acquired de novo the correct cell fates to replace missing cells. These observations highlight existence of unknown intercellular positional signaling and demonstrate the capability of specified cells to re-acquire stem cell programs as a crucial part of the plant-specific mechanism of wound healing. AU - Marhavá, Petra AU - Hörmayer, Lukas AU - Yoshida, Saiko AU - Marhavy, Peter AU - Benková, Eva AU - Friml, Jiří ID - 6351 IS - 4 JF - Cell SN - 00928674 TI - Re-activation of stem cell pathways for pattern restoration in plant wound healing VL - 177 ER - TY - JOUR AB - Plants as sessile organisms are constantly under attack by herbivores, rough environmental situations, or mechanical pressure. These challenges often lead to the induction of wounds or destruction of already specified and developed tissues. Additionally, wounding makes plants vulnerable to invasion by pathogens, which is why wound signalling often triggers specific defence responses. To stay competitive or, eventually, survive under these circumstances, plants need to regenerate efficiently, which in rigid, tissue migration-incompatible plant tissues requires post-embryonic patterning and organogenesis. Now, several studies used laser-assisted single cell ablation in the Arabidopsis root tip as a minimal wounding proxy. Here, we discuss their findings and put them into context of a broader spectrum of wound signalling, pathogen responses and tissue as well as organ regeneration. AU - Hörmayer, Lukas AU - Friml, Jiří ID - 6943 JF - Current Opinion in Plant Biology SN - 1369-5266 TI - Targeted cell ablation-based insights into wound healing and restorative patterning VL - 52 ER - TY - JOUR AB - Polar auxin transport plays a pivotal role in plant growth and development. PIN auxin efflux carriers regulate directional auxin movement by establishing local auxin maxima, minima, and gradients that drive multiple developmental processes and responses to environmental signals. Auxin has been proposed to modulate its own transport by regulating subcellular PIN trafficking via processes such as clathrin-mediated PIN endocytosis and constitutive recycling. Here, we further investigated the mechanisms by which auxin affects PIN trafficking by screening auxin analogs and identified pinstatic acid (PISA) as a positive modulator of polar auxin transport in Arabidopsis thaliana. PISA had an auxin-like effect on hypocotyl elongation and adventitious root formation via positive regulation of auxin transport. PISA did not activate SCFTIR1/AFB signaling and yet induced PIN accumulation at the cell surface by inhibiting PIN internalization from the plasma membrane. This work demonstrates PISA to be a promising chemical tool to dissect the regulatory mechanisms behind subcellular PIN trafficking and auxin transport. AU - Oochi, A AU - Hajny, Jakub AU - Fukui, K AU - Nakao, Y AU - Gallei, Michelle C AU - Quareshy, M AU - Takahashi, K AU - Kinoshita, T AU - Harborough, SR AU - Kepinski, S AU - Kasahara, H AU - Napier, RM AU - Friml, Jiří AU - Hayashi, KI ID - 6260 IS - 2 JF - Plant Physiology SN - 0032-0889 TI - Pinstatic acid promotes auxin transport by inhibiting PIN internalization VL - 180 ER - TY - JOUR AB - Cortical microtubule arrays in elongating epidermal cells in both the root and stem of plants have the propensity of dynamic reorientations that are correlated with the activation or inhibition of growth. Factors regulating plant growth, among them the hormone auxin, have been recognized as regulators of microtubule array orientations. Some previous work in the field has aimed at elucidating the causal relationship between cell growth, the signaling of auxin or other growth-regulating factors, and microtubule array reorientations, with various conclusions. Here, we revisit this problem of causality with a comprehensive set of experiments in Arabidopsis thaliana, using the now available pharmacological and genetic tools. We use isolated, auxin-depleted hypocotyls, an experimental system allowing for full control of both growth and auxin signaling. We demonstrate that reorientation of microtubules is not directly triggered by an auxin signal during growth activation. Instead, reorientation is triggered by the activation of the growth process itself and is auxin-independent in its nature. We discuss these findings in the context of previous relevant work, including that on the mechanical regulation of microtubule array orientation. AU - Adamowski, Maciek AU - Li, Lanxin AU - Friml, Jiří ID - 6627 IS - 13 JF - International Journal of Molecular Sciences TI - Reorientation of cortical microtubule arrays in the hypocotyl of arabidopsis thaliana is induced by the cell growth process and independent of auxin signaling VL - 20 ER - TY - CHAP AB - Adventitious roots (AR) are de novo formed roots that emerge from any part of the plant or from callus in tissue culture, except root tissue. The plant tissue origin and the method by which they are induced determine the physiological properties of emerged ARs. Hence, a standard method encompassing all types of AR does not exist. Here we describe a method for the induction and analysis of AR that emerge from the etiolated hypocotyl of dicot plants. The hypocotyl is formed during embryogenesis and shows a determined developmental pattern which usually does not involve AR formation. However, the hypocotyl shows propensity to form de novo roots under specific circumstances such as removal of the root system, high humidity or flooding, or during de-etiolation. The hypocotyl AR emerge from a pericycle-like cell layer surrounding the vascular tissue of the central cylinder, which is reminiscent to the developmental program of lateral roots. Here we propose an easy protocol for in vitro hypocotyl AR induction from etiolated Arabidopsis seedlings. AU - Trinh, Hoang AU - Verstraeten, Inge AU - Geelen, Danny ID - 408 SN - 1064-3745 T2 - Root Development TI - In vitro assay for induction of adventitious rooting on intact arabidopsis hypocotyls VL - 1761 ER - TY - CHAP AB - Immunolocalization is a valuable tool for cell biology research that allows to rapidly determine the localization and expression levels of endogenous proteins. In plants, whole-mount in situ immunolocalization remains a challenging method, especially in tissues protected by waxy layers and complex cell wall carbohydrates. Here, we present a robust method for whole-mount in situ immunolocalization in primary root meristems and lateral root primordia in Arabidopsis thaliana. For good epitope preservation, fixation is done in an alkaline paraformaldehyde/glutaraldehyde mixture. This fixative is suitable for detecting a wide range of proteins, including integral transmembrane proteins and proteins peripherally attached to the plasma membrane. From initiation until emergence from the primary root, lateral root primordia are surrounded by several layers of differentiated tissues with a complex cell wall composition that interferes with the efficient penetration of all buffers. Therefore, immunolocalization in early lateral root primordia requires a modified method, including a strong solvent treatment for removal of hydrophobic barriers and a specific cocktail of cell wall-degrading enzymes. The presented method allows for easy, reliable, and high-quality in situ detection of the subcellular localization of endogenous proteins in primary and lateral root meristems without the need of time-consuming crosses or making translational fusions to fluorescent proteins. AU - Karampelias, Michael AU - Tejos, Ricardo AU - Friml, Jirí AU - Vanneste, Steffen ED - Ristova, Daniela ED - Barbez, Elke ID - 411 T2 - Root Development. Methods and Protocols TI - Optimized whole mount in situ immunolocalization for Arabidopsis thaliana root meristems and lateral root primordia VL - 1761 ER - TY - JOUR AB - Asymmetric auxin distribution is instrumental for the differential growth that causes organ bending on tropic stimuli and curvatures during plant development. Local differences in auxin concentrations are achieved mainly by polarized cellular distribution of PIN auxin transporters, but whether other mechanisms involving auxin homeostasis are also relevant for the formation of auxin gradients is not clear. Here we show that auxin methylation is required for asymmetric auxin distribution across the hypocotyl, particularly during its response to gravity. We found that loss-of-function mutants in Arabidopsis IAA CARBOXYL METHYLTRANSFERASE1 (IAMT1) prematurely unfold the apical hook, and that their hypocotyls are impaired in gravitropic reorientation. This defect is linked to an auxin-dependent increase in PIN gene expression, leading to an increased polar auxin transport and lack of asymmetric distribution of PIN3 in the iamt1 mutant. Gravitropic reorientation in the iamt1 mutant could be restored with either endodermis-specific expression of IAMT1 or partial inhibition of polar auxin transport, which also results in normal PIN gene expression levels. We propose that IAA methylation is necessary in gravity-sensing cells to restrict polar auxin transport within the range of auxin levels that allow for differential responses. AU - Abbas, Mohamad AU - Hernández, García J AU - Pollmann, Stephan AU - Samodelov, Sophia L AU - Kolb, Martina AU - Friml, Jirí AU - Hammes, Ulrich Z AU - Zurbriggen, Matias D AU - Blázquez, Miguel AU - Alabadí, David ID - 203 IS - 26 JF - PNAS TI - Auxin methylation is required for differential growth in Arabidopsis VL - 115 ER - TY - JOUR AB - CLE peptides have been implicated in various developmental processes of plants and mediate their responses to environmental stimuli. However, the biological relevance of most CLE genes remains to be functionally characterized. Here, we report that CLE9, which is expressed in stomata, acts as an essential regulator in the induction of stomatal closure. Exogenous application of CLE9 peptides or overexpression of CLE9 effectively led to stomatal closure and enhanced drought tolerance, whereas CLE9 loss-of-function mutants were sensitivity to drought stress. CLE9-induced stomatal closure was impaired in abscisic acid (ABA)-deficient mutants, indicating that ABA is required for CLE9-medaited guard cell signalling. We further deciphered that two guard cell ABA-signalling components, OST1 and SLAC1, were responsible for CLE9-induced stomatal closure. MPK3 and MPK6 were activated by the CLE9 peptide, and CLE9 peptides failed to close stomata in mpk3 and mpk6 mutants. In addition, CLE9 peptides stimulated the induction of hydrogen peroxide (H2O2) and nitric oxide (NO) synthesis associated with stomatal closure, which was abolished in the NADPH oxidase-deficient mutants or nitric reductase mutants, respectively. Collectively, our results reveal a novel ABA-dependent function of CLE9 in the regulation of stomatal apertures, thereby suggesting a potential role of CLE9 in the stress acclimatization of plants. AU - Zhang, Luosha AU - Shi, Xiong AU - Zhang, Yutao AU - Wang, Jiajing AU - Yang, Jingwei AU - Ishida, Takashi AU - Jiang, Wenqian AU - Han, Xiangyu AU - Kang, Jingke AU - Wang, Xuening AU - Pan, Lixia AU - Lv, Shuo AU - Cao, Bing AU - Zhang, Yonghong AU - Wu, Jinbin AU - Han, Huibin AU - Hu, Zhubing AU - Cui, Langjun AU - Sawa, Shinichiro AU - He, Junmin AU - Wang, Guodong ID - 5830 JF - Plant Cell and Environment SN - 01407791 TI - CLE9 peptide-induced stomatal closure is mediated by abscisic acid, hydrogen peroxide, and nitric oxide in arabidopsis thaliana ER - TY - JOUR AB - The plant hormone gibberellic acid (GA) is a crucial regulator of growth and development. The main paradigm of GA signaling puts forward transcriptional regulation via the degradation of DELLA transcriptional repressors. GA has also been shown to regulate tropic responses by modulation of the plasma membrane incidence of PIN auxin transporters by an unclear mechanism. Here we uncovered the cellular and molecular mechanisms by which GA redirects protein trafficking and thus regulates cell surface functionality. Photoconvertible reporters revealed that GA balances the protein traffic between the vacuole degradation route and recycling back to the cell surface. Low GA levels promote vacuolar delivery and degradation of multiple cargos, including PIN proteins, whereas high GA levels promote their recycling to the plasma membrane. This GA effect requires components of the retromer complex, such as Sorting Nexin 1 (SNX1) and its interacting, microtubule (MT)-associated protein, the Cytoplasmic Linker-Associated Protein (CLASP1). Accordingly, GA regulates the subcellular distribution of SNX1 and CLASP1, and the intact MT cytoskeleton is essential for the GA effect on trafficking. This GA cellular action occurs through DELLA proteins that regulate the MT and retromer presumably via their interaction partners Prefoldins (PFDs). Our study identified a branching of the GA signaling pathway at the level of DELLA proteins, which, in parallel to regulating transcription, also target by a nontranscriptional mechanism the retromer complex acting at the intersection of the degradation and recycling trafficking routes. By this mechanism, GA can redirect receptors and transporters to the cell surface, thus coregulating multiple processes, including PIN-dependent auxin fluxes during tropic responses. AU - Salanenka, Yuliya AU - Verstraeten, Inge AU - Löfke, Christian AU - Tabata, Kaori AU - Naramoto, Satoshi AU - Glanc, Matous AU - Friml, Jirí ID - 428 IS - 14 JF - PNAS TI - Gibberellin DELLA signaling targets the retromer complex to redirect protein trafficking to the plasma membrane VL - 115 ER - TY - JOUR AB - Flowers have a species-specific functional life span that determines the time window in which pollination, fertilization and seed set can occur. The stigma tissue plays a key role in flower receptivity by intercepting pollen and initiating pollen tube growth toward the ovary. In this article, we show that a developmentally controlled cell death programme terminates the functional life span of stigma cells in Arabidopsis. We identified the leaf senescence regulator ORESARA1 (also known as ANAC092) and the previously uncharacterized KIRA1 (also known as ANAC074) as partially redundant transcription factors that modulate stigma longevity by controlling the expression of programmed cell death-associated genes. KIRA1 expression is sufficient to induce cell death and terminate floral receptivity, whereas lack of both KIRA1 and ORESARA1 substantially increases stigma life span. Surprisingly, the extension of stigma longevity is accompanied by only a moderate extension of flower receptivity, suggesting that additional processes participate in the control of the flower's receptive life span. AU - Gao, Zhen AU - Daneva, Anna AU - Salanenka, Yuliya AU - Van Durme, Matthias AU - Huysmans, Marlies AU - Lin, Zongcheng AU - De Winter, Freya AU - Vanneste, Steffen AU - Karimi, Mansour AU - Van De Velde, Jan AU - Vandepoele, Klaas AU - Van De Walle, Davy AU - Dewettinck, Koen AU - Lambrecht, Bart AU - Nowack, Moritz ID - 280 IS - 6 JF - Nature Plants TI - KIRA1 and ORESARA1 terminate flower receptivity by promoting cell death in the stigma of Arabidopsis VL - 4 ER - TY - JOUR AB - The angiosperm seed is composed of three genetically distinct tissues: the diploid embryo that originates from the fertilized egg cell, the triploid endosperm that is produced from the fertilized central cell, and the maternal sporophytic integuments that develop into the seed coat1. At the onset of embryo development in Arabidopsis thaliana, the zygote divides asymmetrically, producing a small apical embryonic cell and a larger basal cell that connects the embryo to the maternal tissue2. The coordinated and synchronous development of the embryo and the surrounding integuments, and the alignment of their growth axes, suggest communication between maternal tissues and the embryo. In contrast to animals, however, where a network of maternal factors that direct embryo patterning have been identified3,4, only a few maternal mutations have been described to affect embryo development in plants5–7. Early embryo patterning in Arabidopsis requires accumulation of the phytohormone auxin in the apical cell by directed transport from the suspensor8–10. However, the origin of this auxin has remained obscure. Here we investigate the source of auxin for early embryogenesis and provide evidence that the mother plant coordinates seed development by supplying auxin to the early embryo from the integuments of the ovule. We show that auxin response increases in ovules after fertilization, due to upregulated auxin biosynthesis in the integuments, and this maternally produced auxin is required for correct embryo development. AU - Robert, Hélène AU - Park, Chulmin AU - Gutièrrez, Carla AU - Wójcikowska, Barbara AU - Pěnčík, Aleš AU - Novák, Ondřej AU - Chen, Junyi AU - Grunewald, Wim AU - Dresselhaus, Thomas AU - Friml, Jirí AU - Laux, Thomas ID - 158 IS - 8 JF - Nature Plants TI - Maternal auxin supply contributes to early embryo patterning in Arabidopsis VL - 4 ER - TY - JOUR AB - AtNHX5 and AtNHX6 are endosomal Na+,K+/H+ antiporters that are critical for growth and development in Arabidopsis, but the mechanism behind their action remains unknown. Here, we report that AtNHX5 and AtNHX6, functioning as H+ leak, control auxin homeostasis and auxin-mediated development. We found that nhx5 nhx6 exhibited growth variations of auxin-related defects. We further showed that nhx5 nhx6 was affected in auxin homeostasis. Genetic analysis showed that AtNHX5 and AtNHX6 were required for the function of the ER-localized auxin transporter PIN5. Although AtNHX5 and AtNHX6 were co-localized with PIN5 at ER, they did not interact directly. Instead, the conserved acidic residues in AtNHX5 and AtNHX6, which are essential for exchange activity, were required for PIN5 function. AtNHX5 and AtNHX6 regulated the pH in ER. Overall, AtNHX5 and AtNHX6 may regulate auxin transport across the ER via the pH gradient created by their transport activity. H+-leak pathway provides a fine-tuning mechanism that controls cellular auxin fluxes. AU - Fan, Ligang AU - Zhao, Lei AU - Hu, Wei AU - Li, Weina AU - Novák, Ondřej AU - Strnad, Miroslav AU - Simon, Sibu AU - Friml, Jirí AU - Shen, Jinbo AU - Jiang, Liwen AU - Qiu, Quan ID - 462 JF - Plant, Cell and Environment TI - NHX antiporters regulate the pH of endoplasmic reticulum and auxin-mediated development VL - 41 ER - TY - JOUR AB - The phytohormone auxin is the information carrier in a plethora of developmental and physiological processes in plants(1). It has been firmly established that canonical, nuclear auxin signalling acts through regulation of gene transcription(2). Here, we combined microfluidics, live imaging, genetic engineering and computational modelling to reanalyse the classical case of root growth inhibition(3) by auxin. We show that Arabidopsis roots react to addition and removal of auxin by extremely rapid adaptation of growth rate. This process requires intracellular auxin perception but not transcriptional reprogramming. The formation of the canonical TIR1/AFB-Aux/IAA co-receptor complex is required for the growth regulation, hinting to a novel, non-transcriptional branch of this signalling pathway. Our results challenge the current understanding of root growth regulation by auxin and suggest another, presumably non-transcriptional, signalling output of the canonical auxin pathway. AU - Fendrych, Matyas AU - Akhmanova, Maria AU - Merrin, Jack AU - Glanc, Matous AU - Hagihara, Shinya AU - Takahashi, Koji AU - Uchida, Naoyuki AU - Torii, Keiko U AU - Friml, Jirí ID - 192 IS - 7 JF - Nature Plants TI - Rapid and reversible root growth inhibition by TIR1 auxin signalling VL - 4 ER - TY - JOUR AB - The intercellular transport of auxin is driven by PIN-formed (PIN) auxin efflux carriers. PINs are localized at the plasma membrane (PM) and on constitutively recycling endomembrane vesicles. Therefore, PINs can mediate auxin transport either by direct translocation across the PM or by pumping auxin into secretory vesicles (SVs), leading to its secretory release upon fusion with the PM. Which of these two mechanisms dominates is a matter of debate. Here, we addressed the issue with a mathematical modeling approach. We demonstrate that the efficiency of secretory transport depends on SV size, half-life of PINs on the PM, pH, exocytosis frequency and PIN density. 3D structured illumination microscopy (SIM) was used to determine PIN density on the PM. Combining this data with published values of the other parameters, we show that the transport activity of PINs in SVs would have to be at least 1000× greater than on the PM in order to produce a comparable macroscopic auxin transport. If both transport mechanisms operated simultaneously and PINs were equally active on SVs and PM, the contribution of secretion to the total auxin flux would be negligible. In conclusion, while secretory vesicle-mediated transport of auxin is an intriguing and theoretically possible model, it is unlikely to be a major mechanism of auxin transport inplanta. AU - Hille, Sander AU - Akhmanova, Maria AU - Glanc, Matous AU - Johnson, Alexander J AU - Friml, Jirí ID - 14 IS - 11 JF - International Journal of Molecular Sciences TI - Relative contribution of PIN-containing secretory vesicles and plasma membrane PINs to the directed auxin transport: Theoretical estimation VL - 19 ER - TY - JOUR AB - Wheat (Triticum ssp.) is one of the most important human food sources. However, this crop is very sensitive to temperature changes. Specifically, processes during wheat leaf, flower, and seed development and photosynthesis, which all contribute to the yield of this crop, are affected by high temperature. While this has to some extent been investigated on physiological, developmental, and molecular levels, very little is known about early signalling events associated with an increase in temperature. Phosphorylation-mediated signalling mechanisms, which are quick and dynamic, are associated with plant growth and development, also under abiotic stress conditions. Therefore, we probed the impact of a short-term and mild increase in temperature on the wheat leaf and spikelet phosphoproteome. In total, 3822 (containing 5178 phosphosites) and 5581 phosphopeptides (containing 7023 phosphosites) were identified in leaf and spikelet samples, respectively. Following statistical analysis, the resulting data set provides the scientific community with a first large-scale plant phosphoproteome under the control of higher ambient temperature. This community resource on the high temperature-mediated wheat phosphoproteome will be valuable for future studies. Our analyses also revealed a core set of common proteins between leaf and spikelet, suggesting some level of conserved regulatory mechanisms. Furthermore, we observed temperature-regulated interconversion of phosphoforms, which probably impacts protein activity. AU - Vu, Lam AU - Zhu, Tingting AU - Verstraeten, Inge AU - Van De Cotte, Brigitte AU - Gevaert, Kris AU - De Smet, Ive ID - 36 IS - 19 JF - Journal of Experimental Botany TI - Temperature-induced changes in the wheat phosphoproteome reveal temperature-regulated interconversion of phosphoforms VL - 69 ER - TY - JOUR AB - Land plants evolved from charophytic algae, among which Charophyceae possess the most complex body plans. We present the genome of Chara braunii; comparison of the genome to those of land plants identified evolutionary novelties for plant terrestrialization and land plant heritage genes. C. braunii employs unique xylan synthases for cell wall biosynthesis, a phragmoplast (cell separation) mechanism similar to that of land plants, and many phytohormones. C. braunii plastids are controlled via land-plant-like retrograde signaling, and transcriptional regulation is more elaborate than in other algae. The morphological complexity of this organism may result from expanded gene families, with three cases of particular note: genes effecting tolerance to reactive oxygen species (ROS), LysM receptor-like kinases, and transcription factors (TFs). Transcriptomic analysis of sexual reproductive structures reveals intricate control by TFs, activity of the ROS gene network, and the ancestral use of plant-like storage and stress protection proteins in the zygote. AU - Nishiyama, Tomoaki AU - Sakayama, Hidetoshi AU - De Vries, Jan AU - Buschmann, Henrik AU - Saint Marcoux, Denis AU - Ullrich, Kristian AU - Haas, Fabian AU - Vanderstraeten, Lisa AU - Becker, Dirk AU - Lang, Daniel AU - Vosolsobě, Stanislav AU - Rombauts, Stephane AU - Wilhelmsson, Per AU - Janitza, Philipp AU - Kern, Ramona AU - Heyl, Alexander AU - Rümpler, Florian AU - Calderón Villalobos, Luz AU - Clay, John AU - Skokan, Roman AU - Toyoda, Atsushi AU - Suzuki, Yutaka AU - Kagoshima, Hiroshi AU - Schijlen, Elio AU - Tajeshwar, Navindra AU - Catarino, Bruno AU - Hetherington, Alexander AU - Saltykova, Assia AU - Bonnot, Clemence AU - Breuninger, Holger AU - Symeonidi, Aikaterini AU - Radhakrishnan, Guru AU - Van Nieuwerburgh, Filip AU - Deforce, Dieter AU - Chang, Caren AU - Karol, Kenneth AU - Hedrich, Rainer AU - Ulvskov, Peter AU - Glöckner, Gernot AU - Delwiche, Charles AU - Petrášek, Jan AU - Van De Peer, Yves AU - Friml, Jirí AU - Beilby, Mary AU - Dolan, Liam AU - Kohara, Yuji AU - Sugano, Sumio AU - Fujiyama, Asao AU - Delaux, Pierre Marc AU - Quint, Marcel AU - Theissen, Gunter AU - Hagemann, Martin AU - Harholt, Jesper AU - Dunand, Christophe AU - Zachgo, Sabine AU - Langdale, Jane AU - Maumus, Florian AU - Van Der Straeten, Dominique AU - Gould, Sven B AU - Rensing, Stefan ID - 148 IS - 2 JF - Cell TI - The Chara genome: Secondary complexity and implications for plant terrestrialization VL - 174 ER - TY - JOUR AB - The trafficking of subcellular cargos in eukaryotic cells crucially depends on vesicle budding, a process mediated by ARF-GEFs (ADP-ribosylation factor guanine nucleotide exchange factors). In plants, ARF-GEFs play essential roles in endocytosis, vacuolar trafficking, recycling, secretion, and polar trafficking. Moreover, they are important for plant development, mainly through controlling the polar subcellular localization of PIN-FORMED (PIN) transporters of the plant hormone auxin. Here, using a chemical genetics screen in Arabidopsis thaliana, we identified Endosidin 4 (ES4), an inhibitor of eukaryotic ARF-GEFs. ES4 acts similarly to and synergistically with the established ARF-GEF inhibitor Brefeldin A and has broad effects on intracellular trafficking, including endocytosis, exocytosis, and vacuolar targeting. Additionally, Arabidopsis and yeast (Sacharomyces cerevisiae) mutants defective in ARF-GEF show altered sensitivity to ES4. ES4 interferes with the activation-based membrane association of the ARF1 GTPases, but not of their mutant variants that are activated independently of ARF-GEF activity. Biochemical approaches and docking simulations confirmed that ES4 specifically targets the SEC7 domain-containing ARF-GEFs. These observations collectively identify ES4 as a chemical tool enabling the study of ARF-GEF-mediated processes, including ARF-GEF-mediated plant development. AU - Kania, Urszula AU - Nodzyński, Tomasz AU - Lu, Qing AU - Hicks, Glenn R AU - Nerinckx, Wim AU - Mishev, Kiril AU - Peurois, Francois AU - Cherfils, Jacqueline AU - De, Rycke Riet Maria AU - Grones, Peter AU - Robert, Stéphanie AU - Russinova, Eugenia AU - Friml, Jirí ID - 147 IS - 10 JF - The Plant Cell SN - 1040-4651 TI - The inhibitor Endosidin 4 targets SEC7 domain-type ARF GTPase exchange factors and interferes with sub cellular trafficking in eukaryotes VL - 30 ER - TY - JOUR AB - The root cap protects the stem cell niche of angiosperm roots from damage. In Arabidopsis, lateral root cap (LRC) cells covering the meristematic zone are regularly lost through programmed cell death, while the outermost layer of the root cap covering the tip is repeatedly sloughed. Efficient coordination with stem cells producing new layers is needed to maintain a constant size of the cap. We present a signalling pair, the peptide IDA-LIKE1 (IDL1) and its receptor HAESA-LIKE2 (HSL2), mediating such communication. Live imaging over several days characterized this process from initial fractures in LRC cell files to full separation of a layer. Enhanced expression of IDL1 in the separating root cap layers resulted in increased frequency of sloughing, balanced with generation of new layers in a HSL2-dependent manner. Transcriptome analyses linked IDL1-HSL2 signalling to the transcription factors BEARSKIN1/2 and genes associated with programmed cell death. Mutations in either IDL1 or HSL2 slowed down cell division, maturation and separation. Thus, IDL1-HSL2 signalling potentiates dynamic regulation of the homeostatic balance between stem cell division and sloughing activity. AU - Shi, Chun Lin AU - Von Wangenheim, Daniel AU - Herrmann, Ullrich AU - Wildhagen, Mari AU - Kulik, Ivan AU - Kopf, Andreas AU - Ishida, Takashi AU - Olsson, Vilde AU - Anker, Mari Kristine AU - Albert, Markus AU - Butenko, Melinka A AU - Felix, Georg AU - Sawa, Shinichiro AU - Claassen, Manfred AU - Friml, Jirí AU - Aalen, Reidunn B ID - 146 IS - 8 JF - Nature Plants TI - The dynamics of root cap sloughing in Arabidopsis is regulated by peptide signalling VL - 4 ER - TY - JOUR AB - Strigolactones (SLs) are a relatively recent addition to the list of plant hormones that control different aspects of plant development. SL signalling is perceived by an α/β hydrolase, DWARF 14 (D14). A close homolog of D14, KARRIKIN INSENSTIVE2 (KAI2), is involved in perception of an uncharacterized molecule called karrikin (KAR). Recent studies in Arabidopsis identified the SUPPRESSOR OF MAX2 1 (SMAX1) and SMAX1-LIKE 7 (SMXL7) to be potential SCF–MAX2 complex-mediated proteasome targets of KAI2 and D14, respectively. Genetic studies on SMXL7 and SMAX1 demonstrated distinct developmental roles for each, but very little is known about these repressors in terms of their sequence features. In this study, we performed an extensive comparative analysis of SMXLs and determined their phylogenetic and evolutionary history in the plant lineage. Our results show that SMXL family members can be sub-divided into four distinct phylogenetic clades/classes, with an ancient SMAX1. Further, we identified the clade-specific motifs that have evolved and that might act as determinants of SL-KAR signalling specificity. These specificities resulted from functional diversities among the clades. Our results suggest that a gradual co-evolution of SMXL members with their upstream receptors D14/KAI2 provided an increased specificity to both the SL perception and response in land plants. AU - Moturu, Taraka Ramji AU - Thula, Sravankumar AU - Singh, Ravi Kumar AU - Nodzyński, Tomasz AU - Vařeková, Radka Svobodová AU - Friml, Jiří AU - Simon, Sibu ID - 10881 IS - 9 JF - Journal of Experimental Botany KW - Plant Science KW - Physiology SN - 0022-0957 TI - Molecular evolution and diversification of the SMXL gene family VL - 69 ER -