TY - JOUR A1 - Zhao, Liming A1 - Xia, Yan A1 - Wu, Xiao-Yuan A1 - Schippers, Jos H. M. A1 - Jing, Hai-Chun T1 - Phenotypic analysis and molecular markers of leaf senescence JF - Plant Senescence: Methods and Protocols N2 - The process of leaf senescence consists of the final stage of leaf development. It has evolved as a mechanism to degrade macromolecules and micronutrients and remobilize them to other developing parts of the plant; hence it plays a central role for the survival of plants and crop production. During senescence, a range of physiological, morphological, cellular, and molecular events occur, which are generally referred to as the senescence syndrome that includes several hallmarks such as visible yellowing, loss of chlorophyll and water content, increase of ion leakage and cell death, deformation of chloroplast and cell structure, as well as the upregulation of thousands of so-called senescence-associated genes (SAGs) and downregulation of photosynthesis-associated genes (PAGs). This chapter is devoted to methods characterizing the onset and progression of leaf senescence at the morphological, physiological, cellular, and molecular levels. Leaf senescence normally progresses in an age-dependent manner but is also induced prematurely by a variety of environmental stresses in plants. Focused on the hallmarks of the senescence syndrome, a series of protocols is described to asses quantitatively the senescence process caused by developmental cues or environmental perturbations. We first briefly describe the senescence process, the events associated with the senescence syndrome, and the theories and methods to phenotype senescence. Detailed protocols for monitoring senescence in planta and in vitro, using the whole plant and the detached leaf, respectively, are presented. For convenience, most of the protocols use the model plant species Arabidopsis and rice, but they can be easily extended to other plants. KW - Leaf senescence KW - Visible yellowing KW - Chlorophyll KW - Ion leakage KW - Cell death KW - Senescence-associated genes (SAGs) KW - Arabidopsis KW - Rice Y1 - 2018 SN - 978-1-4939-7672-0 SN - 978-1-4939-7670-6 U6 - https://doi.org/10.1007/978-1-4939-7672-0_3 SN - 1064-3745 SN - 1940-6029 VL - 1744 SP - 35 EP - 48 PB - Humana Press Inc. CY - Totowa ER - TY - JOUR A1 - Watanabe, Mutsumi A1 - Tohge, Takayuki A1 - Balazadeh, Salma A1 - Erban, Alexander A1 - Giavalisco, Patrick A1 - Kopka, Joachim A1 - Mueller-Roeber, Bernd A1 - Fernie, Alisdair A1 - Hoefgen, Rainer T1 - Comprehensive Metabolomics Studies of Plant Developmental Senescence JF - Plant Senescence: Methods and Protocols N2 - Leaf senescence is an essential developmental process that involves diverse metabolic changes associated with degradation of macromolecules allowing nutrient recycling and remobilization. In contrast to the significant progress in transcriptomic analysis of leaf senescence, metabolomics analyses have been relatively limited. A broad overview of metabolic changes during leaf senescence including the interactions between various metabolic pathways is required to gain a better understanding of the leaf senescence allowing to link transcriptomics with metabolomics and physiology. In this chapter, we describe how to obtain comprehensive metabolite profiles and how to dissect metabolic shifts during leaf senescence in the model plant Arabidopsis thaliana. Unlike nucleic acid analysis for transcriptomics, a comprehensive metabolite profile can only be achieved by combining a suite of analytic tools. Here, information is provided for measurements of the contents of chlorophyll, soluble proteins, and starch by spectrophotometric methods, ions by ion chromatography, thiols and amino acids by HPLC, primary metabolites by GC/TOF-MS, and secondary metabolites and lipophilic metabolites by LC/ESI-MS. These metabolite profiles provide a rich catalogue of metabolic changes during leaf senescence, which is a helpful database and blueprint to be correlated to future studies such as transcriptome and proteome analyses, forward and reverse genetic studies, or stress-induced senescence studies. KW - Senescence KW - Metabolomics KW - Arabidopsis KW - GC/MS KW - LC/MS KW - HPLC KW - IC Y1 - 2018 SN - 978-1-4939-7672-0 SN - 978-1-4939-7670-6 U6 - https://doi.org/10.1007/978-1-4939-7672-0_28 SN - 1064-3745 SN - 1940-6029 VL - 1744 SP - 339 EP - 358 PB - Humana Press CY - Totowa ER - TY - THES A1 - Wang, Yang T1 - Role of the actin cytoskeleton in cellular morphogenesis at the shoot apical meristem of Arabidopsis thaliana N2 - The morphogenesis of sessile plants is mainly driven by directional cell growth and cell division. The organization of their cytoskeleton and the mechanical properties of the cell wall greatly influence morphogenetic events in plants. It is well known that cortical microtubules (CMTs) contribute to directional growth by regulating the deposition of the cellulose microfibrils, as major cell wall fortifying elements. More recent findings demonstrate that mechanical stresses existing in cells and tissues influence microtubule organization. Also, in dividing cells, mechanical stress directions contribute to the orientation of the new cell wall. In comparison to the microtubule cytoskeleton, the role of the actin cytoskeleton in regulating shoot meristem morphogenesis has not been extensively studied. This thesis focuses on the functional relevance of the actin cytoskeleton during cell and tissue scale morphogenesis in the shoot apical meristem (SAM) of Arabidopsis thaliana. Visualization of transcriptional reporters indicates that ACTIN2 and ACTIN7 are two highly expressed actin genes in the SAM. A link between the actin cytoskeleton and SAM development derives from the observation that the act2-1 act7-1 double mutant has abnormal cell shape and perturbed phyllotactic patterns. Live-cell imaging of the actin cytoskeleton further shows that its organization correlates with cell shape, which indicates a potential role of actin in influencing cellular morphogenesis. In this thesis, a detailed characterization of the act2-1 act7-1 mutant reveals that perturbation of actin leads to more rectangular cellular geometries with more 90° cell internal angles, and higher incidences of four-way junctions (four cell boundaries intersecting together). This observation deviates from the conventional tricellular junctions found in epidermal cells. Quantitative cellular-level growth data indicates that such differences in the act2-1 act7-1 mutant arise due to the reduced accuracy in the placement of the new cell wall, as well as its mechanical maturation. Changes in cellular morphology observed in the act2-1 act7-1 mutant result in cell packing defects that subsequently compromise the flow of information among cells in the SAM. N2 - Die Morphogenese sessiler Pflanzen wird hauptsächlich durch gerichtetes Zellwachstum und Zellteilung angetrieben. Die Organisation des Zytoskeletts und die mechanischen Eigenschaften der Zellwand haben großen Einfluss auf die morphogenetischen Vorgänge in Pflanzen. Es ist bekannt, dass kortikale Mikrotubuli (CMTs) zum gerichteten Wachstum beitragen, indem sie die Bildung von Zellulose-Mikrofibrillen als wichtige Elemente zur Stärkung der Zellwand regulieren. Neuere Erkenntnisse zeigen, dass mechanische Spannungen in den Zellen und Geweben die Organisation der Mikrotubuli beeinflussen. Bei der Zellteilung tragen auch die mechanischen Belastungsrichtungen zur Ausrichtung der neuen Zellwand bei. Im Vergleich zum Zytoskelett der Mikrotubuli ist die Rolle des Aktinzytoskeletts bei der Regulierung der Morphogenese des Sprossmeristems noch nicht umfassend untersucht worden. Diese Arbeit befasst sich mit der funktionellen Bedeutung des Aktinzytoskeletts bei der Zell- und Gewebemorphogenese im Sprossapikalmeristem (SAM) von Arabidopsis thaliana. Die Visualisierung von Transkriptionsreportern zeigt, dass ACTIN2 und ACTIN7 zwei stark exprimierte Aktingene im SAM sind. Eine Verknüpfung zwischen dem Aktinzytoskelett und der Entwicklung des SAMs ergibt sich aus der Beobachtung, dass die act2-1 act7-1 Doppelmutante eine abnorme Zellform und ein gestörtes phyllotaktisches Muster aufweist. Die Lebend-Zell Aufnahmen des Aktinzytoskeletts zeigt außerdem, dass seine Organisation mit der Zellform korreliert, was auf eine mögliche Rolle des Aktins bei der Beeinflussung der zellulären Morphogenese hinweist. In dieser Arbeit wird anhand einer detaillierten Charakterisierung der act2-1 act7-1 Mutante gezeigt, dass eine Störung des Aktins zu rechteckigeren Zellgeometrien mit mehr 90°-Zellinnenwinkeln und einem höheren Vorkommen von Vierfach-Verbindungen (vier sich kreuzende Zellgrenzen) führt. Diese Beobachtung weicht von den konventionellen trizellulären Verbindungen der Epidermiszellen ab. Quantitative Wachstumsdaten auf zellulärer Ebene deuten darauf hin, dass diese Unterschiede in der act2-1 act7-1 Mutante auf die geringere Präzision bei der Platzierung der neuen Zellwand sowie auf ihre mechanische Reifung zurückzuführen sind. Die bei der act2-1 act7-1 Mutante beobachteten Veränderungen der Zellmorphologie führen zu Defekten in der Zellanordnung, die in der weiteren Folge den Informationsfluss zwischen den Zellen im SAM beeinträchtigen. KW - Arabidopsis KW - shoot apical meristem KW - actin KW - cell division KW - cell shape KW - morphogenesis Y1 - 2022 U6 - https://doi.org/10.25932/publishup-55908 ER - TY - JOUR A1 - Streubel, Susanna A1 - Fritz, Michael Andre A1 - Teltow, Melanie A1 - Kappel, Christian A1 - Sicard, Adrien T1 - Successive duplication-divergence mechanisms at the RCO locus contributed to leaf shape diversity in the Brassicaceae JF - Development : Company of Biologists N2 - Gene duplication is a major driver for the increase of biological complexity. The divergence of newly duplicated paralogs may allow novel functions to evolve, while maintaining the ancestral one. Alternatively, partitioning the ancestral function among paralogs may allow parts of that role to follow independent evolutionary trajectories. We studied the REDUCED COMPLEXITY (RCO) locus, which contains three paralogs that have evolved through two independent events of gene duplication, and which underlies repeated events of leaf shape evolution within the Brassicaceae. In particular, we took advantage of the presence of three potentially functional paralogs in Capsella to investigate the extent of functional divergence among them. We demonstrate that the RCO copies control growth in different areas of the leaf. Consequently, the copies that are retained active in the different Brassicaceae lineages contribute to define the leaf dissection pattern. Our results further illustrate how successive gene duplication events and subsequent functional divergence can increase trait evolvability by providing independent evolutionary trajectories to specialized functions that have an additive effect on a given trait. KW - Plant development KW - Gene duplication KW - Leaf shape KW - Morphological evolution KW - Capsella KW - Arabidopsis Y1 - 2018 U6 - https://doi.org/10.1242/dev.164301 SN - 0950-1991 SN - 1477-9129 VL - 145 IS - 8 PB - Company of Biologists CY - Cambridge ER - TY - JOUR A1 - Singh, Aakanksha A1 - Compart, Julia A1 - AL-Rawi, Shadha Abduljaleel A1 - Mahto, Harendra A1 - Ahmad, Abubakar Musa A1 - Fettke, Jörg T1 - LIKE EARLY STARVATION 1 alters the glucan structures at the starch granule surface and thereby influences the action of both starch-synthesizing and starch-degrading enzymes JF - The plant journal N2 - For starch metabolism to take place correctly, various enzymes and proteins acting on the starch granule surface are crucial. Recently, two non-catalytic starch-binding proteins, pivotal for normal starch turnover in Arabidopsis leaves, namely, EARLY STARVATION 1 (ESV1) and its homolog LIKE EARLY STARVATION 1 (LESV), have been identified. Both share nearly 38% sequence homology. As ESV1 has been found to influence glucan phosphorylation via two starch-related dikinases, alpha-glucan, water dikinase (GWD) and phosphoglucan, water dikinase (PWD), through modulating the surface glucan structures of the starch granules and thus affecting starch degradation, we assess the impact of its homolog LESV on starch metabolism. Thus, the 65-kDa recombinant protein LESV and the 50-kDa ESV1 were analyzed regarding their influence on the action of GWD and PWD on the surface of the starch granules. We included starches from various sources and additionally assessed the effect of these non-enzymatic proteins on other starch-related enzymes, such as starch synthases (SSI and SSIII), starch phosphorylases (PHS1), isoamylase and beta-amylase. The data obtained indicate that starch phosphorylation, hydrolyses and synthesis were affected by LESV and ESV1. Furthermore, incubation with LESV and ESV1 together exerted an additive effect on starch phosphorylation. In addition, a stable alteration of the glucan structures at the starch granule surface following treatment with LESV and ESV1 was observed. Here, we discuss all the observed changes that point to modifications in the glucan structures at the surface of the native starch granules and present a model to explain the existing processes. KW - starch KW - starch metabolism KW - starch surface structure KW - Arabidopsis KW - thaliana Y1 - 2022 U6 - https://doi.org/10.1111/tpj.15855 SN - 0960-7412 SN - 1365-313X VL - 111 IS - 3 SP - 819 EP - 835 PB - Wiley-Blackwell CY - Oxford ER - TY - JOUR A1 - Shahnejat-Bushehri, Sara A1 - Nobmann, Barbara A1 - Allu, Annapurna Devi A1 - Balazadeh, Salma T1 - JUB1 suppresses Pseudomonas syringae-induced defense responses through accumulation of DELLA proteins JF - Journal of trace elements in medicine and biology N2 - Phytohormones act in concert to coordinate plant growth and the response to environmental cues. Gibberellins (GAs) are growth-promoting hormones that recently emerged as modulators of plant immune signaling. By regulating the stability of DELLA proteins, GAs intersect with the signaling pathways of the classical primary defense hormones, salicylic acid (SA) and jasmonic acid (JA), thereby altering the final outcome of the immune response. DELLA proteins confer resistance to necrotrophic pathogens by potentiating JA signaling and raise the susceptibility to biotrophic pathogens by attenuating the SA pathway. Here, we show that JUB1, a core element of the GA - brassinosteroid (BR) - DELLA regulatory module, functions as a negative regulator of defense responses against Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) and mediates the crosstalk between growth and immunity. KW - Arabidopsis KW - defense KW - DELLA proteins KW - gibberellin KW - jasmonic acid KW - pathogens KW - salicylic acid KW - transcription factor Y1 - 2016 U6 - https://doi.org/10.1080/15592324.2016.1181245 SN - 1559-2316 SN - 1559-2324 VL - 11 PB - Elsevier CY - Philadelphia ER - TY - JOUR A1 - Shahnejat-Bushehri, Sara A1 - Allu, Annapurna Devi A1 - Mehterov, Nikolay A1 - Thirumalaikumar, Venkatesh P. A1 - Alseekh, Saleh A1 - Fernie, Alisdair A1 - Mueller-Roeber, Bernd A1 - Balazadeh, Salma T1 - Arabidopsis NAC Transcription Factor JUNGBRUNNEN1 Exerts Conserved Control Over Gibberellin and Brassinosteroid Metabolism and Signaling Genes in Tomato JF - Frontiers in plant science N2 - The Arabidopsis thaliana NAC transcription factor JUNGBRUNNEN1 (AtJUB1) regulates growth by directly repressing GA3ox1 and DWF4, two key genes involved in gibberellin (GA) and brassinosteroid (BR) biosynthesis, respectively, leading to GA and BR deficiency phenotypes. AtJUB1 also reduces the expression of PIF4, a bHLH transcription factor that positively controls cell elongation, while it stimulates the expression of DELLA genes, which are important repressors of growth. Here, we extend our previous findings by demonstrating that AtJUB1 induces similar GA and BR deficiency phenotypes and changes in gene expression when overexpressed in tomato (Solanum lycopersicum). Importantly, and in accordance with the growth phenotypes observed, AtJUB1 inhibits the expression of growth-supporting genes, namely the tomato orthologs of GA3ox1, DWF4 and PIF4, but activates the expression of DELLA orthologs, by directly binding to their promoters. Overexpression of AtJUB1 in tomato delays fruit ripening, which is accompanied by reduced expression of several ripeningrelated genes, and leads to an increase in the levels of various amino acids (mostly proline, beta-alanine, and phenylalanine), gamma-aminobutyric acid (GABA), and major organic acids including glutamic acid and aspartic acid. The fact that AtJUB1 exerts an inhibitory effect on the GA/BR biosynthesis and PIF4 genes but acts as a direct activator of DELLA genes in both, Arabidopsis and tomato, strongly supports the model that the molecular constituents of the JUNGBRUNNEN1 growth control module are considerably conserved across species. KW - Arabidopsis KW - tomato KW - fruit KW - growth KW - transcription factor KW - gibberellic acid KW - brassinosteroid KW - DELLA proteins Y1 - 2017 U6 - https://doi.org/10.3389/fpls.2017.00214 SN - 1664-462X VL - 8 PB - Frontiers Research Foundation CY - Lausanne ER - TY - JOUR A1 - Sedaghatmehr, Mastoureh A1 - Thirumalaikumar, Venkatesh P. A1 - Kamranfar, Iman A1 - Marmagne, Anne A1 - Masclaux-Daubresse, Celine A1 - Balazadeh, Salma T1 - A regulatory role of autophagy for resetting the memory of heat stress in plants JF - Plant, cell & environment : cell physiology, whole-plant physiology, community physiology N2 - As sessile life forms, plants are repeatedly confronted with adverse environmental conditions, which can impair development, growth, and reproduction. During evolution, plants have established mechanisms to orchestrate the delicate balance between growth and stress tolerance, to reset cellular biochemistry once stress vanishes, or to keep a molecular memory, which enables survival of a harsher stress that may arise later. Although there are several examples of memory in diverse plants species, the molecular machinery underlying the formation, duration, and resetting of stress memories is largely unknown so far. We report here that autophagy, a central self-degradative process, assists in resetting cellular memory of heat stress (HS) in Arabidopsis thaliana. Autophagy is induced by thermopriming (moderate HS) and, intriguingly, remains high long after stress termination. We demonstrate that autophagy mediates the specific degradation of heat shock proteins at later stages of the thermorecovery phase leading to the accumulation of protein aggregates after the second HS and a compromised heat tolerance. Autophagy mutants retain heat shock proteins longer than wild type and concomitantly display improved thermomemory. Our findings reveal a novel regulatory mechanism for HS memory in plants. KW - Arabidopsis KW - heat shock proteins KW - priming KW - resetting Y1 - 2019 U6 - https://doi.org/10.1111/pce.13426 SN - 0140-7791 SN - 1365-3040 VL - 42 IS - 3 SP - 1054 EP - 1064 PB - Wiley CY - Hoboken ER - TY - JOUR A1 - Schroeder, Florian A1 - Lisso, Janina A1 - Obata, Toshihiro A1 - Erban, Alexander A1 - Maximova, Eugenia A1 - Giavalisco, Patrick A1 - Kopka, Joachim A1 - Fernie, Alisdair A1 - Willmitzer, Lothar A1 - Muessig, Carsten T1 - Consequences of induced brassinosteroid deficiency in Arabidopsis leaves JF - BMC plant biology N2 - Background: The identification of brassinosteroid (BR) deficient and BR insensitive mutants provided conclusive evidence that BR is a potent growth-promoting phytohormone. Arabidopsis mutants are characterized by a compact rosette structure, decreased plant height and reduced root system, delayed development, and reduced fertility. Cell expansion, cell division, and multiple developmental processes depend on BR. The molecular and physiological basis of BR action is diverse. The BR signalling pathway controls the activity of transcription factors, and numerous BR responsive genes have been identified. The analysis of dwarf mutants, however, may to some extent reveal phenotypic changes that are an effect of the altered morphology and physiology. This restriction holds particularly true for the analysis of established organs such as rosette leaves. Results: In this study, the mode of BR action was analysed in established leaves by means of two approaches. First, an inhibitor of BR biosynthesis (brassinazole) was applied to 21-day-old wild-type plants. Secondly, BR complementation of BR deficient plants, namely CPD (constitutive photomorphogenic dwarf)-antisense and cbb1 (cabbage1) mutant plants was stopped after 21 days. BR action in established leaves is associated with stimulated cell expansion, an increase in leaf index, starch accumulation, enhanced CO2 release by the tricarboxylic acid cycle, and increased biomass production. Cell number and protein content were barely affected. Conclusion: Previous analysis of BR promoted growth focused on genomic effects. However, the link between growth and changes in gene expression patterns barely provided clues to the physiological and metabolic basis of growth. Our study analysed comprehensive metabolic data sets of leaves with altered BR levels. The data suggest that BR promoted growth may depend on the increased provision and use of carbohydrates and energy. BR may stimulate both anabolic and catabolic pathways. KW - Brassinosteroids KW - Arabidopsis KW - Tricarboxylic acid cycle KW - Biomass KW - Cell expansion KW - Growth Y1 - 2014 U6 - https://doi.org/10.1186/s12870-014-0309-0 SN - 1471-2229 VL - 14 PB - BioMed Central CY - London ER - TY - JOUR A1 - Ruprecht, Colin A1 - Mutwil, Marek A1 - Saxe, Friederike A1 - Eder, Michaela A1 - Nikoloski, Zoran A1 - Persson, Staffan T1 - Large-scale co-expression approach to dissect secondary cell wall formation across plant species JF - Frontiers in plant science N2 - Plant cell walls are complex composites largely consisting of carbohydrate-based polymers, and are generally divided into primary and secondary walls based on content and characteristics. Cellulose microfibrils constitute a major component of both primary and secondary cell walls and are synthesized at the plasma membrane by cellulose synthase (CESA) complexes. Several studies in Arabidopsis have demonstrated the power of co-expression analyses to identify new genes associated with secondary wall cellulose biosynthesis. However, across-species comparative co-expression analyses remain largely unexplored. Here, we compared co-expressed gene vicinity networks of primary and secondary wall CESAsin Arabidopsis, barley, rice, poplar, soybean, Medicago, and wheat, and identified gene families that are consistently co-regulated with cellulose biosynthesis. In addition to the expected polysaccharide acting enzymes, we also found many gene families associated with cytoskeleton, signaling, transcriptional regulation, oxidation, and protein degradation. Based on these analyses, we selected and biochemically analyzed T-DNA insertion lines corresponding to approximately twenty genes from gene families that re-occur in the co-expressed gene vicinity networks of secondary wall CESAs across the seven species. We developed a statistical pipeline using principal component analysis and optimal clustering based on silhouette width to analyze sugar profiles. One of the mutants, corresponding to a pinoresinol reductase gene, displayed disturbed xylem morphology and held lower levels of lignin molecules. We propose that this type of large-scale co-expression approach, coupled with statistical analysis of the cell wall contents, will be useful to facilitate rapid knowledge transfer across plant species. KW - secondary cell wall KW - comparative co-expression analysis KW - Arabidopsis KW - cellulose Y1 - 2011 U6 - https://doi.org/10.3389/fpls.2011.00023 SN - 1664-462X VL - 2 PB - Frontiers Research Foundation CY - Lausanne ER - TY - JOUR A1 - Rocchetti, Alessandra A1 - Sharma, Tripti A1 - Wulfetange, Camilla A1 - Scholz-Starke, Joachim A1 - Grippa, Alexandra A1 - Carpaneto, Armando A1 - Dreyer, Ingo A1 - Vitale, Alessandro A1 - Czempinski, Katrin A1 - Pedrazzini, Emanuela T1 - The putative K+ channel subunit AtKCO3 forms stable dimers in arabidopsis JF - Frontiers in plant science N2 - The permeation pore of K+ channels is formed by four copies of the pore domain. AtKCO3 is the only putative voltage-independent K+ channel subunit of Arabidopsis thaliana with a single pore domain. KCO3-like proteins recently emerged in evolution and, to date, have been found only in the genus Arabidopsis (A. thaliana and A. lyrata). We show that the absence of KCO3 does not cause marked changes in growth under various conditions. Only under osmotic stress we observed reduced root growth of the kco3-1 null-allele line. This phenotype was complemented by expressing a KCO3 mutant with an inactive pore, indicating that the function of KCO3 under osmotic stress does not depend on its direct ability to transport ions. Constitutively overexpressed AtKCO3 or AtKCO3::G FP are efficiently sorted to the tonoplast indicating that the protein is approved by the endoplasmic reticulum quality control. However, vacuoles isolated from transgenic plants do not have significant alterations in current density. Consistently, both AtKCO3 and AtKCO3::GFP are detected as homodimers upon velocity gradient centrifugation, an assembly state that would not allow for activity. We conclude that if AtKCO3 ever functions as a K+ channel, active tetramers are held by particularly weak interactions, are formed only in unknown specific conditions and may require partner proteins. KW - Arabidopsis KW - membrane proteins KW - potassium channels KW - protein assembly KW - tonoplast Y1 - 2012 U6 - https://doi.org/10.3389/fpls.2012.00251 SN - 1664-462X VL - 3 PB - Frontiers Research Foundation CY - Lausanne ER - TY - JOUR A1 - Olas, Justyna Jadwiga A1 - Wahl, Vanessa T1 - Tissue-specific NIA1 and NIA2 expression in Arabidopsis thaliana JF - Plant Signaling & Behavior N2 - Nitrogen (N) is an essential macronutrient for optimal plant growth and ultimately for crop productivity Nitrate serves as the main N source for most plants. Although it seems a well-established fact that nitrate concentration affects flowering, its molecular mode of action in flowering time regulation was poorly understood. We recently found how nitrate, present at the shoot apical meristem (SAM), controls flowering time In this short communication, we present data on the tissue-specific expression patterns of NITRATE REDUCTASE 1 (NIA1) and NIA2 in planta. We show that transcripts of both genes are present throughout the life cycle of Arabidopsis thaliana plants with NIA1 being predominantly active in leaves and NIA2 in meristematic tissues. KW - Arabidopsis KW - NIA1 KW - NIA2 KW - nitrate assimilation KW - plant development KW - RNA in situ hybridization KW - expression KW - cell KW - and tissue-specificity Y1 - 2019 U6 - https://doi.org/10.1080/15592324.2019.1656035 SN - 1559-2316 SN - 1559-2324 VL - 14 IS - 11 PB - Taylor & Francis Group CY - Philadelphia ER - TY - JOUR A1 - Nietzsche, Madlen A1 - Schiessl, Ingrid A1 - Börnke, Frederik T1 - The complex becomes more complex: protein-protein interactions of SnRK1 with DUF581 family proteins provide a framework for cell and stimulus type-specific SnRK1 signaling in plants JF - Frontiers in plant science N2 - In plants, SNF1-related kinase (SnRK1) responds to the availability of carbohydrates as well as to environmental stresses by down-regulating ATP consuming biosynthetic processes, while stimulating energy-generating catabolic reactions through gene expression and post-transcriptional regulation. The functional SnRK1 complex is a heterotrimer where the catalytic alpha subunit associates with a regulatory beta subunit and an activating gamma subunit. Several different metabolites as well as the hormone abscisic acid (ABA) have been shown to modulate SnRK1 activity in a cell- and stimulus-type specific manner. It has been proposed that tissue- or stimulus-specific expression of adapter proteins mediating SnRK1 regulation can at least partly explain the differences observed in SnRK1 signaling. By using yeast two-hybrid and in planta bi-molecular fluorescence complementation assays we were able to demonstrate that proteins containing the domain of unknown function (DUF) 581 could interact with both isoforms of the SnRK1 alpha subunit (AKIN10/11) of Arabidopsis. A structure/function analysis suggests that the DUF581 is a generic SnRK1 interaction module and co-expression with DUF581 proteins in plant cells leads to reallocation of the kinase to specific regions within the nucleus. Yeast two-hybrid analyses suggest that SnRK1 and DUF581 proteins share common interaction partners inside the nucleus. The analysis of available microarray data implies that expression of the 19 members of the DUF581 encoding gene family in Arabidopsis is differentially regulated by hormones and environmental cues, indicating specialized functions of individual family members. We hypothesize that DUF581 proteins could act as mediators conferring tissue- and stimulus-type specific differences in SnRK1 regulation. KW - Arabidopsis KW - SnRK1 KW - DUF581 KW - protein-protein interaction KW - stress signaling KW - ABA Y1 - 2014 U6 - https://doi.org/10.3389/fpls.2014.00054 SN - 1664-462X VL - 5 PB - Frontiers Research Foundation CY - Lausanne ER - TY - THES A1 - Montulet, Orianne T1 - Functional characterization of putative interactors of the Cellulose Synthase Complex T1 - Funktionelle Charakterisierung von mutmaßlichen Interaktoren des Cellulose-Synthase-Komplexes N2 - The plant cell wall plays several crucial roles during plant development with its integrity acting as key signalling component for growth regulation during biotic and abiotic stresses. Cellulose microfibrils, the principal load-bearing components is the major component of the primary cell wall, whose synthesis is mediated by microtubule-associated CELLULOSE SYNTHASE (CESA) COMPLEXES (CSC). Previous studies have shown that CSC interacting proteins COMPANION OF CELLULOSE SYNTHASE (CC) facilitate sustained cellulose synthesis during salt stress by promoting repolymerization of cortical microtubules. However, our understanding of cellulose synthesis during salt stress remains incomplete. In this study, a pull-down of CC1 protein led to the identification of a novel interactor, termed LEA-like. Phylogenetic analysis revealed that LEA-like belongs to the LATE EMBRYOGENESIS ABUNDANT (LEA) protein family, specifically to the LEA_2 subgroup, showing a close relationship with the CC proteins. Roots of the double mutants lea-like and its closest homolog emb3135 exhibited hypersensitivity when grown on cellulose synthesis inhibitors. Further analysis of higher-order mutants of lea-like, emb3135, and cesa6 demonstrated a genetic interaction between them indicating a significant role in cellulose synthesis. Live-cell imaging revealed that both LEA-like and EMB3135 migrated with the CSC at the plasma membrane along microtubule tracks in control and oryzalin-treated conditions which destabilize microtubules, suggesting a tight interaction. Investigation of fluorescently labeled lines of different domains of the LEA-like protein revealed that the N-terminal cytosolic domain of LEA-like colocalizes with microtubules, suggesting a physical association between the two. Considering the established role of LEA proteins in abiotic stress tolerance, we performed phenotypic analysis of the mutant under various stresses. Growth of double mutants of lea-like and emb3135 on NaCl containing media resulted in swelling of root cell indicating a putative role in salt stress tolerance. Supportive of this the quadruple mutant, lacking LEA-like, EMB3135, CC1, and CC2 proteins, exhibited a severe root growth defect on NaCl media compared to control conditions. Live-cell imaging revealed that under salt stress, the LEA-like protein forms aggregates in the plasma membrane. In conclusion, this study has unveiled two novel interactors of the CSC that act with the CC proteins that regulate plant growth in response to salt stress providing new insights into the intricate regulation of cellulose synthesis, particularly under such conditions. N2 - Die pflanzliche Zellwand spielt während der Pflanzenentwicklung mehrere entscheidende Rollen, wobei ihre Integrität als zentrale Signalkomponente für die Wachstumsregulierung bei biotischem und abiotischem Stress fungiert. Zellulose-Mikrofibrillen, die wichtigsten tragenden Komponenten, sind der Hauptbestandteil der primären Zellwand, deren Synthese durch Mikrotubuli assoziierte CELLULOSE SYNTHASE (CESA) Komplexe (CSC) vermittelt wird. Frühere Studien haben gezeigt, dass die mit den CSC interagierenden Proteinen COMPANION OF CELLULOSE SYNTHASE (CC) die anhaltende Zellulosesynthese bei Salzstress erleichtern, indem sie die Repolymerisation der kortikalen Mikrotubuli fördern. Unser Verständnis der Zellulosesynthese bei Salzstress ist jedoch noch unvollständig. In dieser Studie führte ein Pull-down des CC1-Proteins zur Identifizierung eines neuen Interaktors, der als LEA-like bezeichnet wird. Eine phylogenetische Analyse ergab, dass LEA-like zur Late Embryogenesis Abundant (LEA)-Proteinfamilie gehört, insbesondere zur LEA_2-Untergruppe, die eine enge Beziehung zu den CC-Proteinen aufweist. Die Wurzeln der Doppelmutanten lea-like und seines engsten Homologen emb3135 zeigten eine Überempfindlichkeit, wenn sie auf Zellulose-Synthese-Inhibitoren wuchsen. Weitere Analysen von Mutanten höherer Ordnung von lea-like, emb3135 und cesa6 zeigten eine genetische Interaktion zwischen ihnen, die auf eine bedeutende Rolle bei der Zellulosesynthese hinweist. Die Bildgebung in lebenden Zellen zeigte, dass sowohl LEA-like als auch EMB3135 mit dem CSC an der Plasmamembran entlang von Mikrotubuli-Spuren wandern, und zwar sowohl unter Kontrollbedingungen als auch unter Oryzalin-Behandlung, die die Mikrotubuli destabilisiert, was auf eine enge Interaktion hindeutet. Die Untersuchung von fluoreszenzmarkierten Linien verschiedener Domänen des LEA-like-Proteins ergab, dass die N-terminale zytosolische Domäne von LEA-like mit Mikrotubuli kolokalisiert, was auf eine physische Verbindung zwischen den beiden hindeutet. In Anbetracht der bekannten Rolle der LEA-Proteine bei der abiotischen Stresstoleranz haben wir eine phänotypische Analyse der Mutante unter verschiedenen Stressbedingungen durchgeführt. Das Wachstum von Doppelmutanten von lea-like und emb3135 auf NaCl-haltigen Medien führte zu einem Anschwellen der Wurzelzellen, was auf eine mutmaßliche Rolle bei der Salzstresstoleranz hindeutet. Die Vierfachmutante, der die Proteine LEA-like, EMB3135, CC1 und CC2 fehlen, wies im Vergleich zu den Kontrollbedingungen auf NaCl-Medien einen schweren Wachstumsdefekt der Wurzeln auf. Die Bildgebung in lebenden Zellen zeigte, dass das LEA-like-Protein unter Salzstress Aggregate in der Plasmamembran bildet. Zusammenfassend lässt sich sagen, dass diese Studie zwei neue Interaktoren des CSC aufgedeckt hat, die mit den CC-Proteinen zusammenwirken und das Pflanzenwachstum als Reaktion auf Salzstress regulieren. KW - cell wall KW - cellulose KW - salt stress KW - cellulose synthase complex KW - Arabidopsis KW - Zellwand KW - zellulose, Salzstress KW - Cellulose-Synthese-Complex KW - Arabidopsis Y1 - 2024 ER - TY - JOUR A1 - Liu, Qinsong A1 - Vain, Thomas A1 - Viotti, Corrado A1 - Doyle, Siamsa M. A1 - Tarkowska, Danuse A1 - Novak, Ondrej A1 - Zipfel, Cyril A1 - Sitbon, Folke A1 - Robert, Stephanie A1 - Hofius, Daniel T1 - Vacuole integrity maintained by DUF300 proteins is required for brassinosteroid signaling regulation JF - Molecular plant N2 - Brassinosteroid (BR) hormone signaling controls multiple processes during plant growth and development and is initiated at the plasma membrane through the receptor kinase BRASSINOSTEROID INSENSITIVE1 (BRI1) together with co-receptors such as BRI1-ASSOCIATED RECEPTOR KINASE1 (BAK1). BRI1 abundance is regulated by endosomal recycling and vacuolar targeting, but the role of vacuole-related proteins in BR receptor dynamics and BR responses remains elusive. Here, we show that the absence of two DUF300 domain-containing tonoplast proteins, LAZARUS1 (LAZ1) and LAZ1 HOMOLOG1 (LAZ1H1), causes vacuole morphology defects, growth inhibition, and constitutive activation of BR signaling. Intriguingly, tonoplast accumulation of BAK1 was substantially increased and appeared causally linked to enhanced BRI1 trafficking and degradation in laz1 laz1h1 plants. Since unrelated vacuole mutants exhibited normal BR responses, our findings indicate that DUF300 proteins play distinct roles in the regulation of BR signaling by maintaining vacuole integrity required to balance subcellular BAK1 pools and BR receptor distribution. KW - brassinosteroid signaling KW - vacuole integrity KW - DUF300 proteins KW - tonoplast KW - Arabidopsis Y1 - 2018 U6 - https://doi.org/10.1016/j.molp.2017.12.015 SN - 1674-2052 SN - 1752-9867 VL - 11 IS - 4 SP - 553 EP - 567 PB - Cell Press CY - Cambridge ER - TY - JOUR A1 - Kiefer, Christian S. A1 - Claes, Andrea R. A1 - Nzayisenga, Jean-Claude A1 - Pietra, Stefano A1 - Stanislas, Thomas A1 - Ikeda, Yoshihisa A1 - Grebe, Markus T1 - Arabidopsis AIP1-2 restricted by WER-mediated patterning modulates planar polarity JF - Development N2 - The coordination of cell polarity within the plane of the tissue layer (planar polarity) is crucial for the development of diverse multicellular organisms. Small Rac/Rho-family GTPases and the actin cytoskeleton contribute to planar polarity formation at sites of polarity establishment in animals and plants. Yet, upstream pathways coordinating planar polarity differ strikingly between kingdoms. In the root of Arabidopsis thaliana, a concentration gradient of the phytohormone auxin coordinates polar recruitment of Rho-of-plant (ROP) to sites of polar epidermal hair initiation. However, little is known about cytoskeletal components and interactions that contribute to this planar polarity or about their relation to the patterning machinery. Here, we show that ACTIN7 (ACT7) represents a main actin isoform required for planar polarity of root hair positioning, interacting with the negative modulator ACTIN-INTERACTING PROTEIN1-2 (AIP1-2). ACT7, AIP1-2 and their genetic interaction are required for coordinated planar polarity of ROP downstream of ethylene signalling. Strikingly, AIP1-2 displays hair cell file-enriched expression, restricted by WEREWOLF (WER)-dependent patterning and modified by ethylene and auxin action. Hence, our findings reveal AIP1-2, expressed under control of the WER-dependent patterning machinery and the ethylene signalling pathway, as a modulator of actin-mediated planar polarity. KW - AIP1 KW - Actin KW - Arabidopsis KW - Patterning KW - Planar polarity Y1 - 2015 UR - http://dev.biologists.org/content/142/1/151.long U6 - https://doi.org/doi: 10.1242/dev.111013 IS - 142 SP - 151 EP - 161 ER - TY - JOUR A1 - Kiefer, Christian S. A1 - Claes, Andrea R. A1 - Nzayisenga, Jean-Claude A1 - Pietra, Stefano A1 - Stanislas, Thomas A1 - Hueser, Anke A1 - Ikeda, Yoshihisa A1 - Grebe, Markus T1 - Arabidopsis AIP1-2 restricted by WER-mediated patterning modulates planar polarity JF - Development : Company of Biologists N2 - The coordination of cell polarity within the plane of the tissue layer (planar polarity) is crucial for the development of diverse multicellular organisms. Small Rac/Rho-family GTPases and the actin cytoskeleton contribute to planar polarity formation at sites of polarity establishment in animals and plants. Yet, upstream pathways coordinating planar polarity differ strikingly between kingdoms. In the root of Arabidopsis thaliana, a concentration gradient of the phytohormone auxin coordinates polar recruitment of Rho-of-plant (ROP) to sites of polar epidermal hair initiation. However, little is known about cytoskeletal components and interactions that contribute to this planar polarity or about their relation to the patterning machinery. Here, we show that ACTIN7 (ACT7) represents a main actin isoform required for planar polarity of root hair positioning, interacting with the negative modulator ACTIN-INTERACTING PROTEIN1-2 (AIP1-2). ACT7, AIP1-2 and their genetic interaction are required for coordinated planar polarity of ROP downstream of ethylene signalling. Strikingly, AIP1-2 displays hair cell file-enriched expression, restricted by WEREWOLF (WER)-dependent patterning and modified by ethylene and auxin action. Hence, our findings reveal AIP1-2, expressed under control of the WER-dependent patterning machinery and the ethylene signalling pathway, as a modulator of actin-mediated planar polarity. KW - AIP1 KW - Arabidopsis KW - WEREWOLF KW - Actin KW - Patterning KW - Planar polarity Y1 - 2015 U6 - https://doi.org/10.1242/dev.111013 SN - 0950-1991 SN - 1477-9129 VL - 142 IS - 1 SP - 151 EP - 161 PB - Company of Biologists Limited CY - Cambridge ER - TY - JOUR A1 - Kamranfar, Iman A1 - Xue, Gang-Ping A1 - Tohge, Takayuki A1 - Sedaghatmehr, Mastoureh A1 - Fernie, Alisdair A1 - Balazadeh, Salma A1 - Mueller-Roeber, Bernd T1 - Transcription factor RD26 is a key regulator of metabolic reprogramming during dark-induced senescence JF - New phytologist : international journal of plant science N2 - Leaf senescence is a key process in plants that culminates in the degradation of cellular constituents and massive reprogramming of metabolism for the recovery of nutrients from aged leaves for their reuse in newly developing sinks. We used molecular-biological and metabolomics approaches to identify NAC transcription factor (TF) RD26 as an important regulator of metabolic reprogramming in Arabidopsis thaliana. RD26 directly activates CHLOROPLAST VESICULATION (CV), encoding a protein crucial for chloroplast protein degradation, concomitant with an enhanced protein loss in RD26 over-expressors during senescence, but a reduced decline of protein in rd26 knockout mutants. RD26 also directly activates LKR/SDH involved in lysine catabolism, and PES1 important for phytol degradation. Metabolic profiling revealed reduced c-aminobutyric acid (GABA) in RD26 overexpressors, accompanied by the induction of respective catabolic genes. Degradation of lysine, phytol and GABA is instrumental for maintaining mitochondrial respiration in carbon-limiting conditions during senescence. RD26 also supports the degradation of starch and the accumulation of mono-and disaccharides during senescence by directly enhancing the expression of AMY1, SFP1 and SWEET15 involved in carbohydrate metabolism and transport. Collectively, during senescence RD26 acts by controlling the expression of genes across the entire spectrum of the cellular degradation hierarchy. KW - Arabidopsis KW - fatty acid KW - primary metabolism KW - protein and amino acid degradation KW - respiration KW - senescence Y1 - 2018 U6 - https://doi.org/10.1111/nph.15127 SN - 0028-646X SN - 1469-8137 VL - 218 IS - 4 SP - 1543 EP - 1557 PB - Wiley CY - Hoboken ER - TY - JOUR A1 - Frescatada-Rosa, Marcia A1 - Stanislas, Thomas A1 - Backues, Steven K. A1 - Reichardt, Ilka A1 - Men, Shuzhen A1 - Boutte, Yohann A1 - Juergens, Gerd A1 - Moritz, Thomas A1 - Bednarek, Sebastian York A1 - Grebe, Markus T1 - High lipid order of Arabidopsis cell-plate membranes mediated by sterol and Dynamin-Related Protein 1A function JF - The plant journal N2 - Membranes of eukaryotic cells contain high lipid-order sterol-rich domains that are thought to mediate temporal and spatial organization of cellular processes. Sterols are crucial for execution of cytokinesis, the last stage of cell division, in diverse eukaryotes. The cell plate of higher-plant cells is the membrane structure that separates daughter cells during somatic cytokinesis. Cell-plate formation in Arabidopsis relies on sterol- and DYNAMIN-RELATED PROTEIN1A (DRP1A)-dependent endocytosis. However, functional relationships between lipid membrane order or lipid packing and endocytic machinery components during eukaryotic cytokinesis have not been elucidated. Using ratiometric live imaging of lipid order-sensitive fluorescent probes, we show that the cell plate of Arabidopsis thaliana represents a dynamic, high lipid-order membrane domain. The cell-plate lipid order was found to be sensitive to pharmacological and genetic alterations of sterol composition. Sterols co-localize with DRP1A at the cell plate, and DRP1A accumulates in detergent-resistant membrane fractions. Modifications of sterol concentration or composition reduce cell-plate membrane order and affect DRP1A localization. Strikingly, DRP1A function itself is essential for high lipid order at the cell plate. Our findings provide evidence that the cell plate represents a high lipid-order domain, and pave the way to explore potential feedback between lipid order and function of dynamin-related proteins during cytokinesis. KW - membrane order KW - sterol KW - cytokinesis KW - DRP1A KW - Arabidopsis Y1 - 2014 U6 - https://doi.org/10.1111/tpj.12674 SN - 0960-7412 SN - 1365-313X VL - 80 IS - 5 SP - 745 EP - 757 PB - Wiley-Blackwell CY - Hoboken ER - TY - JOUR A1 - Eldridge, Tilly A1 - Langowski, Lukasz A1 - Stacey, Nicola A1 - Jantzen, Friederike A1 - Moubayidin, Laila A1 - Sicard, Adrien A1 - Southam, Paul A1 - Kennaway, Richard A1 - Lenhard, Michael A1 - Coen, Enrico S. A1 - Ostergaard, Lars T1 - Fruit shape diversity in the Brassicaceae is generated by varying patterns of anisotropy JF - Development : Company of Biologists N2 - Fruits exhibit a vast array of different 3D shapes, from simple spheres and cylinders to more complex curved forms; however, the mechanism by which growth is oriented and coordinated to generate this diversity of forms is unclear. Here, we compare the growth patterns and orientations for two very different fruit shapes in the Brassicaceae: the heart-shaped Capsella rubella silicle and the near-cylindrical Arabidopsis thaliana silique. We show, through a combination of clonal and morphological analyses, that the different shapes involve different patterns of anisotropic growth during three phases. These experimental data can be accounted for by a tissue level model in which specified growth rates vary in space and time and are oriented by a proximodistal polarity field. The resulting tissue conflicts lead to deformation of the tissue as it grows. The model allows us to identify tissue-specific and temporally specific activities required to obtain the individual shapes. One such activity may be provided by the valve-identity gene FRUITFULL, which we show through comparative mutant analysis to modulate fruit shape during post-fertilisation growth of both species. Simple modulations of the model presented here can also broadly account for the variety of shapes in other Brassicaceae species, thus providing a simplified framework for fruit development and shape diversity. KW - Brassicaceae KW - Capsella KW - Arabidopsis KW - Fruit shape KW - Modelling KW - Anisotropic growth Y1 - 2016 U6 - https://doi.org/10.1242/dev.135327 SN - 0950-1991 SN - 1477-9129 VL - 143 SP - 3394 EP - 3406 PB - Company of Biologists Limited CY - Cambridge ER -