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Publikation

Wasternack, C.; Hause, B. Jasmonsäure – ein universelles Pflanzenhormon: Blütenduft, Abwehr, Entwicklung Biologie in unserer Zeit 44, 164 - 171, (2014) DOI: 10.1002/biuz.201410535

Jasmonsäure (JA) und ihre Metaboliten kommen in allen niederen und höheren Pflanzen vor. Sie sind universell wirksame, aus Lipiden gebildete Signalstoffe bei der Abwehr von biotischem und abiotischem Stress sowie in der pflanzlichen Entwicklung. Rezeptor und Komponenten von JA–Signalketten wurden identifiziert. In der Entwicklung von Blüten, Früchten, Samen, Trichomen oder in der Abwehr von Insekten und Pathogenen treten ähnliche JA-vermittelte Signalproteine auf, die eine Feinregulation der Prozesse erlauben und eine Verbindung (cross-talk) zu anderenPflanzenhormonen aufweisen.
Bücher und Buchkapitel

Wasternack, C. Jasmonates in plant growth and stress responses. (Tran, L.-S.; Pal, S.). Springer, 221-264, (2014) ISBN: 978-1-4939-0490-7 (hardcover) 978-1-4939-4814-7 (softcover) DOI: 10.1007/978-1-4939-0491-4_8

Abiotic and biotic stresses adversely affect plant growth and productivity. The phytohormones regulate key physiological events under normal and stressful conditions for plant development. Accumulative research efforts have discovered important roles of phytohormones and their interactions in regulation of plant adaptation to numerous stressors. Intensive molecular studies have elucidated various plant hormonal pathways; each of which consist of many signaling components that link a specific hormone perception to the regulation of downstream genes. Signal transduction pathways of auxin, abscisic acid, cytokinins, gibberellins and ethylene have been thoroughly investigated. More recently, emerging signaling pathways of brassinosteroids, jasmonates, salicylic acid and strigolactones offer an exciting gateway for understanding their multiple roles in plant physiological processes.At the molecular level, phytohormonal crosstalks can be antagonistic or synergistic or additive in actions. Additionally, the signal transduction component(s) of one hormonal pathway may interplay with the signaling component(s) of other hormonal pathway(s). Together these and other research findings have revolutionized the concept of phytohormonal studies in plants. Importantly, genetic engineering now enables plant biologists to manipulate the signaling pathways of plant hormones for development of crop varieties with improved yield and stress tolerance.This book, written by internationally recognized scholars from various countries, represents the state-of-the-art understanding of plant hormones’ biology, signal transduction and implications. Aimed at a wide range of readers, including researchers, students, teachers and many others who have interests in this flourishing research field, every section is concluded with biotechnological strategies to modulate hormone contents or signal transduction pathways and crosstalk that enable us to develop crops in a sustainable manner. Given the important physiological implications of plant hormones in stressful environments, our book is finalized with chapters on phytohormonal crosstalks under abiotic and biotic stresses. 
Publikation

Budiharjo, A.; Chowdhury, S.M.; Dietel, C.; Beator, B.; Dolgova, O.; Fan, B.; Bleiss, W.; Ziegler, J.; Schmid, M.; Hartmann, A.; Borris, R. Transposon Mutagenesis of the Plant-Associated Bacillus amyloliquefaciens ssp. plantarum FZB42 Revealed That the nrfA and RBAM17410 Genes Are Involved in Plant-Microbe-Interactions PLOS ONE 9, e98267, (2014) DOI: 10.1371/journal.pone.0098267

Bacillus amyloliquefaciens ssp. plantarum FZB42 represents the prototype of Gram-positive plant growth promoting and biocontrol bacteria. In this study, we applied transposon mutagenesis to generate a transposon library, which was screened for genes involved in multicellular behavior and biofilm formation on roots as a prerequisite of plant growth promoting activity. Transposon insertion sites were determined by rescue-cloning followed by DNA sequencing. As in B. subtilis, the global transcriptional regulator DegU was identified as an activator of genes necessary for swarming and biofilm formation, and the DegU-mutant of FZB42 was found impaired in efficient root colonization. Direct screening of 3,000 transposon insertion mutants for plant-growth-promotion revealed the gene products of nfrA and RBAM_017140 to be essential for beneficial effects exerted by FZB42 on plants. We analyzed the performance of GFP-labeled wild-type and transposon mutants in the colonization of lettuce roots using confocal laser scanning microscopy. While the wild-type strain heavily colonized root surfaces, the nfrA mutant did not colonize lettuce roots, although it was not impaired in growth in laboratory cultures, biofilm formation and swarming motility on agar plates. The RBAM17410 gene, occurring in only a few members of the B. subtilis species complex, was directly involved in plant growth promotion. None of the mutant strains were affected in producing the plant growth hormone auxin. We hypothesize that the nfrA gene product is essential for overcoming the stress caused by plant response towards bacterial root colonization.
Publikation

Erickson, J. l.; Ziegler, J.; Guevara, D.; Abel, S.; Klösgen, R. B.; Mathur, J.; Rothstein, S. J.; Schattat, M.H. Agrobacterium-derived cytokinin influences plastid morphology and starch accumulation inNicotiana benthamiana during transient assays BMC Plant Biol. 14, 127, (2014) DOI: 10.1186/1471-2229-14-127

Background: Agrobacterium tumefaciens-based transient assays have become a common tool for answering questions related to protein localization and gene expression in a cellular context. The use of these assays assumes that the transiently transformed cells are observed under relatively authentic physiological conditions and maintain ‘normal’ sub-cellular behaviour. Although this premise is widely accepted, the question of whether cellular organization andorganelle morphology is altered in Agrobacterium-infiltrated cells has not been examined in detail. The first indications of an altered sub-cellular environment came from our observation that a common laboratory strain, GV3101(pMP90), caused a drastic increase in stromule frequency. Stromules, or ‘stroma-filled-tubules’ emanate fromthe surface of plastids and are sensitive to a variety of biotic and abiotic stresses. Starting from this observation, the goal of our experiments was to further characterize the changes to the cell resulting from short-term bacterial infestation, and to identify the factor responsible for eliciting these changes.Results: Using a protocol typical of transient assays we evaluated the impact of GV3101(pMP90) infiltration on chloroplast behaviour and morphology in Nicotiana benthamiana. Our experiments confirmed that GV3101(pMP90) consistently induces stromules and alters plastid position relative to the nucleus. These effects were found to be the result of strain-dependant secretion of cytokinin and its accumulation in the plant tissue. Bacterial production of the hormone was found to be dependant on the presence of a trans-zeatin synthase gene (tzs) located on the Ti plasmidof GV3101(pMP90). Bacteria-derived cytokinins were also correlated with changes to both soluble sugar level and starch accumulation.Conclusion: Although we have chosen to focus on how transient Agrobacterium infestation alters plastid based parameters, these changes to the morphology and position of a single organelle, combined with the measured increases in sugar and starch content, suggest global changes to cell physiology. This indicates that cells visualized during transient assays may not be as ‘normal’ as was previously assumed. Our results suggest that the impact of the bacteria can be minimized by choosing Agrobacterium strains devoid of the tzs gene, but that the alterations to sub-cellular organization and cell carbohydrate status cannot be completely avoided using this strategy.Keywords: Agrobacterium tumefaciens, Nicotiana benthamiana, Transient assays, GV3101(pMP90), LBA4404, Plastid,Stromules, Bacteria-derived, Cytokinin, Trans-zeatin synthase
Publikation

Wasternack, C. Perception, signaling and cross-talk of jasmonates and the seminal contributions of the Daoxin Xie´s lab and the Chuanyou Li´s lab Plant Cell Rep 33, 707-718, (2014) DOI: 10.1007/s00299-014-1608-5

Jasmonates (JAs) are lipid-derived signals in plant responses to biotic and abiotic stresses and in development. The most active JA compound is (+)-7-iso-JA-Ile, a JA conjugate with isoleucine. Biosynthesis, metabolism and key components of perception and signal transduction have been identified and numerous JA-induced gene expression data collected. For JA-Ile perception, the SCFCOI1–JAZ co-receptor complex has been identified and crystalized. Activators such as MYC2 and repressors such as JAZs including their targets were found. Involvement of JA-Ile in response to herbivores and pathogens and in root growth inhibition is among the most studied aspects of JA-Ile signaling. There are an increasing number of examples, where JA-Ile shows cross-talk with other plant hormones. Seminal contributions in JA/JA-Ile research were given by Daoxin Xie’s lab and Chuanyou Li’s lab, both in Beijing. Here, characterization was done regarding components of the JA-Ile receptor, such as COI1 (JAI1) and SCF, regarding activators (MYCs, MYBs) and repressors (JAV1, bHLH IIId’s) of JA-regulated gene expression, as well as regarding components of auxin biosynthesis and action, such as the transcription factor PLETHORA active in the root stem cell niche. This overview reflects the work of both labs in the light of our present knowledge on biosynthesis, perception and signal transduction of JA/JA-Ile and its cross-talk to other hormones.
Publikation

Song, S.; Qi, T.; Wasternack, C.; Xie, D. Jasmonate signaling and crosstalk with gibberellin and ethylene Curr Opin Plant Biol. 21 , 112-119, (2014) DOI: 10.1016/j.pbi.2014.07.005

The phytohormone jasmonate (JA) plays essential roles in plant growth, development and defense. In response to the JA signal, the CORONATINE INSENSITIVE 1 (COI1)-based SCF complexes recruit JASMONATE ZIM-domain (JAZ) repressors for ubiquitination and degradation, and subsequently regulate their downstream signaling components essential for various JA responses. Tremendous progress has been made in understanding the JA signaling pathway and its crosstalk with other phytohormone pathways during the past two decades. Recent studies have revealed that a variety of positive and negative regulators act as targets of JAZs to control distinctive JA responses, and that JAZs and these regulators function as crucial interfaces to mediate synergy and antagonism between JA and other phytohormones. Owing to different regulatory players in JA perception and JA signaling, a fine-tuning of JA-dependent processes in plant growth, development and defense is achieved. In this review, we will summarize the latest progresses in JA signaling and its crosstalk with gibberellin and ethylene.
Publikation

Maldonado-Bonilla, L.D.; Eschen-Lippold, L.; Gago-Zachert, S.; Tabassum, N.; Bauer, N.; Scheel, D.; Lee, J. The Arabidopsis tandem zinc finger 9 protein binds RNA and mediates pathogen-associated molecular pattern-triggered immune responses Plant Cell Physiol. 55, 412-425, (2014) DOI: 10.1093/pcp/pct175

Recognition of pathogen-associated molecular patterns (PAMPs) induces multiple defense mechanisms to limit pathogen growth. Here, we show that the Arabidopsis thaliana tandem zinc finger protein 9 (TZF9) is phosphorylated by PAMP-responsive mitogen-activated protein kinases (MAPKs) and is required to trigger a full PAMP-triggered immune response. Analysis of a tzf9 mutant revealed attenuation in specific PAMP-triggered reactions such as reactive oxygen species accumulation, MAPK activation and, partially, the expression of several PAMP-responsive genes. In accordance with these weaker PAMP-triggered responses, tzf9 mutant plants exhibit enhanced susceptibility to virulent Pseudomonas syringae pv. tomato DC3000. Visualization of TZF9 localization by fusion to green fluorescent protein revealed cytoplasmic foci that co-localize with marker proteins of processing bodies (P-bodies). This localization pattern is affected by inhibitor treatments that limit mRNA availability (such as cycloheximide or actinomycin D) or block nuclear export (leptomycin B). Coupled with its ability to bind the ribohomopolymers poly(rU) and poly(rG), these results suggest involvement of TZF9 in post-transcriptional regulation, such as mRNA processing or storage pathways, to regulate plant innate immunity. 
Publikation

Jayaweera, T.; Siriwardana, C.; Dharmasiri, S.; Quint, M.; Gray, W. M.; Dharmasiri, N. Alternative Splicing of Arabidopsis IBR5 Pre-mRNA Generates Two IBR5 Isoforms with Distinct and Overlapping Functions PLoS ONE 9, e102301, (2014) DOI: 10.1371/journal.pone.0102301

The INDOLE-3-BUTYRIC ACID RESPONSE5 (IBR5) gene encodes a dual specificity phosphatase that regulates plant auxinresponses. IBR5 has been predicted to generate two transcripts through alternative splicing, but alternative splicing of IBR5has not been confirmed experimentally. The previously characterized ibr5-1 null mutant exhibits many auxin related defectssuch as auxin insensitive primary root growth, defective vascular development, short stature and reduced lateral rootdevelopment. However, whether all these defects are caused by the lack of phosphatase activity is not clear. Here wedescribe two new auxin insensitive IBR5 alleles, ibr5-4, a catalytic site mutant, and ibr5-5, a splice site mutant.Characterization of these new mutants indicates that IBR5 is post-transcriptionally regulated to generate two transcripts,AT2G04550.1 and AT2G04550.3, and consequently two IBR5 isoforms, IBR5.1 and IBR5.3. The IBR5.1 isoform exhibitsphosphatase catalytic activity that is required for both proper degradation of Aux/IAA proteins and auxin-induced geneexpression. These two processes are independently regulated by IBR5.1. Comparison of new mutant alleles with ibr5-1indicates that all three mutant alleles share many phenotypes. However, each allele also confers distinct defects implicatingIBR5 isoform specific functions. Some of these functions are independent of IBR5.1 catalytic activity. Additionally, analysis ofthese new mutant alleles suggests that IBR5 may link ABP1 and SCFTIR1/AFBs auxin signaling pathways.
Publikation

Flores, R.; Gago-Zachert, S.; Serra, P.; Sanjuán, R.; Elena, S. F. Viroids: Survivors from the RNA World? Annual Rev Microbiol 68, 395 - 414, (2014) DOI: 10.1146/annurev-micro-091313-103416

Because RNA can be a carrier of genetic information and a biocatalyst, there is a consensus that it emerged before DNA and proteins, which eventually assumed these roles and relegated RNA to intermediate functions. If such a scenario—the so-called RNA world—existed, we might hope to find its relics in our present world. The properties of viroids that make them candidates for being survivors of the RNA world include those expected for primitive RNA replicons: (a) small size imposed by error-prone replication, (b) high G+ C content to increase replication fidelity, (c) circular structure for assuring complete replication without genomic tags, (d) structural periodicity for modular assembly into enlarged genomes, (e) lack of protein-coding ability consistent with a ribosome-free habitat, and (f) replication mediated in some by ribozymes, the fingerprint of the RNA world. With the advent of DNA and proteins, those protoviroids lost some abilities and became the plant parasites we now know.
Publikation

Floková, K.; Tarkowská, D.; Miersch, O.; Strnad, M.; Wasternack, C.; Novak, O. UHPLC-MS/MS based target profiling of stress-induced phytohormones Phytochemistry 105, 147-157, (2014) DOI: 10.1016/j.phytochem.2014.05.015

Stress-induced changes in phytohormone metabolite profiles have rapid effects on plant metabolic activity and growth. The jasmonates (JAs) are a group of fatty acid-derived stress response regulators with roles in numerous developmental processes. To elucidate their dual regulatory effects, which overlap with those of other important defence-signalling plant hormones such as salicylic acid (SA), abscisic acid (ABA) and indole-3-acetic acid (IAA), we have developed a highly efficient single-step clean-up procedure for their enrichment from complex plant matrices that enables their sensitive quantitative analysis using hyphenated mass spectrometry technique. The rapid extraction of minute quantities of plant material (less than 20 mg fresh weight, FW) into cold 10% methanol followed by one-step reversed-phase polymer-based solid phase extraction significantly reduced matrix effects and increased the recovery of labile JA analytes. This extraction and purification protocol was paired with a highly sensitive and validated ultra-high performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) method and used to simultaneously profile sixteen stress-induced phytohormones in minute plant material samples, including endogenous JA, several of its biosynthetic precursors and derivatives, as well as SA, ABA and IAA.
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