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Publikationen - Molekulare Signalverarbeitung

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Publikation

Grubb, C. D.; Zipp, B. J.; Ludwig-Müller, J.; Masuno, M. N.; Molinski, T. F.; Abel, S.; Arabidopsis glucosyltransferase UGT74B1 functions in glucosinolate biosynthesis and auxin homeostasis Plant J. 40, 893-908, (2004) DOI: 10.1111/j.1365-313X.2004.02261.x

Glucosinolates are a class of secondary metabolites with important roles in plant defense and human nutrition. Here, we characterize a putative UDP‐glucose:thiohydroximate S‐glucosyltransferase, UGT74B1, to determine its role in the Arabidopsis glucosinolate pathway. Biochemical analyses demonstrate that recombinant UGT74B1 specifically glucosylates the thiohydroximate functional group. Low K m values for phenylacetothiohydroximic acid (approximately 6 μ m ) and UDP‐glucose (approximately 50 μm ) strongly suggest that thiohydroximates are in vivo substrates of UGT74B1. Insertional loss‐of‐function ugt74b1 mutants exhibit significantly decreased, but not abolished, glucosinolate accumulation. In addition, ugt74b1 mutants display phenotypes reminiscent of auxin overproduction, such as epinastic cotyledons, elongated hypocotyls in light‐grown plants, excess adventitious rooting and incomplete leaf vascularization. Indeed, during early plant development, mutant ugt74b1 seedlings accumulate nearly threefold more indole‐3‐acetic acid than the wild type. Other phenotypes, however, such as chlorosis along the leaf veins, are likely caused by thiohydroximate toxicity. Analysis of UGT74B1 promoter activity during plant development reveals expression patterns consistent with glucosinolate metabolism and induction by auxin treatment. The results are discussed in the context of known mutations in glucosinolate pathway genes and their effects on auxin homeostasis. Taken together, our work provides complementary in vitro and in vivo evidence for a primary role of UGT74B1 in glucosinolate biosynthesis.
Publikation

Groß, N.; Wasternack, C.; Köck, M.; Wound-induced RNaseLE expression is jasmonate and systemin independent and occurs only locally in tomato (Lycopersicon esculentum cv. Lukullus) Phytochemistry 65, 1343-1350, (2004) DOI: 10.1016/j.phytochem.2004.04.036

Tomato RNaseLE is induced by phosphate deficiency and wounding and may play a role in macromolecular recycling as well as wound healing. Here, we analyzed the role of jasmonate and systemin in the wound-induced RNaseLE activation. The rapid expression of RNaseLE upon wounding of leaves leading to maximal RNase activity within 10 h, appeared only locally. Jasmonic acid (JA) or its molecular mimic ethyl indanoyl isoleucine conjugate did not induce RNaseLE expression. Correspondingly, RNaseLE was expressed upon wounding of 35S::allene oxide cyclase antisense plants known to be JA deficient. RNaseLE was not expressed upon systemin treatment, but was locally expressed in the spr1 mutant which is affected in systemin perception. In tomato plants carrying a PromLE::uidA construct, GUS activity could be detected upon wounding, but not following treatment with JA or systemin. The data indicate a locally acting wound-inducible systemin- and JA-independent signaling pathway for RNaseLE expression.RNaseLE expression was analyzed by pharmacological studies of different tomato lines and upon wounding of leaves. The gene is only locally activated via a new type of wound-induced signaling pathway in a jasmonate/systemin-independent manner.
Publikation

Frisch, M.; Quint, M.; Lübberstedt, T.; Melchinger, A. E.; Duplicate marker loci can result in incorrect locus orders on linkage maps Theor. Appl. Genet. 109, 305-316, (2004) DOI: 10.1007/s00122-003-1578-4

Genetic linkage maps, constructed from multi-locus recombination data, are the basis for many applications of molecular markers. For the successful employment of a linkage map, it is essential that the linear order of loci on a chromosome is correct. The objectives of this theoretical study were to (1) investigate the occurrence of incorrect locus orders caused by duplicate marker loci, (2) develop a statistical test for the detection of duplicate markers, and (3) discuss the implications for practical applications of linkage maps. We derived conditions, under which incorrect locus orders do or do not occur with duplicate marker loci for the general case of n markers on a chromosome in a BC1 mapping population. We further illustrated these conditions numerically for the special case of four markers. On the basis of the extent of segregation distortion, an exact test for the presence of duplicate marker loci was suggested and its power was investigated numerically. Incorrect locus orders caused by duplicate marker loci can (1) negatively affect the assignment of target genes to chromosome regions in a map-based cloning experiment, (2) hinder indirect selection for a favorable allele at a quantitative trait locus, and (3) decrease the efficiency of reducing the length of the chromosome segment attached to a target gene in marker-assisted backcrossing.
Publikation

Flores, R.; Delgado, S.; Gas, M.-E.; Carbonell, A.; Molina, D.; Gago, S.; De la Peña, M.; Viroids: the minimal non-coding RNAs with autonomous replication FEBS Lett. 567, 42-48, (2004) DOI: 10.1016/j.febslet.2004.03.118

Viroids are small (246–401 nucleotides), non‐coding, circular RNAs able to replicate autonomously in certain plants. Viroids are classified into the families Pospiviroidae and Avsunviroidae , whose members replicate in the nucleus and chloroplast, respectively. Replication occurs by an RNA‐based rolling‐circle mechanism in three steps: (1) synthesis of longer‐than‐unit strands catalyzed by host DNA‐dependent RNA polymerases forced to transcribe RNA templates, (2) processing to unit‐length, which in family Avsunviroidae is mediated by hammerhead ribozymes, and (3) circularization either through an RNA ligase or autocatalytically. Disease induction might result from the accumulation of viroid‐specific small interfering RNAs that, via RNA silencing, could interfere with normal developmental pathways.
Publikation

Bücking, H.; Förster, H.; Stenzel, I.; Miersch, O.; Hause, B.; Applied jasmonates accumulate extracellularly in tomato, but intracellularly in barley FEBS Lett. 562, 45-50, (2004) DOI: 10.1016/S0014-5793(04)00178-4

Jasmonic acid (JA) and its derivatives are well‐characterized signaling molecules in plant defense and development, but the site of their localization within plant tissue is entirely unknown. To address the question whether applied JA accumulates extracellularly or intracellularly, leaves of tomato and barley were fed with 14C‐labeled JA and the label was localized in cryofixed and lyophilized leaf tissues by microautoradiography. In tomato the radioactivity was detectable within the apoplast, but no label was found within the mesophyll cells. By contrast, in barley leaf tissues, radioactivity was detected within the mesophyll cells suggesting a cellular uptake of exogenously applied JA. JA, applied to leaves of both plants as in the labeling experiments, led in all leaf cells to the expression of JA‐inducible genes indicating that the perception is completed by JA signal transduction.
Publikation

Wasternack, C.; Introductory Remarks on Biosynthesis and Diversity in Actions J. Plant Growth Regul. 23, 167-169, (2004) DOI: 10.1007/s00344-004-0051-1

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Publikation

Ticconi, C. A.; Abel, S.; Short on phosphate: plant surveillance and countermeasures Trends Plant Sci. 9, 548-555, (2004) DOI: 10.1016/j.tplants.2004.09.003

Metabolism depends on inorganic phosphate (Pi) as reactant, allosteric effector and regulatory moiety in covalent protein modification. To cope with Pi shortage (a common situation in many ecosystems), plants activate a set of adaptive responses to enhance Pi recycling and acquisition by reprogramming metabolism and restructuring root system architecture. The physiology of Pi starvation responses has become well understood, and so current research focuses on the initial molecular events that sense, transmit and integrate information about external and internal Pi status. Recent studies have provided evidence for Pi as a signaling molecule and initial insight into the coordination of Pi deficiency responses at the cellular and molecular level.
Publikation

Ticconi, C. A.; Delatorre, C. A.; Lahner, B.; Salt, D. E.; Abel, S.; Arabidopsis pdr2 reveals a phosphate-sensitive checkpoint in root development Plant J. 37, 801-814, (2004) DOI: 10.1111/j.1365-313x.2004.02005.x

Plants have evolved complex strategies to maintain phosphate (Pi) homeostasis and to maximize Pi acquisition when the macronutrient is limiting. Adjustment of root system architecture via changes in meristem initiation and activity is integral to the acclimation process. However, the mechanisms that monitor external Pi status and interpret the nutritional signal remain to be elucidated. Here, we present evidence that the Pi deficiency response , pdr2 , mutation disrupts local Pi sensing. The sensitivity and amplitude of metabolic Pi‐starvation responses, such as Pi‐responsive gene expression or accumulation of anthocyanins and starch, are enhanced in pdr2 seedlings. However, the most conspicuous alteration of pdr2 is a conditional short‐root phenotype that is specific for Pi deficiency and caused by selective inhibition of root cell division followed by cell death below a threshold concentration of about 0.1 mm external Pi. Measurements of general Pi uptake and of total phosphorus (P) in root tips exclude a defect in high‐affinity Pi acquisition. Rescue of root meristem activity in Pi‐starved pdr2 by phosphite (Phi), a non‐metabolizable Pi analog, and divided‐root experiments suggest that pdr2 disrupts sensing of low external Pi availability. Thus, PDR2 is proposed to function at a Pi‐sensitive checkpoint in root development, which monitors environmental Pi status, maintains and fine‐tunes meristematic activity, and finally adjusts root system architecture to maximize Pi acquisition.
Publikation

Schwechheimer, C.; Villalobos, L. I. A. C.; Cullin-containing E3 ubiquitin ligases in plant development Curr. Opin. Plant Biol. 7, 677-686, (2004) DOI: 10.1016/j.pbi.2004.09.009

In eukaryotes, the ubiquitin–proteasome system participates in the control of signal transduction events by selectively eliminating regulatory proteins. E3 ubiquitin ligases specifically bind degradation substrates and mediate their poly-ubiquitylation, a prerequisite for their degradation by the 26S proteasome. On the basis of the analysis of the Arabidopsis genome sequence, it is predicted that there are more than 1000 E3 ubiquitin ligases in plants. Several types of E3 ubiquitin ligases have already been characterized in eukaryotes. Recently, some of these E3 enzymes have been implicated in specific plant signaling pathways.
Publikation

Schüler, G.; Mithöfer, A.; Baldwin, I. T.; BERGER, S.; Ebel, J.; Santos, J. G.; Herrmann, G.; Hölscher, D.; Kramell, R.; Kutchan, T. M.; Maucher, H.; Schneider, B.; Stenzel, I.; Wasternack, C.; Boland, W.; Coronalon: a powerful tool in plant stress physiology FEBS Lett. 563, 17-22, (2004) DOI: 10.1016/S0014-5793(04)00239-X

Coronalon, a synthetic 6‐ethyl indanoyl isoleucine conjugate, has been designed as a highly active mimic of octadecanoid phytohormones that are involved in insect and disease resistance. The spectrum of biological activities that is affected by coronalon was investigated in nine different plant systems specifically responding to jasmonates and/or 12‐oxo‐phytodienoic acid. In all bioassays analyzed, coronalon demonstrated a general strong activity at low micromolar concentrations. The results obtained showed the induction of (i) defense‐related secondary metabolite accumulation in both cell cultures and plant tissues, (ii) specific abiotic and biotic stress‐related gene expression, and (iii) root growth retardation. The general activity of coronalon in the induction of plant stress responses together with its simple and efficient synthesis suggests that this compound might serve as a valuable tool in the examination of various aspects in plant stress physiology. Moreover, coronalon might become employed in agriculture to elicit plant resistance against various aggressors.
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