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Carbohydrates are synthesised in photosynthetically active source tissues and exported, in most species in the form of sucrose, to photosynthetically less active or inactive sink tissues. Sucrose hydrolysis at the site of utilisation contributes to phloem unloading. This phenomenon links sink metabolism with phloem transport to, and partitioning between, sinks. Invertases catalyse the irreversible hydrolysis of sucrose and thus are expected to contribute to carbohydrate partitioning. Different invertase isoenzymes may be distinguished based on their intracellular location, their isoelectric points and pH optima. Extracellular, cell-wall-bound invertase is uniquely positioned to supply carbohydrates to sink tissues via an apoplasmic pathway, and links the transport sugar sucrose to hexose transporters. A number of studies demonstrate an essential function of this invertase isoenzyme for phloem unloading, carbohydrate partitioning and growth of sink tissues. Extracellular invertases were shown to be specifically expressed under conditions that require a high carbohydrate supply to sink tissues. Further, their expression is upregulated by a number of stimuli that affect source–sink relations. Substrate and reaction products of invertases are not only nutri-ents, but also signal molecules. Like hormones and in combination with hormones and other stimuli, they can regu-late many aspects of plant development from gene expression to long-distance nutrient allocation. Based on studies in Chenopodium rubrum, tomato (Lycopersicon esculentum) and tobacco (Nicotiana tabacum), the regulation of extracellular invertase and its function in assimilate partitioning, defence reactions and sugar signal transduction pathways are discussed.
Publications
Sugarcane mosaic virus (SCMV) is an important disease in maize, which is emerging in Germany since 1983. Using this pest as a model for the inheritance of oligogenic traits, we clarified the genetic basis for resistance in early maturing European maize germplasm. Screening of 122 adapted European inbred lines identified three completely resistant lines, which were used for further analyses. The genetics of SCMV resistance was investigated by allelism tests in field experiments combined with QTL and bulked segregant analyses (BSA) on the marker level. QTL analyses revealed the presence of two major genes Scm1 and Scm2 plus three minor QTL. Involvement of Scm1 and Scm2 in the inheritance of SCMV resistance could be confirmed by BSA in a second cross. Breeders can make use of tightly linked STS markers for marker-assisted selection (MAS) as well as our SCMV resistant flint lines to improve their elite germplasm. Currently, recurrent backcrossing with phenotypic selection is the most appropriate and cost effective breeding method. With decreasing costs of DNA chip technology, MAS can be competitive with phenotypic selection in the near future. Further objectives of our research are the isolation and cloning of Scm1 and Scm2. To achieve this goal we follow two different approaches. (1) Positional cloning based on more than 500 AFLP primer combinations resulted in Scm1/Scm2 specific markers with a resolution of approximately 0.2 cM in the respective regions. (2) Resistance gene analogues (RGAs), cosegregating with the target genes are used to identify further candidate genes for transformation experiments.
Publications
From Fissistigma bracteolatum Chatt. (Annonaceae), a Vietnamese folk medicinal plant, the novel type natural products fissistigmatin A–D (1–4), being composed of a flavonoid and a sesquiterpene moiety, were isolated. Their structures were elucidated by extensive NMR techniques, MS, CD, molecular modelling calculations and X-ray analysis.
Publications
A galactose-specific and a mannose-specific lectin of the family of the jacalin-related lectins have been localized by immunofluorescence microscopy. The present localization studies provide for the first time unambiguous evidence for the cytoplasmic location of the mannose-specific jacalin-related lectin from rhizomes of Calystegia sepium, which definitely differs from the vacuolar location of the galactose-specific jacalin from Artocarpus integrifolia. These observations support the hypothesis that the galactose-specific jacalin-related lectins evolved from their mannose-specific homologues through the acquisition of vacuolar targeting sequences.
Publications
Treatment of parsley cell cultures with a fungal elicitor triggered the induction of a lipoxygenase isoform which may be involved in the de novo synthesis of defence-response inducers, such as jasmonic acid or 12-oxo-phytodienoic acid.
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A cDNA encoding a UDP-glucose:sinapate glucosyltransferase (SGT) that catalyzes the formation of 1-O-sinapoylglucose, was isolated from cDNA libraries constructed from immature seeds and young seedlings of rape (Brassica napus L.). The open reading frame encoded a protein of 497 amino acids with a calculated molecular mass of 55,970 Da and an isoelectric point of 6.36. The enzyme, functionally expressed in Escherichia coli, exhibited broad substrate specificity, glucosylating sinapate, cinnamate, ferulate, 4-coumarate and caffeate. Indole-3-acetate, 4-hydroxybenzoate and salicylate were not conjugated. The amino acid sequence of the SGT exhibited a distinct sequence identity to putative indole-3-acetate glucosyltransferases from Arabidopsis thaliana and a limonoid glucosyltransferase from Citrus unshiu, indicating that SGT belongs to a distinct subgroup of glucosyltransferases that catalyze the formation of 1-O-acylglucosides (β-acetal esters).
Publications
A mutant of Synechocystis sp. strain PCC6803 was obtained by random cartridge mutagenesis, which could not grow at low sodium concentrations. Genetic analyses revealed that partial deletion of the sll 0273 gene, encoding a putative Na+ /H+ exchanger, was responsible for this defect. Physiological characterization indicated that the sll 0273 mutant exhibited an increased sensitivity towards K+ , even at low concentrations, which was compensated for by enhanced concentrations of Na+ . This enhanced Na+ demand could also be met by Li+ . Furthermore, addition of monensin, an ionophore mediating electroneutral Na+ /H+ exchange, supported growth of the mutant at unfavourable Na+ /K+ ratios. Measurement of internal Na+ and K+ contents of wild‐type and mutant cells revealed a decreased Na+ /K+ ratio in mutant cells pre‐incubated at a low external Na+ /K+ ratio, while it remained at the level of the wild type after pre‐incubation at a high external Na+ /K+ ratio. We conclude that the Sll0273 protein is required for Na+ influx, especially at low external Na+ concentrations or low Na+ /K+ ratios. This system may be part of a sodium cycle and may permit re‐entry of Na+ into the cells, if nutrient/Na+ symporters are not functional or operating.
Publications
Jasmonates and their precursors, the octadecanoids, are signals in stress-induced alteration of gene expression. Several mRNAs coding for enzymes of jasmonic acid (JA) biosynthesis are up-regulated upon JA treatment or endogenous increase of the JA level. Here we investigated the positive feedback of endogenous JA on JA formation, as well as its β-oxidation steps. JA-responsive gene expression was recorded in terms of proteinase inhibitor2 (pin2) mRNA accumulation. JA formed upon treatment of tomato (Lycopersicon esculentum cv. Moneymaker) leaves with JA derivatives carrying different lengths of the carboxylic acid side chain was quantified by gas chromatography-mass spectrometry (GC-MS). The data revealed that β-oxidation of the side chain occurs up to a butyric acid moiety. The amount of JA formed from side-chain modified JA derivatives correlated with pin2-mRNA accumulation. JA derivatives with a carboxylic side chain of 3, 5 or 7 carbon atoms were unable to form JA and to express on pin2, whereas evennumbered derivatives were active.After treatment of tomato leaves with (10-2H)-(–)-12-oxophytoenoic acid, (4-2H)-(–)-JA and its methyl ester were formed and could be quantified separately from the endogenously nonlabeled JA pool by GC-MS analysis via isotopic discrimination. The level of 8 nmol per g fresh weight JA and its methyl ester originated exclusively from labeled 12-oxophytoenic acid. This and further data indicate that endogenous synthesis of the JA precursor 12-oxophytodienoic acid, as well as of JA and its methyl ester, are not induced in tomato leaves, suggesting that positive feedback in JA biosynthesis does not function in vivo.