Phloem sap exudates as a criterion for sink strength appreciation in Vitis vinifera cv. Pinot noir grapevines

The temporal evolution of the main compounds present in the phloem sap feeding the cluster of Vitis vinifera Pinot noir has been determined from the beginning of flowering until fruit set, after improvement of the facilitated exudation technique. The retained composition for the dipping solution was: HEPES (10 mM, pH 7.5), EDTA (10 mM). The first ramification of the cluster, maintained in situ, was sectionned then immersed in the dipping solution in order to favour the phloem exudation. The major organic components of the phloem sap were carbohydrates, amino acids and organic acids (i.e. sucrose, glutamine and tartrate, respectively). For each metabolic group, the mean exuded quantities correspond to 300 nmol per cluster in 4 h. The sharp increase in both organic compounds and potassium released over the flowering time-course reflects the rise of the mass flow supplying the cluster and underline the increasing sink strength of this organ. Moreover, the increasing contents of glutamine and hexoses in the exudate suggest a regulation in the allocation of assimilates to the reproductive organs.Les exsudats de sève phloémique comme critère d'appreciation de la force de puits de la grappe chez Vitis vinifera cv. Pinot noirL'évolution temporelle des principaux constituants de la sève libérienne alimentant la grappe de Vitis vinifera Pinot noir a été étudiée au cours de la floraison, après adaptation d'une technique de prélèvement par exsudation facilitée. La composition de la solution d'exsudation retenue est la suivante: HEPES (10 mM, pH 7,5), EDTA (10 mM). Sur la grappe maintenue in situ, l'extrémité de la première ramification est sectionnée puis immergée dans le milieu précédemment défini pour permettre la récupération des assimilats. Les composés organiques prédominants dans la sève libérienne sont les glucides solubles, les acides aminés et les acides organiques (saccharose, glutamine et tartrate respectivement). Pour chacun de ces groupes métaboliques, les quantités moyennes exsudées sont voisines de 300 nmol par grappe en 4 heures. Les quantités croissantes de glucides, d'acides aminés et de potassium collectées entre Je début de l'anthèse et la nouaison reflètent l'augmentation de flux de masse parvenant à la grappe et soulignent l'évolution de la force d'appel de ce puits. De plus, la part croissante de la glutamine et des hexoses dans les exsudats suggère une régulation dans la distribution des assimilats aux organes reproducteurs

Design Constraints on a Synthetic Metabolism

A metabolism is a complex network of chemical reactions that converts sources of energy and chemical elements into biomass and other molecules. To design a metabolism from scratch and to implement it in a synthetic genome is almost within technological reach. Ideally, a synthetic metabolism should be able to synthesize a desired spectrum of molecules at a high rate, from multiple different nutrients, while using few chemical reactions, and producing little or no waste. Not all of these properties are achievable simultaneously. We here use a recently developed technique to create random metabolic networks with pre-specified properties to quantify trade-offs between these and other properties. We find that for every additional molecule to be synthesized a network needs on average three additional reactions. For every additional carbon source to be utilized, it needs on average two additional reactions. Networks able to synthesize 20 biomass molecules from each of 20 alternative sole carbon sources need to have at least 260 reactions. This number increases to 518 reactions for networks that can synthesize more than 60 molecules from each of 80 carbon sources. The maximally achievable rate of biosynthesis decreases by approximately 5 percent for every additional molecule to be synthesized. Biochemically related molecules can be synthesized at higher rates, because their synthesis produces less waste. Overall, the variables we study can explain 87 percent of variation in network size and 84 percent of the variation in synthesis rate. The constraints we identify prescribe broad boundary conditions that can help to guide synthetic metabolism design

C. PRESL) at the transcriptional level.

This paper investigates differences in gene expression among the two Thlaspi caerulescens ecotypes La Calamine (LC) and Lellingen (LE) that have been shown to differ in metal tolerance and metal uptake. LC originates from a metalliferous soil and tolerates higher metal concentrations than LE which originates from a non-metalliferous soil. The two ecotypes were treated with different levels of zinc in solution culture, and differences in gene expression were assessed through application of a cDNA microarray consisting of 1,700 root and 2,700 shoot cDNAs. Hybridisation of root and shoot cDNA from the two ecotypes revealed a total of 257 differentially expressed genes. The regulation of selected genes was verified by quantitative reverse transcriptase polymerase chain reaction. Comparison of the expression profiles of the two ecotypes suggests that LC has a higher capacity to cope with reactive oxygen species and to avoid the formation of peroxynitrite. Furthermore, increased transcripts for the genes encoding for water channel proteins could explain the higher Zn tolerance of LC compared to LE. The higher Zn tolerance of LC was reflected by a lower expression of the genes involved in disease and defence mechanisms. The results of this study provide a valuable set of data that may help to improve our understanding of the mechanisms employed by plants to tolerate toxic concentrations of metal in the soil

Evidence for the Glutamine Synthetase/Glutamate Synthase Pathway during the Photorespiratory Nitrogen Cycle in Spinach Leaves

Spinach leaf (Spinacia oleracea L.) discs infiltrated with [(15)N]glycine were incubated at 25°C in the light and in darkness for 0, 30, 60 and 90 minutes. The kinetics of (15)N-incorporation into glutamine, glutamate, asparagine, aspartate, and serine from [(15)N]glycine was determined. At the beginning of the experiment, just after infiltration (0 min incubation) serine, and the amido-N of glutamine and asparagine were the only compounds significantly labeled in both light- and dark-treated leaf discs. Incorporation of (15)N-label into the other amino acids was observed at longer incubation time. The per cent (15)N-enrichment in all amino acids was found to increase with incubation. However, serine and the amido-N of glutamine remained the most highly labeled products in all treatments. The above pattern of (15)N-labeling suggests that glutamine synthetase was involved in the initial refixation of (15)NH(3) derived from [(15)N]glycine oxidation in spinach leaf discs. The (15)N-enrichment of the amino-N of glutamine was found to increase rapidly from 0 to 19% during incubation in the light. There was a comparatively smaller increase (4-9%) in the (15)N-label of the amino-N of glutamine in tissue incubated in darkness. Furthermore the total flux of (15)N-label into each of the amino acids examined was found to be greater in tissue incubated in the light than those in the dark. The above evidence indicates the involvement of the glutamine synthetase/glutamate synthase pathway in the recycling of photorespiratory NH(3) during glycine oxidation in spinach leaves