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Responses to successive drought and re-watering cycles (1-3 cycles) were compared in greenhouse-grown cuttings of Populus x canadensis (Moench) clones, 'Luisa_ Avanzo' and 'Dorskamp.' Total leaf number increment rate, duration of leaf expansion, total and individual leaf area expansion rates and stomatal conductance were recorded periodically during the experiment. Soil water content (SWC) and predawn leaf water potential (Psi(WP)) were measured four times during each drought cycle. In parallel, relative leaf water content (RWC) and specific leaf area (SLA) were estimated on leaves collected from the top to bottom of each cutting. Under well-watered conditions, 'Luisa_Avanzo' and 'Dorskamp' differed in their patterns of leaf area expansion. Although duration of leaf expansion was similar between clones, 'Luisa_ Avanzo' exhibited higher total leaf number increment rates and individual leaf area increases than 'Dorskamp.' As a result, 'Luisa_Avanzo' cuttings reached larger individual and total leaf areas than 'Dorskamp.' 'Dorskamp' leaves had lower SLA than 'Luisa_Avanzo' leaves. In response to successive drought cycles, both clones underwent decreases in total leaf number increment rates and in total leaf area expansion rates, but both whole-plant and individual leaf areas were drastically reduced only in 'Luisa_Avanzo.' 'Dorskamp' maintained a constant leaf area as a result of an increase in the duration of leaf expansion during drought and a significant stimulation of individual leaf area expansion rate and total leaf number increment rate in response to re-watering. Drought caused a greater decrease in SLA in 'Luisa_Avanzo' than in 'Dorskamp.' Expanded leaves of 'Dorskamp' were constitutively dense or thick, or both, whereas leaves of 'Luisa_Avanzo' became dense or thick, or both, only in response to drought. In both clones, re-watering caused partial recovery of SLA to control values. Our data confirm previous field and greenhouse observations that 'Dorskamp' is more tolerant to successive drought and re-watering cycles than 'Luisa_Avanzo.' We conclude that this difference is mainly attributable to the plasticity of 'Dorskamp' after re-watering, rather than to clonal differences in drought responses.

Expression of Agrobacterium tumefaciens virulence genes and transformation of dicots by this organism are dependent upon host plant phenolic compounds. Several alkylsyringamides have recently been shown to be powerful inducers of these vir-genes. These synthetic amides, and especially ethylsyringamide, are much stronger inducers than syringic acid. In this work, four alkylamides derived from ferulic or sinapic acids were synthesized by a dicyclohexylcarbodiimide method and tested for their potential to induce vir-gene expression on A. tumefaciens strains harbouring virB::lacZ or virE::lacZ fusion plasmids. Their effectiveness was compared to that of ethylsyringamide and tyraminylferulamide, a naturally occurring amide in plants. Whatever the amine moiety of the amide (ethylamine, propylamine, tyramine or beta-alanine ethyl ester) conjugation of the acid functional group clearly diminished the toxicity to the bacteria of the respective acid at high concentration and thereby increased the vir-inducing potential. However, none of the inducers tested exhibited higher activity than acetosyringone, the reference compound for vir-gene induction, with the exception of ethylsyringamide at concentrations above 1mM. When tested on Agrobacterium tumefaciens strain A348(pSM243cd), ethylferulamide and ethylsinapamide were more efficient than the corresponding phenolic acids but only above 100 microM.

International audience; Lectins are present on the surface of many cells. Many lectins actively recycle from membrane to endosomes and efficiently take up glycoconjugates in a sugar-dependent manner. On this basis, glycoconjugates, specially those obtained by chemical means, are good candidates as carriers of drugs, oligonucleotides or genes. In this paper, we present a panel of methods suitable to transform unprotected reducing oligosaccharides into glycosynthons designed to be easily linked to therapeutic agents. All the glycosynthons presented here are glycosylamines or derivatives, mainly glyco-amino acids or glycopeptides. Glycosylamines are easy to obtain, but they are very labile in slightly acidic or neutral medium; they must be stabilized, by acylation for instance. The coupling efficiency of a reducing sugar with ammonia as well as an alkylamine or an arylamine is higher at high temperature, however, because of the Amadori rearrangement, special conditions have to be selected to prepare the expected glycosylamine derivative with a high yield. Glycosylamines are easily acylated by N-protected amino acids, or by halogeno acids which can then be transformed into amino acids. Alternatively, unprotected reducing oligosaccharides may very efficiently be transformed into N-glycosyl-amino acids and then protected by N-acylation. With a glutamyl derivative having both the alpha-amino and the gamma-carboxylic groups free, the coupling and the acylation, which is intramolecular, are roughly quantitative. N-oligosaccharyl-amino acid derivatives are interesting glycosynthons, because their sugar moiety bears the specificity towards membrane lectins while the amino acid part has the capacity to easily substitute a therapeutic agent.

The virulence genes of Agrobacterium tumefaciens are specifically activated by plant phenolic compounds and allow this organism to genetically transform plant cells. New types of phenolic compounds, three phenol amides derived from syringic acid, were synthesized. Introduction of an amide group in syringic acid strongly enhances its vir gene inducing activity.

Coniferyl alcohol β-glycosides were synthesized by coupling coniferyl alcohol sodium salt in aqueous acetone with peracetylated glycosyl halides of d -glucopyranose, d -galactopyranose and l -fucopyranose. After O -deacetylation by sodium methoxide, the glycosides were tested for their potential vir gene-inducing activities in Agrobacterium tumefaciens A348/pSM358 and A348/pSM243cd harbouring, respectively, virE::lacZ and virB::lacZ fusion plasmids. In the large range of concentrations tested, these synthetic derivatives had vir -inducing activity on strain A348/pSM358: d -Gal > d -Glc. The dose-response curves of these glycosylated compounds were completely different from those of free coniferyl alcohol; the β-galactopyranoside at 500,μM exhibited interestingly a greater activity than the free aglycone after a 12 hr incubation period. The coniferyl alcohol β-glucopyranoside was a better inducer than the related β-galactopyranoside on strain A348/pSM243cd. The β- l -fucopyranoside was inactive with both A. tumefaciens strains. The activities of these glycosylated inducers were correlated with the presence of intracellular glycosylhydrolases from the various A. tumefaciens strains studied. It is noteworthy that the vir -gene induction activity of the coniferyl alcohol β-galactopyranoside on strain A348/pSM358 was due to a high basal level of expression of the virE::lacZ fusion gene.

Lectins are present on the surface of many cells. Many lectins actively recycle from membrane to endosomes and efficiently take up glycoconjugates in a sugar-dependent manner. On this basis, glycoconjugates, specially those obtained by chemical means, are good candidates as carriers of drugs, oligonucleotides or genes. In this paper, we present a panel of methods suitable to transform unprotected reducing oligosaccharides into glycosynthons designed to be easily linked to therapeutic agents. All the glycosynthons presented here are glycosylamines or derivatives, mainly glyco-amino acids or glycopeptides. Glycosylamines are easy to obtain, but they are very labile in slightly acidic or neutral medium; they must be stabilized, by acylation for instance. The coupling efficiency of a reducing sugar with ammonia as well as an alkylamine or an arylamine is higher at high temperature, however, because of the Amadori rearrangement, special conditions have to be selected to prepare the expected glycosylamine derivative with a high yield. Glycosylamines are easily acylated by N-protected amino acids, or by halogeno acids which can then be transformed into amino acids. Alternatively, unprotected reducing oligosaccharides may very efficiently be transformed into N-glycosyl-amino acids and then protected by N-acylation. With a glutamyl derivative having both the alpha-amino and the gamma-carboxylic groups free, the coupling and the acylation, which is intramolecular, are roughly quantitative. N-oligosaccharyl-amino acid derivatives are interesting glycosynthons, because their sugar moiety bears the specificity towards membrane lectins while the amino acid part has the capacity to easily substitute a therapeutic agent.

Aryl β-glycopyranosides have been synthesized by coupling syringaldehyde (3-methoxyvanillin) with l -fucose, d -galactose, and maltose; acetosyringone (4-hydroxy-3,5-dimethoxyacetophenone) with l -fucose and maltose; acetovanillone (4-hydroxy-3-methoxyacetophenone) with l -fucose; and syringic acid (4-hydroxy-3,5-dimethoxybenzoic acid) with d -galactose. The procedure using peracetylated glycosyl halides and sodium phenolates in aqueous acetone afforded the acetylated β-glycosides, which were deacetylated.

International audience; Genotypic variability for productivity, wateruse efficiency and leaf traits in 33 genotypes selected from an F1 progeny of Populus deltoides Bartr. ex Marsh • Populus trichocarpa L. was explored under optimal and moderate water-deficit conditions. Saplings of the 33 genotypes were grown in a two-plot open field at INRA Orle´ ans (France) and coppiced every year. A moderate water deficit was induced during two successive years on one plot by withholding irrigation, while the second one remained irrigated (control). Stem biomass and leaf structure (e.g., specific leaf area and leaf area) were measured in 2004 and 2005 and functional leaf traits (e.g., carbon isotope discrimination, D) were measured only in 2004. Tolerance to water deficit was estimated at genotype level as the ability to limit losses in biomass production in water deficit versus control trees. Stem biomass, leaf structure and D displayed a significant genotypic variability whatever the irrigation regime. For all traits, genotype ranks remained stable across years for similar irrigation conditions. Carbon isotope discrimination scaled negatively with productivity and leaf nitrogen content in controls. The most productive genotypes were the least tolerant to moderate water deficit. No relationship was evidenced between D and the level of tolerance to water deficit. The relationships between traits evidenced in this collection of P. deltoides • P. trichocarpa F1 genotypes contrast with the ones that were previously detected in a collection of P. deltoides • Populus nigra L. cultivars tested in the same field trial.

International audience; • We examined the relationships among roductivity, water use efficiency (WUE) and drought tolerance in 29 genotypes of Populus × euramericana (Populus deltoides × Populus nigra), and investigated whether some leaf traits could be used as predictors for productivity, WUE and drought tolerance. • At Orléans, France, drought was induced on one field plot by withholding water, while a second plot remained irrigated and was used as a control. Recorded variables included stem traits (e.g. biomass) and leaf structural (e.g. leaf area) and functional traits [e.g. intrinsic water use efficiency (Wi) and carbon isotope discrimination (∆)]. • Productivity and ∆ displayed large genotypic variability and were not correlated. ∆ scaled negatively with Wi and positively with stomatal conductance under moderate drought, suggesting that the diversity for ∆ was mainly driven by stomatal conductance. • Most of the productive genotypes displayed a low level of drought tolerance (i.e. a large reduction of biomass), while the less productive genotypes presented a large range of drought tolerance. The ability to increase WUE in response to water deficit was necessary but not sufficient to explain the genotypic diversity of drought tolerance.

To test if some leaf parameters are predictors of productivity in a range of Populus deltoides (Bartr.) Marsh. x P. nigra L. clones, we assessed leaf traits and productivity in 2-month-old rooted cuttings from 31 clones growing in 4-l pots in a greenhouse, under conditions of controlled temperature and optimal irrigation. We evaluated four groups of variables describing (1) productivity (total biomass), (2) leaf growth (total leaf number increment and total leaf area increment rate), (3) leaf structure (specific leaf area and nitrogen and carbon contents) and (4) carbon isotope discrimination (delta), which is negatively correlated with time-integrated water-use efficiency. High-yielding clones did not necessarily display high leaf growth rates, but they displayed a larger total leaf area, lower specific leaf area and lower leaf nitrogen concentration than clones with low productivity. Total leaf area was mainly controlled by maximal individual leaf area and total leaf area increment rate (r = 0.51 and 0.56, respectively). Carbon isotope discrimination did not correlate with total biomass, but it was associated with total number of leaves and total leaf area increment rate (r = 0.39 and 0.45, respectively). Therefore, leaf area and specific leaf area were better indicators of productivity than leaf growth traits. The observed independence of delta from biomass production provides opportunities for selecting poplar clones combining high productivity and high water-use efficiency.

International audience; • Here we tested whether some leaf traits could be used as predictors for productivity in a range of Populus deltoides × P. nigra clones. These traits were assessed in 3-yr-old rooted cuttings from 29 clones growing in an open field trial, in a five randomized complete block design, under optimal irrigation. • Variables were assigned to four groups describing productivity (above-ground biomass, total leaf area), leaf growth (total number of leaves increment rate), leaf structure (area of the largest leaf, specific leaf area, carbon and nitrogen contents), and carbon isotope discrimination in the leaves (∆). • High-yielding clones displayed larger total leaf area and individual leaf area, while no correlation could be detected between productivity and either leaf structure or ∆. By contrast, ∆ was negatively correlated with number of leaves increment rate and leaf N content. • Our study shows that there is a potential to improve water-use efficiency in poplar without necessarily reducing the overall productivity.

International audience; We compared responses to drought and re-watering of greenhouse-grown cuttings of Populus × euramericana (Dode) Guinier clones, Luisa Avanzo and Dorskamp. Total leaf area, leaf number, leaf area increment and stomatal conductance were evaluated periodically during a 29-day drought period and for 16 days after re-watering. Soil water content and predawn leaf water potential (Øwp) were measured on Days 29 and 45. On the same days, relative water content (RWC), specific leaf area (SLA), nitrogen, chlorophyll, soluble sugars, total phenols, flavanols and antioxidant activity were determined for leaves taken from the bottom to the top of each cutting. Leaves of Luisa Avanzo cuttings grew more rapidly than leaves of Dorskamp and exhibited higher SLA, but lower concentrations of nitrogen, chlorophyll and soluble sugars and lower antioxidant activity per unit area. On Day 29, after withholding water, both clones had closed their stomata, reduced rates of leaf growth, and lower Øwp and RWC; however, the clones differed in their responses to soil water depletion. Compared to Dorskamp, Luisa Avanzo closed its stomata earlier and maintained higher Øwp, but lower RWC and leaf sugar concentrations. Antioxidant activity of leaf methanolic extracts decreased in response to water stress only in Luisa Avanzo. Leaf physiology and its modulation by water stress were age dependent in Luisa Avanzo.

An original procedure which preserves the structure of the sugar ring is described to link a plant hormone as N6-isopent-2-enyladenosine [( 9R]iP) onto a protein carrier to prepare a more specific immunogen. This cytokinin is bound to bovine serum albumin (BSA) and ovalbumin by a five-step procedure. These [9R]iP-protein conjugates have a maximal absorption at 269 nm and show molar ratios of hormone bound to proteins in the range of 12:1 and 18:1 for BSA and ovalbumin, respectively. Polyclonal antibodies were raised in rabbits against [9R]iP-BSA and were purified by affinity chromatography. Titers and specificity of the antisera and purified antibodies were determined by ELISA and RIA. These antibodies are highly specific for [9R]iP and do not cross-react with zeatin and ribosylzeatin. An immunoaffinity matrix was prepared with a capacity of 1 microgram of [9R]iP/mL of gel.

Alpha- and beta-D-galactosidases were characterized from a hydroalcoholic extract of wheat germ (Triticum vulgare). Kinetic constants (Vmax and KM) and the optimal pHs for the hydrolysis of p-nitrophenyl galactopyranosides by both enzymes were determined. These enzymes presented a high stability in hydroalcoholic medium and were inhibited by iodoacetamide and sodium p-hydroxy-mercuribenzoate.

Expression of the Agrobacterium tumefaciens virulence ( vir ) gene is known to be dependent on host plant phenolic compounds. The A. tumefaciens strain A348 (pSM358) harbouring a vir E:: lac Z fusion plasmid was used to detect the ability of 13 synthetic acetosyringone, acetovanillone, syringaldehyde and syringic acid β-glycosides to induce virulence. The activity of the reporter β-galactosidase was detected by spectrofluorimetry using 4-methylumbelliferyl β-galactopyranoside as substrate. Acetosyringonyl β- l -fucopyranoside was the most active monoglycoside tested; even at high concentration this compound was devoid of toxic effects. However, monoglycosides were less active vir inducers than free acetosyringone. In contrast, the β-maltoside of syringaldehyde showed higher activity than the free phenol at high concentration. The activity of such glycosylated inducers may be related to specific sugar receptors on the bacterial cell surface.

The β-L-fucopyranosyl chloride 2,3,4-tri(chlorosulfate) (1) has been prepared by reaction of sulfuryl chloride with L-fucose at low temperature (yield, 47%). This β-L-fucopyranosyl chloride (1) gave high yields of the α-L-fucoside by reaction with alcohols. The p-nitrobenzyl α-L-fucoside has been prepared (yield, 35%). From L-fucose, the α-L-fucoside is obtained in three steps.