Isabelle S. Lefèvre

Discrimination between the ionic and osmotic components of salt stress in relation to free polyamine level in rice (Oryza sativa)

In order to precise the relative importance of ionic and osmotic components of salt stress on modification of free polyamine level, seedling of two rice cultivars (I Kong Pao (IKP): salt-sensitive and Pokkali: salt-resistant) were exposed to isoosmotic concentrations of NaCl, KCl (50 and 100 mM) or polyethylene glycol (PEG 6000, 16 and 26%) for a maximal period of 12 h in daylight conditions, Both ion and polyamine concentrations were already modified after 3 h of exposure to ionic stresses. At the shoot level. this response occurred independently of any change in the plant water status. Conversely, exposure to 26% PEG during 12 h induced a strong decrease in shoot osmotic potential and water content but had only a limited impact on endogenous free polyamine level. It was thus suggested that the ionic component by itself might trigger short-term polyamine accumulation. This effect might be attributed to monovalent ions, although K+ and Na+ accumulation had different effects on polyamine content. Putrescine may assume differential role in non-photosynthetic organs versus photosynthetic ones since it accumulated to high amounts in the roots of the salt-resistant cultivar Pokkali comparatively to salt-sensitive cultivar IKP. while an opposite trend was recorded in the shoots. Tyramine was also present at hi.-her concentrations in the roots of Pokkali;, its level clearly increased in response to ionic stresses while cadaverine level increased in water stress conditions only. No clear relationship was observed between the mean level of salinity resistance and the endogenous concentrations of spermidine or spermine

Putrescine differently influences the effect of salt stress on polyamine metabolism and ethylene synthesis in rice cultivars differing in salt resistance

Effects of salt stress on polyamine metabolism and ethylene production were examined in two rice (Oryza sativa L.) cultivars [I Kong Pao (IKP), salt sensitive; and Pokkali, salt resistant] grown for 5 d and 12 d in nutrient solution in the presence or absence of putrescine (1 mM) and 0, 50, and 100 mM NaCl. The salt-sensitive (IKP) and salt-resistant (Pokkali) cultivars differ not only in their mean levels of putrescine, but also in the physiological functions assumed by this molecule in stressed tissues. Salt stress increased the proportion of conjugated putrescine in salt-resistant Pokkali and decreased it in the salt-sensitive IKP, suggesting a possible protective function in response to NaCl. Activities of the enzymes ornithine decarboxylase (ODC; EC 4.1.1.17) and arginine decarboxylase (ADC; EC 4.1.1.19) involved in putrescine synthesis were higher in salt-resistant Pokkali than in salt-sensitive IKP. Both enzymes were involved in the response to salt stress. Salt stress also increased diamine oxidase (DAO; 1.4.3.6) and polyamine oxidase (PAO EC 1.5.3.11) activities in the roots of salt-resistant Pokkali and in the shoots of salt-sensitive IKP. Gene expression followed by reverse transcription-PCR suggested that putrescine could have a post-translational impact on genes coding for ADC (ADCa) and ODC (ODCa and ODCb) but could induce a transcriptional activation of genes coding for PAO (PAOb) mainly in the shoot of salt-stressed plants. The salt-resistant cultivar Pokkali produced higher amounts of ethylene than the salt-sensitive cultivar IKP, and exogenous putrescine increased ethylene synthesis in both cultivars, suggesting no direct antagonism between polyamine and ethylene pathways in rice.

Identification of drought-responsive compounds in potato through a combined transcriptomic and targeted metabolite approach

Two potato clones (Solanum tuberosum L.) of the Andean cultivar group, called Sullu and SS2613, with different drought-tolerance phenotypes were exposed to a continuously increasing drought stress in a field trial. At the physiological level, while relative leaf water contents were similar in both clones, osmotic potential was lower in Sullu and declined more strongly during drought compared with SS2613. In the drought-stressed plants, tuber yield was reduced by about 70% compared with control plants in both clones. Potato cDNA microarrays and target metabolite analysis were performed on leaves sampled at several time-points after the onset of drought. At the transcriptomic level, photosynthesis-related genes were already strongly repressed in Sullu after 28 d of withholding irrigation and even more strongly after a longer stress duration, whereas, in SS2613, repression occurred only after 49 d of soil drying; similarly, a strong perturbation of carbohydrate-related genes was observed in Sullu. At the metabolite level, differential accumulation of osmotically active solutes was observed between the two cultivars; indeed, in Sullu, contents of galactose, inositol, galactinol, proline, and proline analogues were higher upon drought stress compared with SS2613. These results point to different drought responses in the cultivars at the leaf level, with, however, similar tuber yield reductions. The previously shown tolerant clone Sullu lost part of its tolerance under the experimental conditions used here; it was, however, able to maintain an absolute yield three times higher than SS2613.

Gene expression changes related to the production of phenolic compounds in potato tubers grown under drought stress.

Polyphenols represent a large family of plant secondary metabolites implicated in the prevention of various diseases such as cancers and cardiovascular diseases. The potato is a significant source of polyphenols in the human diet. In this study, we examined the expression of thirteen genes involved in the biosynthesis of polyphenols in potato tubers using real-time RT-PCR. A selection of five field grown native Andean cultivars, presenting contrasting polyphenol profiles, was used. Moreover, we investigated the expression of the genes after a drought exposure. We concluded that the diverse polyphenolic profiles are correlated to variations in gene expression profiles. The drought-induced variations of the gene expression was highly cultivar-specific. In the three anthocyanin-containing cultivars, gene expression was coordinated and reflected at the metabolite level supporting a hypothesis that regulation of gene expression plays an essential role in the potato polyphenol production. We proposed that the altered sucrose flux induced by the drought stress is partly responsible for the changes in gene expression. This study provides information on key polyphenol biosynthetic and regulatory genes, which could be useful in the development of potato varieties with enhanced health and nutritional benefits.

How can we take advantage of halophyte properties to cope with heavy metal toxicity in salt-affected areas?

BACKGROUND Many areas throughout the world are simultaneously contaminated by high concentrations of soluble salts and by high concentrations of heavy metals that constitute a serious threat to human health. The use of plants to extract or stabilize pollutants is an interesting alternative to classical expensive decontamination procedures. However, suitable plant species still need to be identified for reclamation of substrates presenting a high electrical conductivity. SCOPE Halophytic plant species are able to cope with several abiotic constraints occurring simultaneously in their natural environment. This review considers their putative interest for remediation of polluted soil in relation to their ability to sequester absorbed toxic ions in trichomes or vacuoles, to perform efficient osmotic adjustment and to limit the deleterious impact of oxidative stress. These physiological adaptations are considered in relation to the impact of salt on heavy metal bioavailabilty in two types of ecosystem: (1) salt marshes and mangroves, and (2) mine tailings in semi-arid areas. CONCLUSIONS Numerous halophytes exhibit a high level of heavy metal accumulation and external NaCl may directly influence heavy metal speciation and absorption rate. Maintenance of biomass production and plant water status makes some halophytes promising candidates for further management of heavy-metal-polluted areas in both saline and non-saline environments.

Differential cadmium and zinc distribution in relation to their physiological impact in the leaves of the accumulating Zygophyllum fabago L

Cadmium and zinc share many similar physiochemical properties, but their compartmentation, complexation and impact on other mineral element distribution in plant tissues may drastically differ. In this study, we address the impact of 10 μm Cd or 50 μm Zn treatments on ion distribution in leaves of a metallicolous population of the non-hyperaccumulating species Zygophyllum fabago at tissue and cell level, and the consequences on the plant response through a combined physiological, proteomic and metabolite approach. Micro-proton-induced X-ray emission and laser ablation inductively coupled mass spectrometry analyses indicated hot spots of Cd concentrations in the vicinity of vascular bundles in response to Cd treatment, essentially bound to S-containing compounds as revealed by extended X-ray absorption fine structure and non-protein thiol compounds analyses. A preferential accumulation of Zn occurred in vascular bundle and spongy mesophyll in response to Zn treatment, and was mainly bound to O/N-ligands. Leaf proteomics and physiological status evidenced a protection of photosynthetically active tissues and the maintenance of cell turgor through specific distribution and complexation of toxic ions, reallocation of some essential elements, synthesis of proteins involved in photosynthetic apparatus or C-metabolism, and metabolite synthesis with some specificities regarding the considered heavy metal treatment.

Combined transcriptomic and physiological approaches reveal strong differences between short‐ and long‐term response of rice (Oryza sativa) to iron toxicity

Ferrous iron toxicity is a mineral disorder frequently occurring under waterlogged soils where rice is cultivated. To decipher the main metabolic pathways involved in rice response to iron excess, seedlings have been exposed to 125 mg L(-1) FeSO(4) for 3 weeks. A combined transcriptomic, biochemical and physiological study has been performed after short-term (3 d) or long-term (3 weeks) exposure to iron in order to elucidate the strategy of stress adaptation with time. Our results showed that short- and long-term exposure involved a very different response in gene expression regarding both the number and function. A larger number of genes were up- or down-regulated after 3 d than after 3 weeks of iron treatment; these changes also occurred in shoot even though no significant difference in iron concentration was recorded. Those modifications in gene expression after 3 d affected not only genes involved in hormonal signalling but also genes involved in C-compound and carbohydrate metabolism, oxygen and electron transfer, oxidative stress, and iron homeostasis and transport. Modification in some gene expression can be followed by modification in corresponding metabolic products and physiological properties, or differed in time for some others, underlying the importance of an integrated study.

Differential impact of chronic ozone exposure on expanding and fully expanded poplar leaves.

Populus tremula L. × Populus alba L. (Populus ×c anescens (Aiton) Smith) - clone INRA 717-1-B4 saplings (50 cm apex to base and carrying 19 leaves on average) - were followed for 28 days. Half of the trees were grown in charcoal-filtered air while the other half were exposed to 120 ppb ozone for 11 h a day during the light period. The expanding leaf number 4 was tagged at the beginning of the experiment and finished expansion between 7 and 14 days. These leaves were harvested weekly for biochemical and proteome analyses using quantitative bidimensional electrophoresis (DiGE). Independent of the ozone treatment, all the analyses allowed a distinction between expanding and adult leaves. The results indicate that during the expansion phase (Days 0-7) the enzymatic machinery of the leaves is set up, and remains dynamically stable in the adult leaves (Days 14-28). Although ozone had no apparent effect on expanding leaves, the metabolic stability in fully expanded leaves observed in ozone-free plants was disturbed after 2 weeks of exposure and a stress-induced response became apparent.

Antioxidant enzyme activities and hormonal status in response to Cd stress in the wetland halophyte Kosteletzkya virginica under saline conditions

Salt marshes constitute major sinks for heavy metal accumulation but the precise impact of salinity on heavy metal toxicity for halophyte plant species remains largely unknown. Young seedlings of Kosteletzkya virginica were exposed during 3 weeks in nutrient solution to Cd 5 µM in the presence or absence of 50 mM NaCl. Cadmium (Cd) reduced growth and shoot water content and had major detrimental effect on maximum quantum efficiency (F(v) /F(m) ), effective quantum yield of photosystem II (Y(II)) and electron transport rates (ETRs). Cd induced an oxidative stress in relation to an increase in O(2) (•-) and H(2) O(2) concentration and lead to a decrease in endogenous glutathione (GSH) and α-tocopherol in the leaves. Cd not only increased leaf zeatin and zeatin riboside concentration but also increased the senescing compounds 1-aminocyclopropane-1-carboxylic acid (ACC) and abscisic acid (ABA). Salinity reduced Cd accumulation already after 1 week of stress but was unable to restore shoot growth and thus did not induce any dilution effect. Salinity delayed the Cd-induced leaf senescence: NaCl reduced the deleterious impact of Cd on photosynthesis apparatus through an improvement of F(v) /F(m) , Y(II) and ETR. Salt reduced oxidative stress in Cd-treated plants through an increase in GSH, α-tocopherol and ascorbic acid synthesis and an increase in glutathione reductase (EC 1.6.4.2) activity. Additional salt reduced ACC and ABA accumulation in Cd+NaCl-treated leaves comparing to Cd alone. It is concluded that salinity affords efficient protection against Cd to the halophyte species K. virginica, in relation to an improved management of oxidative stress and hormonal status.

Comparative evaluation of the Versant HIV-1 RNA 1.0 kinetic PCR molecular system (kPCR) for the quantification of HIV-1 plasma viral load.

BACKGROUND New automated and ultrasensitive assays are becoming available to monitor HIV-1 plasma viral load, which is an essential marker for the clinical follow-up. OBJECTIVES To evaluate the performances of the VERSANT HIV-1 RNA 1.0 (kPCR) automated assay in a clinical laboratory setting. STUDY DESIGN Frozen plasma samples from various HIV-1 subtypes, previously analysed with the VERSANT HIV-1 RNA 3.0 (bDNA) in clinical routine, were retested with the new VERSANT kPCR assay. A comparison was also done with two other commercial assays (NucliSens EasyQ HIV-1 and Abbott real time HIV-1). RESULTS We observed a good correlation between the viral load measurements obtained with the kPCR assay and the other techniques. Nevertheless, in terms of absolute quantification, we observed discrepancies of more than 0.5 log cop/ml plasma with 36%, 35% and 0% of the samples respectively with NucliSens EasyQ, VERSANT bDNA 3.0 and Abbott real time. No HIV-1 negative sample was amplified by the kPCR. Tenfold dilutions of samples from HIV-1 subtypes A-D, F-H, K, CRF01, CRF02 and CRF06 were analysed to evaluate the kPCR efficiency: the amplification had an efficiency close to the maximum of 2 for each of the subtypes tested. CONCLUSIONS The VERSANT HIV-1 RNA 1.0 assay (kPCR) is suitable for use in a clinical setting with various HIV-1 subtypes. The plasma viral load quantifications obtained with the kPCR assay were close to those obtained with the Abbott real time HIV-1 assay.