Walid Zorrig

Identification of three relationships linking cadmium accumulation to cadmium tolerance and zinc and citrate accumulation in lettuce.

Lettuce (Lactuca sativa) is a plant species that shows high accumulation of cadmium, a toxic heavy metal. Lettuce is therefore a good model both for identifying determinants controlling cadmium accumulation in plant tissues and for developing breeding strategies aimed at limiting cadmium accumulation in edible tissues. In this work, 14-day-old plants from three lettuce varieties were grown for 8 days on media supplemented with cadmium concentrations ranging from 0 to 50 microM. Growth, as well as Cd(2+), Zn(2+), K(+), Ca(2+), NO(3)(-), SO(4)(2-), Cl(-), phosphate, malate and citrate root an shoot contents were analyzed. The three lettuce varieties Paris Island Cos, Red Salad Bowl and Kordaat displayed differential abilities to accumulate cadmium in roots and shoots, Paris Island Cos displaying the lowest cadmium content and Kordaat the highest. From the global analysis of the three varieties, three main trends were identified. First, a common negative correlation linked cadmium tissue content and relative dry weight reduction in response to cadmium treatments in the three varieties. Second, increasing cadmium concentration in the culture medium resulted in a parallel increase in zinc tissue content in all lettuce varieties. A common strong positive correlation between cadmium and zinc contents was observed for all varieties. This suggested that systems enabling zinc and cadmium transport were induced by cadmium. Finally, the cadmium treatments had a contrasting effect on anion contents in tissues. Interestingly, citrate content in shoots was correlated with cadmium translocation from roots to shoots, suggesting that citrate might play a role in cadmium transport in the xylem vessels. Altogether, these results shed light on three main strategies developed by lettuce to cope with cadmium, which could help to develop breeding strategies aimed at limiting cadmium accumulation in lettuce.

Lettuce (Lactuca sativa): a species with a high capacity for cadmium (Cd) accumulation and growth stimulation in the presence of low Cd concentrations

Summary Cadmium (Cd) is a metal pollutant that accumulates in cultivated soils and has detrimental consequences in terms of food safety. Lettuce (Lactuca sativa) can be characterised as having a high capacity to accumulate Cd in its tissues. An analysis of Cd tolerance and Cd accumulation was carried out using two varieties of lettuce (‘Divina’ and ‘Melina’). A wide range of CdCl2 concentrations was used (0.0, 0.1, 0.6, 3.0, and 15.0 µM CdCl2). The lowest concentration (0.1 µM CdCl2) stimulated growth, while the two highest concentrations resulted in a reduction in biomass. Cadmium concentrations were found to be twice as high in roots as in shoots. ‘Divina’ displayed lower concentrations of Cd than ‘Melina’ in nearly all treatments. A strong negative correlation was observed between Cd concentration and Cd tolerance in the roots and shoots (R2 > 0.87) of both ‘Melina’ and ‘Divina’. Lettuce grown in the presence of 15.0 µM CdCl2 had leaf Cd concentrations that were 100-fold higher than the legal maximum level for vegetable products marketed for human consumption, but showed no symptoms of dehydration, chlorosis, or necrosis. This result represents an important alert for lettuce consumers and growers.

PHYTODESALINATION OF A MODERATELY-SALT-AFFECTED SOIL BY SULLA CARNOSA

The aim of this investigation was to evaluate the ability of the indifferent halophyte Sulla carnosa Desf. to desalinize a moderately-salt-affected soil. Seeds were sown on a fertile soil added or not with 1.5 g NaCl. kg−1. Analogous treatments without plantation (control and salinized) were also used. Plant culture was performed under greenhouse conditions in non-perforated pots containing 10 kg soil each and irrigated with non-saline tap water. After 80 days of treatment, shoots were harvested. Soil samples were also collected after division of soil column in each pot into two horizons. Our results showed that salt addition increased electrical conductivity of saturation paste extract (ECe) from 3.3 to 8.4 dS. m−1 and soluble sodium concentration from 0.32 to 1.15 g. kg−1 soil in the upper horizon. In the lower horizon however, Na+ concentration was quasi-constant and then ECe was less increased. Plant culture inversed this pattern of sodium accumulation and salinity. Its productivity and phytodesalination capacity in 80 days were 5.0 t DW. ha−1 and 0.3 t Na+. ha−1 (24% of the added quantity), respectively. Interestingly, sodium dilution within biomass (41.5–45.6 mg. g−1 DW) and the non-altered nutrition make this plant suitable for forage as second use after phytodesalination.

Effects of magnesium deficiency on photosynthesis and carbohydrate partitioning

Magnesium nutrition is often forgotten, while its absence adversely affects numerous functions in plants. Magnesium deficiency is a growing concern for crop production frequently observed in lateritic and leached acid soils. Competition with other cations (Ca2+, Na+, and K+) is also found to be an essential factor, inducing magnesium deficiency in plants. This nutrient is required for chlorophyll formation and plays a key role in photosynthetic activity. Moreover, it is involved in carbohydrate transport from source-to-sink organs. Hence, sugar accumulation in leaves that results from the impairment of their transport in phloem is considered as an early response to Mg deficiency. The most visible effect is often recorded in root growth, resulting in a significant reduction of root/shoot ratio. Carbohydrate accumulation in source leaves is attributed to the unique chemical proprieties of magnesium. As magnesium is a nutrient with high mobility in plants, it is preferentially transported to source leaves to prevent severe declines in photosynthetic activity. In addition, Mg is involved in the source-to-sink transport of carbohydrates. Hence, an inverse relationship between Mg shortage and sugar accumulation in leaves is often observed. We hereby review all these aspects with a special emphasis on the role of Mg in photosynthesis and the structural and functional effects of its deficiency on the photosynthetic apparatus.

The phylogenetic tree gathering the plant Zn/Cd/Pb/Co P1B-ATPases appears to be structured according to the botanical families.

Plant Zn/Cd/Pb/Co P1B-ATPases (HMAs) play different roles, among which are the control of metal transport from the roots to the shoot and/or from the cytoplasm into the cell vacuole. Transferring the knowledge acquired on HMAs from model species to HMAs from other species requires one to identify orthologues in these other species. Through an extensive screening of the public sequence databases, 96 plant P1B-ATPases showing orthology to any of the AtHMA1, AtHMA2, AtHMA3 or AtHMA4 isoforms were identified from 32 plant species belonging to 15 botanical families. The number of paralogues within a species varied greatly from species to species, even within a specific botanical family, suggesting that gene duplication events occurred after speciation. The phylogenetic tree gathering the Zn/Cd/Pb/Co P1B-ATPases was strongly structured according to the botanical family to which the sequences could be related to. In particular, no strict orthology relationship links the Brassicaceae HMAs to the non-Brassicaceae or the Poaceae ones. Recent data showed that the sole rice HMA characterised to date displays different functional properties from the Arabidopsis HMAs. Altogether, data suggest that it might be risky to directly transfer the knowledge acquired through the study of HMAs in model plant species to HMAs from other species.

Differential performance of two forage species, Medicago truncatula and Sulla carnosa, under water-deficit stress and recovery

Abstract. The response patterns during water deficit stress and subsequent recovery of two forage species, Medicago truncatula and Sulla carnosa, were studied. After germination and pre-treatment, seedlings were individually cultivated for two months under two irrigation modes: 100% and 33% of field capacity. Measured parameters were plant growth, water relations, leaf osmotic potential, lipid peroxidation, and leaf inorganic (Na+ and K+) and organic (proline and soluble sugars) solute contents, as well as delta-1-pyrroline-5-carboxylate synthase (P5CS) and proline dehydrogenase (PDH) activities. Our results showed that under control conditions, and in contrast to roots, no significant differences were observed in shoot biomass production between the two species. However, when subjected to water-deficit stress, M. truncatula appeared to be more tolerant than S. carnosa (reduction by 50 and 70%, respectively). In the two studied species, water-deficit stress led to an increase in root/shoot ratio and leaf proline and soluble sugar contents, and a decrease in leaf osmotic potential. Enzymatic assay revealed that in the two species, P5CS activity was stimulated whereas that of PDH was inhibited under stress conditions. Despite greater accumulation of proline, sugar, and potassium in leaves of S. carnosa, M. truncatula was more tolerant to water deficit. This was essentially due to its capacity to control tissue hydration and water-use efficiency, in addition to its greater ability to protect membrane integrity. Following stress relief, M. truncatula and S. carnosa showed partial re-establishment of growth capacity.

Nutrient uptake and use efficiencies in Medicago ciliaris under salinity

ABSTRACT Salt-induced responses of Medicago ciliaris was studied under controlled conditions. Twenty-two-day old seedlings were cultivated for one month in a nutrient medium added or not with 75 mM sodium chloride (NaCl). Our results showed that this species is relatively salt-tolerant since the whole biomass production of salt-treated plants was affected a little (−30%) as compared to the control. The slight salt effect was mainly nutritional and concerned both macro potassium, calcium and magnesium (K, Ca, and Mg) and micro-nutrients iron (Fe). K and Fe uptake efficiencies were more affected than those of Ca and Mg. Nevertheless, M. ciliaris was able to counterbalance this impact by increasing both K and Fe use efficiencies. The enhancement of K use efficiency could be due in part to the plant aptitude to accumulate sodium (Na+) ions within its shoot tissues and to use them for osmotic adjustment. This “includer” behavior allowed M. ciliaris to maintain an adequate water status under saline conditions.

Photosynthetic behaviour of Arabidopsis thaliana (Pa-1 accession) under salt stress

The growth reduction observed in many plants caused by salinity is often associated with a decrease in their photosynthetic capacity. This effect could be associated with the partial stomatal closure and/or the non-stomatal limitation which involves the decrease in ribulose-1,5-bisphosphate carboxylase oxygenase (RUBISCO) activity. The objective of this study was to explore the mechanisms of inhibited photosynthesis in Arabidopsis thaliana ( Pa-1 accession) under salt stress. Pa-1 seeds grown on a solid substrate for 25 days on standard medium were challenged with 50 mM NaCl for 15 days. Harvests were carried out every five days by separating the rosette leaves and roots. Salt stress reduced growth by limiting the number of the rosette leaves and not their biomass. Accumulation of Na+ and Cl- increased during the treatment period, whereas K+ and Ca2+ accumulation were reduced in salt treatment. RUBISCO and phosphoenolpyruvate carboxylase (PEPC) activities were increased with the age of the leaves to a maximum after 10 days of treatment then later decreased. We concluded that the sensitivity of Pa-1 to salinity may be due to a reduction in number of leaves, in the photosynthetic assimilation with stomatal closure and damage of the RUBISCO and PEPC activities. Keywords : Arabidopsis thaliana , photosynthetic parameters, salinity, ribulose-1,5-bisphosphate carboxylase oxygenase (RUBISCO), phosphoenolpyruvate carboxylase (PEPC). African Journal of Biotechnology Vol. 12(29), pp. 4594-4602

The grapevine VvWRKY2 gene enhances salt and osmotic stress tolerance in transgenic Nicotiana tabacum

Our study aims to assess the implication of WRKY transcription factor in the molecular mechanisms of grapevine adaptation to salt and water stresses. In this respect, a full-length VvWRKY2 cDNA, isolated from a Vitis vinifera grape berry cDNA library, was constitutively over-expressed in Nicotiana tabacum seedlings. Our results showed that transgenic tobacco plants exhibited higher seed germination rates and better growth, under both salt and osmotic stress treatments, when compared to wild type plants. Furthermore, our analyses demonstrated that, under stress conditions, transgenic plants accumulated more osmolytes, such as soluble sugars and free proline, while no changes were observed regarding electrolyte leakage, H2O2, and malondialdehyde contents. The improvement of osmotic adjustment may be an important mechanism underlying the role of VvWRKY2 in promoting tolerance and adaptation to abiotic stresses. Principal component analysis of our results highlighted a clear partition of plant response to stress. On the other hand, we observed a significant adaptation behaviour response for transgenic lines under stress. Taken together, all our findings suggest that over-expression of VvWRKY2 gene has a compelling role in abiotic stress tolerance and, therefore, would provide a useful strategy to promote abiotic stress tolerance in grape via molecular-assisted breeding and/or new biotechnology tools.