Nathalie Verbruggen

Proline accumulation in plants: a review

Proline (Pro) accumulation is a common physiological response in many plants in response to a wide range of biotic and abiotic stresses. Controversy has surrounded the possible role(s) of proline accumulation. In this review, knowledge on the regulation of Pro metabolism during development and stress, results of genetic manipulation of Pro metabolism and current debate on Pro toxicity in plants are presented.

Molecular mechanisms of metal hyperaccumulation in plants

Metal hyperaccumulator plants accumulate and detoxify extraordinarily high concentrations of metal ions in their shoots. Metal hyperaccumulation is a fascinating phenomenon, which has interested scientists for over a century. Hyperaccumulators constitute an exceptional biological material for understanding mechanisms regulating plant metal homeostasis as well as plant adaptation to extreme metallic environments.Our understanding of metal hyperaccumulation physiology has recently increased as a result of the development of molecular tools. This review presents key aspects of our current understanding of plant metal – in particular cadmium (Cd),nickel (Ni) and zinc (Zn) – hyperaccumulation.

Small heat shock proteins and stress tolerance in plants.

Small heat shock proteins (sHsps) are produced ubiquitously in prokaryotic and eukaryotic cells upon heat. The special importance of sHsps in plants is suggested by unusual abundance and diversity. Six classes of sHsps have been identified in plants based on their intracellular localization and sequence relatedness. In addition to heat stress, plant sHsps are also produced under other stress conditions and at certain developmental stages. Induction of sHsp gene expression and protein accumulation upon environmental stresses point to the hypothesis that these proteins play an important role in stress tolerance. The function of sHsps as molecular chaperones is supported by in vitro and in vivo assays. This review summarizes recent knowledge about plant sHsp gene expression, protein structure and functions.

Mechanisms to cope with arsenic or cadmium excess in plants.

The metalloid arsenic and the heavy metal cadmium have no demonstrated biological function in plants. Both elements are highly toxic and of major concern with respect to their accumulation in soils, in the food-chain or in drinking water. Arsenate is taken up by phosphate transporters and rapidly reduced to arsenite, As(III). In reducing environments, As(III) is taken up by aquaporin nodulin 26-like intrinsic proteins. Cd(2+) enters the root via essential metal uptake systems. As(III) and Cd(2+) share some similarity between their toxicology and sequestration machineries. Recent progress in understanding the mechanisms of As and Cd uptake and detoxification is presented, including the elucidation of why rice takes up so much arsenic from soil and of mechanisms of As and Cd hypertolerance.

Plant science: the key to preventing slow cadmium poisoning

Practically all human populations are environmentally exposed to cadmium (Cd), mostly through plant-derived food. A growing body of epidemiological evidence suggests that there is no margin of safety between current Cd exposure levels and the threshold for adverse health effects and, hence, there is an urgent need to lower human Cd intake. Here we review recent studies on rice (Oryza sativa) and Cd-hyperaccumulating plants that have led to important insights into the processes controlling the passage of Cd from the soil to edible plant organs. The emerging molecular understanding of Cd uptake, root retention, root-to-shoot translocation and grain loading will enable the development of low Cd-accumulating crops.

A Major Quantitative Trait Locus for Cadmium Tolerance in Arabidopsis halleri Colocalizes with HMA4, a Gene Encoding a Heavy Metal ATPase

Cadmium (Cd) tolerance seems to be a constitutive species-level trait in Arabidopsis halleri sp. halleri. Therefore, an interspecific cross was made between A. halleri and its closest nontolerant interfertile relative, Arabidopsis lyrata sp. petraea, and a first-generation backcross population (BC1) was used to map quantitative trait loci (QTL) for Cd tolerance. Three QTL were identified, which explained 43%, 24%, and 16% of the phenotypic variation in the mapping population. Heavy metal transporting ATPases4 (HMA4), encoding a predicted heavy metal ATPase, colocalized with the peak of the major QTL Cdtol-1 and was consequently further studied. HMA4 transcripts levels were higher in the roots and the shoots of A. halleri than in A. lyrata sp. petraea. Furthermore, HMA4 was also more highly expressed in all BC1 genotypes harboring the HMA4 A. halleri allele at the QTL Cdtol-1, independently of the presence of an A. halleri allele at the two other QTL. Overexpression of AhHMA4 in yeast (Saccharomyces cerevisiae) supported a role of HMA4 in zinc (Zn) and Cd transport by reducing the Cd and Zn contents of the yeast cells. In epidermal tobacco (Nicotiana tabacum) cells, AhHMA4:green fluorescent protein was clearly localized in the plasma membrane. Taken together, all available data point to the elevated expression of HMA4 P1B-type ATPase as an efficient mechanism for improving Cd/Zn tolerance in plants under conditions of Cd/Zn excess by maintaining low cellular Cd2+ and Zn2+ concentrations in the cytoplasm.

Response to copper excess in Arabidopsis thaliana: Impact on the root system architecture, hormone distribution, lignin accumulation and mineral profile

Growth, in particular reorganization of the root system architecture, mineral homeostasis and root hormone distribution were studied in Arabidopsis thaliana upon copper excess. Five-week-old Arabidopsis plants growing in hydroponics were exposed to different Cu(2+) concentrations (up to 5 muM). Root biomass was more severely inhibited than shoot biomass and Cu was mainly retained in roots. Cu(2+) excess also induced important changes in the ionome. In roots, Mg, Ca, Fe and Zn concentrations increased, whereas K and S decreased. Shoot K, Ca, P, and Mn concentrations decreased upon Cu(2+) exposure. Further, experiments with seedlings vertically grown on agar were carried out to investigate the root architecture changes. Increasing Cu(2+) concentrations (up to 50 muM) reduced the primary root growth and increased the density of short lateral roots. Experiment of split-root system emphasized a local toxicity of Cu(2+) on the root system. Observations of GUS reporter lines suggested changes in auxin and cytokinin accumulations and in mitotic activity within the primary and secondary root tips treated with Cu(2+). At toxic Cu(2+) concentrations (50 muM), these responses were accompanied by higher root apical meristem death. Contrary to previous reports, growth on high Cu(2+) did not induce an ethylene production. Finally lignin deposition was detected in Cu(2+)-treated roots, probably impacting on the translocation of nutrients. The effects on mineral profile, hormonal status, mitotic activity, cell viability and lignin deposition changes on the Cu(2+)-induced reorganization of the root system architecture are discussed.

Genetic basis of Cd tolerance and hyperaccumulation in Arabidopsis halleri

The genetic basis of Cd tolerance and hyperaccumulation was investigated in Arabidopsis halleri. The study was conducted in hydroponic culture with a backcross progeny, derived from a cross between A. halleri and a non-tolerant and non-accumulating related species Arabidopsis lyrata ssp. petraea, as well as with the parents of the backcross. The backcross progeny segregates for both cadmium (Cd) tolerance and accumulation. The results support that (i) Cd tolerance may be governed by more than one major gene, (ii) Cd tolerance and Cd accumulation are independent characters, (iii) Cd and Zn tolerances co-segregate suggesting that they are under pleiotropic genetic control, at least to a certain degree, (iv) the same result was obtained for Cd and Zn accumulation.

Isolation, characterization, and chromosomal location of a gene encoding the Δ1-pyrroline-5-carboxylate synthetase in Arabidopsis thaliana

A full‐length cDNA and the corresponding At‐P5S gene encoding the first enzyme of the proline biosynthetic pathway, the Δ 1‐pyrroline‐5‐carboxylate (P5C) synthetase, were isolated in Arabidopsis thaliana. The At‐P5S cDNA encodes a protein of 717 amino acids showing high identity with the P5C synthetase of Vigna aconitifolia. Strong homology is also found at the N‐terminus to bacterial and yeast γ‐glutamyl kinase and at the C‐terminus to bacterial γ‐glutamyl phosphate reductase. Putative ATP‐ and NAD(P)H‐binding sites are suggested in the At‐P5S protein. The transcribed region of the At‐P5S gene is 4.8 kb long and contains 20 exons. Southern analysis suggests the presence of only one At‐P5S gene in the A. thaliana genome mapped at the bottom of the chromosome two. Expression analysis of At‐P5S in different organs reveals abundant At‐P5S transcripts in mature flowering plant. Rapid induction of the At‐P5S gene followed by accumulation of proline was observed in NaCl‐treated seedlings suggesting that At‐P5S is osmoregulated.

Expression of cell cycle regulatory genes and morphological alterations in response to salt stress in Arabidopsis thaliana

Abstract. Hyperosmotic stress severely affects plant growth and development. To examine the effect of salt stress on cell cycle activity in Arabidopsis thaliana (L.) Heynh., the transcriptional regulation of a cyclin-dependent kinase, CDC2aAt, and two mitotic cyclins, Arath;CycB1;1 and Arath;CycA2;1, was studied by using the β-glucuronidase (gus) reporter gene. Moreover, the mRNA abundance of these cell cycle genes as well as CDC2bAt were monitored during salt stress. Upon NaCl treatment, the promoter activities and transcript levels of all cell cycle genes diminished initially in the shoot apex and were subsequently induced during salt-stress adaptation. Additionally, the promoter activities of CDC2aAt and CycA2;1 decreased in the vascular cylinder of the root in correlation with reduced lateral root formation. In the root tips, a regression of CDC2aAt, CycA2;1, and CycB1;1:gus expression was observed, concomitant with a shrinkage of the root meristem and inhibition of root growth. Our data indicate that salt stress interferes with cell cycle regulation at the transcriptional level, resulting in an adaptive growth response.