Analysis of rice plants exposed to a broad range of relatively low and environmentally realistic Cd concentrations showed that the root capacity to retain Cd ions rose from 49 to 79%, corresponding to increases in the external Cd

ZmST1;1, a putative high-affinity sulfate transporter gene expressed in maize (Zea mays) roots, was functionally characterized and its expression patterns were analyzed in roots of plants exposed to different heavy metals (Cd, Zn, and Cu) interfering with thiol metabolism. The ZmST1;1 cDNA was expressed in the yeast (Saccharomyces cerevisiae) sulfate transporter mutant CP154-7A. Kinetic analysis of sulfate uptake isotherm, determined on complemented yeast cells, revealed that ZmST1;1 has a high affinity for sulfate (K m value of 14.6 6 0.4 mM). Cd, Zn, and Cu exposure increased both ZmST1;1 expression and root sulfate uptake capacity. The metal-induced sulfate uptakes were accompanied by deep alterations in both thiol metabolism and levels of compounds such as reduced glutathione (GSH), probably involved as signals in sulfate uptake modulation. Cd and Zn exposure strongly increased the level of nonprotein thiols of the roots, indicating the induction of additional sinks for reduced sulfur, but differently affected root GSH contents that decreased or increased following Cd or Zn stress, respectively. Moreover, during Cd stress a clear relation between the ZmST1;1 mRNA abundance increment and the entity of the GSH decrement was impossible to evince. Conversely, Cu stress did not affect nonprotein thiol levels, but resulted in a deep contraction of GSH pools. Our data suggest that during heavy metal stress sulfate uptake by roots may be controlled by both GSH-dependent or -independent signaling pathways. Finally, some evidence suggesting that root sulfate availability in Cd-stressed plants may limit GSH biosynthesis and thus Cd tolerance are discussed.To minimize the detrimental effects of heavy metal accumulation, plants have evolved detoxification mechanisms, mainly based on chelation and subcellular compartmentalization (Clemens, 2001). The efficiency of these processes might result in the natural heavy metals tolerance and their basic understanding might be crucial for improving plant performances in phytoextraction of heavy metals from polluted soils (Salt et al., 1998;Pilon-Smits, 2005).Chelation of heavy metals is a ubiquitous detoxification strategy described in a wide variety of plants (Zenk, 1996;Clemens, 2001). One of the principal classes of heavy metal chelators known in plants is phytochelatins (PCs), a family of Cys-rich peptides with the general structure (g-Glu-Cys) n -Gly (n 5 2-11). PCs are synthesized nontranslationally from reduced glutathione (GSH) in a transpeptidation reaction catalyzed by the enzyme PC synthase (PCS; Rea et al., 2004). Their synthesis is induced within minutes following exposure to different metals or metalloids; among these, Cd is the strongest inducer, whereas other metals such as Cu, Zn, Pb, and Ni are less effective and require higher external levels for induction (Grill et al., 1987;Maitani et al., 1996). Interestingly, some inducers, such as Zn, do not seem to serve as substrates for chelation (Clemens, 2001;Souza and Rauser, 2003).The mechanism of heavy metal detoxif...

The effect of cadmium (Cd) on high-affinity sulfate transport of maize (Zea mays) roots was studied and related to the changes in the levels of sulfate and nonprotein thiols during Cd-induced phytochelatin (PC) biosynthesis. Ten micromolar CdCl 2 in the nutrient solution induced a 100% increase in sulfate uptake by roots. This was not observed either for potassium or phosphate uptake, suggesting a specific effect of Cd 2ϩ on sulfate transport. The higher sulfate uptake was not dependent on a change in the proton motive force that energizes it. In fact, in Cd-treated plants, the transmembrane electric potential difference of root cortical cells was only slightly more negative than in the controls, the external pH did not change, and the activity of the plasma membrane H ϩ -ATPase did not increase. Kinetics analysis showed that in the range of the high-affinity sulfate transport systems, 10 to 250 m, Cd exposure did not influence the K m value (about 20 m), whereas it doubled the V max value with respect to the control. Northern-blot analysis showed that Cd-induced sulfate uptake was related to a higher level of mRNA encoding for a putative high-affinity sulfate transporter in roots. Cd-induced sulfate uptake was associated to both a decrease in the contents of sulfate and glutathione and synthesis of a large amount of PCs. These results suggest that Cd-induced sulfate uptake depends on a pretranslational regulation of the high-affinity sulfate transporter gene and that this response is necessary for sustaining the higher sulfur demand during PC biosynthesis.

Summary• The changes in endopolygalacturonase (endo-PG) levels and endo-PG expression in nonmelting flesh (NMF) and melting flesh (MF) peach fruits ( Prunus persica ) during softening were studied. The endo-PG gene was analysed to identify polymorphisms exploitable for early marker-assisted selection (MAS) of flesh texture.• The role of endo-PG in softening was assessed by western and northern blotting and by biochemical analyses. Polymorphisms in the endo-PG gene were revealed by reverse transcription-polymerase chain reaction (RT-PCR) and sequencing.• An endo-PG protein was detected in both NMF and MF fruits. The levels of this endo-PG protein were higher and increased with softening in MF fruits, but remained lower and were constant in NMF fruits. The different levels of endo-PG appeared to be caused by the differential expression of an endo-PG gene, whose open-reading frame (ORF) showed five single nucleotide polymorphisms (SNPs) in NMF 'Oro A' compared with MF 'Bolero'. One of these SNPs allowed us to determine the allelic configuration at the melting flesh ( M ) locus and also seemed to be exploitable for early MAS in other NMF/MF phenotypes.• The NMF phenotype does not seem to be caused by a large deletion of the endo-PG gene.

This work presents a simple in vitro system to study physiological, biochemical and molecular changes occurring in a pear callus (Pyrus communis L., cv. Beurré Bosc) grown in close proximity to spatially separated undifferentiated homologous (pear) or heterologous (quince; Cydonia oblonga Mill., East Malling clone C) cells in its neighboring environment. After a 7-day co-culture period, the presence of heterologous cells produced negative effects on the pear callus, whose relative weight increase and adenylate energy charge decreased by 30 and 24%, respectively. Such behavior was associated with a higher O(2) consumption rate (+125%) which did not seem to be coupled to adenosine triphosphate synthesis. Analyses of alternative oxidase and enzymatic activities involved in reactive oxygen species (ROS) detoxification strongly suggested that the higher O(2) consumption rate, measured in the pear callus grown in the heterologous combination, may probably be ascribed to extra-respiratory activities. These, in turn, might contribute to generate metabolic scenarios where ROS-induced oxidative stresses may have the upper hand. The increase in the levels of 2-thiobarbituric acid reactive metabolites, considered as diagnostic indicators of ROS-induced lipid peroxidation, seemed to confirm this hypothesis. Moreover, reverse transcription polymerase chain reaction analysis revealed that the expression levels of a few senescence-associated genes were higher in the pear callus grown in the heterologous combination than in the homologous one. Taken as a whole, physiological and molecular data strongly suggest that undifferentiated cells belonging to a pear graft-incompatible quince clone may induce an early senescence-like status in a closely co-cultured pear callus.