Proteomic and ecophysiological responses of soybean (Glycine max L.) root nodules to Pb and hg stress.

BACKGROUND: Lead (Pb) and mercury (Hg) are persistent hazardous metals in industrially polluted soils which can be toxic in low quantities. Metal toxicity can cause changes at cellular and molecular level which should be studied for better understanding of tolerance mechanism in plants. Soybean (Glycine max L.) is an important oilseed crop of the world including India. Indian soils growing soybean are often contaminated by Pb and Hg. The aim of this study was to explore how soybean root nodule responds to Pb and Hg through proteomic and ecophysiological alterations in order to enhance tolerance to metal stress. RESULTS: Soybean plants were exposed to Pb (30 ppm PbCl2) and Hg (0.5 ppm HgCl2) to study histological, histochemical, biochemical and molecular response of N2-fixing symbiotic nodules. Both Pb and Hg treatment increased the level of oxidative stress in leaves and nodules. Chlorosis in leaves and morphological/anatomical changes in nodules were observed. Activities of ascorbate peroxidase, glutathione reductase and catalase were also modulated. Significant changes were observed in abundance of 76 proteins by Pb and Hg. Pb and Hg influenced abundance of 33 proteins (17 up and 16 down) and 43 proteins (33 up and 10 down), respectively. MS/MS ion search identified 55 proteins which were functionally associated with numerous cellular functions. Six crucial proteins namely catalase (CAT), allene oxide synthase (AOS), glutathione S-transferase (GST), calcineurin B like (CBL), calmodulin like (CML) and rapid alkalinisation factor (RAF) were selected for transcript abundance estimation. The qRT-PCR based real time expression exhibited a positive correlation with proteomics expression except for GST and RAF. CONCLUSION: Soybean root nodule responds to metal stress by increased abundance of defence, development and repair related proteins. An efficient proteomic modulation might lead to metal-induced stress tolerance in N2-fixing nodules. Although concentrations of Pb and Hg used in the study cannot be considered equimolar, yet Hg seems to induce more changes in nodule proteomic profile, and higher damage to both bacteroides and root anatomy.

Impact of Ferrous Sulfate on Thylakoidal Multiprotein Complexes, Metabolism and Defence of Brassica juncea L. under Arsenic Stress.

Forty-day-old Brassica juncea (var. Pusa Jai Kisan) plants were exposed to arsenic (As, 250 µM Na2HAsO4·7H2O) stress. The ameliorative role of ferrous sulfate (2 mM, FeSO4·7H2O, herein FeSO4) was evaluated at 7 days after treatment (7 DAT) and 14 DAT. Whereas, As induced high magnitude oxidative stress, FeSO4 limited it. In general, As decreased the growth and photosynthetic parameters less when in the presence of FeSO4. Furthermore, components of the antioxidant system operated in better coordination with FeSO4. Contents of non-protein thiols and phytochelatins were higher with the supply of FeSO4. Blue-Native polyacrylamide gel electrophoresis revealed an As-induced decrease in almost every multi-protein-pigment complex (MPC), and an increase in PSII subcomplex, LHCII monomers and free proteins. FeSO4 supplication helped in the retention of a better stoichiometry of light-harvesting complexes and stabilized every MPC, including supra-molecular complexes, PSI/PSII core dimer/ATP Synthase, Cytochrome b6/f dimer and LHCII dimer. FeSO4 strengthened the plant defence, perhaps by channelizing iron (Fe) and sulfur (S) to biosynthetic and anabolic pathways. Such metabolism could improve levels of antioxidant enzymes, and the contents of glutathione, and phytochelatins. Important key support might be extended to the chloroplast through better supply of Fe-S clusters. Therefore, our results suggest the importance of both iron and sulfur to combat As-induced stress in the Indian mustard plant at biochemical and molecular levels through enhanced antioxidant potential and proteomic adjustments in the photosynthetic apparatus.