Expression analysis of bZIP transcription factor encoding genes in response to water deficit stress in rice.

In plants, basic region/leucine zipper motif (bZIP) transcription factors regulate several developmental processes and activate genes in response to biotic and abiotic stresses. Role of stress responsive bZIP transcription factors was studied in paddy in relation to different stages of development and water deficit stress (WDS) in a drought tolerant cultivar N22 and susceptible IR 64. Further, relative water content (RWC), membrane stability index (MSI) and abscisic acid (ABA) content were measured as indices of WDS at different stages of development and levels of stress. Expression of stress responsive bZIP transcription factors was directly correlated to developmental stage and WDS and indirectly to RWC, MSI and ABA content.

Induced defence responses of contrasting bread wheat genotypes under differential salt stress imposition.

Plants, being sessile in nature, have developed mechanisms to cope with high salt concentrations in the soil. In this study, the effects of NaCl (50-200 mM) on expression of high-affinity potassium transporters (HKTs), antioxidant enzymes and their isozyme profiles were investigated in two contrasting bread wheat (Triticum aestivum L.) genotypes viz., HD2329 (salt-sensitive) and Kharchia65 (salt-tolerant). Kharchia65 can successfully grow in salt affected soils, while HD2329 cannot tolerate salt stress. Differential expression studies of two HKT genes (TaHKT2;1.1 and TaHKT2;3.1) revealed their up-regulated expression (-1.5-fold) in the salt-sensitive HD2329 and down-regulated (-5-fold) inducible expression in the salt-tolerant genotype (Kharchia65). Specific activity of antioxidant enzymes, viz. superoxide dismutase (SOD), peroxidase (POX), ascorbate peroxidase (APX), catalase (CAT) and glutathione reductase (GR) was found to be higher in the salt-tolerant genotype. Isozyme profile of two (POX and GR) antioxidant enzymes showed polymorphism between salt-tolerant and salt-sensitive genotypes. A new gene TaHKT2;3.1 was also identified and its expression profile and role in salt stress tolerance in wheat was also studied. Partial sequences of the TaHKT2;1.1 and TaHKT2;3.1 genes from bread wheat were submitted to the EMBL GenBank database. Our findings indicated that defence responses to salt stress were induced differentially in contrasting bread wheat genotypes which provide evidences for functional correlation between salt stress tolerance and differential biochemical and molecular expression patterns in bread wheat.

Role of gamma-oryzanol in drought-tolerant and susceptible cultivars of rice (Oryza sativa L.).

Drought-tolerant cultivars and their phytochemical composition, which has a role in providing drought tolerance are gaining importance. In this study, rice bran oil and semi-purified oryzanol (SPO) obtained from five rice (Oryza sativa L.) cultivars, namely P1401 and PB1 (drought-susceptible) and N22, PNR381 and APO (drought-tolerant) were analyzed for the gamma-oryzanol content, an antioxidant present in considerable amount in the rice bran. The higher level of gamma-oryzanol and its antioxidant activity was observed in drought-tolerant cultivars (N22, PNR381 and APO) as compared to drought-susceptible (PB1 and P1401), suggesting the role of gamma-oryzanol in drought tolerance, as antioxidants are known to play an important role by scavenging free radicals. The total antioxidant activity of gamma-oryzanol might be attributed to 24-methylene cycloartanyl ferulate, a major component of gamma-oryzanol. By enhancing the level of active oryzanol components identified in this study by genetic and molecular means could impart increased drought tolerance.

Characterization and expression of codon optimized soybean phytase gene in E. coli.

Phytic acid, the major storage form of phosphorus in plant seeds is degraded by the phytases to yield inositol and free phosphate, contributing thereby to the improved bioavailability of phytate phosphorus and essential minerals in plant foods and simultaneous reduction in phosphorus pollution of the terrestrial and aquatic ecosystems. As a possible strategy for altering seed phytate levels, the approach involving reduction of phytate content by ectopically expressing endogenous phytase gene during seed development of soybean (Glycine max L. cv. Pusa-20) was attempted in the present study. Semi-quantitative RT-PCR revealed the maximum expression of phytase gene transcripts in germinating cotyledons (approximately 10 days after germinations), compared to other vegetative tissues. A full-length phytase cDNA was amplified from the germinating seedlings by splicing by overlap extension (SOE)-PCR and its sequence analysis revealed an open-reading-frame of 1644 bp, including an N terminal signal peptide of 28 amino acids. Predicted amino acid sequence (547-aa) of molecular mass 62 kDa on alignment with related purple acid phosphatases in other plants shared five conserved domains and seven invariant amino acids involved in coordination of the metals in the binuclear center of purple acid phosphatases. Owing to a large number of E. coli low-usage codons in soybean phytase gene, the modified gene was cloned into a prokaryotic expression vector pET-28a (+) and its expression in E. coli was confirmed by SDS-PAGE and Western blot analysis. Bioassay of the crude expression product in E. coli revealed a functional phytase gene, showing a great potential for developing low phytate transgenic soybean through its seed-specific overexpression in the early stages of seed development.