Archive - Accomplishments

  • Genes of novel and conserved transcription factor (HSF), heat shock proteins (HSP90, HSP70, HSP26, HSP23 and HSP17), antioxidant enzymes (SOD, APX) were cloned from different wheat cultivars and submitted in NCBI Gen Bank.

  • Quantitative real time expression profiling of SOD showed abundance of transcript in thermotolerant wheat cultivar compared to susceptible under heat stress. 2 fold (C-306) and 1.4 fold (PBW343) increase in SOD transcript was observed at seed hardening stage, whereas 2.2 fold (C-306) and 1.8 fold (PBW343) increase in transcript was observed against heat stress of 40°C for 4h.

  • Differential protein profile of thermotolerant (C-306) and susceptible (HD2329) wheat cultivars were carried out using 2DE. Numerous spots (encircled) showed the differentially expressed proteins in C-306 and HD2329 under HS of 42°C for 4h.These spots are prominent in case of C-306.

 

Differential protein profile of thermotolerant and susceptible cultivars of wheat by 2DE

  • Three different gene sequences associated with drought stress (two from Arabidopsis thaliana and one from rice) have been isolated and submitted in Gen Bank.
  • An AP2/ERF transcription factor encoding gene associated with drought stress has been isolated, cloned, sequenced and re-cloned in a binary vector (PBI121). Recombinant plasmid has been mobilized in Agrobacterium and transferred to Arabidopsis.
  • Activities of immobilizing enzymes (protease, invertase and sucrose phosphate synthase) were progressively enhanced in 14 DAT of salt stress.
  • Q-PCR confirmed that the four MIPS genes are both temporally and spatially expressed with maximum transcript levels of MIPS1 in early developing cotyledons.
  • A full length MIPS1 cDNA was amplified from developing seeds (4-6mm) and in silico analysis revealed   an ORF of 1533 nucleotides encoding 510 amino acids with a molecular mass 52 kDa (HM46196.9). The MIPS amplicon was cloned and expressed in the prokaryotic expression vector.
  • Gene silencing constructs (ihp, AS) for down regulating the expression of MIPS gene in developing seeds of soybean were prepared for Agrobacterium mediated transformation into soybean for validating their efficacy.


Transformation vector for down regulating the expression of MIPS gene in developing seeds of soybean
    • Screening of EMS treated and gamma irradiated mutants generated from bulked populations of 34 genotypes revealed only five plants with relatively low phytate levels in the M10 generation.
    • The enzyme lipoxygenase has been isolated and purified from defatted soybean seeds to 105 fold and purified enzyme on SDS - PAGE exhibited single band about 98 kDa. When this purified fraction was run on IEF and improved SDS-PAGE three isozymes of Lox could be separated. Enzyme kinetics is being studied.
    • The volatile compounds generated via lipoxygenase mediated reaction were studied using GC-MS analysis and eleven compounds, predominantly aldehydes, ketones and their alcoholic counterparts were identified; hexanal was the major volatile for linoleic acid whereas linolenic acid releases trans-2-hexenal. A reciprocal relationship was present between PUFA ratios with relative amount of hexanal to trans-2-hexenal ratio.
    • Fourteen lines of soybean varying in storage ability analyzed for two major enzymes in the oxidative pathway namely LOX and HPL along with their total antioxidant capacity, and it was observed that storage attributes of seeds are influenced by LOX pathway enzymes.
    • Gamma radiation treatment resulted in significant enhancement in anthocyanin contents in the dark-colored soybean varieties which led to the reduction in the levels of parameters taken as indices of off-flavour.
    • A decrease in percent solubility and changes in protein profiles were observed after gamma radiation treatment suggesting changes in the composition of oligomeric proteins.


Scanning electron microscopy of seeds of C-306 cultivar of wheat, (a)Starch granule synthesized under normal temperature, (b) Starch granules synthesize in response to heat stress (40°C), (c) Structure of aleuronic membrane under normal temperature and (d) Aleuronic membrane synthesized under heat stress condition (40°C).




Scanning electron microscopy of seeds of PBW343 cultivar of wheat, (a) Starch granule synthesized under normal temperature, (b) Starch granules synthesize in response