Patel G.K.,Indian Institute of Technology Roorkee |
Patel G.K.,Graphic Era University |
Gupta A.K.,CSIR - Central Electrochemical Research Institute |
Gupta A.,Indian Institute of Technology Roorkee |
And 4 more authors.
Protein and Peptide Letters | Year: 2014
A thermotolerant protein with trypsin inhibitory activity designated as CaTI was purified to homogeneity from seeds of Cassia absus. Gel filtration chromatography and SDS-PAGE analysis showed the apparent molecular mass of ∼20 kDa. Partial internal sequences indicate that CaTI belongs to Kunitz-inhibitor family. CaTI inhibits the bovine trypsin in 1:1 molar ratio and exhibited a competitive-type inhibitory activity with Ki = 5.6 × 10-9 M. The inhibitory activity was retained over a broad pH range (2-12). Thermal stability study showed that it is stable up to 80 °C and inhibition activity reduced at and above 90 °C which might be due to the presence of predominantly β-sheets revealed by the CD study. The proteolysis studies of CaTI exhibited strong resistance to proteolysis by different proteases tested. The studies show that CaTI can be used as potential candidates for the development of the transgenic plant against the microbes and insect pests. © 2014 Bentham Science Publishers.
Koul B.,CSIRNational Botanical Research Institute |
Yadav R.,CSIRNational Botanical Research Institute |
Sanyal I.,CSIRNational Botanical Research Institute |
Sawant S.,CSIRNational Botanical Research Institute |
And 2 more authors.
Plant Physiology and Biochemistry | Year: 2012
Efficacy of artificial synthetic expression modules was compared with native CaMV35S and DECaMV35S promoter in transgenic tomato developed by Agrobacterium-mediated transformation. The promoters under trial were CaMV35S-mec (PcamI), CaMV35S (PcamII), DECaMV35S (PcamIII), synthetic minimal expression cassette (Pmec), complete expression cassette (Pcec), double enhancer expression cassette (Pdec) and triple enhancer expression cassette (Ptec) for driving the uidA gene for β-glucuronidase (GUS) activity. The promoter efficiency based on average of GUS expression in T0 and T1 transgenic tomato was in the order Pcec > Pdec > PcamIII > PcamII > PcamI > Ptec > Pmec. The two promoters Pcec and PcamIII were deployed for development of insect-resistant transgenic tomato with optimal expression of modified cry1Ac insecticidal toxin gene from Bacillus thuringiensis (Bt). The transgenic status and copy number of the cry1Ac in T0 transgenic tomato was confirmed through PCR, Southern hybridization, RT-PCR and Western immunoassay, while toxin expression was monitored by DAS-ELISA. The expression level of Cry1Ac toxin driven by Pcec in T0 population ranged from 0.08 to 0.8% of total soluble protein (TSP) that was significantly higher to PcamIII which ranged from 0.02 to 0.13% of TSP. The outcome of insect mortality bioassay with Helicoverpa armigera correlated well with the results of DAS-ELISA. The higher expression of cry1Ac gene driven by Pcec promoter in transgenic tomato did not show any yield penalty and reflected complete protection, while low recovery of promising transgenics expressing Cry1Ac toxin driven by PcamIII was a major limitation for complete protection against the fruit borer insect. © 2012 Elsevier Masson SAS.
Pandey V.C.,CSIRNational Botanical Research Institute |
Sahu N.,CSIRNational Botanical Research Institute |
Behera S.K.,CSIRNational Botanical Research Institute |
Singh N.,CSIRNational Botanical Research Institute
Ecological Engineering | Year: 2016
The aim of the present study was to measure in-situ fly ash (FA) CO2 flux from naturally vegetated and non-vegetated sites of FA dumps for identifying potential plant species for carbon sequestration by using an automated soil CO2 flux system. The FA CO2 flux was found to be higher in vegetated site than non-vegetated site due to higher root density and respiration. The presence of organic carbon, microbial activity and root biomass are important indicators for sequestration of atmospheric CO2 in naturally vegetated site of FA dumps because of the fresh FA dumps are supposed to be initially free of organic carbon. Furthermore, in the naturally vegetated site, the FA CO2 efflux rates were least in Saccharum spontaneum (lower by 84.29%) and Prosopis juliflora (lower by 92.09%) association as compared to Typha latifolia association. Thus, the field results proved that S. spontaneum and P. juliflora associations are potentially suitable for sequestering atmospheric CO2 in the fresh FA deposited sites. © 2016 Elsevier B.V.
Singh R.,CSIRNational Botanical Research Institute |
Pandey N.,CSIRNational Botanical Research Institute |
Kumar A.,CSIRNational Botanical Research Institute |
Shirke P.A.,CSIRNational Botanical Research Institute
Protoplasma | Year: 2015
Root growth in drying soil is generally limited by a combination of mechanical impedance and water stress. As the major function of root tissue is water and nutrient uptake, so it imparts an important role in plant growth and stress management. Previously, we have studied physiological performance and expression profiling of gene associated with drought tolerance in leaf tissue of four cotton varieties. Here, we have further continued our studies with the root tissue of these varieties. The Gossypium hirsutum species JKC-770 is drought-tolerant and KC-2 is drought-sensitive, while Gossypium herbaceum species JKC-717 is drought-tolerant and RAHS-187 is drought-sensitive. JKC-770 and JKC-717 the drought-tolerant varieties showed a comparatively high glutathione-S-transferase, superoxide dismutase, proline along with their gene expression, and low malondialdehyde content indicating low membrane damage and better antioxidative defense under drought condition. The expression levels of cellulose synthase, xyloglucan:xyloglucosyl transferase, and glycosyl hydrolases suggest modulation in cell wall structure and partitioning of sugars towards osmoprotectants instead of cell wall biosynthesis in tolerant varieties. Heat shock proteins and serine/threonine protein phosphotases show upregulation under drought condition, which are responsible for temperature tolerance and protein phosphorylation, respectively. These effects many metabolic processes and may be playing a key role in drought tolerance and adaptability of JKC-770 towards drought tolerance. The long-term water use efficiency (WUE) estimated in terms of carbon isotope discrimination (∆13C) in the root tissues showed maximum depletion in the ∆13C values in JKC-770 variety, while minimum in RAHS-187 under drought stress with reference to their respective control, suggesting a high WUE in JKC-770 variety. © 2015 Springer-Verlag Wien