Punia Himani, Tokas Jayanti, Bhadu Surina, Mohanty Ashok K, Rawat Preeti, Malik Anurag
Department of Biochemistry, College of Basic Sciences & Humanities, CCS Haryana Agricultural University, Hisar, Haryana 125 004 India.
Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana 132001 India.
3 Biotech. 2020 Sep;10(9):412. doi: 10.1007/s13205-020-02392-1. Epub 2020 Aug 29.
Sorghum is a C cereal grain crop which is well adapted to harsh environment. It is a potential model for gaining better understanding of the molecular mechanism due to its wider adaptability to abiotic stresses. In this study, protein extraction was standardized using different methods to study the electrophoretic pattern of sorghum leaves under different salinity levels. The extraction of soluble protein with lysis buffer, followed by its clean-up was found to be the most effective method. The different profiles of salt-responsive proteins were analyzed in G-46 and CSV 44F sorghum genotypes based on their tolerance behavior towards salinity. The kafirin level also changed depending upon the concentration and exposure time to salts suggesting the stored proteins as energy source under stress conditions. The relative expression of salt-responsive genes was studied using Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) which might be used as a molecular screening tool for identification of salt-tolerant genotypes in affected areas. The validated responses were examined in terms of metabolic changes and the expression of stress-induced proteins-viz. heat shock proteins () via immunoblotting assay. The results showed that the two sorghum genotypes adopted distinct approaches in response to salinity, with G-46 performing better in terms of leaf function. Also, we have standardized different protein extraction methods followed by their clean-up for electrophoretic profiling.
高粱是一种C4谷物作物，能很好地适应恶劣环境。由于其对非生物胁迫具有更广泛的适应性，它是深入了解分子机制的潜在模型。在本研究中，使用不同方法对蛋白质提取进行了标准化，以研究不同盐度水平下高粱叶片的电泳图谱。发现用裂解缓冲液提取可溶性蛋白质并随后进行纯化是最有效的方法。基于G-46和CSV 44F高粱基因型对盐度的耐受行为，分析了盐响应蛋白的不同谱型。醇溶蛋白水平也随盐浓度和暴露时间而变化，这表明在胁迫条件下储存的蛋白质作为能量来源。使用逆转录聚合酶链反应（RT-PCR）研究了盐响应基因的相对表达，该方法可作为在受影响地区鉴定耐盐基因型的分子筛选工具。通过免疫印迹分析，从代谢变化和应激诱导蛋白（即热休克蛋白）的表达方面对验证的反应进行了检查。结果表明，两种高粱基因型对盐度的反应采用了不同的方法，G-46在叶片功能方面表现更好。此外，我们已经对不同的蛋白质提取方法及其纯化进行了标准化，用于电泳分析。
