Department of Molecular Biology and Genetic Engineering, College of Basic Sciences & Humanities, G.B. Pant University of Agriculture and Technology, Pantnagar, 263 145, India,
Mol Biol Rep. 2012 Apr;39(4):4949-60. doi: 10.1007/s11033-011-1291-3. Epub 2011 Dec 14.
Fifty-two genotypes of Eleusine coracana collected from Uttarakhand hills were subjected to simple sequence repeat (SSR), random amplified polymorphic DNA (RAPD)-PCR and protein profiling analysis to investigate the variation in protein content. The main objective of the present study was to detect variability among E. coracana and also assess the discriminating ability of these three molecular methods. A total of 21 RAPD and 24 SSR primers were assayed for their specificity in detecting genetic variability in E. coracana, of which 20 RAPD and 21 SSR primers were highly reproducible and were found suitable for use in PCR analysis. Assessing genetic diversity among E. coracana genotypes by RAPD-PCR using 20 polymorphic primers yielded 56 different RAPD markers which clustered the genotypes into different groups on the basis of protein content. Similarly, SSR-PCR with 21 polymorphic primers clustered the genotypes into different groups. On the other hand, biochemical typing of E. coracana using whole seed proteins generated profiles that showed no major difference indicating the technique to be not useful in typing genotypes of this crop. However, a few of the genotypes showed the presence of a unique band of 32 kDa that needs to be further investigated to understand the role of the protein from nutritional point of view, if any. In the present study, significant negative correlation (r = -0.69*) was found between the protein and calcium content of finger millet genotypes. Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis based seed storage proteins generated profiles showed no major differences in banding pattern among 52 finger millet genotypes while quantitative estimation of seed storage protein fractions using Lowry method revealed that glutelin was highest followed by prolamin, globulin and albumin.
从北阿坎德邦山区采集的 52 种珍珠粟基因型进行简单序列重复（SSR）、随机扩增多态性 DNA（RAPD）-PCR 和蛋白质谱分析，以研究蛋白质含量的变化。本研究的主要目的是检测珍珠粟之间的变异性，并评估这三种分子方法的鉴别能力。总共测试了 21 个 RAPD 和 24 个 SSR 引物，以检测其在检测珍珠粟遗传变异性方面的特异性，其中 20 个 RAPD 和 21 个 SSR 引物具有高度的重现性，适合用于 PCR 分析。使用 20 个多态性引物的 RAPD-PCR 评估珍珠粟基因型之间的遗传多样性，产生了 56 个不同的 RAPD 标记，根据蛋白质含量将基因型聚类为不同的组。同样，使用 21 个多态性引物的 SSR-PCR 将基因型聚类为不同的组。另一方面，使用整个种子蛋白质对珍珠粟进行生化分型生成的图谱没有显示出主要差异，表明该技术在对该作物的基因型进行分型方面没有用。然而，一些基因型显示出存在 32 kDa 的独特带，需要进一步研究，以了解从营养角度来看该蛋白质的作用，如果有的话。在本研究中，发现手指小米基因型的蛋白质和钙含量之间存在显著负相关（r=-0.69*）。基于十二烷基硫酸钠聚丙烯酰胺凝胶电泳的种子贮藏蛋白生成的图谱在 52 个手指小米基因型之间没有显示出主要的带型差异，而使用 Lowry 法对种子贮藏蛋白分数进行定量估计显示，谷蛋白最高，其次是醇溶蛋白、球蛋白和白蛋白。
