Chakdar Hillol, Pabbi Sunil
Centre for Conservation and Utilisation of Blue Green Algae (CCUBGA), Division of Microbiology, Indian Agricultural Research Institute (IARI), New Delhi, 110012, India.
National Bureau of Agriculturally Important Microorganisms (NBAIM), Kushmaur, Maunath Bhanjan, UP, 275103, India.
3 Biotech. 2017 Jun;7(2):125. doi: 10.1007/s13205-017-0779-5. Epub 2017 Jun 1.
Nostoc is a diverse genus of filamentous cyanobacteria with tremendous potential for agricultural and industrial applications. Morphometric methods and routine 16S rDNA-based identification undermines the genetic diversity and impedes strain-level differentiation. A comparative study to deduce the discriminatory power of random amplified polymorphic DNA (RAPD) and amplified ribosomal DNA restriction analysis (ARDRA) for analyzing the genetic diversity of 20 Nostoc strains of diverse geographical origin was carried out. The RAPD primer used in the study generated 100% polymorphic profile. HIP TG primer produced the highest number of bands and fragments. Five primers, viz. OPA 08, OPA 11, HIP GC, OPAH 02 and OPF 05 could produce unique bands for 11 strains. Cluster analysis using the RAPD profile showed 12.5-25% similarity among the strains. Following in silico restriction analysis, two restriction enzymes, viz. HaeIII and HinfI were selected for ARDRA. However, clustering based on the restriction pattern showed 22.5-100% similarity. Results of the present study clearly indicate higher resolution of RAPD which can be reliably used for strain-level differentiation of Nostoc strains.
念珠藻属是丝状蓝细菌的一个多样化属，在农业和工业应用方面具有巨大潜力。形态测量方法和基于常规16S rDNA的鉴定方法会破坏遗传多样性并阻碍菌株水平的区分。开展了一项比较研究，以推断随机扩增多态性DNA（RAPD）和扩增核糖体DNA限制性分析（ARDRA）对分析20株不同地理来源的念珠藻菌株遗传多样性的鉴别能力。该研究中使用的RAPD引物产生了100%的多态性图谱。HIP TG引物产生的条带和片段数量最多。5种引物，即OPA 08、OPA 11、HIP GC、OPAH 02和OPF 05可为11株菌株产生独特条带。使用RAPD图谱进行的聚类分析显示，菌株之间的相似性为12.5%至25%。经过电子限制性分析后，选择了两种限制性内切酶，即HaeIII和HinfI用于ARDRA。然而，基于限制性图谱的聚类显示相似性为22.5%至100%。本研究结果清楚地表明，RAPD具有更高的分辨率，可可靠地用于念珠藻菌株的菌株水平区分。
