Department of Biotechnology, Deenbandhu Chhotu Ram University of Science and Technology, Murthal, Sonepat, 131 039 Haryana India ; Department of Biotechnology and Molecular Biology, College of Basic Sciences and Humanities, CCS Haryana Agricultural University, Hisar, 125 004 Haryana India.
Indian J Microbiol. 2010 Mar;50(1):27-32. doi: 10.1007/s12088-009-0011-3. Epub 2009 Mar 5.
Seventy isolates of Bacillus thuringiensis were isolated from soil samples collected from cotton fields. These isolates were characterized by randomly amplified poylmorphic DNA (RAPD) markers to determine their genetic diversity pattern based on their source of origin. Different random decamer primers were used for RAPD amplification, which generated a total of 1935 fragments; of these 1865 were polymorphic and 68 monomorphic. The primers OPA03, OPA08, OPD14, OPD19, OPD20, OPE17 and OPD19 produced 100% polymorphic fragments, whereas primers OPC06, OPC20 and OPD17 produced 20, 31 and 17 monomorphic fragments, respectively. When the RAPD banding pattern data was subjected to dendrogram construction, the 70 isolates fell into two separate clusters, cluster I and cluster II, which includes 26 and 44 B. thuringiensis isolates, respectively. These two main clusters were further divided into four subclusters at Eucledian distance of 150 and 80% similarity index. All primers showed amplification and indicated the good diversity of B. thuringiensis isolates. The RAPD pattern showed 4-10 bands per isolate, with MWt in the range of 0.4-3.5 Kb and an average of 193.5 fragments were produced per primer. The primer OPE17 was found to be the most discriminatory as it produced 286 polymorphic bands.
从棉田土壤样本中分离出 70 株苏云金芽孢杆菌。这些分离株通过随机扩增多态性 DNA（RAPD）标记进行特征分析，以确定其遗传多样性模式，这些标记基于其来源。使用不同的随机十聚体引物进行 RAPD 扩增，共产生了 1935 个片段；其中 1865 个是多态性的，68 个是单态性的。引物 OPA03、OPA08、OPD14、OPD19、OPD20、OPE17 和 OPD19 产生了 100%的多态性片段，而引物 OPC06、OPC20 和 OPD17 分别产生了 20、31 和 17 个单态性片段。当 RAPD 带型数据用于构建聚类树时，70 个分离株分为两个单独的聚类，聚类 I 和聚类 II，分别包含 26 和 44 个 B. thuringiensis 分离株。这两个主要聚类进一步在欧几里得距离为 150 和 80%相似性指数处分为四个亚聚类。所有引物均显示出扩增效果，表明 B. thuringiensis 分离株具有良好的多样性。RAPD 模式显示每个分离株有 4-10 条带，MWt 范围为 0.4-3.5 Kb，每个引物平均产生 193.5 个片段。引物 OPE17 是最具区分性的，因为它产生了 286 个多态性带。
