Ground Floor Laboratory, Department of Microbiology and Biotechnology Center, The Maharaja Sayajirao University of Baroda, Vadodara, Gujarat, India.
Appl Biochem Biotechnol. 2013 May;170(1):58-66. doi: 10.1007/s12010-013-0145-y. Epub 2013 Mar 7.
New or more efficient methodologies having different principles are needed, as one method could not be suitable for isolation of organisms from samples of diverse types and from various environments. In present investigation, growth kinetics study revealed a higher germination rate, a higher growth rate, and maximum sporulation of Bacillus thuringiensis (Bt) compared to other Bacillus species. Considering these facts, a simple and efficient enrichment method was devised which allowed propagation of spores and vegetative cells of Bt and thereby increased Bt cell population proportionately. The new enrichment method yielded Bt from 44 out of 58 samples. Contrarily, Bt was isolated only from 16 and 18 samples by sodium acetate selection and dry heat pretreatment methods, respectively. Moreover, the percentages of Bt colonies isolated by the enrichment method were higher comparatively. Vegetative whole cell protein profile analysis indicated isolation of diverse population of Bt from various samples. Bt strains isolated by the enrichment method represented novel serovars and possibly new cry2 gene.
需要新的或更有效的方法，因为一种方法不可能适用于从不同类型和不同环境的样本中分离出生物体。在本研究中，与其他芽孢杆菌属相比，生长动力学研究显示苏云金芽孢杆菌（Bt）具有更高的发芽率、更高的生长速度和最大的孢子形成率。考虑到这些事实，设计了一种简单有效的富集方法，允许 Bt 的孢子和营养细胞繁殖，从而使 Bt 细胞群体按比例增加。新的富集方法从 58 个样本中的 44 个中得到了 Bt。相比之下，通过醋酸钠选择法和干热处理预处理法，分别仅从 16 个和 18 个样本中分离到了 Bt。此外，富集法分离的 Bt 菌落在比例上更高。营养体全细胞蛋白图谱分析表明，从各种样本中分离出了不同种群的 Bt。通过富集法分离出的 Bt 菌株代表了新型血清型，可能还有新的 cry2 基因。
