Department of Food Science, National Penghu University of Science and Technology, No. 300, Liuhe Rd., Magong City, Penghu County 880, Taiwan.
Department of Food Science and Technology, Hung Kuang University, No. 1018, Sec. 6, Taiwan Boulevard, Shalu District, Taichung City 43302, Taiwan.
Toxins (Basel). 2014 Aug 19;6(8):2453-70. doi: 10.3390/toxins6082453.
The use of Bacillus thuringiensis (Bt) strains with high insecticidal activity is essential for the preparation of bioinsecticide. In this study, for 60 Bt strains isolated in Taiwan, their genotypes and the correlation of some cry genes as well as the expression levels of cry1 genes, with their insecticidal activities against Plutella xylostella, were investigated. Pulsed field gel electrophoresis (PFGE) and random amplified polymorphic DNA (RAPD) results revealed that the genotypes of these Bt strains are highly diversified. Also, a considerable number of the Bt strains isolated in Taiwan were found to have high insecticidal activities. Since strains that showed individual combined patterns of PFGE and RAPD exhibited distinct insecticidal activities against P. xylostella, thus, these genotypes may be useful for the identification of the new Bt strains and those which have been used in bioinsecticides. In addition, although the presence of cry2Aa1 may have a greater effect on the insecticidal activity of Bt strains in bioassay than other cry genes, only high expression level of cry1 genes plays a key role to determine the insecticidal activity of Bt strains. In conclusion, both RAPD and PFGE are effective in the differentiation of Bt strains. The presence of cry2Aa1 and, especially, the expression level of cry1 genes are useful for the prediction of the insecticidal activities of Bt strains against P. xylostella.
利用杀虫活性高的苏云金芽孢杆菌(Bt)菌株对于制备生物杀虫剂至关重要。在这项研究中,对台湾分离的 60 株 Bt 菌株进行了研究,分析了它们的基因型以及某些 cry 基因的相关性,以及它们对小菜蛾的杀虫活性。脉冲场凝胶电泳(PFGE)和随机扩增多态性 DNA(RAPD)结果表明,这些 Bt 菌株的基因型高度多样化。此外,还发现台湾分离的大量 Bt 菌株具有较高的杀虫活性。由于表现出 PFGE 和 RAPD 单独组合模式的菌株对小菜蛾表现出不同的杀虫活性,因此这些基因型可能有助于鉴定新的 Bt 菌株和已用于生物杀虫剂的菌株。此外,尽管 cry2Aa1 的存在可能比其他 cry 基因对生物测定中 Bt 菌株的杀虫活性有更大的影响,但只有 cry1 基因的高表达水平才对决定 Bt 菌株的杀虫活性起关键作用。总之,RAPD 和 PFGE 均可有效区分 Bt 菌株。cry2Aa1 的存在,特别是 cry1 基因的表达水平,可用于预测 Bt 菌株对小菜蛾的杀虫活性。