Nazarian Amin, Jahangiri Rosa, Jouzani Gholamreza Salehi, Seifinejad Ali, Soheilivand Saeed, Bagheri Omolbanin, Keshavarzi Mansoureh, Alamisaeid Khalil
Department of Microbial Biotechnology and Biosafety, Agricultural Biotechnology Research Institute of Iran, Mahdasht Road, PO Box 31535-1897, Karaj, Iran.
J Invertebr Pathol. 2009 Oct;102(2):101-9. doi: 10.1016/j.jip.2009.07.009. Epub 2009 Jul 22.
The characterization of the strains containing Coleopteran-specific and also putative novel cry genes in Iranian native Bacillus thuringiensis collection is presented. Characterization was based on PCR analysis using 31 general and specific primers for cry1B, cry1I, cry3A, cry3B, cry3C, cry7A, cry8A, cry8B, cry8C, cry14, cry18, cry26, cry28, cry34 and cry35 genes, protein band patterns as well as their insecticidal activity on Xanthogaleruca luteola Mull. larvae. Forty six isolates (65.7%) contained minimum one Coleopteran-active cry gene. Based on universal primers, strains containing cry18 and cry26 genes were the most abundant and represent 27.1% and 24% of the isolates, respectively, whereas cry14, cry3, cry28, cry34, cry35, cry7, cry8 genes were less abundant, found in 14.2, 12.5, 10, 7, 7 and 5.6% of the strains, respectively. Based on specific primers, isolates containing cry1I were the most abundant (48.5%). Two strains containing Coleopteran-active cry genes showed higher activity against X. luteola larvae than B. thuringiensis subsp. morrisoni pathovar tenebrionis. Thirty isolates, when assayed for cry1C, cry5, cry6, cry8b, cry9, cry10, cry11, cry18, cry24 and cry35 genes, showed unexpected size bands. Cloning and sequencing of the amplicons allowed both the identification of known cry genes and the detection of putative novel cry1C sequences.
本文介绍了对伊朗苏云金芽孢杆菌本土菌株库中含有鞘翅目特异性以及推测的新型cry基因的菌株的特性分析。特性分析基于PCR分析，使用了31种针对cry1B、cry1I、cry3A、cry3B、cry3C、cry7A、cry8A、cry8B、cry8C、cry14、cry18、cry26、cry28、cry34和cry35基因的通用引物和特异性引物，蛋白质条带模式以及它们对黄斑叶甲幼虫的杀虫活性。46株分离株（65.7%）至少含有一个鞘翅目活性cry基因。基于通用引物，含有cry18和cry26基因的菌株最为丰富，分别占分离株的27.1%和24%，而cry14、cry3、cry28、cry34、cry35、cry7、cry8基因含量较少，分别在14.2%、12.5%、10%、7%、7%和5.6%的菌株中发现。基于特异性引物，含有cry1I的分离株最为丰富（48.5%）。两株含有鞘翅目活性cry基因的菌株对黄斑叶甲幼虫的活性高于苏云金芽孢杆菌莫里森亚种致黑变种。对30株分离株检测cry1C、cry5、cry6、cry8b、cry9、cry10、cry11、cry18、cry24和cry35基因时，显示出意外大小的条带。扩增子的克隆和测序既能够鉴定已知的cry基因，也能够检测推测的新型cry1C序列。
