Characterization of cry Genes in a Mexican Bacillus thuringiensis Strain Collection
Characterization of cry Genes in a Mexican Bacillus thuringiensis Strain Collection
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DOI：
10.1128/aem.64.12.4965-4972.1998
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发表时间：
1998-12
影响因子：
4.4
通讯作者：
A. Bravo;Sergio Sarabia;L. López;H. Ontiveros;C. Abarca;A. Ortiz;M. Ortiz;L. Lina;F. Villalobos;Guadalupe Peña;M. Nuñez-Valdez;M. Soberón;R. Quintero
中科院分区：
文献类型：
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作者：
A. Bravo;Sergio Sarabia;L. López;H. Ontiveros;C. Abarca;A. Ortiz;M. Ortiz;L. Lina;F. Villalobos;Guadalupe Peña;M. Nuñez-Valdez;M. Soberón;R. Quintero
摘要墨西哥位于新北区和新热带区之间的过渡地带，拥有丰富而独特的生物多样性。从全国5个大区采集的503份土样中分离到496株苏云金芽孢杆菌。菌株收集的特征提供了有用的信息，生态模式的分布B。苏云金芽孢杆菌和选择菌株的机会，以开发新的生物杀虫产品。采用新型通用引物和特异引物进行多重PCR扩增，可检测cry1、cry3、cry5、cry7、cry8、cry9、cry11、cry12、cry13、cry14、cry21和cyt基因。属于Cry1和Cry9组的蛋白质对鳞翅目昆虫是有毒的。Cry3、Cry7和Cry8蛋白对鞘翅目昆虫具有活性。Cry5、Cry12、Cry13和Cry14蛋白是杀线虫的。Cry11、Cry21和Cyt蛋白对双翅目昆虫具有毒性。该方法使用了6对通用引物。使用通用引物获得独特PCR产物谱的菌株通过使用特异性引物的额外PCR进一步表征。含cry1基因的菌株在我们收集的菌株中最丰富（49.5%）。鉴定了33种不同的cry1型谱。B。含cry3基因的菌株占21.5%，含cry11和cyt基因的菌株占7.9%，含cry7、cry8和cry9基因的菌株分别占0.6%、2.4%和2.6%。未发现携带cry5、cry12、cry13、cry14和cry21基因的菌株，14%的菌株不产生任何PCR产物，也不与任何多克隆抗血清反应。我们的研究结果表明，存在可能含有潜在的新型Cry蛋白的菌株，以及具有较少观察到的cry基因组合的菌株。
ABSTRACT Mexico is located in a transition zone between the Nearctic and Neotropical biogeographical regions and contains a rich and unique biodiversity. A total of 496 Bacillus thuringiensis strains were isolated from 503 soil samples collected from the five macroregions of the country. The characterization of the strain collection provided useful information on the ecological patterns of distribution of B. thuringiensis and opportunities for the selection of strains to develop novel bioinsecticidal products. The analysis of the strains was based on multiplex PCR with novel general and specific primers that could detect the cry1,cry3, cry5, cry7, cry8,cry9, cry11, cry12,cry13, cry14, cry21, andcyt genes. The proteins belonging to the Cry1 and Cry9 groups are toxic for lepidopteran insects. The Cry3, Cry7, and Cry8 proteins are active against coleopteran insects. The Cry5, Cry12, Cry13, and Cry14 proteins are nematocidal. The Cry11, Cry21, and Cyt proteins are toxic for dipteran insects. Six pairs of general primers are used in this method. Strains for which unique PCR product profiles were obtained with the general primers were further characterized by additional PCRs with specific primers. Strains containingcry1 genes were the most abundant in our collection (49.5%). Thirty-three different cry1-type profiles were identified. B. thuringiensis strains harboringcry3 genes represented 21.5% of the strains, and 7.9% of the strains contained cry11 and cyt genes.cry7, cry8, and cry9 genes were found in 0.6, 2.4, and 2.6% of the strains, respectively. No strains carrying cry5, cry12, cry13,cry14, or cry21 genes were found. Finally, 14% of the strains did not give any PCR product and did not react with any polyclonal antisera. Our results indicate the presence of strains that may harbor potentially novel Cry proteins as well as strains with combinations of less frequently observed cry genes.