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筛选和鉴定具有大而高效杀虫活性的苏云金芽孢杆菌 S1/4 菌株。

Screening and identification of a Bacillus thuringiensis strain S1/4 with large and efficient insecticidal activities.

机构信息

Biopesticides Team (LPAP), Centre of Biotechnology of Sfax, University of Sfax, Sfax, Tunisia.

出版信息

J Basic Microbiol. 2013 Jun;53(6):539-48. doi: 10.1002/jobm.201100653. Epub 2012 Aug 23.

Abstract

The bacterium Bacillus thuringiensis was recognized for its entomopathogenic activities related to Cry and Cyt proteins forming the δ-endotoxins and some extracellular activities like the vegetative insecticidal proteins (VIP) and Cry1I. These activities may act specifically against diverse organisms and some of them typically characterize each strain. Here, we screened a set of 212 B. thuringiensis strains to search the higher insecticidal activities. These strains had bipyramidal and cubic crystal morphologies and 30% of them showed PCR amplification of vip3 internal region, from which five isolates (S1/4, S17, S122, S123, and S144) showed plasmid profile variability. These five strains contained the cry1I, cry1Aa and/or cry1Ac, cry1Ab and cry2 genes, and S1/4 harbored in addition the cry1C, vip1, and vip2 genes. They produced from 25 to 46 µg δ-endotoxin per 10(7) spores. Their δ-endotoxins displayed distinct lethal concentrations 50% against either Spodoptera littoralis or Ephestia kuehniella larvae with the lowest one for S1/4, which was also active against Tuta absoluta. Fortunately, the analysis of the culture supernatants revealed that S1/4 had the higher toxicity towards these lepidopteron but it did not show any toxicity against the Tribolium castaneum coleopteran larvae; additionally, S1/4 displayed an antibacterial activity. S1/4 is a good candidate for agricultural pest control, as it is more efficient than the reference strain HD1.

摘要

苏云金芽孢杆菌因其与 Cry 和 Cyt 蛋白形成 δ-内毒素的昆虫病原活性而被认可,同时还具有一些细胞外活性,如植物杀虫蛋白 (VIP) 和 Cry1I。这些活性可能特异性地作用于不同的生物体,其中一些活性通常是每种菌株所特有的。在这里,我们筛选了一组 212 株苏云金芽孢杆菌,以寻找更高的杀虫活性。这些菌株具有双锥形和立方体形晶体形态,其中 30%的菌株显示 VIP3 内部区域的 PCR 扩增,其中 5 个分离株(S1/4、S17、S122、S123 和 S144)显示出质粒谱的可变性。这 5 个菌株含有 cry1I、cry1Aa 和/或 cry1Ac、cry1Ab 和 cry2 基因,S1/4 还含有 cry1C、vip1 和 vip2 基因。它们每 10(7)个孢子产生 25 到 46 µg δ-内毒素。它们的 δ-内毒素对斯氏棉铃虫或地中海实蝇幼虫的致死浓度 50%各不相同,其中对 S1/4 的致死浓度最低,S1/4 对桃小食心虫也具有活性。幸运的是,对培养液上清液的分析表明,S1/4 对这些鳞翅目昆虫具有更高的毒性,但它对鞘翅目步甲科幼虫没有任何毒性;此外,S1/4 还具有抗菌活性。S1/4 是农业害虫防治的一个很好的候选者,因为它比参考菌株 HD1 更有效。

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