Molecular characterization of Brevibacillus laterosporus and its potential use in biological control
Molecular characterization of Brevibacillus laterosporus and its potential use in biological control
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DOI：
10.1128/aem.70.11.6657-6664.2004
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发表时间：
2004-11-01
影响因子：
4.4
通讯作者：
Zahner, V
中科院分区：
文献类型：
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作者：
de Oliveira, EJ;Rabinovitch, L;Zahner, V
33株侧孢短芽孢杆菌，包括3个新的菌株分离自巴西土壤样品，进行了研究，通过使用不同的PCR为基础的方法的遗传变异。研究了可以表征细菌菌株致病潜力的分子标记物。此外，还使用鳞翅目和鞘翅目昆虫以及软体动物光滑双脐螺评估了毒性。在所测试的靶标中，光滑双脐螺对B表现出最高程度的敏感性。侧孢型，一些菌株诱导90至100%的死亡率在蜗牛年龄3和12天后关闭。鞘翅目Anthonomus grandis的幼虫也是敏感的，呈现33和63%之间的死亡率水平。还注意到对鳞翅目黎豆夜蛾的毒性。相反，黄粉虫或草地贪夜蛾的测试种群中没有死亡率记录。应用基因间转录间隔区PCR和BOX-PCR分别产生15和17个不同的基因型。没有一种分子技术可以鉴定出与任何昆虫病原表型相关的方便标记。然而，所有B菌株均检测到1，078-bp扩增子。当使用用于扩增BOXA 1 R区域的引物时，类似地，通过使用引物OPA-11从所有分离株产生900-bp扩增子用于随机扩增多态性DNA分析。在其他表型相关的短芽孢杆菌属物种中未检测到这些扩增子，表明它们代表了对B具有特异性的标记。laterosporus，这可能证明有用的分离和鉴定该物种的新菌株。
Thirty-three strains of Brevibacillus laterosporus, including three novel strains isolated from Brazilian soil samples, were examined for genetic variability by the use of different PCR-based methods. Molecular markers that could characterize bacterial strains with regards to their pathogenic potential were investigated. In addition, toxicity was assessed by the use of insects belonging to the orders Lepidoptera and Coleoptera and the mollusk Biomphalaria glabrata. Among the targets tested, Biomphalaria glabrata demonstrated the highest degree of sensitivity to B. laterosporus, with some strains inducing 90 to 100% mortality in snails aged 3 and 12 days posteclosion. Larvae of the coleopteron Anthonomus grandis were also susceptible, presenting mortality levels of between 33 and 63%. Toxicity was also noted towards the lepidopteron Anticarsia gemmatalis. In contrast, no mortality was recorded among test populations of Tenebrio molitor or Spodoptera frugiperda. The application of intergenic transcribed spacer PCR and BOX-PCR generated 15 and 17 different genotypes, respectively. None of the molecular techniques allowed the identification of a convenient marker that was associated with any entomopathogenic phenotype. However, a 1,078-bp amplicon was detected for all strains of B. laterosporus when a primer for amplification of the BOXA1R region was used. Similarly, a 900-bp amplicon was generated from all isolates by use of the primer OPA-11 for randomly amplified polymorphic DNA analysis. These amplicons were not detected for other phenotypically related Brevibacillus species, indicating that they represent markers that are specific for B. laterosporus, which may prove useful for the isolation and identification of new strains of this species.