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首页> 外文期刊>Applied Microbiology >Isolation and Characterization of Burkholderia rinojensis sp. nov., a Non-Burkholderia cepacia Complex Soil Bacterium with Insecticidal and Miticidal Activities
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Isolation and Characterization of Burkholderia rinojensis sp. nov., a Non-Burkholderia cepacia Complex Soil Bacterium with Insecticidal and Miticidal Activities

机译：伯克霍尔德菌rinojensis sp。的分离和鉴定。 nov。，具有杀虫和杀螨活性的非伯克霍尔德菌洋葱复合土壤细菌

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Isolate A396, a bacterium isolated from a Japanese soil sample demonstrated strong insecticidal and miticidal activities in laboratory bioassays. The isolate was characterized through biochemical methods, fatty acid methyl ester (FAME) analysis, sequencing of 16S rRNA, multilocus sequence typing and analysis, and DNA-DNA hybridization. FAME analysis matched A396 to Burkholderia cenocepacia , but this result was not confirmed by 16S rRNA or DNA-DNA hybridization. 16S rRNA sequencing indicated closest matches with B. glumae and B. plantarii . DNA-DNA hybridization experiments with B. plantarii , B. glumae , B. multivorans , and B. cenocepacia confirmed the low genetic similarity (11.5 to 37.4%) with known members of the genus. PCR-based screening showed that A396 lacks markers associated with members of the B. cepacia complex. Bioassay results indicated two mechanisms of action: through ingestion and contact. The isolate effectively controlled beet armyworms ( Spodoptera exigua ; BAW) and two-spotted spider mites ( Tetranychus urticae ; TSSM). In diet overlay bioassays with BAW, 1% to 4% (vol/vol) dilution of the whole-cell broth caused 97% to 100% mortality 4 days postexposure, and leaf disc treatment bioassays attained 75% ± 22% mortality 3 days postexposure. Contact bioassays led to 50% larval mortality, as well as discoloration, stunting, and failure to molt. TSSM mortality reached 93% in treated leaf discs. Activity was maintained in cell-free supernatants and after heat treatment (60°C for 2 h), indicating that a secondary metabolite or excreted thermostable enzyme might be responsible for the activity. Based on these results, we describe the novel species Burkholderia rinojensis , a good candidate for the development of a biocontrol product against insect and mite pests.
机译：从日本土壤样品中分离出的细菌分离物A396在实验室生物测定中显示出强大的杀虫和杀螨活性。通过生化方法，脂肪酸甲酯（FAME）分析，16S rRNA测序，多基因座序列分型和分析以及DNA-DNA杂交对分离株进行了表征。 FAME分析将A396匹配到伯克霍尔德菌，但16S rRNA或DNA-DNA杂交未证实该结果。 16S rRNA测序表明与B. glumae和B. plantarii最匹配。用植物芽孢杆菌，小球芽孢杆菌，多孢芽孢杆菌和新芽孢杆菌的DNA-DNA杂交实验证实与已知属的遗传相似性低（11.5％至37.4％）。基于PCR的筛选显示，A396缺乏与洋葱伯克霍尔德菌复合体成员相关的标记。生物测定结果表明了两种作用机制：通过摄入和接触。该分离物有效地控制了甜菜夜蛾（Spodoptera exigua; BAW）和两个斑点的蜘蛛螨（Tetranychus urticae; TSSM）。在使用BAW的饮食覆盖生物测定法中，全细胞肉汤稀释1％至4％（体积/体积）可导致暴露后4​​天的死亡率为97％至100％，而叶盘处理生物测定法在暴露后3天的死亡率达到75％±22％ 。接触式生物测定法可导致50％的幼虫死亡率以及变色，发育迟缓和蜕皮失败。经处理的叶盘TSSM死亡率达到93％。在无细胞的上清液中以及在热处理（60°C 2 h）后均保持活性，这表明次级代谢产物或分泌的热稳定酶可能是造成这种活性的原因。基于这些结果，我们描述了新种Burkholderia rinojensis，它是开发针对昆虫和螨虫的生物防治产品的良好候选者。

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