Division of Plant Pathology, ICAR-Indian Agricultural Research Institute, New Delhi, India.
Division of Agricultural Chemicals, ICAR-Indian Agricultural Research Institute, New Delhi, India.
PLoS One. 2022 Nov 15;17(11):e0277124. doi: 10.1371/journal.pone.0277124. eCollection 2022.
A desirable substitute for chemical pesticides is mycopesticides. In the current investigation, rDNA-ITS (Internal transcribed spacer) and TEF (Transcriptional Elongation Factor) sequencing were used for molecular identification of six Beauveria bassiana strains. Both, leaf discs and potted plant bioassaye were carried out to study their pathogenicity against the cassava mite, Tetranychus truncatus. LC50 and LC90 values of potential B. bassiana strains were estimated. We also discovered a correlation between intraspecific B. bassiana strains pathogenicity and comprehensive metabolome profiles. Bb5, Bb6, Bb8, Bb12, Bb15, and Bb21 strains were identified as B. bassiana by sequencing of rDNA-ITS and TEF segments and sequence comparison to NCBI (National Center for Biotechnology Information) GenBank. Out of the six strains tested for pathogenicity, Bb6, Bb12, and Bb15 strains outperformed against T. truncatus with LC50 values 1.4×106, 1.7×106, and 1.4×106 and with a LC90 values 7.3×107, 1.4×108, and 4.2×108 conidia/ml, respectively, at 3 days after inoculation and were considered as potential strains for effective mite control. Later, Gas Chromatography-Mass Spectrometry (GC-MS) analysis of the above six B. bassiana strains was done on secondary metabolites extracted with ethyl acetate revealed that the potential B. bassiana strains (Bb6, Bb12, and Bb15) have higher levels of acaricidal such as Bis(dimethylethyl)-phenol: Bb6 (5.79%), Bb12 (6.15%), and Bb15 (4.69%). Besides, insecticidal (n-Hexadecanoic acid), and insect innate immunity overcoming compound (Nonadecene) were also identified; therefore, the synergistic effect of these compounds might lead toa higher pathogenicity of B. bassiana against T. truncatus. Further, these compounds also exhibited two clusters, which separate the potential and non-potential strains in the dendrogram of Thin Layer Chromatography. These results clearly demonstrated the potentiality of the B. bassiana strains against T. truncatus due to the occurrence of their bioactive volatile metabolome.
一种理想的化学农药替代品是真菌杀虫剂。在本研究中,使用 rDNA-ITS(内部转录间隔区)和 TEF(转录延伸因子)测序对六种球孢白僵菌菌株进行了分子鉴定。通过叶圆盘和盆栽植物生物测定法研究了它们对木薯螨虫(Tetranychus truncatus)的致病性。估计了潜在的球孢白僵菌菌株的 LC50 和 LC90 值。我们还发现了种内球孢白僵菌菌株致病性与综合代谢组谱之间的相关性。通过 rDNA-ITS 和 TEF 片段的测序以及与 NCBI(美国国家生物技术信息中心)GenBank 的序列比较,Bb5、Bb6、Bb8、Bb12、Bb15 和 Bb21 菌株被鉴定为球孢白僵菌。在测试致病性的六种菌株中,Bb6、Bb12 和 Bb15 菌株对 T. truncatus 的表现优于其他菌株,LC50 值分别为 1.4×106、1.7×106 和 1.4×106,LC90 值分别为 7.3×107、1.4×108 和 4.2×108 个孢子/ml,分别在接种后 3 天。这些菌株被认为是有效控制螨虫的潜在菌株。随后,对上述六种球孢白僵菌菌株进行了气相色谱-质谱(GC-MS)分析,用乙酸乙酯提取的次生代谢产物表明,潜在的球孢白僵菌菌株(Bb6、Bb12 和 Bb15)具有更高水平的杀螨活性,如双(二甲乙基)-苯酚:Bb6(5.79%)、Bb12(6.15%)和 Bb15(4.69%)。此外,还鉴定出杀虫(正十六烷酸)和昆虫先天免疫克服化合物(正十九烯);因此,这些化合物的协同作用可能导致球孢白僵菌对 T. truncatus 的更高致病性。此外,这些化合物在薄层色谱的系统发育树中也显示出两个聚类,将潜在菌株和非潜在菌株分开。这些结果清楚地表明,由于其生物活性挥发代谢物的存在,球孢白僵菌菌株对 T. truncatus 具有潜力。