The entomopathogenic fungus T011, parasitizing on nymph of Cicada, collected in the coffee garden in Dak Lak Province, Vietnam, was preliminarily morphologically identified as Isaria cicadae, belonged to order Hypocreales and family Clavicipitaceae. To ensure the authenticity of T011, phylogenetic analysis of the concatenated set of multiple genes including ITS, nrLSU, nrSSU, Rpb1, and Tef1 was applied to support the identification. Genomic DNA was isolated from dried sample T011. The PCR assay sequencing was applied to amplify ITS, nrLSU, nrSSU, Rpb1, and Tef1 gene. For phylogenetic analysis, the concatenated data of both target gens were constructed with MEGAX with a 1,000 replicate bootstrap based on the neighbor-joining, maximum likelihood, maximum parsimony method. As the result, the concatenated data containing 62 sequences belonged to order Hypocreales, families Clavicipitaceae, and 2 outgroup sequences belonged to order Hypocreales, genus Verticillium. The phylogenetic analysis results indicated that T011 was accepted at subclade Cordyceps and significantly formed the monophyletic group with referent Cordyceps cicadae (Telemorph of Isaria cicadae) with high bootstrap value. The phylogenetically analyzed result was strongly supported by our morphological analysis described as the Isaria cicadae. In summary, phylogenetic analyses based on the concatenated dataset were successfully applied to strengthen the identification of T011 as Isaria cicadae.

中文翻译：

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首次鉴定越南蝉的分子记录

越南多乐省咖啡园采集的寄生于蝉若虫的昆虫病原真菌T011初步形态鉴定为蝉蝉，属于Hypocreales目和Clavicipitaceae科。为确保 T011 的真实性，对串联的多个基因组进行系统发育分析，包括它的,nrLSU,nrSSU,rpb1， 和铁氟龙用于支持识别。从干燥样品 T011 中分离基因组 DNA。应用 PCR 测定测序来扩增它的,nrLSU,nrSSU,rpb1， 和铁氟龙基因。对于系统发育分析，两个目标基因的级联数据是使用 MEGAX 构建的，基于邻接、最大似然、最大简约法，具有 1,000 次重复引导。结果，包含62个序列的级联数据属于Hypocreales目，Clavicipitaceae科，2个外群序列属于Hypocreales，属黄萎病菌. 系统发育分析结果表明T011在亚进化枝上被接受虫草并与参照物显着形成单系群虫草蝉（变形金刚蝉蝉) 具有高引导值。系统发育分析的结果得到了我们描述为蝉蝉. 总之，基于级联数据集的系统发育分析已成功应用于加强 T011 的识别蝉蝉.