一株灰茶尺蠖高毒力金龟子绿僵菌的筛选、鉴定与培养条件

摘要/Abstract

摘要： 绿僵菌（Metarhizium）是一类重要的虫生真菌,在农林害虫生物防治中被广泛应用。为从7个保存的菌种中筛选出对灰茶尺蠖毒力最强的菌株,并验证该菌株在不同剂量下的毒力效果,通过形态学特征观察和多基因系统进化树分析方法确定其分类地位,并对菌株生长的最适培养基、温度、pH进行了研究。结果表明,供试的7个菌株对灰茶尺蠖3龄幼虫具有不同的防治效果,菌株Ma54的毒力最强,处理10 d后的校正死亡率达81.67%,半致死时间（LT₅₀）为4.93 d;灰茶尺蠖的死亡率与菌株Ma54的孢子浓度呈剂量效应,高浓度（1.0×10⁸个·mL^-1）处理7 d后,死亡率达98.33%,LT₅₀为4.09 d;菌株Ma54感染灰茶尺蠖7 d后,虫尸长满菌丝、呈黑绿色,在PDA培养基上生长14 d后,分生孢子向外扩散、呈深绿色;菌株Ma54的（Internal transcribed spacer,ITS）、β微管蛋白（β-tubulin）、RNA聚合酶Ⅱ第一大亚基RPB1基因的系统进化树都与金龟子绿僵菌（Metarhizium anisopliae）处在相同分支;菌株Ma54的菌落直径、单位面积产孢量、孢子萌发率在PPDA培养基、温度25 ℃、pH 7.0条件下最为理想。该研究获取了对灰茶尺蠖具有高毒力的菌株Ma54,鉴定为金龟子绿僵菌,初步明确了菌株的生长条件,为后续灰茶尺蠖的生物防治奠定了基础。

关键词: 灰茶尺蠖, 金龟子绿僵菌, 高毒力, 分类地位, 生长条件

Abstract: As an important entomogenous fungus, Metarhizium has been extensively utilized in the biological control of agricultural and forestry pests. This study aimed to screen the most infective strain against E. grisescens from 7 preserved strains and verify its infectivity at different doses. Subsequently, the taxonomic status of the strain was determined through observation of morphological characteristics and methods for multigene phylogenetic tree analysis. Finally, the optimal culture medium, temperature and pH value for strain growth were investigated. The results indicate that the seven tested strains exhibited different control effects on the 3rd instar larvae of E. grisescens, with strain Ma54 exhibiting the highest virulence, achieving a corrected mortality rate of 81.67% after 10 d of treatment and an LT₅₀ (lethal time for 50% mortality) of 4.93 d. The mortality rate of E. grisescens shows a dose-response relationship with the spore concentration of strain Ma54. After 7 d of treatment under a high concentration of Metarhizium (1.0×10⁸spores·mL^-1), the mortality rate reached 98.33%, and the LT₅₀value was 4.09 days. Seven days after strain Ma54 infected E. grisescens, the cadavers were covered with mycelia and turned dark green. After 14 days of growth on PDA medium, the conidia further spread and appeared dark green. The phylogenetic trees of the ribosomal transcription factor, β-tubulin, and the largest subunit of RNA polymerase Ⅱ of strain Ma54 all clustered with Metarhizium anisopliae. The colony diameter, spore production per unit area, and spore germination rate of strain Ma54 were optimal under conditions of a PPDA medium, a temperature of 25 ℃, and a pH value of 7.0. This study obtained the Ma54 strain with high virulence against E. grisescens, authenticated it as M. anisopliae, and preliminarily clarified its growth conditions of the strain, laying a foundation for its subsequent utilization in the biological control of E. grisescens.

Key words: Ectropis grisescens, Metarhizium anisopliae, high virulence, taxonomic status, growth conditions

中图分类号:

李良德, 李金玉, 李慧玲, 张辉, 王定锋. 一株灰茶尺蠖高毒力金龟子绿僵菌的筛选、鉴定与培养条件[J]. 茶叶科学, 2025, 45(3): 427-438.

LI Liangde, LI Jinyu, LI Huiling, ZHANG Hui, WANG Dingfeng. Screening, Identification and Culture Conditions of a Highly Virulent Metarhizium anisopliae Strain against Ectropis grisescens Warre[J]. Journal of Tea Science, 2025, 45(3): 427-438.

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