冬虫夏草菌和昆虫激素对小金蝠蛾幼虫僵化、血淋巴和肠道微生物以及血淋巴化学物质的影响  

Effects of injection with Ophiocordyceps sinensis spores and insect hormones on larval mummification rate, hemolymph, gut microorganisms and hemolymph chemicals of Thitarodes xiaojinensis

作　　者：吴华 曹莉 饶中臣 刘桂清 唐睿 韩日畴 WU Hua;CAO Li;RAO Zhong-Chen;LIU Gui-Qing;TANG Rui;HAN Ri-Chou(Guangdong Key Laboratory of Animal Conservation and Resource Utilization,Guangdong Public Laboratory of Wild Animal Conservation and Utilization,Institute of Zoology,Guangdong Academy of Sciences,Guangzhou 510260,China)

机构地区：[1]广东省科学院动物研究所,广东省动物保护与资源利用重点实验室,广东省野生动物保护与利用公共实验室,广州510260

出　　处：《应用昆虫学报》2021年第6期1257-1276,共20页Chinese Journal of Applied Entomology

基　　金：广东省科学院千名博士(后)计划引进专项(2020GDASYL20200103097和2019GDASYL-0103056);广东省重点领域研发计划项目(2020B1111580001);国家自然科学基金项目(31900368);青海省重大科技专项(2021-SF-A4-1)。

摘　　要：【目的】冬虫夏草是由冬虫夏草菌Ophiocordycepssinensis侵染蝙蝠蛾幼虫形成的一种独特而有价值的虫菌复合体。人工栽培冬虫夏草时,将蝙蝠蛾幼虫血淋巴中的冬虫夏草菌芽生孢子诱导成为菌丝,是被感染幼虫僵化的先决条件。为探讨冬虫夏草菌和昆虫激素对小金蝠蛾Thitarodes xiaojinensis幼虫僵化、血淋巴和肠道微生物以及血淋巴化学物质的影响。【方法】将冬虫夏草菌芽生孢子(KD菌株)、芽生孢子和蜕皮激素(KD+20E)、芽生孢子和保幼激素(甲氧普林)(KD+M)、20E和M分别注射小金蝠蛾6龄幼虫,以注射无菌PBS、35%DMSO以及未注射任何物质的幼虫作为对照,测定幼虫的僵化率、血淋巴和肠道可培养微生物,以及血淋巴的关键物质(法尼醇、酪醇、麦角甾醇、蜕皮激素、保幼激素、葡萄糖、一氧化氮、海藻糖、N-乙酰葡糖胺)的含量。【结果】注射KD+M 120 d后小金蝠蛾幼虫的僵虫率显著提高。注射KD+20E的幼虫血淋巴中分离到更多细菌种类,而注射KD+M幼虫血淋巴中分离得到更多真菌种类。Pseudomonasmucidolens、Serratiaproteamaculans和Tsukammurella standjordii细菌出现于所有处理和注射PBS的幼虫血淋巴中(除了T. standjordii未出现于注射甲氧普林的幼虫血淋巴中);高频率出现的真菌种类为Cladosporium sp.、Isaria farinosa和Talaromyces sp.,这些菌株均未出现在未注射幼虫的血淋巴中。7种细菌(Carnobacteriummaltaromaticum、Flavobacterium frigidimaris、Microbacterium oxydans、P. mucidolens、Rahnella aquatilis、S. proteamaculans、T. strandjordii)和1种真菌(Apiotrichum porosum)出现于所有幼虫肠道中;C. maltaromaticum是幼虫肠道优势种;注射KD+M增加幼虫肠道真菌的可培养种类;高频率出现的真菌种类为I. farinosa和Penicilliumsp.;从幼虫肠道组织中未分离到Ophiocordycepssinensis。未注射芽生孢子、注射KD+20E的小金蝠蛾幼虫血淋巴中均未检测到N-乙酰葡萄�[Objectives] To investigate the effect of the fungus Ophiocordyceps sinensis(a unique and valuable parasitic complex formed by the infection and subsequent mummification of Thitarodes larvae by O. sinensis) and insect hormones on the blastospore-hypha conversion in Thitarodes larvae(the formation of hyphae from the blastospores of O. sinensis in the hemolymph of Thitarodes larvae is a prerequisite for the mummification of infected larvae). [Methods] One of five treatments;blastospores of O. sinensis(KD strain), a mixture of blastospores and 20-hydroxyecdysone(KD+20 E), a mixture of blastospores and methoprene(KD+M), 20-hydroxyecdysone(20 E) or Methoprene, were injected into sixth instar T.xiaojinensis larvae and the subsequent larval mummification rate, hemolymph and intestinal microorganisms and amounts of related substances(farnesol, ergosterol, ecdysterone, juvenile hormone III, N-acetylglucosamine, glucose, trehalose, nitric oxide) in larval hemolymph were compared. [Results] The mixture of blastospores and methoprene significantly increased the mummification rate of larvae after 120 days. More bacterial species were isolated from larval hemolymph injected with KD+20 E, whereas more fungi were isolated from larvae injected with KD+M. Three bacterial species including Pseudomonas mucidolens, Serratia proteamaculans and Tsukammurella standjordii, were detected in all treatments, except for the absence of T. standjordii in the hemolymph of larvae injected with Methoprene. Cladosporium sp., Isaria farinosa and Talaromyces sp.were the most frequently observed fungal species, none of which were found in the hemolymph of larvae that had not been injected. Seven bacterial species, including Carnobacterium maltaromaticum, Flavobacterium frigidimaris, Microbacterium oxydans, P. mucidolens, Rahnella aquatilis, S. proteamaculans and T. strandjordii, as well as one fungal species, Apiotrichum porosum were detected in the guts of all treated larvae. C. maltaromaticum was the dominant bacterial species found in larval guts.

关 键 词：冬虫夏草 蜕皮激素 保幼激素 芽生孢子-菌丝转化 僵化 微生物菌群 

分 类 号：S567.35[农业科学—中草药栽培]