Wu Hua, Rao Zhong-Chen, Cao Li, De Clercq Patrick, Han Ri-Chou
Department of Plants and Crops, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium.
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, China.
Front Microbiol. 2020 Dec 3;11:577268. doi: 10.3389/fmicb.2020.577268. eCollection 2020.
The Chinese cordyceps is a unique and valuable parasitic complex of / ghost moths and the fungus for medicine and health foods from the Tibetan Plateau. During artificial cultivation of Chinese cordyceps, the induction of blastospores into hyphae is a prerequisite for mummification of the infected larvae. To explore the microbial involvement in the induction of mycelia-blastospore transition, the microbiota of the hemolymph and gut from larvae with or without injected blastospores were investigated by culture-dependent and -independent methods. Twenty-five culturable bacterial species and 14 fungal species, together with 537 bacterial operational taxonomic units (OTUs) and 218 fungal OTUs, were identified from the hemolymph and gut of samples from five stages including living larvae without injected fungi (A) or with high blastospore load (B), mummifying larvae without mycelia coating (C), freshly mummifying larvae coated with mycelia (D), and completely mummified larvae with mycelia (E). Two culturable bacterial species (), and 47 bacterial and 15 fungal OTUs were considered as shared species. The uninfected larval hemolymph contained 13 culturable bacterial species but no fungal species, together with 164 bacterial and 73 fungal OTUs. To our knowledge, this is the first study to detect large bacterial communities from the hemolymph of healthy insect larvae. When the living larvae contained high blastospore load, the culturable bacterial community was sharply inhibited in the hemolymph but the bacterial and fungal community greatly increased in the gut. In general, high blastospore load increased bacterial diversity but sharply decreased fungal diversity in the hemolymph and gut by OTUs. The bacterial loads of four culturable species ( sp., , ) increased significantly and . and spp. became dominant microbes, when the infected larvae became mummified, indicating their possible involvement in the larval mummification process. The discovery of many opportunistic pathogenic bacteria in the hemolymph of the healthy larvae, the larval microbial diversity influenced by challenge and the involvement of dominant bacteria during larval mummification process provide new insight into the infection and mummification mechanisms of in its hosts.
中华虫草是青藏高原上一种独特且珍贵的虫草复合体,是蝙蝠蛾与真菌形成的用于医药和保健食品的复合体。在中华虫草人工培育过程中,将芽生孢子诱导为菌丝是被感染幼虫木乃伊化的前提条件。为探究微生物在菌丝-芽生孢子转变诱导过程中的作用,采用依赖培养和不依赖培养的方法,对注射或未注射芽生孢子的幼虫血淋巴和肠道的微生物群进行了研究。从五个阶段的样本(包括未注射真菌的活幼虫(A)或芽生孢子负载量高的活幼虫(B)、无菌丝覆盖的木乃伊化幼虫(C)、有菌丝覆盖的新鲜木乃伊化幼虫(D)和有菌丝的完全木乃伊化幼虫(E))的血淋巴和肠道中,鉴定出25种可培养细菌物种和14种真菌物种,以及537个细菌操作分类单元(OTU)和218个真菌OTU。两种可培养细菌物种()以及47个细菌OTU和15个真菌OTU被视为共有物种。未感染幼虫的血淋巴中含有13种可培养细菌物种,但无真菌物种,以及164个细菌OTU和73个真菌OTU。据我们所知,这是首次从健康昆虫幼虫血淋巴中检测到大量细菌群落的研究。当活幼虫芽生孢子负载量高时,血淋巴中可培养细菌群落受到显著抑制,但肠道中的细菌和真菌群落大幅增加。总体而言,高芽生孢子负载量通过OTU增加了血淋巴和肠道中的细菌多样性,但大幅降低了真菌多样性。当被感染幼虫木乃伊化时,四种可培养物种(种、、)的细菌载量显著增加, 菌属和 菌属成为优势微生物,表明它们可能参与了幼虫木乃伊化过程。在健康幼虫血淋巴中发现许多机会性病原菌、受 挑战影响的幼虫微生物多样性以及优势细菌在幼虫木乃伊化过程中的参与,为 在其宿主中的感染和木乃伊化机制提供了新的见解。