Studies on the cloning of few-coiled type of spore in entomopathogenic microsporidia

昆虫病原微孢子虫少卷曲型孢子的克隆研究

• 批准号: 10660056
• 负责人: IWANO Hidetoshi
• 金额: $ 1.92万
• 依托单位: Nihon University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10660056/
• 关键词: microsporidia; spore dimorphism; cloning; Nosema bombycis; micromanipulator; Vairimorpha necatrix; isolation; cell culture; Nosema; Vairimorpha

The purpose of this study was to clone entomogenous microsporidia by infecting an insect cultured cell with a single spore. In an attempt to isolate the octospore of Vairimorha necatrix that had been propagated in silkworm larvea reared at 16゜C. In the genus Nusema, one of the two types of the spore, the few-coiled type was recovered from cultured Helicoverpa zea cells infected with Nosema furnacalis and cultured Antheraea eucalypti cells infected with N. bombycis.A single spore was recovered from each suspension of few-coiled type spores and transferred to a cell culture using a micromanupulator.Although the developmental stages of N. furnacalis were detected in cultured H. zea cells after 30-day incubation, those of N. bombycis were not. When a single spore of many-coiled type of N. bombycis spore was transferred to cell cultures after alkaline activation, various developmental stages were detected in cultured A. eucalypti cells after 30-day incubation. The cloned microsporidia from a single spore could be subcultured by supplementing the culture with the healthy cells.

本研究的目的是通过用单个孢子感染昆虫培养细胞来克隆虫生微孢子虫。试图分离在 16°C 饲养的蚕幼虫中繁殖的 Vairimorha necatrix 八孢子。在两种类型的孢子之一的 Nusema 属中，从培养的感染了玉米微孢子虫的玉米夜蛾细胞和培养的感染了家蚕的桉树蚕细胞中回收了少卷曲型。从每个少卷曲型孢子悬浮液中回收了单个孢子，并使用显微操作器将其转移到细胞培养物中。 培养 30 天后，在培养的玉蜀黍细胞中检测到了亚洲玉米螟，而家蚕则未检测到。当多卷曲型家蚕孢子的单个孢子在碱活化后转移到细胞培养物中时，在培养的桉树细胞中在培养30天后检测到不同的发育阶段。来自单个孢子的克隆微孢子虫可以通过用健康细胞补充培养物进行传代培养。

项目成果

E. U. Canning, A. Curry, S. A. Cheney, N. J. Lafranchi-Tristem, Y. Kawakami, Y. Hatakeyama, H. Iwano and R. Ishihara: "Nosema tyriae n.sp. And Nosema sp., microporidian parasites of cinnabar moth Tyria jacobaeae"J. Invertebrate Pathology. 74. 29-38 (1999)

E. U. Canning、A. Curry、S. A. Cheney、N. J. Lafranchi-Tristem、Y. Kawakami、Y. Hatakeyama、H. Iwano 和 R. Ishihara：“Nosema tyriae n.sp. 和 Nosema sp.，朱砂蛾 Tyria jacobaaee 的微孔寄生虫

Elizabeth U. Canning: "Nosema tyriae n.sp. and Nosema sp., microsporidian parasites of cinnabar moth Tyria jacobaeae"Journal of Invertebrate Pathology. 74. 29-38 (1999)

Elizabeth U. Canning：“Nosema tyriae n.sp. 和 Nosema sp.，朱砂蛾 Tyria jacobaaee 的微孢子虫寄生虫”无脊椎动物病理学杂志。