橘小实蝇无菌品系的构建方法
Method for constructing axenic strain of Bactrocera dorsalis
谢俊飞1** 何文峰2 邱 敏2 舒浩然3 林嘉欣2 刘磊磊1***
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DOI：10.7679/j.issn.2095-1353.2026.073
作者单位：1. 武汉生物工程学院生命科学与技术学院，武汉 430415；2. 华中农业大学植物科学技术学院，武汉 430070； 3. 湖北工业大学生命科学与健康工程学院，武汉 430068
中文关键词：橘小实蝇；肠道微生物；无菌培养，品系构建
英文关键词：Bactrocera dorsalis; gut microbiota; axenic cultivation; strain establishment
中文摘要：
【目的】 建立橘小实蝇Bactrocera dorsalis无菌品系的标准化培育方法，为探究其肠道微生物与宿主之间的互作关系提供有力的技术支撑。【方法】 操作流程包括环境控制、材料灭菌、胚胎脱菌和无菌培养四个环节。利用平板涂布结合细菌16S rDNA和真菌ITS1区PCR扩增与电泳检测，综合评价无菌培育效果。【结果】 脱菌后的卵可稳定发育至蛹期。平板涂布培养及分子检测（16S rDNA与ITS1区PCR扩增电泳）结果一致表明，所获幼虫体内外均无外源微生物污染。【结论】 本研究通过优化操作流程与环境控制，建立了稳定、可重复的橘小实蝇无菌品系培育体系，可为肠道共生菌等功能研究提供标准化的实验模型。
英文摘要：
[Aim] To establish a standardized method for
cultivating germ-free strains of Bactrocera dorsalis, providing robust
technical support for investigating the interactions between gut microbiota and
host. [Methods] The operational
procedure comprised four key steps: Environmental control, material
sterilization, embryo surface sterilization, and axenic cultivation. The
effectiveness of the axenic rearing was comprehensively evaluated using plate
culture combined with PCR amplification and electrophoretic detection of
bacterial 16S rDNA and fungal ITS1 regions. [Results] Following the above procedure,
surface-sterilized embryos consistently developed to the pupal stage. Both
plate culture and molecular detection (PCR amplification and electrophoresis of
16S rDNA and ITS1 regions) yielded consistent results, indicating no detectable
exogenous microbial contamination inside or outside the obtained larvae. [Conclusion] By optimizing operational procedures and
environmental controls, this study established a stable and reproducible system
for cultivating axenic strain of B. dorsalis, providing a standardized
experimental model for functional studies on gut symbionts and other aspects.