昆虫学报 ›› 2025, Vol. 68 ›› Issue (5): 607-618.doi: 10.16380/j.kcxb.2025.05.007

• 研究论文 • 上一篇    下一篇

wMelPop侵染对雄性黑腹果蝇肠道细菌的影响

钟子倩#, 马倩#, 李栋, 汪家林, 王玉凤*   

  1. (华中师范大学生命科学学院, 湖北省遗传调控与整合生物学重点实验室, 武汉 430079)
  • 出版日期:2025-05-20 发布日期:2025-06-06

Effects of wMelPop infection on the gut bacteria in male Drosophila melanogaster

ZHONG Zi-Qian#, MA Qian#, LI Dong, WANG Jia-Lin, WANG Yu-Feng*   

  1.  (Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Sciences, Central China Normal University, Wuhan 430079, China)
  • Online:2025-05-20 Published:2025-06-06

摘要: 7【目的】沃尔巴克氏体Wolbachia是一类母系遗传的革兰氏阴性菌,是昆虫中最常见的胞内共生菌之一,可通过多种方式影响宿主的生殖和行为,但其影响机制还不甚清楚。本研究旨在探讨一种毒力较强的Wolbachia菌株wMelPop侵染对雄性黑腹果蝇Drosophila melanogaster肠道细菌的影响。【方法】解剖感染wMelPop的黑腹果蝇(Pop+)和未感染的黑腹果蝇(Pop-)雄成虫完整肠道,提取总DNA,利用Illumina NovaSeq平台对肠道细菌16S rDNA基因的V3-V4区进行测序,比较分析Pop+和Pop-肠道细菌的物种组成、丰度及α多样性,并通过qPCR验证表达量具有显著差异的3个属的6种肠道微生物高通量测序数据的有效性。【结果】α多样性分析显示,wMelPop侵染提高了雄性黑腹果蝇肠道细菌丰度,降低了肠道菌的多样性。Pop-肠道中,优势菌属是不动杆菌属Acinetobacter、假单胞菌属Pseudomonas、醋杆菌属Acetobacter和嗜盐单胞菌属Halomonas。与Pop-相比,Pop+肠道中假单胞菌属的相对丰度显著下调,而醋杆菌属和乳杆菌属Lactobacillus的相对丰度显著上调。qPCR验证结果显示,与Pop-相比, Pop+肠道中总细菌16S rDNA基因表达量极显著上调,Pop+肠道中番茄醋酸杆菌A. pomorum的16S rDNA基因表达量下调至Pop-的0.7倍,Pop+肠道中热带醋酸杆菌A. tropicalis的16S rDNA基因表达量比Pop-的上调约3 700倍,果状乳杆菌L. frutivorans、植物乳杆菌L. plantarum和短乳杆菌L. brevis的16S rDNA基因表达量均显著上调,其中果状乳杆菌16S rDNA基因表达量上调的倍数最高,约为Pop-的23 000倍,产碱假单胞菌P. alcaligenes的16S rDNA基因表达量下调至Pop-的0.6倍,这些结果与16S rDNA测序结果的趋势基本一致。【结论】wMelPop的侵染显著影响了雄性黑腹果蝇肠道细菌的相对丰度。其中,醋杆菌属和乳杆菌属的肠道菌载量显著升高,这可能是wMelPop感染影响昆虫宿主生殖和行为的机制之一。

关键词:  黑腹果蝇, wMelPop, 雄性, 肠道细菌, 醋杆菌, 乳杆菌

Abstract: 【Aim】 Wolbachia is a genus of maternally inherited Gram-negative bacteria, which is one of the most common symbiotic bacteria in insects and can affect host reproduction and behavior in various ways. But the mechanisms of Wolbachia’s influences on hosts are still unclear. This study aims to characterize the effects of the infection of wMelPop, a high toxic Wolbachia strain, on the gut bacteria in male Drosophila melanogaster. 【Methods】 The whole guts of male adults of wMelPop-infected (Pop+) and uninfected (Pop-) D. melanogaster were dissected, from which the total DNA was extracted, respectively. The V3-V4 regions of the 16S rDNA genes of the gut bacteria were sequenced using Illumina NovaSeq platform. The composition, abundance, and α-diversity of the gut bacteria of Pop+ and Pop- were analyzed and compared, and the validity of high-throughput sequencing data of six gut microbial species from three genera with significant differences in expression level was confirmed by qPCR. 【Results】 α-Diversity analysis indicated that wMelPop infection resulted in increased abundance, but decreased diversity of gut bacteria in male D. melanogaster. In the gut of Pop-, the dominant bacterial genera were Acinetobacter, Pseudomonas, Acetobacter and Halomonas. In the gut of Pop+, the relative abundance of Pseudomonas was significantly decreased, while that of Acetobacter and Lactobacillus was significantly increased as compared to those of Pop-. qPCR analysis result revealed that the expression level of the 16S rDNA gene of the total gut bacteria of Pop+ was extremely significantly upregulated as compared to that of Pop-. The expression level of the 16S rDNA gene of A. pomorum in the gut of Pop+ was downregulated to 0.7-fold of that of Pop-, and that of A. tropicalis in the gut of Pop+ was upregulated by about 3 700-fold of that of Pop-. The expression levels of the 16S rDNA genes of L. frutivorans, L. plantarum and L. brevis in the gut of Pop+ were significantly higher than those of Pop-, with that of L. frutivorans showing the highest upregulation level of about 23 000-fold of that of Pop- and that of P. alkaligenes showing downregulation level of about 0.6-fold of that of Pop-. These results were consistent with the trends of the 16S rDNA sequencing results.【Conclusion】The infection of wMelPop significantly affects the relative abundance of gut bacteria in male D. melanogaster, with a significant increase in gut microbiota load of Acetobacter and Lactobacillus. This may be at least one of the mechanisms by which wMelPop infection affects the reproduction and behavior of insect hosts.

Key words: Drosophila melanogasterwMelPop, males, gut bacteria; Acetobacter, Lactobacillus