昆虫学报 ›› 2021, Vol. 64 ›› Issue (4): 460-470.doi: 10.16380/j.kcxb.2021.04.005

• 研究论文 • 上一篇    下一篇

植物乳杆菌、苹果醋酸杆菌和酿酒酵母三种微生物对黑腹果蝇行为和发育的影响

王露, 魏博帆, 李苗苗, 李晓哲, 王博, 阚云超, 乔惠丽   

  1.  (南阳师范学院, 河南省伏牛山昆虫生物学重点实验室, 河南南阳 473061)
  • 出版日期:2021-04-20 发布日期:2021-04-25

Effects of three microbes, Lactobacillus plantarum, Acetobacter malorum, and Saccharomyces cerevisiae, on the behavior and development of Drosophila melanogaster

 WANG Lu, WEI Bo-Fan, LI Miao-Miao, LI Xiao-Zhe, WANG Bo, KAN Yun-Chao, QIAO Hui-Li*   

  1. (Henan Provincial Key Laboratory of Insect Biology in Funiu Mountain, Nanyang Normal University, Nanyang, Henan 473061, China)
  • Online:2021-04-20 Published:2021-04-25

摘要:

【目的】肠道微生物在宿主的多种生命活动中发挥重要作用。本研究旨在通过研究植物乳杆菌Lactobacillus plantarum, 苹果醋酸杆菌Acetobacter malorum和酿酒酵母Saccharomyces cerevisiae 3种微生物对黑腹果蝇Drosophila melanogaster觅食、产卵和发育的影响,进一步阐明肠道微生物与宿主的互作调控机制。【方法】采用引诱实验检测体外培养的植物乳杆菌、苹果醋酸杆菌和酿酒酵母对交配前后黑腹果蝇成虫的引诱力以及这3种微生物的单一微生物和其中2种或3种微生物的混合物对非处女蝇的引诱力。采用产卵选择实验检测黑腹果蝇非处女蝇对单一微生物和其中2种或3种微生物的混合物的产卵偏好。将黑腹果蝇卵在接种分别含有植物乳杆菌、苹果醋酸杆菌和酿酒酵母的培养基和正常培养基(对照)上培养,测量幼虫体重;同时将卵在分别含有活菌或灭活的这3种单一微生物的培养基上和正常培养基上培养,统计化蛹数,以检测不同微生物对黑腹果蝇幼虫发育的影响。将黑腹果蝇幼虫在分别含有植物乳杆菌、苹果醋酸杆菌和酿酒酵母的培养基及正常培养基中培养72 h,qRT-PCR检测胰岛素信号通路的关键基因InR在黑腹果蝇幼虫中的相对表达量变化。【结果】苹果醋酸杆菌主要影响黑腹果蝇的产卵,酿酒酵母和植物乳杆菌则可同时影响黑腹果蝇觅食和产卵,且与单一微生物相比,非处女黑腹果蝇更趋向于含有2种或3种微生物的混合物。产卵选择实验表明,3种单一供试微生物及其中2种或3种微生物的混合物对黑腹果蝇的产卵选择具有显著的吸引力,其中混合物的影响最大,然后依次是苹果醋酸杆菌、酿酒酵母、植物乳杆菌。3种微生物均可促进黑腹果蝇幼虫的发育,活菌在接种早期加快幼虫到蛹的发育,灭活菌的促进作用则相对滞后。与对照比较,InR在接种苹果醋酸杆菌的培养基上培养的黑腹果蝇幼虫中的表达量显著降低,而在分别接种植物乳杆菌和酿酒酵母的培养基上培养的幼虫中则被显著促进表达。【结论】苹果醋酸杆菌可引诱黑腹果蝇的产卵,酿酒酵母和植物乳杆菌对黑腹果蝇的觅食和产卵都有具有引诱力;同时微生物的多样性可增加黑腹果蝇的觅食和产卵偏好。植物乳杆菌、苹果醋酸杆菌和酿酒酵母可能通过不同的调控机制影响黑腹果蝇幼虫的生长发育。

关键词: 黑腹果蝇, 肠道微生物, 植物乳杆菌, 苹果醋酸杆菌, 酿酒酵母, 引诱力, 产卵

Abstract: 【Aim】 Gut microbes play very important roles in various life processes of host. This study aims to further clarify the mechanism of interaction between gut microbes and host by exploring the effects of three microbes, Lactobacillus plantarum, Acetobacter malorum, and Saccharomyces cerevisiae, on the foraging, oviposition and development of Drosophila melanogaster. 【Methods】 Attraction assays were performed to detect the attractiveness of L. plantarum, A. malorum or S. cerevisiae in the culture to the unmated and mated adults of D. melanogaster, and the attractiveness of the single microbe or the mixture of the two or three microbes to the non-virgin D. melanogaster. Oviposition assays were performed to detect the oviposition preference of non-virgin D. melanogaster to the single microbe or the mixture of the two or three microbes. The eggs of D. melanogaster were transferred to the media inoculated with L. plantarum, A. malorum and S. cerevisiae, respectively, and the normal medium (control), and the larval body weight was measured; meanwhile, the eggs of D. melanogaster were transferred to the media inoculated with live and inactivated single microbe of the three microbes, respectively, and the normal medium, and the number of pupae was counted to ascertain the effects of different microbes on the development of larvae of D. melanogaster. The relative expression levels of InR, a key gene of insulin signaling pathway, in D. melanogaster larvae raised in the media containing L. plantarum, A. malorum and S. cerevisiae, respectively, and the normal medium for 72 h were detected by qRT-PCR. 【Results】 A. malorum mainly affected the oviposition of D. melanogaster, while L. plantarum and S. cerevisiae could affect both its foraging and oviposition. Compared with single microbe, the mixture of the two or three microbes had stronger attractiveness to non-virgin D. melanogaster. The oviposition preference assays showed that L. plantarum, A. malorum, S. cerevisiae and the mixture of the two or three microbes had significant attractiveness to D. melanogaster for oviposition in the order of mixture>A. malorum>S. cerevisiae>L. plantarum. In addition, L. plantarum, A. malorum and S. cerevisiae could promote the development of D. melanogaster larvae. The live microbes accelerated the development of D. melanogaster from larvae to pupae in the early stage of inoculation, but the promotion effect of the inactivated microbes lagged behind. The expression level of InR in the larvae of D. melanogaster cultured on the medium inoculated with A. malorum decreased significantly as compared to that in the control, while those in the larvae cultured on the media inoculated with L. plantarum and S. cerevisiae, respectively, significantly increased. 【Conclusion】 A. malorum can induce D. melanogaster to lay eggs, and L. plantarum and S. cerevisiae are attractive to D. melanogaster for foraging and oviposition. Meanwhile, microbial diversity can increase the foraging and oviposition preference of D. melanogaster. L. plantarum, A. malorum and S. cerevisiae may affect the growth and development of D. melanogaster larvae via different regulatory mechanisms.

Key words: Drosophila melanogaster, gut microbes, Lactobacillus plantarum; Acetobacter malorum, Saccharomyces cerevisiae, attractiveness, oviposition