苯菌灵胁迫对蜜蜂肠道微生物菌群结构的影响

doi:10.11924/j.issn.1000-6850.casb20190900692

摘要/Abstract

摘要：

旨在探明蜜蜂主要肠道菌群在药物胁迫下的变化规律,为进一步开展蜜蜂肠道共生菌群的生理功能及蜜蜂的健康养殖等研究提供理论参考。以意大利蜜蜂为材料,用PCR-DGGE分析方法对添食苯菌灵前后蜜蜂肠道微生物菌群结构进行研究。基于微生物的rDNA不同可变区经PCR后呈现出一条明亮的条带,表明蜜蜂肠道中有细菌和真菌的驻留。其中细菌的16S rDNA的V3可变区的PCR-DGGE显示有5条带,2条主带和3条副带;真菌18S rDNA V1+V2可变区的PCR-DGGE显示有3条带,1条主带和2条副带,表明蜜蜂肠道微生物较为丰富。在苯菌灵药物胁迫下,条带b、e、f、g逐渐变弱甚至消失,预示着蜜蜂肠道中的真菌种类和数目都有所减少,说明添食苯菌灵对肠道微生物具有抑制作用。苯菌灵药物胁迫会造成蜜蜂肠道微生态平衡失调,从而破坏机体正常的生理功能,严重影响到蜜蜂的健康。

关键词: 蜜蜂, 肠道微生物, 菌群结构, 苯菌灵胁迫, DGGE技术

Abstract:

The objectives of this study are to determine the change rule of the main intestinal microbial community of honeybee under drug stress, which can provide theoretical references for further investigating the physiological function of intestinal symbiotic community and healthy cultivation of honeybee. The worker bees of Apis mellifera ligustica are used as experimental materials. The intestinal microflora structure was investigated by PCR-DGGE analysis before and after adding benomyl. A specific bright band emerged from the gel after PCR based on different variable regions of rDNA, which indicated that bacteria and fungi were resident in bee intestine. Five bands (2 main belts and 3 sub belts) of the V3 region of 16S rDNA of bacteria and three bands (1 main belt and 2 sub belts) of the 18S rDNA V1+V2 region of fungi were separated by PCR-DGGE, which indicated that there were abundant intestinal microorganisms in honeybee intestine. The bands of b, e, f, g gradually weakened or even disappeared, which indicated that the species and amount of bacteria and fungi in the intestines of honeybees decreased, so the benomyl stress had inhibitory effect on intestinal microorganisms. The benomyl stress can cause the imbalance of intestinal micro-ecology of honeybee, which will damage the normal physiological function of the body and seriously affect the health of honeybee.

Key words: bee, intestinal microorganism, microbial community structures, benomyl stress, DGGE technology

中图分类号:

S895

代君君, 舒蕊, 刘健, 章玉萍, 张丽丽, 陈明, 赵萍, 刘震. 苯菌灵胁迫对蜜蜂肠道微生物菌群结构的影响[J]. 中国农学通报, 2020, 36(26): 136-140.

Dai Junjun, Shu Rui, Liu Jian, Zhang Yuping, Zhang Lili, Chen Ming, Zhao Ping, Liu Zhen. Benomyl Stress: Effects on the Microbial Community Structure in the Intestine of Honey Bees[J]. Chinese Agricultural Science Bulletin, 2020, 36(26): 136-140.

图/表 3

图1. 蜜蜂16S rDNA V3区扩增结果 M是Marker,1是对照组,2、3、4组分别是添食24、48、72 h苯菌灵后的扩增条带

图2. 蜜蜂18S rDNA V1+V2区的扩增结果 M是Marker,1是对照组,2、3、4组分别是添食24、48、72 h苯菌灵后的扩增条带

图3. 不同添食时间的蜜蜂中肠道微生物群落结构DGGE图谱（A凝胶图、B示意图) 1是未添食的16S rDNA V3区PCR DGGE（对照组）;2、3、4分别是添食24、48、72 h后16S rDNA V3区PCRDGGE;5是未添食18S rDNA V1+V2区PCRDGGE（对照组）,6、7、8分别是添食24、48、72 h后18S rDNA V1+V2区PCRDGGE

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