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蜜蜂成虫工蜂肠道可培养细菌群落结构分析
引用本文:王洪秀,靳亮,陈庆隆,魏云辉,姚健,马吉平,张诚. 蜜蜂成虫工蜂肠道可培养细菌群落结构分析[J]. 微生物学通报, 2017, 44(3): 620-630
作者姓名:王洪秀  靳亮  陈庆隆  魏云辉  姚健  马吉平  张诚
作者单位:1. 江西省农业科学院农业应用微生物研究所 江西 南昌 330200,2. 江西省科学院微生物研究所 江西 南昌 330029,1. 江西省农业科学院农业应用微生物研究所 江西 南昌 330200,1. 江西省农业科学院农业应用微生物研究所 江西 南昌 330200,1. 江西省农业科学院农业应用微生物研究所 江西 南昌 330200,1. 江西省农业科学院农业应用微生物研究所 江西 南昌 330200,1. 江西省农业科学院农业应用微生物研究所 江西 南昌 330200
基金项目:国家自然科学基金项目(No. 31460490);中国科学院农业与环境微生物学重点实验室开放研究基金项目(No. 2014AEM003);江西省青年科学基金项目(No. 20142BAB214021);江西省农业科学院创新基金博士启动项目(No. 2012CBS006);江西省农业科学院协同创新项目(No. JXXTCX2015003-010)
摘    要:【目的】研究2种蜜蜂(健康意大利蜜蜂和健康中华蜜蜂)成虫工蜂肠道可培养细菌的群落结构组成。【方法】利用16S r RNA基因的聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)分析技术,结合菌落形态观察和生理生化特征鉴定细菌种类。【结果】从2种蜜蜂成虫工蜂肠道200株可培养细菌得到18种不同细菌遗传型,分属于肠杆菌科(Enterobacteriaceae)、弧菌科(Vibrionaceae)和肠球菌科(Enterococcaceae)3个科。其中肠杆菌科是肠道可培养细菌最优势的细菌种类。同样以序列相似性大于97%的菌株归为相同细菌种类为标准,找到了2种蜜蜂可培养细菌的共有菌种,结合菌落形态观察和生理生化特征鉴定,确定肠道可培养细菌为肠杆菌属8株,克雷伯氏菌属1株,肠球菌属2株,以及气单胞菌属1株。【结论】通过研究健康意大利蜜蜂和中华蜜蜂成虫工蜂肠道可培养细菌群落结构组成,可为开展蜜蜂的微生态研究提供基础性资料。

关 键 词:蜜蜂,16S rRNA基因,肠道细菌,变性梯度凝胶电泳,群落结构

Diversity and community structure of intestinal cultivable bacteria in adult worker-bee
WANG Hong-Xiu,JIN Liang,CHEN Qing-Long,WEI Yun-Hui,YAO Jian,MA Ji-Ping and ZHANG Cheng. Diversity and community structure of intestinal cultivable bacteria in adult worker-bee[J]. Microbiology China, 2017, 44(3): 620-630
Authors:WANG Hong-Xiu  JIN Liang  CHEN Qing-Long  WEI Yun-Hui  YAO Jian  MA Ji-Ping  ZHANG Cheng
Affiliation:1. Institute of Agricultural Applied Microbiology, Jiangxi Agricultural Academy of Sciences, Nanchang, Jiangxi 330200, China,2. Institute of Microbiology, Jiangxi Academy of Sciences, Nanchang, Jiangxi 330029, China,1. Institute of Agricultural Applied Microbiology, Jiangxi Agricultural Academy of Sciences, Nanchang, Jiangxi 330200, China,1. Institute of Agricultural Applied Microbiology, Jiangxi Agricultural Academy of Sciences, Nanchang, Jiangxi 330200, China,1. Institute of Agricultural Applied Microbiology, Jiangxi Agricultural Academy of Sciences, Nanchang, Jiangxi 330200, China,1. Institute of Agricultural Applied Microbiology, Jiangxi Agricultural Academy of Sciences, Nanchang, Jiangxi 330200, China and 1. Institute of Agricultural Applied Microbiology, Jiangxi Agricultural Academy of Sciences, Nanchang, Jiangxi 330200, China
Abstract:[Objective] In order to identify the diversity and community structure of the cultivable bacteria inhabiting the intestinal tract in worker adults of honeybee from herlthy Apis mellifera ligustica and Apis cerana cerana Fabricius populations. [Methods] 16S rRNA gene PCR-DGGE (Denaturing gradient gel electrophoresis) molecular methods was employed to study the bacterial diversity. In addition, the cultivable bacteria were identified by colony morphology, physiological and biochemical characteristics tests. [Results] A total of 200 culturable bacteria from aboved two populations were classified into 18 unique phylotypes. All sequenced bacteria strains were grouped into three families: Enterobacteriaceae, Vibrionaceae and Enterococcaceae. The Enterobacteriaceae was dominated in all the populations. The closely related sequences (>97% sequence similarity) which had been retrieved from culturable bacteria of two populations were grouped as one common species. We identified the cultivable bacteria: eight strains of bacteria as Enterobacter, one strain of Klebsiella, two strains of Enterococcus, and one strain of Aeromonas. [Conclusion] The present study significantly contributes to the available information on bacterial isolates from worker adults of honeybee.
Keywords:Honeybee   16S rRNA gene   Intestine cultivable bacteria   Denaturing gradient gel electrophoresis   Community structure
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