氟化物对家蚕肠道微生物菌群的影响  

Effect of fluoride on gut microflora of silkworm( Bombyx mori)

作　　者：李冠楠[1] 夏雪娟[2] Parfait Sendegeya 赵欢欢[1] 隆耀航[1] 朱勇[1] 

机构地区：[1]西南大学生物技术学院,重庆400716 [2]西南大学食品科学学院,重庆400716

出　　处：《微生物学报》2015年第7期926-934,共9页Acta Microbiologica Sinica

基　　金：国家农业科技成果转化资金(2012GB2F100376);西南大学基本科研业务费专项资金(XDJK2014D039)~~

摘　　要：【目的】探讨氟化物对家蚕肠道微生物菌群的影响,开发具有潜在应用价值的益生菌以提高家蚕对氟化物的抗性。【方法】PCR扩增家蚕肠道内细菌16S rRNA基因并构建克隆文库;用核糖体DNA限制性分析(Amplified ribosomal DNA restriction analysis,ARDRA)方法对克隆子进行分型。从家蚕T6和734肠道样品中共获得14种分类操作单元(Operational taxonomic unit,OTUs),以16S rRNA基因为基础构建系统发育树进行分析。再经巢式PCR-DGGE技术检测家蚕肠道内优势菌群的变化。【结果】结果表明:家蚕氟中毒后肠道内肠球菌属Enterococcus和芽孢杆菌属Bacillus细菌菌群减少,而葡萄球菌属Staphylococcus的细菌增多。【结论】氟化物能通过改变家蚕肠道内细菌的多样性和比例,从而破坏家蚕肠道微生物菌群平衡,且对家蚕734的影响作用大于T6。[Objective] We examined the effect of fluoride on gut microflora of silkworm. [Methods] After DNA extraction and PCR amplification,clone libraries of 16 S rRNA gene fragment were constructed. Amplified ribosomal DNA restriction analysis（ ARDRA） was performed by digestion of the 16 S rRNA gene,and each unique restriction fragment polymorphism pattern was designated as an operational taxonomic unit（ OTU）. A total of 14 OTUs were identified from intestinal samples of both T6 and 734. Phylogenetic trees of bacterial 16 S rRNA nucleotide sequences were constructed and analyzed. Furthermore,the dominant bacteria were studied by the nested polymerase chain reaction denaturing gradient gel electrophoresis（ PCR-DDGE） technology. [Results] After fluorosis,the flora of Enterococcus and Bacillus reduced. However,the flora of Staphylococcus increased. [Conclusion]Fluoride can destroy the balance of microflora in the gut of silkworm by changing the bacteria diversity and proportion,which has bigger effect to 734 than T6.

关 键 词：家蚕 氟化物 肠道菌群 克隆文库 巢式PCR-DGGE 

分 类 号：S881.2[农业科学—特种经济动物饲养]