16S rDNA克隆文库方法分析中华通草蛉共生细菌组成

doi:10.16409/j.cnki.2095-039x.2017.06.019

中国生物防治学报 ›› 2017, Vol. 33 ›› Issue (6): 849-856.DOI: 10.16409/j.cnki.2095-039x.2017.06.019

• 研究简报 • 上一篇

16S rDNA克隆文库方法分析中华通草蛉共生细菌组成

赵辉^1,2, 张帅², 雒珺瑜², 张利娟², 王爱英¹, 崔金杰²

1. 石河子大学生命科学学院, 石河子 832000;
2. 中国农业科学院棉花研究所/棉花生物学国家重点实验室, 安阳 455000

收稿日期:2017-06-14 出版日期:2017-12-08 发布日期:2017-12-16
通讯作者: 王爱英,教授,E-mail:way-sh@126.com;崔金杰,研究员,E-mail:cuijj163@.com
作者简介:赵辉,硕士研究生,E-mali:zh1364223027@126.com
基金资助:
转基因生物新品种培育重大专项（2016ZX08011-002）

Analysis of Endophytic Symbiotic Bacterial Composition in Chrysoperla sinica (Tjeder)Adults with 16S rDNA Clone Library

ZHAO Hui^1,2, ZHANG Shuai², LUO Junyu², ZHANG Lijuan², WANG Aiying¹, CUI Jinjie²

1. College of life science, Shihezi University, Shehezi 832000, China;
2. State Key Laboratory of Cotton Biology/Institute of Cotton Research, Chinese Academy of Agriculture Sciences, Anyang 455000, China

Received:2017-06-14 Online:2017-12-08 Published:2017-12-16

摘要/Abstract

摘要： 中华通草蛉chrysoperla sinica（Tjeder）是农田生态系统中重要的捕食性天敌，为探索内生菌与宿主取食和生殖生理关系，本研究采用16S rDNA克隆文库的方法探究了中华通草蛉成虫内生菌的组成，分析了雌、雄成虫体内内生菌菌群结构是否存在差异。研究结果表明，中华通草蛉内生菌主要分3大类群：γ变形菌γ-Proteobacteria、α变形菌α-Proteobacteria和β变形菌β-Proteobacteria。按照属划分，中华通草蛉成虫共生细菌主要包含沃尔巴克氏体Wolbachia sp.、立克次氏体Rickettsia sp.、肠杆菌属Enterobacter sp.、免培养细菌Methyloversatilis sp.、沙雷氏菌Serratieae sp.、泛生菌属Pantoea sp.和欧文氏菌Erwinia sp.。仅在雌虫中存在Cosenzaea sp.，雄虫中存在醋酸杆菌属Acetobacter sp.、Asaia sp.、Rosenbergiella sp.、柠檬酸杆菌属Citrobacter sp.。得到中华通草蛉内生菌的优势菌群为沃尔巴克氏体Wolbachia pipientis，在雌、雄成虫中分别占23.2%和18.4%。结果证明中华通草蛉存在多种内生菌，雌、雄群体间存在差异。

关键词: 中华通草蛉, 16S rDNA文库, 内生菌

Abstract: Chrysoperla sinica (Tjeder) is an important predator in farmland ecosystem. In order to explore the effects of endophytic bacteria on host feeding and reproductive physiology, the composition of endophyte in adults C. sinica was studied by the construction and analysis of a 16S rDNA clone library. The results indicated that the bacterial could be divided into three major groups, i.e., Gamma Proteobacteria, Alpha Proteobacteria and Beta Proteobacteria. The endophyte in C. sinica adults are mainly composed of Wolbachia sp., Rickettsia sp., Enterobacter sp., uncultured Methyloversatilis sp., Serratieae sp., Pantoea sp.and Erwinia sp.according to genus. Females and males differed in that Cosenzaea sp.occurs in female only, while Acetobacter sp., Asaia sp., Rosenbergiella sp., and Citrobacter sp.exist in males only. The dominant endophytes in C. sinica adults are Wolbachia pipientis, accounting for 23.3% in males and 18.4% in females. Therefore a diversity of bacteria exists in C. sinica,and there are sexual differences in composition of the bacteria.

Key words: Chrysoperla sinica, 16S rDNA library, endophyte

中图分类号:

S476.2

赵辉, 张帅, 雒珺瑜, 张利娟, 王爱英, 崔金杰. 16S rDNA克隆文库方法分析中华通草蛉共生细菌组成[J]. 中国生物防治学报, 2017, 33(6): 849-856.

ZHAO Hui, ZHANG Shuai, LUO Junyu, ZHANG Lijuan, WANG Aiying, CUI Jinjie. Analysis of Endophytic Symbiotic Bacterial Composition in Chrysoperla sinica (Tjeder)Adults with 16S rDNA Clone Library[J]. journal1, 2017, 33(6): 849-856.

参考文献

[1] Saeed R, Razaq M. Effect of prey resource on the fitness of the predator, Chrysoperla carnea (Neuroptera:Chrysopidae)[J]. Pakistan Journal of Zoology, 2015, 47(1):103-109.
[2] Mansoor M M, Abbas N, Shad S A, et al. Increased fitness and realized heritability in emamectin benzoate-resistant Chrysoperla carnea (Neuroptera:Chrysopidae)[J]. Ecotoxicology, 2013, 22(8):1232-1240.
[3] 陈列忠, 俞晓平, 陈建明, 等. 内生菌在褐飞虱防治中的应用[J]. 农药, 2006(11):726-729.
[4] 谭周进, 肖启明, 谢丙炎, 等. 昆虫内内生菌研究概况[J]. 微生物学通报, 2005, 32(4):140-143.
[5] 薛宝燕, 程新胜, 陈树仁, 等. 昆虫内生菌研究进展[J]. 中国微生态学杂志, 2004, 16(3):189-191.
[6] Sasaki T, Kawamura M, Ishikawa H. Nitrogen recycling in the brown planthopper, Nilaparvata lugens:involvement of yeast-like endosymbionts in uric acid metabolism[J]. Journal of Insect Physiology, 1996, 42(2):125-129.
[7] 李献辉, 李保平. 蚜虫与体内布赫纳氏菌及其次生内生菌的相互关系[J]. 应用昆虫学报, 2006, 43(4):443-447.
[8] 徐红星, 郑许松, 刘淑平, 等. 昆虫内内生菌在昆虫防御中的作用[J]. 应用昆虫学报, 2009, 46(3):350-354.
[9] 杨义婷. 烟粉虱隐种内内生菌种类多样性分析及其优势内生菌对烟粉虱生长发育的影响[D]. 重庆:西南大学, 2015.
[10] 付海滨, 丛斌, 戴秋慧. 赤眼蜂内生菌沃尔巴克氏体及其对宿主影响[J]. 中国生物防治学报, 2005, 21(2):70-73.
[11] Sinkins S P, Braig H R, O'Neill S L. Wolbachia pipientis:bacterial density and unidirectional cytoplasmic incompatibility between infected populations of Aedes albopictus[J]. Experimental Parasitology, 1995, 81(3):284-291.
[12] 朱卓琳. 日本弓背蚁室内饲养种群消化道细菌组成及其多样性研究[D]. 杨凌:西北农林科技大学, 2015.
[13] Douglas A E. Mycetocyte symbiosis in insects[J]. Biological Reviews, 1989, 64(4):409-434.
[14] Franzini P Z, Ramond J B, Scholtz C H, et al. The gut microbiomes of two pachysoma macleay desert dung beetle species (Coleoptera:Scarabaeidae:Scarabaeinae) feeding on different diets[J]. PLoS ONE, 2016, 11(8):e0161118.
[15] 王甸洪, 吴伟坚, 符悦冠. 螺旋粉虱成虫体内细菌群落多样性的PCR-DGGE和16S rRNA文库序列分析[J]. 昆虫学报, 2012, 55(7):772-781.
[16] Huang S W, Sheng P, Zhang H Y. Isolation and identification of cellulolytic bacteria from the gut of Holotrichia parallela larvae (Coleoptera:Scarabaeidae)[J]. International Journal of Molecular Sciences, 2012, 13(3):2563-2577.
[17] Dillon R J, Webster G A. Diversity of gut microbiota increases with aging and starvation in the desert locust[J]. Antonie Van Leeuwenhoek, 2010, 97(1):69-77.
[18] 刘宁, 刘玉升, 付卫东, 等. 中华通草蛉越冬成虫肠道可培养好氧细菌多样性分析[J]. 应用昆虫学报, 2012, 49(5):1287-1292.
[19] 李建婷, 纪树兰, 刘志培, 等. 16S rDNA克隆文库方法分析好氧颗粒污泥细菌组成[J]. 环境科学研究, 2009, 22(10):1218-1223.
[20] Brown B P, Wernegreen J J. Deep divergence and rapid evolutionary rates in gut-associated Acetobacteraceae of ants[J]. BMC Microbiology, 2016, 16(1):1-17.
[21] 相辉, 黄勇平. 肠道微生物与昆虫的共生关系[J]. 应用昆虫学报, 2008, 45(5):687-693.
[22] 王斌, 陈福生. 醋酸菌的分类进展[J]. 中国酿造, 2014, 33(12):1-10.
[23] Kommanee J, Akaracharanya A, Tanasupawat S, et al. Identification of Acetobacter, strains isolated in Thailand based on 16S-23S rRNA gene ITS restriction and 16S rRNA gene sequence analyses[J]. Annals of Microbiology, 2008, 58(2):741-747.
[24] Chouaia B, Rossi P, Montagna M, et al. Molecular evidence for multiple infections as revealed by typing of Asia bacterial symbionts of four mosquito species[J]. Applied and Environmental Microbiology, 2010, 76(22):7444-7450.
[25] Crotti E, Rizzi A, Chouaia B, et al. Acetic acid bacteria, newly emerging symbionts of insects[J]. Applied and Environmental Microbiology, 2010, 76(21):6963-6970.
[26] Alvarez-Perez S, Lievens B, Jacquemyn H, et al. Acinetobacter nectaris sp. nov. and Acinetobacter boissieri sp. nov. isolated from floral nectar of wild Mediterranean insect-pollinated plants[J]. International Journal of Systematic and Evolutionary Microbiology, 2013, 63(4):1532.
[27] Ventorino V, Robertiello A, Viscardi S, et al. Bio-based chemical production from Arundo donax feedstock fermentation using Cosenzaea myxofaciens BPM1[J]. Bioresources, 2016, 11(3):6566-6581.
[28] 向芸庆, 王晓强, 冯伟, 等. 不同饲料饲养家蚕其肠道微生态优势菌群类型的组成及差异性[J]. 生态学报, 2010, 30(14):3875-3882.
[29] 刘莉, 王中康, 俞和韦, 等. 贡嘎蝠蛾幼虫肠道细菌多样性分析[J]. 微生物学报, 2008, 48(5):616-622.
[30] 何正波, 殷幼平, 曹月青, 等. 桑粒肩天牛幼虫肠道菌群的研究[J]. 微生物学报, 2001, 41(6):741-744.
[31] Behar A, Yuval B, Jurkevitch E. Enterobacteria-mediated nitrogen fixation in natural populations of the fruit fly Ceratitis capitata[J]. Molecular Ecology, 2005, 14(9):2637-2643.
[32] 陈虹, 梅建凤, 闵航. 白蚁肠道微生物[J]. 微生物学杂志, 2005(2):75-79.
[33] Hongoh Y, Sasaki T, Ishikawa H. Cloning, sequence analysis and expression in Escherichia coli of the gene encoding a uricase from the yeast-like symbiont of the brown planthopper, Nilaparvata lugens[J]. Insect Biochemistry and Molecular Biology, 2000, 30(2):173-182.
[34] Hertig M, Wolbach S B. Studies on rickettsia-like micro-organisms in insects[J]. Journal of Medical Research, 1924, 44(3):329.
[35] Werren J H, Windsor D, Guo L. Distribution of wolbachia among neotropical arthropods[J]. Proceedings of the Royal Society B Biological Sciences, 1995, 262(1364):197-204.
[36] 廖珊, 康琳, 陈小爱, 等. Wolbachia在灰飞虱体内的分布[J]. 复旦学报(自然科学版), 2001, 40(5):539-543.

16S rDNA克隆文库方法分析中华通草蛉共生细菌组成

Analysis of Endophytic Symbiotic Bacterial Composition in Chrysoperla sinica (Tjeder)Adults with 16S rDNA Clone Library

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 10

编辑推荐

Metrics

本文评价

联系我们

[1]	孙艺昕, 门兴元, 李超, 于毅, 吕素洪, 孙廷林, 叶保华, 李丽莉. 高抗虫玉米内生真菌对玉米蚜生长、繁殖及取食选择的影响[J]. 中国生物防治学报, 2020, 36(2): 184-188.
[2]	孙艺昕, 门兴元, 李超, 于毅, 吕素洪, 孙廷林, 叶保华, 李丽莉. 抗玉米蚜玉米内生菌的筛选[J]. 中国生物防治学报, 2019, 35(4): 570-575.
[3]	孙艺昕, 门兴元, 李超, 于毅, 夏小菊, 吕素洪, 叶保华, 李丽莉. 双委夜蛾高致病力球孢白僵菌的筛选及在玉米内定殖率的测定[J]. 中国生物防治学报, 2019, 35(1): 70-74.
[4]	畅涛, 冯中红, 杨成德, 薛莉. 一株抗马铃薯坏疽病生防细菌B-401产抑菌物质条件优化及稳定性[J]. 中国生物防治学报, 2016, 32(3): 365-371.
[5]	赵龙飞, 徐亚军, 常佳丽. 轮纹病菌拮抗性大豆根瘤内生菌的筛选、抗性和促生作用[J]. 中国生物防治学报, 2016, 32(3): 396-405.
[6]	肖军, 杨镇, 陈珣, 王红, 肇莹, 刘国丽, 杨涛. 内生菌醇提取物对水稻抗稻瘟病的诱导作用[J]. , 2015, 31(3): 433-438.
[7]	杨洪凤1 , 余向阳2 , 薛雅蓉1 , 刘常宏1 . 内生解淀粉芽孢杆菌CC09在小麦根部定殖的电镜观察及防病效果[J]. , 2014, 30(6): 839-844.
[8]	王国平1 , 王丽薇1,2 . 一株罗汉松内生真菌及其活性代谢产物[J]. , 2014, 30(2): 287-292.
[9]	林吉恒, 王睿之, 马风兰, 陈美龄, 周陈伟, 徐娟, 阮晖, 何国庆. 植物益生菌对植物的益生效应及其应用[J]. , 2010, 26(增刊): 100-105.
[10]	尹华群1 ;易有金1 ;罗宽1 ;匡传富2 ;邓正平2 . 烟草青枯病内生拮抗细菌的鉴定及小区防效的初步测定 [J]. , 2004, 20(3): 219-220.