›› 2017, Vol. 60 ›› Issue (6): 632-641.doi: 10.16380/j.kcxb.2017.06.003

• 研究论文 • 上一篇    下一篇

饲喂玉米秸秆的白星花金龟幼虫肠道细菌多样性

田小燕1,2, 宋福平2, 张杰2, 刘荣梅3, 张兴鹏3, 段江燕1,*, 束长龙2,*   

  1. (1. 山西师范大学生命科学学院, 山西临汾 041004; 2. 中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193; 3. 东北农业大学生命科学学院, 哈尔滨 150030)
  • 出版日期:2017-06-20 发布日期:2017-06-20

Diversity of gut bacteria in larval Protaetia brevitarsis (Coleoptera: Scarabaedia) fed on corn stalk

TIAN Xiao-Yan1, 2, SONG Fu-Ping2, ZHANG Jie2, LIU Rong-Mei3, ZHANG Xing-Peng3, DUAN Jiang-Yan1, *, SHU Chang-Long 2, *   

  1. (1. School of Life Sciences, Shanxi Normal University, Linfen, Shanxi 041004, China; 2. State Key Laboratory of Biology for Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China; 3. School of Life Sciences, Northeast Agricultural University, Harbin 150030, China)
  • Online:2017-06-20 Published:2017-06-20

摘要: 【目的】分析比较饲喂玉米Zea mays L.秸秆的白星花金龟Protaetia brevitarsis幼虫中肠、后肠中细菌群落结构,挖掘肠道细菌在白星花金龟消化秸秆中的功能。【方法】分别提取健康白星花金龟3龄幼虫10个样品(5个中肠内含物、5个后肠内含物)肠道细菌总DNA,利用Illumina HiSeq技术对白星花金龟幼虫中肠和后肠细菌的16S rDNA V4区序列进行测序,统计样品操作分类单元(operational taxonomic units, OTUs)数量,分析物种丰度、α多样性和β多样性,并将细菌群落信息映射到KEGG基因组数据库,进行细菌群落功能挖掘。【结果】共获得白星花金龟3龄幼虫肠道细菌1 062 897高质量reads,聚类为2 441个OTUs,总共注释到27个门,51个纲,77目,168科,326属。在属的分类阶元上,中肠中以假单胞菌属Pseudomonas为最优势属,后肠中以脱硫弧菌属Desulfovibrio为最优势属。α多样性分析表明,后肠群落多样性更丰富;β多样性分析结果显示,后肠微生物群落更稳定。细菌群落功能分析结果显示,中肠、后肠细菌群落都含有降解秸秆相关的酶系,其中,中肠降解木质素相关的通路更丰富,后肠降解纤维素的酶更多。【结论】采用Illumina HiSeq测序技术明确了饲喂玉米秸秆的白星花金龟幼虫中肠、后肠细菌群落结构与特点,进一步发现了肠道细菌在白星花金龟秸秆消化过程中发挥的作用,相关研究结果将为白星花金龟消化秸秆的机制深入研究提供参考,为我国秸秆资源化利用提供新思路。

关键词: 白星花金龟, 肠道细菌, 16S rDNA, V4区, 细菌群落, 群落多样性, 功能分析

Abstract: 【Aim】 To analyze and compare the structure of the bacterial communities in the midgut and hindgut of Protaetia brevitarsis larvae fed on Zea mays stalk, and to further explore the function of these gut bacteria in stalk digestion of this insect. 【Methods】 The total DNA was extracted from 10 samples of healthy gut bacteria (5 midgut contents and 5 hindgut contents) of the 3rd instar larvae of P. brevitarsis. The V4 region of 16S rDNA of the bacterial communities in the midgut and hindgut of P. brevitarsis was sequenced by Illumina HiSeq techniques, and the number of operational taxonomic units (OTUs), species composition abundance, alpha diversity and beta diversity were analyzed. The gut bacteria community data were mapped to KEGG genome database for further bacterial community function discovery. 【Results】 Totally, 1 062 897 high quality reads of gut bacteria of the 3rd instar larvae of P. brevitarsis were produced and clustered to 2 441 OTUs, which were annotated into 27 phylums, 51 classes, 77 orders, 168 families and 326 genera. At the genus level, Pseudomonas and Desulfovibrio were the most dominant genus in the midgut and hindgut, respectively. Alpha diversity data showed that the bacterial communities in the hindgut were more abundant than those in the midgut, and the beta diversity analysis indicated that the bacterial communities in the hindgut were more stable than those in the midgut. Function analysis of bacterial communities showed that both the bacterial communities in the midgut and hindgut contained stalk degradation enzymes, and the lignin degradation pathways were more abundant in the midgut while the cellulose degradation enzymes more abundant in hindgut. 【Conclusion】 The structure and characteristics of the bacterial communities in the midgut and hindgut of P. brevitarsis larvae fed on corn stalk were clarified, and their functions in helping P. brevitarsis digest stalk were discovered. These results will provide references for the further study on stalk digestion mechanism of this insect, and offer a new idea for utilization of crop straw resources in China.

Key words: Protaetia bevitasis, gut bacteria, 16S rDNA, V4 region, bacterial community, community diversity, function analysis