›› 2018, Vol. 61 ›› Issue (2): 200-208.doi: 10.16380/j.kcxb.2018.02.006

• 研究论文 • 上一篇    下一篇

基于16S rRNA高通量测序的灰飞虱体内细菌群落结构及多样性分析

杨晓晴, 王正亮, 王天召, 俞晓平*   

  1. (中国计量大学生命科学学院, 浙江省生物计量及检验检疫技术重点实验室, 杭州 310018)
  • 出版日期:2018-02-20 发布日期:2018-02-20

Analysis of the bacterial community structure and diversity in the small brown planthopper, Laodelphax striatellus (Hemiptera: Delphacidae) by 16S rRNA high-throughput sequencing

YANG Xiao-Qing, WANG Zheng-Liang, WANG Tian-Zhao, YU Xiao-Ping*   

  1. (Zhejiang Provincial Key Laboratory of Biometrology and Inspection and Quarantine, College of Life Sciences, China Jiliang University, Hangzhou 310018, China)
  • Online:2018-02-20 Published:2018-02-20

摘要: 【目的】探明灰飞虱Laodelphax striatellus体内细菌的群落结构和种类多样性。【方法】采用Illumina MiSeq测序平台对新羽化24 h的灰飞虱雌雄成虫体内细菌16S rRNA的V3-V4 变异区序列进行高通量测序,应用USEARCH 和Silva等软件和数据库来统计样品序列数目和操作分类单元(operational taxonomic unit, OTU) 数量,分析灰飞虱雌雄成虫体内细菌的种类组成、丰度、Alpha 多样性及其差异。【结果】灰飞虱雌、雄成虫样本分别获得29 333和25 919条有效序列,根据序列相似性进行聚类分析分别获得55和57个OTUs。其中,雌、雄成虫两类样本共有OTU数目20个,特有的OTU数目分别为35和37个。基于OTU物种分类分析,雌雄成虫样本中细菌种类一共覆盖7个门、15个纲、23个目、33个科、56个属和73个种。在门、纲、目和科分类阶元上,雌雄成虫两类样本均以变形菌门(Proteobacteria, 雌99.96%/雄99.16%)、α-变形菌纲(Alphaproteobacteria, 雌97.76%/雄97.84%)、红螺菌目(Rhodospirillales, 雌83.92%/雄53.21%)和醋酸杆菌科(Acetobacteraceae, 雌83.90%/雄53.17%)中的细菌为优势菌。在属分类阶元上,灰飞虱雌雄成虫体内优势菌为醋酸杆菌科未分类属(unclassified Acetobacteraceae),沃尔巴克氏体属Wolbachia次之,后者在雌、雄成虫体内丰度分别为13.81%和44.52%。在种水平上,灰飞虱雌、雄成虫体内特有细菌分别为21和32个种,但两类样本中性别特有种的丰度普遍极低,均不足0.5%。【结论】灰飞虱雌雄成虫间细菌群落组成和种类多样性存在差异。本研究为进一步挖掘灰飞虱体内关键微生物并解析其生物学功能及其利用等方面奠定了基础。

关键词:  灰飞虱, 16S rRNA, 高通量测序, 细菌, 多样性

Abstract: 【Aim】 To reveal the bacterial community structure and diversity in the small brown planthopper (SBPH), Laodelphax striatellus. 【Methods】 The Illumina MiSeq high-throughput sequencing platform was used to sequence the V3-V4 regions of the bacterial 16S rRNA gene in the newly emerged female and male adults (24 h after emergence) of L. striatellus. The numbers of sequences and operational taxonomic units (OTUs) and the abundance and diversity of the bacterial communities were analyzed using the related softwares and databases including USEARCH and Silva. 【Results】 A total of 29 333 valid tags and 55 OTUs were obtained from female adults of L. striatellus, while the numbers of valid tags and OTUs from male adults were 25 919 and 57, respectively. Among them, 20 OTUs were shared between the two samples of female and male adults, and the unique OTU numbers for female and male adults were 35 and 37, respectively. The total OTUs of the two samples were annotated into 7 phyla, 15 classes, 23 orders, 33 families, 56 genera, and 73 species. At the levels of phylum, class, order and family, the most dominant bacteria in both samples of female and male adults were from Proteobacteria (99.96% for female and 99.16% for male), Alphaproteobacteria (97.76% for female and 97.84% for male), Rhodospirillales (83.92% for female and 53.21% for male), and Acetobacteraceae (83.90% for female and 53.17% for male), respectively. At the genus level, the dominant bacteria with the highest abundance belonged to an undetermined genus in Acetobacteraceae (unclassified Acetobacteraceae). The second dominant genus was Wolbachia, accounting for 13.81% and 44.52% in female and male adults, respectively. At the species level, the numbers of specific bacterial species in male and female adults were 21 and 32, respectively. However, the abundance of the sex-specific species in the two samples was generally low (less than 0.5%). 【Conclusion】 The results indicate that the bacteria in L. striatellus adults are diverse and the bacterial community structure and diversity are different between female and male adults. This study lays the foundation for further studies on the excavation and utilization of microorganism resources for the biocontrol of L. striatellus.  

Key words:  Laodelphax striatellus, 16S rRNA, high-throughput sequencing, bacteria, diversity