应用Illumina MiSeq高通量测序技术分析玉米内生细菌多样性

为了调查玉米内生细菌种类多样性。应用高通量测序技术测定玉米内生细菌的16S rDNA-V4变异区序列,应用Qiime和Mothur等软件整理和统计样品序列数目和操作分类单元(OTUs)数量,分析物种的丰度、分布和Alpha多样性,以及物种丰富度的差异。本研究获得用于分析的有效序列和OTU数为96334/156;稀疏曲线表明测序深度充分,OTU的数量接近于饱和。玉米(样品名YM)的Chao1指数为156.0,Shanno多样性指数为1.211。玉米内生细菌分布于以下6个属:鞘氨醇单胞菌属(Sphingomonas,37.78%)、盐单胞菌属(Halomonas,33.33%)、假单胞菌属(Pseudomonas,13.33%)、希瓦氏菌属(Shewanella,6.67%)、甲基杆菌属(Methylobacterium,4.44%)、土地杆菌属(Pedobacter,4.44%);玉米内生细菌优势菌属为鞘氨醇单胞菌属(Sphingomonas,37.78%)、盐单胞菌属(Halomonas,33.33%)。Illumina Mi Seq高通量测序技术为植物内生细菌的研究提供了更加准确、科学的数据资源。

Study on the Diversity of Endophytic Bacteria in Maize using Illumina MiSeq High-throughput Sequencing System

To investigate the diversity of the endophytic bacterial communities, high-throughput sequencing technique was used to measure the sequence of 16S rDNA-V4 variable region of endophytic bacteria in maize. QIIME and Mothur software programs were employed to sort and calculate the number of sequences and operational taxonomic units (OTUs) for each sample. Thereafter, the abundance, distribution, and alpha diversity index of species and the differences of species abundance among the samples were analyzed. The number of effective sequences and OTUs for each sample was 96334/156; the rarefaction curves showed a sufficient sequencing depth, and the number of OTUs was close to saturation. The Chao1 and Shannon indices of sample YM were 156.0 and 1.211, respectively. The endophytic bacteria in maize belonged to the following six genera: Sphingomonas (37.78%), Halomonas (33.33%), Pseudomonas (13.33%), Shewanella (6.67%), Methylobacterium (4.44%), and Pedobacter (4.44%). The dominant species were Sphingomonas and Halomonas, and accounted for 37.78% and 33.33%, of the total bacteria identified, respectively. Illumina MiSeq high-throughput sequencing system provides a more accurate andscientific data resource for the study of endophytic bacteria in plants.