基于高通量测序的不同轮作方式下烟田土壤细菌群落组成分析

在自然界中,土壤微生物与植物之间存在着密切的联系,土壤微生物依赖植物提供的养分而生存,同时对植物的生长发育起着促进作用。本研究利用Illumina Hi Seq高通量测序技术,提取西双版纳烟田烟草连作、田菁轮作和青蒿轮作3种种植模式下土壤样品的总DNA,通过PCR方法扩增构建16S r RNA基因文库。结合生物信息学方法分析土壤细菌16S r RNA基因V3~V4区的多样性指数、群落结构等,以确定烟田更好的种植模式。所有样本共测序分析获得4 898个细菌分类单元OTU,经琢多样性和茁多样性分析表明3个处理下细菌群落多样性存在显著差异,并且在对照组中的细菌群落丰富度最高,青蒿轮作中的细菌群落丰富度最低。样本中优势门类为变形菌门(Proteobacteria)、酸杆菌门(Acidobacteria)、拟杆菌门(Bacteroidetes)和绿弯菌门(Chloroflexi),其中变形菌门(Proteobacteria)相对丰度33.49%占绝对优势。优势属类为硝化螺旋菌属、Kaistobacter、红游动菌属和黄杆菌属,可促进氮素转化的硝化螺旋菌属在田菁中的相对丰度最高,烟株青枯病致病菌罗尔斯通菌属在青蒿轮作中相对丰度最低;拮抗黑胫病的假单胞杆菌属在青蒿轮作中相对丰度最高。本研究将对烟草轮作方式和轮作作物的选择提供分子层面的支持。

Analysis of Bacterial Community Composition in Tobacco Field under Different Rotation Patterns Based on High-Throughput Sequencing

In nature,there is a close relationship between soil microbes and plants.Soil microbes depend on the nutrients provided byplants to survive,and at the same time,play a role in promoting plant growth and development.In this study,Illumina Hi Seq high-throughput sequencing technology was used to extract the total DNA of soil samples from three cropping models of tobacco continuous cropping,Sesbania rotation and Artemisia rotation in Xishuangbanna,and the 16 S r RNA gene library was amplified by PCR(Polymerase chain reaction).Combined with bioinformatics methods,the diversity index and community structure of soil bacteria 16 S r RNA gene V3~V4 region were analyzed to determine the better planting pattern in tobacco fields.Through high-throughput sequenced and analyzed,a total of 4898 bacterial taxonomic units were obtained.The alpha diversity and beta diversity analysis showed that there were significant differences in bacterial community diversity under the three treatments,and the bacterial community richness was the highest in the control group,while the lowest in the Artemisia rotation.The dominant species in the sample were Proteobacteria,Acidobacteria,Bacteroidetes and Chloroflexi,and the relative abundance of Proteobacteria was 33.49%.The dominant genus were Nitrospira,Kaistobacter,Rhodoplanes and Flavobacterium.Nitrospira had the highest relative abundance in Sesbania rotation,which promotes nitrogen conversion.The genus Ralstonia had the lowest relative abundance in Artemisia rotation,which could cause bacterial wilt.Pseudomonas was the most abundant in Artemisia rotation,which could antagonize black shank disease.The results of this study will provide molecular support and theoretical basis for tobacco crop rotation and crop rotation selection.