基于高通量测序的裕民红花根际土壤细菌群落特征分析

High throughput sequencing analysis of bacterial communities in Yumin safflower

摘要:

【背景】新疆裕民县是我国最大的红花种植基地，其独特的地理环境和气候条件适合红花生长且有利于红花良好品质的形成。【目的】探究新疆裕民县山地栽培红花不同生育期的根际土壤细菌群落及其与环境因子之间的关系，为揭示道地药材“道地性”的本质与规律提供科学依据。【方法】在3块裕民栽培红花样地中分别采取营养生长期与生殖生长期的根际、非根际土壤，测定其土壤性质，并提取基因组DNA对16S rRNA基因的V4高变区进行PCR扩增构建文库，经过Illumina HiSeq高通量测序平台测序后使用生物信息学技术分析土壤细菌的丰富度、多样性指数和群落结构，然后分析多样性指数与环境因子之间的相关性。【结果】从36个红花土壤样品中共得到10 303个细菌操作分类单元(Operational taxonomic unit，OTU)，分属于47门102纲201目381科738属405种，其中放线菌门(Actinobacteria，32.9%)和变形菌门(Proteobacteria，28.7%)是优势菌群。方差分析显示丰富度指数Chao1与多样性指数Shannon在4个样品分组的细菌群落中无显著差异，而主坐标分析(Principal coordinates analysis，PCoA)和属水平的物种丰度分布情况表明各样地之间的细菌群落构成差异明显。【结论】经分析，红花根际土壤中存在大量适应环境变化的细菌，这些细菌的存在可能对红花道地性的形成有积极作用。此外，土壤性质、植物的生长阶段和根系分泌物可能通过影响红花根际细菌群落来致力于红花道地性的形成。

Abstract:

[Background] Xinjiang Yumin county is the largest safflower planting base of China, its unique geographical environment and climatic conditions suitable for the growth and it is beneficial to formation of quality of safflower. [Objective] The purpose of this study was to explore the relationship between the bacterial community of safflower rhizosphere soil and its environment factors in different growth periods in Xinjiang Yumin hilly area, which may help to provide scientific basis for revealing the nature and low of geoherbalism to genuine regional drugs. [Methods] Took the rhizosphere soil and bulk soil of the vegetative growth stage and reproductive growth stage in the 3 sample plots of Yumin cultivated safflower respectively, determined the soil properties and extracted the genomic DNA, and then established libraries through PCR amplification of 16S rRNA V4 region. Abundance, diversity and structure of soil bacteria were analyzed by bioinformatics technology after sequenced by Illumina HiSeq high-throughput sequencing platform. [Results] A total of 10 303 operational taxonomic units (OTUs) were obtained from 36 soil samples of safflower, and it have identified belonging to 405 species, 738 genera, 381 families, 201 orders, 102 classes, 47 phyla, in which the Actinobacteria (32.9%) and Proteobacteria (28.7%) were predominant. ANOVA analysis showed that there was no significant difference between the richness index Chao1 and the diversity index Shannon in the bacterial communities of the four group samples. However, the principal coordinate analysis (PCoA) and the distribution of species abundances at the level of bacterial genus revealed that there were significant differences in the composition of bacteria communities between different sample plots. [Conclusion] There are a lot to adapt to the changing environment of bacteria in rhizosphere soil of safflower, the presence of these bacteria may have positive effects on forming geoherbalism of the safflower. In addition, soil properties, plant growth stage and root exudates are the possible factors who committed to form the geoherbalism of safflower by effects the rhizosphere bacterial community.