基于核糖体RNA高通量测序研究转Bt基因棉花对根际土壤微生物群落结构的影响

随着转Bt基因棉花的大规模种植，转Bt基因棉花对生态系统的影响受到高度重视，这其中一个重要研究领域即是对土壤生态系统特别是对土壤微生物群落结构的影响。在前期工作的基础上，本项目拟通过基于核糖体RNA的Illumina MiSeq高通量测序技术，结合荧光定量PCR技术，分析不同微生境（主根和侧根）、不同品种、不同年限、不同生长期和不同土壤类型的转Bt基因棉花对根际土壤细菌、真菌和古菌的群落结构组成和群体数量变化情况，研究群落内优势菌群和稀有菌群的多度和丰富度对转Bt基因棉花种植的响应规律，结合Bt蛋白在根系和根际土壤中表达的时空差异及不同环境因子变化情况，采用Pearson相关性和冗余分析（RDA）（或典范对应分析（CCA）），分析根际土壤微生物群落结构变化与环境因子的相关性，较全面、系统的了解转Bt基因棉花对根际土壤微生物群落结构的影响，为转Bt基因棉花的生物安全评价提供更充足的数据支持。

The rapid development of transgenic Bt cotton has raised concerns over the potential impacts of these crops on environment. The unpredicted and complex impacts of transgenic Bt cotton on soil ecosystem, especially microbial community structure, has become one of the hot research directions. In this study, using ribosomal RNA MiSeq sequencing and real-time PCR, we will evaluate the effect of transgenic Bt cotton on soil microbial community structure and population size under the condition of different microhabitats (taproot and lateral root), different varieties, different years, different growth stages and different soil types. Different response of species richness and abundance of dominant and rare taxa to transgenic Bt cottons will be analyzed, considering temporal and spatial variation of Bt protein released from transgenic Bt cotton, and different environmental factors. We will analyze the correlation between rhizosphere soil microbial community structure and environmental factors, using Pearson correlation and redundancy analysis (RDA) (or canonical correspondence analysis (CCA)), which provides more comprehensive understanding of the effect of transgenic Bt cotton on the microbial community structure, and provides more adequate data support to the soil ecological security assessment.