基于高通量培养分析稻虾共作模式土壤的细菌群落

Comparison of Soil Bacterial Communities in Rice-Crayfish Cocropping Based on High-Throughput Culturable Method

高通量测序技术的发展发现并鉴定了土壤微生物的未培养物种,然而,仅依赖非培养的高通量测序无法获得发挥关键功能的微生物培养物,限制了对其功能的验证及其应用,这对关键功能微生物的机理性研究和应用具有一定的局限性.本研究以稻虾共作和水稻单作两种稻作模式的土壤样品为研究对象,分别对比了传统平板涂布、96孔培养板划线分离以及将非培养和培养法相结合的———“96孔高通量培养法”3种方法的土壤细菌分类差异、菌群多样性、组成特异性以及细菌功能特征.结果表明,相对于传统涂布培养法,96孔高通量培养法所获取的细菌数量高约5倍,能鉴定到属的培养物高约4倍,预测功能鉴定提高约2倍.由此可见,“96孔高通量培养法”大大提高了对土壤细菌培养物的获取和鉴定率.本研究弥补了传统培养法的获得率低的不足,拓宽了高通量培养技术的应用领域,同时,也为进一步揭示稻虾共作模式参与土壤生态变化过程的微生物机制提供研究基础.

With the development of high-throughput sequencing technology,more and more unculturable species of soil microorganisms have been discovered and identified.However,high-throughput sequencing cannot isolate and purify a single strain with key functions from massive databases,which has certain limitations on the mechanical research and application of key function microorganisms.In this study,soil samples of rice-crayfish cocropping and rice-single cropping were taken as the research object.The differences were compared in soil bacterial classification,microbial diversity,composition specificity,and microbial functional characteristics of traditional spread plate method,96-well culture plate in situ scribing separation,and the combination of high-throughput and cultivable methods—the“96-well high-throughput cultivable method”.The results showed that comparing with the traditional spread plate method,the number of bacteria obtained by 96-well high-throughput culture method was increased by about 5 times,the identification of genus level was increased by about 4 times,and the identification of function was increased by about 2 times.It can be seen that the 96-well high-throughput culturable method greatly improves the“resolution”of the separation and identification of soil microorganisms.The results of this study made up for the shortage of strains that could not be obtained by traditional spread plate method,expanded the application field of high-throughput culturable technology,and also provided a research basis for further revealing the microbial mechanism involved in the soil ecological change process of rice-crayfish cocropping model.