• 研究简报 •

杨树人工林土壤微生物群落4种提取方法比较

1. 1. 中国林业科学研究院亚热带林业研究所 杭州 311400;
2. 中国科学院遗传与发育生物学研究所 北京 100020
• 收稿日期:2017-05-30 修回日期:2017-11-22 出版日期:2018-09-25 发布日期:2018-09-10
• 基金资助:
中国林业科学研究院中央级公益性科研院所基本科研业务费专项资金（子课题）（重点项目CAFYBB2017ZB001）；2016年度留学回国人员科技活动项目择优资助经费和国家自然科学基金优秀青年项目（31722014）。

Comparison of Four Extraction Methods of Soil Microbiome in Poplar Plantation

Qin Yuan1,2, Pan Xueyu1, Jin Wei1, Chen Lianqing1, Yuan Zhilin1

1. 1. Research Institute of Subtropical Forestry, CAF Hangzhou 311400;
2. Institute of Genetics and Developmental Biology, CAS Beijing 100020
• Received:2017-05-30 Revised:2017-11-22 Online:2018-09-25 Published:2018-09-10

Abstract: [Objective] Efficient extraction of soil microbiome is a crucial step for investigating the physiology and ecology of microbial communities as well as interactions between plant and its associated microbiota. In this work, we compared the microbial diversity and metabolic activity of soil microbiome in poplar plantation through four extraction techniques. The aim is to provide the efficient and reliable means that enables extracted samples to represent for the natural soils. This technique can be further applied in tree rhizosphere microbiome engineering.[Method] The bulk soils in poplar plantation were collected. The soil samples were pretreated with two physical dispersing methods of blending and sonication and two kinds of chemical dispersants of sodium pyrophosphate and MES monohydrate, and then the microbial pellets were obtained through two-step centrifugation. Microbial community structure and diversity, viable counts and the community level physiological profiling in the extracted microbial cell samples were evaluated and compared through the integration of 16S rRNA and ITS high-throughput pyrosequencing, BIOLOG analysis (ECO and FF plates) as well as the colony-forming unit (CFU) assay of culturable fungi and bacteria.[Result] Results showed that, compared to the natural soil microbiome, all treatments had slight negative influence on the species diversity (OTUs and diversity index). When soils were treated with sodium pyrophosphate and blending, the diversity and community structure of the extracted samples were relatively closer to natural bulk soils. In addition, the CFU counts and metabolic activity (carbon utilization capacity) were relatively higher than other treatments. On the contrary, with the sonication treatment the diversity of bacterial species was the lowest, and the number of cultivated fungi was significantly lower than that of two treatments with sodium pyrophosphate and blending ( P <0.05). Moreover, the microbial pellet also showed a lower metabolic activity evidenced by the BIOLOG analysis. The community structure of pellet samples extracted with MES monohydrate differed greatly compared to the natural soils, and the number of culturable bacteria was significantly less than other treatments (P <0.05).[Conclusion] The four extraction methods have different effects on the structure, composition and metabolic activity of soil microbial community, in which the sonication and MES monohydrate treatments maintain a relatively low level of microbial diversity and metabolic activity compared to the natural soils. Therefore, we suggest that the sodium pyrophosphate and physical blending, either alone or in combination, will be good alternatives for extracting the soil microbiome efficiently.