基于荧光定量PCR技术分析滇池沉积物产甲烷菌空间分布特征  

The Spatial Distribution Characteristics of Methanogens in the Sediments of Dianchi Lake by Fluorescence Quantitative PCR

作　　者：陈悦 罗明没 李尚磷 李春艳[1] 冉玉芳 李玮 胡斌[1] CHEN Yue;LUO Mingmo;LI Shanglin;LI Chunyan;RAN Yufang;LI Wei;HU Bin(School of Ecology and Environment,Yunnan University,Key Laboratory of Plateau Ecology and Restoration of Degraded Environment,Kunming 650091,China;Key Laboratory of Plateau Mountain Ecology and Degradation Environmental Remediation of Yunnan Province,Kunming 650091,China)

机构地区：[1]云南大学生态与环境学院,高原湖泊生态与治理研究院,云南昆明650091 [2]云南省高原山地生态与退化环境修复重点实验室,云南昆明650091

出　　处：《云南农业大学学报（自然科学版）》2021年第1期132-139,共8页Journal of Yunnan Agricultural University:Natural Science

基　　金：国家自然科学基金项目(31700411);云南省应用基础研究计划项目(2018FD007)。

摘　　要：【目的】研究滇池沉积物产甲烷菌的空间分布特征。【方法】采集滇池草海和外海的4个不同面源污染区域沉积物,通过沉积物DNA提取、PCR扩增及产物纯化、质粒构建及阳性验证、标准曲线制作和荧光定量PCR方法,定量研究基于mcrA基因的产甲烷菌丰度及其分布特征。【结果】滇池沉积物产甲烷菌丰度范围为5.91×107~7.74×108 copies/g。产甲烷菌的拷贝数在各个采样区域差异显著(F=10.3006,P=0.0012),并具有空间分布特征,即外海中>外海南>草海>外海北,且外海南部和中部的产甲烷菌丰度与草海和外海北部的产甲烷菌丰度具有显著差异(P<0.05)。【结论】滇池沉积物具有较高的产甲烷丰度,且产甲烷菌在滇池的分布空间异质性很大,临近农业面源污染区域的外海中部和临近磷、钾厂的外海南部比草海和外海北部水华密集区具有较高的产甲烷菌丰度。[Purpose]To study the abundance and distribution characteristics of methanogens abundance in the sediments of Dianchi.[Method]The sediments were collected from 4 different non-point source contaminated areas in Caohai and Waihai of Dianchi Lake.The distribution characteristics of methanogen abundance was detected by fluorescence quantitative PCR procedure,of which included the sediment DNA extraction,PCR amplification and product purification,plasmid construction and positive verification,and production of standard curve.[Results]The abundance of methanogenes in the sediments of Dianchi Lake ranged from 5.91×107-7.74×108 copies/g.The copy number of methanogenic was significantly different in each sampling area(F=10.3006,P=0.0012),and their spatial distribution characteristics,that is,the central part of Waihai>southern part of Waihai>Caohai>northern part of Waihai(P<0.05).[Conclusion]Dianchi Lake sediments had a high methaneogen abundance and spatital heterogeneous,the abundances of methanogens in the middle part(adjacent to the agricultural non-point source pollution area)and south part(adjacent to the phosphorus and potassium plant)of the Waihai were significantly higher than that in the Caohai and north of Waihai(algae enriched area).

关 键 词：产甲烷菌 高原湖泊 荧光定量PCR 沉积物 MCRA 

分 类 号：Q939.9[生物学—微生物学]