缙云山森林植被类型对土壤产N2O相关功能微生物的影响

Effects of Different Forest Vegetation Types on Soil Nitrogen-Related Microbial Communities and Functions in Jinyun Mountain

为明确亚热带不同森林植被对土壤涉氮功能基因(amoA、nirS、nosZ)微生物群落和功能的影响,以缙云山5种森林植被类型(针叶林、阔叶林、针-阔混交林、竹林以及荒草地)土壤为研究对象,采用末端限制性片段长度多态性分析(terminal restriction fragment length polymorphism analysis,T-RFLP)和荧光定量PCR(qPCR)等分子技术,结合与氮素循环相关的微生物酶活,研究不同森林植被土壤涉氮微生物丰度、群落结构及其功能的差异.结果表明:①缙云山国家自然保护区土壤反硝化酶活显著高于硝化势,且针叶林土壤2种酶活均显著低于其他植被类型(P<0.05).此外,可溶性有机碳、土壤含水量和总氮是影响酶活的主要环境因子.②qPCR结果显示,竹林土壤涉氮功能微生物的丰度最高,而针叶林土壤最低. 3个涉氮功能微生物的丰度均与可溶性有机碳、总氮、有效氮、总钾以及有效钾显著相关(P<0.01).③基于T-RFLP数据,阔叶林土壤涉氮微生物α-多样性指数最高,而竹林最低.主坐标分析(principal co-ordinates analysis,PCoA)显示:在不同植被间,氨氧化古菌群落结构表现出显著差异,针叶林土壤涉氮功能微生物群落结构最独特.此外,基于距离的冗余分析(distance-based redundancy analysis,db-RDA)表明:涉氮功能微生物群落结构主要受可溶性有机碳(P<0.001)、有效氮(P<0.002)和土壤含水量(P<0.001)调控.④土壤反硝化酶活主要受nirS反硝化菌与氨氧化古菌的丰度和涉氮功能微生物的群落结构的影响;而硝化势主要受氨氧化古菌的丰度影响.由上可知,亚热带森林植被显著影响土壤涉氮功能微生物的丰度和群落结构,进而改变其调控土壤的氮素循环的功能.可为我国亚热带森林土壤微生物与N2O释放的耦合机制提供基础数据.

To clarify the response of a nitrogen-related microbial community and function to different vegetation types in subtropical forest, soil samples were collected for analysis from the topsoil of five vegetation types in the Jinyun Mountain National Nature Reserve, i.e., coniferous forest, broadleaf forest, mixed broadleaf-coniferous forest, P. pubescen forest, and grassland. To analyze the microbial abundance, community, and function discrepancy between different vegetation soils, multiple molecular techniques, such as terminal restriction fragment length polymorphism(T-RFLP) and quantitative polymerase chain reaction(qPCR) analysis, and nitrogen-related microbial enzyme activity procedures were used. The results showed: ① The denitrifying enzyme activity was much higher than nitrifying potential in Jinyun Mountain National Nature Reserve, and the two enzyme activities were much lower in coniferous forest soil than in the other vegetation soils(P<0.05). In addition, dissolved organic carbon, soil water content, and total nitrogen were the key environmental factors controlling enzyme activity. ② The qPCR data showed that the abundance of nitrogen-related microbes was highest in P. pubescen forest, whereas it was lowest in coniferous forest. The abundances of the three nitrogen-related microbes were all significantly correlated with dissolved organic carbon, total nitrogen, available nitrogen, total potassium, and available potassium(P<0.01). ③ Based on T-RFLP data, the α-diversity of nitrogen-related microbes was highest in broadleaf forest, whereas it was lowest in P. pubescen forest. Principal co-ordinates analysis(PCoA) showed that the community structure of ammonia-oxidizing archaea responded significantly to different vegetations, and the community structure of nitrogen-related microbes showed the most difference in coniferous forest. In addition, distance-based redundancy analysis(db-RDA) showed that the community structure of nitrogen-related microbes was mainly shaped by dissolved organic carbon(P<0.001), available nitrogen(P<0.002), and soil water content(P<0.001). ④ Soil-denitrifying enzyme activity was mainly affected by the abundance of nirS-denitrifiers, ammonia-oxidizing archaea, and the community structure of nitrogen-related microbes, whereas nitrifying potential was only controlled by the abundance of ammonia-oxidizing archaea. Above all, subtropical forest vegetation significantly affects the abundance and community structure of soil nitrogen-related microbes, thereby changing their function of controlling the soil nitrogen cycle. This study can provide basic data for the coupling mechanism between soil microbes and N2O release in subtropical forests in China.