典型淹水稻田土壤微生物群落的基因转录活性及其主要生理代谢过程

Microbial metabolism in typical flooded paddy soils

【目的】利用环境转录组技术,研究复杂稻田土壤中微生物群落主要生理代谢过程的基因表达水平及其对长期施氮磷钾肥(Mineral nitrogen,phosphorus,and potassium,NPK)的响应规律。【方法】针对中国科学院常熟农田生态系统长期定位试验的NPK施肥处理和不施肥对照处理(Control check,CK)稻田土壤,淹水培养2周后提取土壤微生物总RNA进行高通量转录组测序,利用MG-RAST网络分析平台(Metagenomics Analysis Server)进行活性微生物组成分析、基因功能注释及基因功能分类。【结果】细菌是CK和NPK处理稻田土壤微生物的优势类群,占比高达95%以上,细菌中的活性基因主要源于变形菌门(Proteobacteria,占细菌的50%以上)。同时也检测到古菌、真核生物和病毒等多种微生物的活性基因,而古菌中的活性基因主要源于奇古菌门(Thaumarchaeota,约占古菌的70%)。酸杆菌门(Acidobacteria)在NPK处理土壤中的转录活性显著高于CK处理土壤,而其他的细菌及古菌类群的转录活性在CK和NPK处理土壤间无显著性差异。CK和NPK处理土壤中表达量最高的基因是ABC transporter编码基因,与物质跨膜运输紧密相关。基于COG(Clusters of Orthologous Genes)、Subsystem、KEGG(Kyoto Encyclopedia of Genes and Genomes)3种基因功能分类数据库,发现CK和NPK处理土壤中微生物的主要代谢活动均为能量产生与转化、碳水化合物代谢、蛋白代谢和氨基酸代谢,而最活跃的代谢路径为氧化磷酸化及氨酰-tRNA合成。【结论】淹水状态下CK和NPK处理稻田土壤中的活性微生物组成较为一致,仅Acidobacteria的转录活性在两者间差异较大;在微生物的主要代谢活动方面,CK和NPK处理土壤之间基本一致,均以能量获取与蛋白代谢为主,长期施用无机化肥对复杂土壤微生物群落水平的主要代谢活动影响较小。

[ Objective] The object of this study is to reveal the composition of active microorganism and their metabolic activities in flooded paddy soils with long-term fertilization （Mineral nitrogen, phosphorus, and potassium, NPK） and without fertilizer （Control check, CK） by environmental transcriptomics. [ Methods ] Flooded soil microcosms were incubated in the laboratory for two weeks, then total RNA were extracted from the soil for transcriptome sequencing. Resulting fastq files were uploaded to the Metagenomics Analysis Server （MG-RAST） for taxonomic analysis, gene annotation and function classification. [ Results] Transcripts from diverse active microorganism, including bacteria （ 〉 95% ） , archaea, eukaryotes and viruses, were detected in both flooded paddy soils of CK and NPK treatments. Most of the transcripts （active genes） of bacteria and archaea were derived from Proteobacteria （more than 50% of total bacterial transcripts） and Thaumarchaaeota （about 70% of total archaeal transcripts ） respectively in both treatments. Transcriptional activity of Acidobacteria in NPK treatment paddy soil was significantly higher than that in CK treatment paddy soil. As for other phyla of bacteria and archaea, there were no significant differences of transcriptional activity of them between CK and NPK treatment paddy soils. The highest expressed gene in both CK and NPK treatment paddy soils is ABC transporter encoding gene which related to the transmembrane transport of substances. Based on gene function category of COG （Clusters of Orthologous Genes） , Subsystem and KEGG （Kyoto Encyclopedia of Genes and Genomes） database, we found that the main metabolic activities of microorganisms in both CK and NPK treatment paddy soils were related to energy production and conversion, carbohydrate metabolism, protein metabolism and amino acid metabolism, and the dominant KEGG pathways were oxidative phosphorylation and aminoacyl-tRNA biosynthesis. [ Conclusion ] Composition of active microorganism in CK and NPK treatment paddy soils was generally similar, except Acidobacteria whose transcriptional activity was significantly different between these two treatment paddy soils. It was also very similar between CK and NPK treatment paddy soils considering the metabolic activities of microorganisms in them, for dominant metabolic processes in these two soils were both related to energy obtaining and protein metabolism. So, dominant metabolic activities of microorganism in flooded paddy soils used in this study were not altered significantly under long - term inorganic fertilization.