文章摘要
枯草芽孢杆菌对氨氮应答的转录组及sRNA分析
Transcriptome and sRNA analysis of Bacillus subtilis response to ammonia nitrogen
投稿时间:2020-02-05  修订日期:2020-03-11
DOI:
中文关键词: 枯草芽孢杆菌  氨氮应答  转录组  差异表达基因  sRNA分析
英文关键词: Bacillus subtilis  the response to ammonia nitrogen  transcriptome  differentially expressed genes  sRNA analysis
基金项目:四川省农业科技成果转化资金项目(2018NZZJ008),四川省科技支撑项目(2016NZ0044),四川肉羊创新团队防疫岗位
作者单位E-mail
郑 姚 西南民族大学 青藏高原动物遗传资源保护与利用教育部和四川省重点实验室 1793946322@qq.com 
吴开年 西南民族大学 青藏高原动物遗传资源保护与利用教育部和四川省重点实验室  
王 利 西南民族大学 青藏高原动物遗传资源保护与利用教育部和四川省重点实验室 qinxin916@aliyun.com 
魏勇 四川畜牧科学研究院  
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中文摘要:
      为探索枯草芽孢杆菌(Bacillus subtilis)脱氮的分子机制,筛选枯草芽孢杆菌对氨氮的分子生态学应答相关候选基因及small RNA分析,本研究对处于富含氨氮环境和对照组的枯草芽孢杆菌R47进行原核链特异性转录组及sRNA分析,并采用Real-time PCR方法检测差异表达基因的相对表达量。结果显示,平均每个测序样本得到约1.40×107条reads。对照组与处理组DESeq2分析得到3918个差异表达基因,并富集在KEGG数据库中的176个信号通路,其中包括8个与适应富含氨氮环境相关的信号通路(细菌双组分系统通路、精氨酸生物合成、嘌呤代谢等),同时发现epsA、tasA、sinR、glnR、glnA、tnrA和ureABC基因可能参与枯草芽孢杆菌对氨氮的应答过程。经sRNA分析获得已注释的枯草芽孢杆菌sRNA 62条。对sRNA靶基因的分析结果显示其有3960个对应的潜在靶基因,主要参与碳水化合物运输和新陈代谢、氨基酸转运和代谢、转录过程,其中发现sRNA2073和sRNA2182对应的靶基因为sinR和tnrA。Real-time PCR结果显示,argH、codY、argG、glnA和glnR基因的相对表达量变化与转录组测序结果一致。这为进一步探究枯草芽孢杆菌污水脱氮的分子机理提供参考数据。
英文摘要:
      To explore the molecular mechanism of denitrification by Bacillus subtilis and screen out candidate genes related to the response of Bacillus subtilis to ammonia nitrogen and analysis of small RNA, transcriptome sequencing and sRNA analysis ware performed on Bacillus subtilis in an ammonia-rich environment and control group. The relative expression changes of differentially expressed genes were analyzed by Real-time PCR. The results showed that each sequencing sample yielded approximately 1.40 × 107 reads on average. There were 3918 differentially expressed genes in the control group and the treatment group by DESeq2 analysis, and which enriched 176 signaling pathways in the KEGG database, including 8 signaling pathways (bacterial two-component system pathway, arginine biosynthesis, purine metabolism and so on) adapted to the ammonia-rich environment. And we found epsA, tasA, SinR, glnR, glnA, tnrA and ureABC genes may be involved in the response of Bacillus subtilis to ammonia nitrogen in water. The annotated 62 strains of Bacillus subtilis sRNA were obtained. The results of prediction and analysis of sRNA target genes revealed that there are 3960 potential target genes involved in carbohydrate transport and metabolism, amino acid transport and metabolism, and transcription processes. Among them, the target genes corresponding to sRNA2073and sRNA2182 are sinR and tnrA. Real-time PCR analysis showed that the relative expression changes of argH, codY, argG, glnA and glnR were consistent with transcriptome sequencing.. It provides reference data for further exploring the molecular mechanism of nitrogen removal by Bacillus subtilis in wastewater
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