生物技术通报 ›› 2020, Vol. 36 ›› Issue (12): 42-53.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0241

• 研究报告 • 上一篇    下一篇

基于转录组测序花烟草响应盐胁迫活性氧清除相关基因的挖掘

鲁琳1(), 杨尚谕2, 刘维东1, 鲁黎明2()   

  1. 1.四川农业大学风景园林学院,成都 611130
    2.四川农业大学农学院,成都 611130
  • 收稿日期:2020-03-09 出版日期:2020-12-26 发布日期:2020-12-22
  • 作者简介:鲁琳,女,副教授,研究方向:园林植物;E-mail:001lulin2003@163.com
  • 基金资助:
    四川省教育厅科技项目(12ZA272)

Mining of Genes Related to Reactive Oxygen Species Scavenging in Response to Salt Stress in Nicotiana alata Based on Transcriptome Sequencing

LU Lin1(), YANG Shang-yu2, LIU Wei-dong1, LU Li-ming2()   

  1. 1. College of Landscape Architecture,Sichuan Agricultural University,Chengdu 611130
    2. College of Agronomy,Sichuan Agricultural University,Chengdu 611130
  • Received:2020-03-09 Published:2020-12-26 Online:2020-12-22

摘要:

为了鉴定参与花烟草盐胁迫响应的活性氧清除相关基因,用200 mmol/L NaCl处理花烟草幼苗,并在处理后12 h采集其幼苗样本,提取总RNA,采用高通量测序技术,进行转录组测序。基于差异表达基因GO及KEGG分析的基础上,对参与其中的活性氧清除基因进行挖掘,利用qRT-PCR的方法验证转录组测序结果。结果表明,盐处理后,花烟草基因表达量变化在2倍以上的基因有7 239个(P<0.01),其中,上调表达基因4 037个,下调表达基因3 162个,将其功能归类于生物过程、细胞组分及分子功能三大类159个GO条目,并显著富集在12条KEGG代谢通路中。同时,在盐胁迫下,花烟草有45个活性氧清除相关基因的表达发生了显著改变,其中,上调表达基因28个,下调表达基因17个。此结果说明,活性氧的清除是花烟草抵御盐胁迫的重要机制之一。

关键词: 花烟草, 转录组, 盐胁迫, 活性氧

Abstract:

In order to identify the genes related to reactive oxygen species(ROS)scavenging involved in salt stress response of Nicotiana tabacum seedlings,tobacco plants were treated with 200 mmol/L NaCl,and the samples of the seedlings were collected at 12 h after the treatment. After total RNA was extracted,high-throughput sequencing technology was carried out to sequence the transcriptome. On the basis of GO and KEGG analysis of the differentially expressed genes,ROS scavenging genes involved were mined,and the results of transcriptome sequencing were verified by qRT-PCR. The results showed that there were 7 239 genes(P < 0.01)with more than two-fold in the expression of tobacco genes after high salt treatment,among which 4 037 genes were up-regulated and 3 162 genes were down-regulated. The functions of these differentially expressed genes were classified into 159 GO items in three categories:biological process,cell components and molecular functions,and were significantly enriched in 12 KEGG metabolic pathways. Meanwhile,the expressions of 45 genes related to ROS scavenging processes changed significantly in tobacco plants under high salt stress,among which 28 genes were up-regulated and 17 genes were down-regulated. The results of this study suggest that the enhanced ROS scavenging ability could be one of the essential mechanisms of N. alata plants in response to salt stress.

Key words: Nicotiana alata, transcriptome, salt stress, reactive oxygen species(ROS)