基于转录组测序的花烟草低温胁迫响应转录因子挖掘

To Explore the Transcription Factor in Response to Low Temperature Stress in Nicotiana alata by Transcriptome Sequencing

为了鉴定参与花烟草低温胁迫的转录因子,对花烟草幼苗进行了4℃低温处理,并在处理后12 h采集其幼苗样本,提取总RNA后,采用高通量测序技术,进行了转录组测序。在对差异表达基因进行GO及KEGG分析的基础上,对参与其中的转录因子进行了挖掘,并采用qRT-PCR的方法,对转录组测序的结果进行了验证。结果表明,低温处理后,花烟草基因表达量变化在2倍以上的基因有8388个(P<0.01),其中,上调表达4229个,下调表达4159个。这些差异表达基因的功能归类于生物过程、细胞组分及分子功能3大类69个GO条目,并显著富集在40条KEGG代谢通路中。同时,在低温胁迫下,花烟草有118个转录因子的表达发生了显著改变,其中,上调表达82个,下调表达36个。这些转录因子属于28个家族,其中,数量最多的为NAC家族19个,其次为ERF家族16个、MYB家族15个、WRKY家族15个。本研究的结果为花烟草低温响应分子机制的研究提供了借鉴。

In order to identify the transcription factors involved in the low temperature stress of Nicotiana alata seedlings,4℃low temperature was carried out,and samples of the seedlings were collected in 12-hour treatment,then the total RNA was extracted,and high-throughput sequencing technology was used to sequence the transcriptome.For differentially expressed genes,GO and KEGG analysis were applied,the transcription factors involved were classified,and the results of transcriptome sequencing were verified by qRT-PCR.There were 8388 genes(P<0.01)whose expression changed more than twice after low temperature treatment,of which 4229 genes were up-regulated and 4159 genes were down regulated.The functions of these differentially expressed genes were classified into three categories:biological process,cell components and molecular functions,69 items in go entries,and were significantly enriched in 40 KEGG metabolic pathways.At the same time,under low temperature stress,the expression of 118 transcription factors in N.alata changed significantly,among which 82 genes were up-regulated and 36 genes were down regulated.These transcription factors belonged to 28 families,of which 19 genes to NAC family,16 genes to ERF family,15 genes to MYB family and 15 genes to WRKY family.The results provided a reference for the study of the molecular mechanism of low temperature response in N.alata.