当归AP2/ERF转录因子家族鉴定及干旱胁迫下的表达分析

Identification of AP2/ERF transcription factor family in Angelica sinensis and its expression analysis under drought stress

目的对当归Angelica sinensis中的AP2/ERF转录因子基因进行鉴定以及生物信息学分析,并分析其在干旱胁迫下的表达差异,以期筛选出与响应干旱胁迫相关的AsAP2/ERF基因,为后续深入研究2品种当归抗逆性差异及抗性育种提供参考依据。方法基于全长转录组测序数据,利用生物信息学方法对AsAP2/ERF转录因子家族成员进行鉴定,并分析该家族特征。基于干旱胁迫转录组数据分析AsAP2/ERF基因的表达模式,并利用qRT-PCR技术进行验证。结果从当归中共鉴定到53个AP2/ERF转录因子,包括28个ERF、18个DREB、5个RAV、2个AP2亚家族成员。AsAP2/ERF转录因子的氨基酸数量是142~440aa,均为亲水性蛋白,亚细胞定位预测结果显示多数成员定位于细胞核。基因表达模式的分析结果表明,在响应干旱胁迫条件下,“岷归2号”(M2)中表达上调的基因数量多于“岷归1号”(M1),其中8个基因在2品种当归D2组表达差异显著,这可能与2品种当归的抗旱性相关。qRT-PCR分析结果表明,6个基因与干旱胁迫转录组数据基本一致。结论鉴定了当归中的AP2/ERF转录因子家族,探究了不同基因在干旱胁迫下的表达模式,最终获得了6个响应干旱胁迫的AsAP2/ERF候选基因,为后期开展当归抗旱功能基因的研究奠定基础。

Objective The AP2/ERF transcription factor genes in two cultivars of Angelica sinensis were identified and analyzed the differences in their expression under drought stress,so as to screen AsAP2/ERF genes related to the response to drought stress,and provide a reference basis for the subsequent in-depth study on the stress resistance differences between the two cultivars of A.sinensis and the breeding for drought resistance.Methods AsAP2/ERF transcription factor family members based on full-length transcriptome sequencing data were identified using bioinformatics approaches and family characterisation was analyzed.The expression patterns of AsAP2/ERF family members were further analyzed based on drought stress transcriptome data and the selected genes were validated by using qRT-PCR.Results A total of 53 AP2/ERF transcription factors were identified from A.sinensis,including 28 ERF,18 DREB,five RAV,and two AP2 subfamily members.The amino acid number of AsAP2/ERF transcription factors ranged from 142—440.These transcription factors were hydrophilic proteins and the prediction of subcellular location was mainly in the nucleus.The results of gene expression patterns analysis showed that more genes were up-regulated in M2 than in M1 in response to drought stress,and eight genes were significantly differentially expressed during the D2 period in the two cultivars of A.sinensis,which might be related to drought resistance in the two cultivars of A.sinensis.The results of qRT-PCR analyses showed that six genes were generally consistent with the drought stress transcriptome data.Conclusion In this study,AP2/ERF family transcription factors were identified in two cultivars of A.sinensis,the expression patterns of different genes under drought stress were preliminarily investigated,and finally six AsAP2/ERF candidate genes in response to drought stress were obtained,which lays the foundation for later studies on drought-resistant functional genes of A.sinensis.