基于转录组数据的半夏AP2/ERF基因家族鉴定及逆境响应分析

Identification and Expression Analysis of AP2/ERF Family in Stress Responses of Pinellia ternata Based on Transcriptome Data

目的:鉴定并研究半夏AP2/ERF转录因子的功能,为推进半夏品种的遗传改良提供理论依据。方法:该文基于半夏三代转录组数据鉴定了半夏中AP2/ERF家族成员,并对其进行系统的生物信息学分析,同时利用实时荧光定量聚合酶链式反应(Real-time PCR)检测半夏AP2/ERF在不同组织及不同胁迫条件下的表达情况。结果:通过转录组数据共筛选出8个全长AP2/ERF转录因子家族成员,归为AP2、ERF和DREB 3个亚家族;半夏AP2/ERF的氨基酸数目为251~512个,等电点为5.29~11.72,不稳定指数为45.90~82.41,且主要定位于细胞核中;半夏AP2/ERF基因的结构域和Motifs相对保守。组织特异性表达模式分析显示,半夏AP2/ERF基因在不同组织部位中具有不同的表达模式,且主要在叶中表达。逆境胁迫应答分析表明,PtERF1主要响应NaCl的胁迫诱导;PtERF2和PtERF4在低温和聚乙二醇(PEG)胁迫下表达量均有较大幅度的上调;PtERF3同时响应低温和NaCl两种逆境的诱导;PtERF5同时响应高温、低温、NaCl、PEG胁迫诱导;PtERF7在高温胁迫下表达量增加;而PtERF8在低温胁迫下表达量的增加最为显著。结论:通过实验初步推测半夏AP2/ERF基因可响应逆境胁迫,可能具有调控半夏光合作用、提高根系抗逆能力等潜在功能。

Objective:To identify the functions of the AP2/ERF family members in Pinellia ternata and promote the genetic improvement of P. ternata varieties. Method:We identified and conducted a systematic bioinformatics analysis of the AP2/ERF family member genes in P. ternata based on the three generations of transcriptome data. Real-time polymerase Chain reaction(Real-time) PCR was employed to determine the expression pattern of AP2/ERF genes in different tissues and under different stress conditions. Result:A total of eight full-length AP2/ERF family members were identified from the transcriptome data, which were classified into three sub-gene families:AP2, ERF, and DREB. The deduced AP2/ERF proteins in P. ternata had the length of 251-512 aa, the theoretical pI of 5.29-11.72, the instability index of 45.90-82.41, subcellular localization in the nucleus, and conserved domains and motifs. AP2/ERF genes were expressed in different tissues of P. ternata, with high expression levels in the leaf. The stress response experiments showed that PtERF1 mainly responded to NaCl stress. The expression of PtERF2 and PtERF4 was significantly up-regulated under low temperature and polyethylene glycol(PEG)-simulated stress. PtERF3 responded to both low temperature and NaCl stress. The expression of PtERF5 was induced by high temperature, low temperature, NaCl and PEG stress. The expression of PtERF7 was up-regulated under high temperature, while that of PtERF8 under low temperature. Conclusion:The AP2/ERF genes in P. ternata can respond to stress and have the potential functions of regulating photosynthesis and improving root stress resistance.