基于转录组金银花AP2基因家族的生物信息学及表达分析

Bioinformatics and Expression Analysis of AP2 Gene Family Based on Transcriptome of Lonicera japonica Thunb.

为研究金银花AP2转录因子参与低温的应答机制,本研究基于低温下金银花转录组测序数据,利用在线生物信息学工具对金银花AP2转录因子的理化性质、亚细胞定位、磷酸化位点、蛋白基序以及进化分析进行鉴定和分析;利用实时荧光定量PCR检测3个DREB基因和1个RAV基因在低温胁迫下的表达情况。结果表明,在金银花中共筛选了34个AP2转录因子,不同转录因子之间的理化性质存在差异,表明其在不同的微环境中发挥不同的功能;亚细胞定位结果显示,仅有2个AP2定位于叶绿体,其余均定位于细胞核,符合其作为转录因子调控下游基因的表达特性;所有AP2的磷酸化位点均依次表现为丝氨酸>苏氨酸>酪氨酸;根据AP2中所含有的AP2数量以及序列同源性,可将34个AP2分为四个大类,20个AP2属于ERF亚家族,3个属于DREB亚家族,RAV亚家族仅有1个,其余10个属于AP2亚家族。实时荧光定量PCR检测结果表明,DREB和RAV基因均能响应低温胁迫,且不同低温处理时间的不同组织的表达量存在差异。本研究为阐明金银花AP2转录因子响应低温胁迫的机制奠定了一定的理论基础。

In order to study the mechanism of AP2 transcription factor of Lonicera japonica Thunb.responding to low temperature.With online bioinformatics,the physicochemical properties the physicochemical,subcellular localization,phosphorylation sites,protein motifs and evolution of AP2 transcription factor of L.japonica Thunb.were identified analyzed based on the transcriptome sequencing data of L.japonica Thunb.at low temperature.The expression of three DREB genes and one RAV gene under low temperature stress was detected by quantitative real-time PCR.Results showed that 34 AP2 transcription factors were screened in L.japonica Thunb.,their physical and chemical properties were different,indicating that they may play different functions in the different micro-environment.Subcellular localization results showed that only two AP2 were localized to chloroplasts,the others were localized to the nucleus,conformed with the properties of transcription factors to regulate downstream genes expression;all AP2 phosphorylation sites were Ser>Thr>Ttry.According to the domain number of AP2 contained in the sequence 34 AP2 were grouped into 4 major categories,20 AP2 belong to ERF subfamily,3 AP2 belong to DREB subfamily,one is RAV subfamily,and the left 10 AP2 belong to AP2 subfamily.The results of quantitative real-time PCR showed that DREB and RAV genes were responded to low temperature stress,and the expression was varying in tissue and treatment time.This experiment provides a theoretical basis for further exploring the mechanism of AP2 transcription factor response to low temperature in L.japonica Thunb..