新疆野苹果AP2/ERF转录因子家族鉴定与响应腐烂病的表达分析  

Identification and Expression Analysis of AP2/ERF Transcription Factor Family in Malus sieversii upon Valsa mali Infection

作　　者：赵明奇 刘晓洁 梁玉青[1,2] 杨瑞瑞 李小双[1,2,3] ZHAO Mingqi;LIU Xiaojie;LIANG Yuqing;YANG Ruirui;LI Xiaoshuang(Xinjiang Key Laboratory of Stress Tolerance Plant Resources Conservation and Resistance Gene Utilization,Urumqi 830011,China;Turpan Desert Botanical Garden,Chinese Academy of Sciences,Turpan,Xinjiang 838008,China;University of Chinese Academy of Sciences,Beijing 100049,China)

机构地区：[1]新疆抗逆植物基因资源保育与利用重点实验室,乌鲁木齐830011 [2]中国科学院吐鲁番沙漠植物园,新疆吐鲁番838008 [3]中国科学院大学,北京100049

出　　处：《西北植物学报》2022年第6期930-942,共13页Acta Botanica Boreali-Occidentalia Sinica

基　　金：中国科学院青年创新促进会(2018478);国家自然科学基金(U1903206)。

摘　　要：AP2/ERF转录因子家族参与了植物生长发育、抵抗胁迫以及植物激素响应等诸多生物过程,是植物中最重要的转录因子家族之一。该研究基于腐烂病菌侵染后的新疆野苹果(Malus sieversii)全长转录组,使用AP2保守结构域的隐马可夫模型PF00847,鉴定新疆野苹果的AP2/ERF家族成员。利用MEGA6、NCBI CDD-batch、MEME、EXPASY、BUSCA对MsAP2/ERF家族成员进行鉴定、分类和结构分析、理化性质和亚细胞定位分析。通过RNA-seq数据和qRT-PCR实验对差异表达的MsAP2/ERFs基因的表达水平进行分析和验证,旨在鉴定新疆野苹果中潜在具有腐烂病抗性的AP2/ERF家族基因资源。结果显示:(1)在新疆野苹果中共鉴定获得106个AP2/ERF基因,涵盖全部AP2(17个)、ERF(57个)、DREB(25个)、RAV(5个)和Soloist(2个)5个亚家族。(2)进一步的细化分类发现MsERF亚家族包括B1-B6 6个组,而MsDREB亚家族中只有A2、A4、A5、A6共4个组,缺少A1和A3组的基因成员。(3)RNA-seq表达模式分析结果表明,29个MsAP2/ERF基因在腐烂病感染过程中差异表达,其中MsERF亚家族中差异表达基因数量最多(19个)。(4)12个MsAP2/ERF代表基因的qRT-PCR结果表明:8个ERF亚家族基因均受腐烂病病菌诱导显著上调表达,其中B4类ERF成员基因(MsERF40)在腐烂病病菌侵染后5 d表达量上调表达倍数最高;4个MsDREB基因中,3个受到腐烂病病原菌诱导显著上调,1个下调表达;此外,含有TIR保守结构域的MsERF05在腐烂病病菌侵染1 d后上调表达69倍,表明ERF亚家族的MsERF40和MsERF05在新疆野苹果抗腐烂病过程中发挥重要作用。该研究结果为新疆野苹果AP2/ERF基因响应腐烂病的功能和机理研究奠定了基础。AP2/ERF transcription factor family is one of the most important transcription factor families in plants, which is involved in plant crucial biological processes such as growth and development, stress resistance and plant hormone signaling pathway. Based on PacBio sequencing database of M. sieversii upon the Valsa mali infection, MsAP2/ERF family members were screened and identified using the hidden Markov model of AP2 domain(PF00847). The MsAP2/ERF transcription factor family were identified and analyzed by MEGA6, NCBI CDD batch, MEME, ExPASY, BUSCA. The gene expression analysis based on RNA-seq data and qRT-PCR assay were performed to screen the potential resistant genes for Valsa canker disease. The main results are as follows:(1) in total, 106 AP2/ERF genes were identified in M. sieversii, including AP2(17 genes), ERF(57 genes), DREB(25 genes), RAV(5 genes), and Soloist(2 genes).(2) Further detailed classification showed that the MsERF subfamily included 6 groups(B1-B6). The MsDREB subfamily contained A2, A4, A5 and A6 groups, and lacked A1 and A3 group genes.(3) Differential expression pattern analysis based on RNA-seq data showed that 29 MsAP2/ERF genes were differentially expressed during the infection of V. mali. Among them, 19 DEGs were MsERF genes.(4) The qRT-PCR results of 12 representative MsAP2/ERF genes showed that 8 MsERF genes were significantly up-regulated after V. mali infection. Particularly, the B4 group of ERF subfamily gene(MsERF40) was remarkably up-regulated with the highest fold increase at 5 d after the infection of V. mali compared with the control. Three MsDREB genes were significantly up-regulated and one MsDREB gene was down-regulated after V. mali infection. In addition, the MsERF gene containing TIR domain(MsERF05) was up-regulated with 69-fold increase at 1 d. Those results show the MsERF40 and MsERF05 play a key role during resisting V. mali in M. sieversii. In conclusion, this study laid a foundation for the further function and mechanism of MsAP2/ERF genes that respond to the

关 键 词：AP2/ERF 新疆野苹果 腐烂病 抗病基因 

分 类 号：Q786[生物学—分子生物学] Q789