‘砀山酥梨’褐皮芽变中两个ERF基因的克隆与表达分析

Cloning and Expression Analysis of Two ERF Genes in Pericarp of Russet Mutant of ‘Dangshansuli' Pear

为了解梨(Pyrus bretschneideri)中ERF基因的功能,采用3′RACE和PCR技术从‘砀山酥梨’中克隆了两个ERF基因,并对其表达进行了分析。克隆的两个ERF基因都具有典型的AP2/ERF结构域,属于ERF基因家族,分别命名为Pb ERF2和Pb ERF4,Gen Bank登录号分别为KJ623716和KJ623718。系统进化分析表明,Pb ERF2与枇杷ERF1,Pb ERF4与黄瓜ERF1的亲缘关系较近。表达分析表明,Pb ERF2和Pb ERF4在叶片中几乎不表达,果皮中的表达量高于果肉;‘锈酥’果皮3个发育时期的Pb ERF2和Pb ERF4表达量均显著高于‘砀山酥梨’,且均呈现先上升后下降的趋势。这为深入研究梨ERF基因家族的作用机理提供了理论依据。

The aim was to understand the function of ethylene responsive factor (ERF) in Pyrus bretschneideri. Two ERF genes were cloned from pericarp of‘Dangshansuli’ and its russet mutant‘Xiusu’ by using 3′RACE and PCR monthods. Both of genes had typical conserved domain of AP2/ERF, named as PbERF2 and PbERF4, with GenBank accession No. of KJ623716 and KJ623718, respectively. The phylogenetic tree showed that PbERF2 had closed correlation with ERF1 in loquat (Eriobotrya japonica) and PbERF4 was closed to ERF1 in cucumber (Cucumis sativus). Semi-quantitative RT-PCR indicated that expressions of PbERF2 and PbERF4 in pericarps were higher than those in sarcocarps, and both hardly expressed in leaves. Real-time RT-PCR showed that expressions of PbERF2 and PbERF4 in pericarps of‘Xiusu’ was signiifcant higher than those in‘Dangshansuli’, and expression trend of them was upgrade at ifrst and then descend. These would provide theory foundation for further study on mechanism of ERF family genes in pear.