油茶CoSAUR32和CoSAUR50基因的克隆及表达分析

Cloning and Expression Analysis of CoSAUR32 and CoSAUR50 Genes in Camellia oleifera

本研究以油茶‘华硕’为试验材料,通过PCR克隆获得两个生长素响应SAUR(Small auxin upregulated RNA)基因家族的基因,命名为CoSAUR32和CoSAUR50。CoSAUR32基因开放阅读框(ORF)为372 bp,编码123个氨基酸;CoSAUR50基因开放阅读框(ORF)为318 bp,编码105个氨基酸。序列分析发现CoSAUR32和CoSAUR50蛋白均无信号肽与跨膜结构,在叶绿体和细胞核中均可表达。对其进行同源性比对及进化树分析发现,油茶两个SAUR与茶树SAUR亲缘关系最近。实时荧光定量PCR结果表明,CoSAUR32和CoSAUR50基因在自交授粉子房中36 h时的表达量显著高于异交子房,在自交授粉花柱中24 h时的表达量显著高于异交花柱。推测CoSAUR32和CoSAUR50可能参与油茶自交不亲和过程。本研究为挖掘SAUR基因的功能及解析油茶自交不亲和分子机制提供参考依据。

In this study,Camellia oleifera’Huashuo’was used as experimental material.Two Samall auxin-up RNAs SAUR family genes were cloned by PCR and named CoSAUR32 and CoSAUR50.CoSAUR32 gene open reading frame(ORF)is 372 bp,encoding 123 amino acids,CoSAUR50 gene open reading frame(ORF)is 318 bp,encoding 105 amino acids.Sequence analysis showed that CoSAUR32 and CoSAUR50 proteins have no signal peptide and transmembrane structure,and can be expressed in chloroplasts and nucleus.Homology comparison and phylogenetic tree analysis showed that the two SAURs in Camellia oleifera have the closest relationship to the SAUR in Camellia sinensis.The results of RT-qPCR showed that the expression of CoSAUR32 and CoSAUR50 genes the expression level in self-pollination ovary was significantly higher than that in cross-pollination ovary at36 h,and in self-pollination styles was significantly higher than that in cross-pollination styles at 24 h.It is speculated that CoSAUR32 and CoSAUR50 participate in the process of self-incompatibility.This study provides a reference for exploring the function of SAUR genes and an alyzing the molecular m echanism of self-incompatibility of Camellia oleifera.