摘要
【目的】克隆中亚杏(Prunus armeniaca)品种自交不亲和花柱S-RNase基因全长序列,为分子手段调控杏自交不亲和性状奠定基础。【方法】以新疆栽培杏品种‘索格佳娜丽’和‘赛买提’为试材,利用RT-PCR克隆2个品种的花柱SRNase基因c DNA片段,RACE技术进行c DNA全长克隆,采用BLAST进行序列比对,Protparam软件分析2个基因的编码蛋白特性,MEGA 5.0构建进化树。【结果】从‘索格佳娜丽’中克隆了S_(52)-RNase(KF951503)基因,从‘赛买提’中克隆了一个新的S_(53)-RNase(KF975455)基因DNA和c DNA全长序列。S_(52)-RNase的DNA全长2 200 bp,c DNA全长765 bp,ORF(开放阅读框)长681 bp,编码226个氨基酸;S_(53)-RNase的DNA全长1 664 bp,c DNA全长907 bp,ORF长732 bp,编码242个氨基酸。BLASTP比对显示:这2个基因都具有保守的RNase-T2基因结构,属于RNase-T2家族。预测相对分子质量分别为26.5 ku和27.5 ku,等电点为9.36和9.03,都属于亲水性蛋白。进化分析表明,S_(52)与李(Prunus salicina,S_7)、S_(53)与大岛樱(Prunus speciosa,S_(13))亲缘关系最近,S_(52)和S_(53)处在2个不同的分支上,表现出较高的序列多态性,表明2个基因亲缘关系较远。【结论】获得了2个中亚杏品种自交不亲和花柱S-RNase基因全长序列。
[Objective] To clone the full length sequence of S-RNase genes involving self-incompatibility in the Central Asian apricots and laid a foundation for the improvement of apricot self-incompatibility by molecular biology methods. [Methods]The two apricots cultivar ＇Suogejianali＇ and ＇Saimaiti＇ in Xinjiang province were tested in this article. The S-RNase gene cDNA fragment was obtained by RT-PCR technology and the full length sequence was obtained by RACE technology. And then using Protparam and MEGA 5.0 software to analyze the characteristics of putative proteins from two genes and construct phylogenetic tree, respectively. [Results]Two full length DNA and eDNA of S52-RNase gene（KF951503） and a new S53-RNase gene （KF975455）were cloned. The full length DNA of S52-RNase was 2 200 bp and the full length eDNA was 765 bp which contained an ORF （open reading frame） of 681 bp and encoded a protein of 226 amino acids. The full length DNA of S53-RNase was 1 664 bp and the full length cDNA was 907 bp which contained an ORF of 732 bp and encoded a protein of 242 amino acids. It is predicted that both of the two genes had the RNase-T2 motif and belonging to RNase-T2 subfamily by BLASTP. The predicted molecular weight was 26.5 ku and 27.5 ku, and isoelectric point was 9.36 and 9.03, respectively, by Protparam. Both of the two RNase were hydrophilic proteins. The phylogenetic analysis indicated that the phylogenetic relationship of S52 and Prunus salicina （S7）, S53 and Prunus speciosa（S13） were closest. S52 and S53 lay different branch and possessing high polymorphism. [Conclusion] Two full length sequence of S-RNase genes involving self-incompatibility in the Central Asian apricots were obtained.
出处
《果树学报》
CAS
CSCD
北大核心
2015年第6期1047-1054,共8页
Journal of Fruit Science
基金
国家自然科学基金面上项目(31272129)
