2个野扁桃自交不亲和S-RNase基因全长的克隆与序列分析  

Cloning and sequence analysis of 2 full-length S-RNase genes in Amygdalus ledebouriana Schlecht

作　　者：关鹏[1] 曾斌[2] 李疆[2] 罗淑萍[1] 王建友[3] 李伟阳[2] 田嘉[2] 李鹏[2] 

机构地区：[1]新疆农业大学农学院,乌鲁木齐830052 [2]新疆农业大学林学与园艺学院.新疆农业大学果树学新疆特色果树研究中心,乌鲁木齐830052 [3]中国林业科学院新疆分院,乌鲁木齐830000

出　　处：《果树学报》2016年第8期905-916,共12页Journal of Fruit Science

基　　金：国家自然科学基金(31260465);国家林业公益性行业科研专项(201304701-1;201304701-5);新疆维吾尔自治区科技计划项目(201130102-1-5);新疆维吾尔自治区果树学重点学科(201007)

摘　　要：【目的】克隆新疆野扁桃(Amygdalus ledebouriana Schlecht.)自交不亲和性花柱特异性决定因子编码基因SRNase全长序列,为自交不亲和性的分子调控奠定基础。【方法】以新疆野扁桃花柱为试材,利用RT-PCR和RACE技术克隆S-RNase基因全长,采用BLAST和ORF Finder对核酸序列进行分析,利用CDD、Prot Param、Tmpred、Signal P、Target P、SOPMA、DANMAN、MEGA6和Prot Fun对推导的氨基酸序列进行分析。【结果】克隆到Pt S16-RNase基因和Pt S17-RNase基因,2者均属于RNase T2基因家族,与其他多种植物的S-RNase基因的序列相似度为83%~98%,序列均具有S-RNas蛋白典型结构。Pt S16-RNase基因ORF长690 bp,编码229个氨基酸,Pt S17-RNase基因ORF长678 bp,编码225个氨基酸。预测2个S-RNase蛋白均为亲水性、不稳定的分泌蛋白,二级结构均以α-螺旋、延伸链和无规卷曲为主,在蔷薇科李属植物中具有较高的系统进化一致性,可能的主要功能为水解酶和激素。【结论】获得2个新疆野扁桃自交不亲和性花柱特异性决定因子编码基因S-RNase全长序列。[ Objective ] Two full- length sequences of S-RNase genes encoding pistil determinant factors of self-incompatibility were cloned, and bioinformatics analyses of the sequences and their decuced amino acid sequences were processed in order to facilitate molecular regulation of self-incompatibility in wild al- mond （Amygdalus ledebouriana Schlecht.）. [Methods] The full-length sequences of S-RNase genes Were cloned by RT-PCR and RACE techniques from the pistils of the wild almond. Regions of local similarity between sequences of the cloned S-RNase genes and those accessioned in GenBank were analysed by the Basic Local Alignment Search Tool （BLAST）.Open reading frames of sequences of the cloned S-RNase genes were analysed by the ORF Finder （Open Reading Frame Finder）. Conserved domains of the deduced amino acid sequences were searched by the Conserved Domain Database （CDD）. Physical and chemical pa- rameters of the deduced amino acide sequences were computed by ProtParam. Membrane-spanning re- gions and their orientation of the deduced amino acid sequences were predicted by TMpred. The presence and location of signal peptide cleavage sites of the deduced amino acid sequences were predicted by Sig- nalP 4.1 Server. The subcellular location of the deduced amino acid sequences were predicted by TargetP 1.1 Server. The secondary structure of the deduced amino acid sequences were predicted by SOPMA. Se- quence alignment between the deduced amino acid sequences and ten homologous proteins of other plant species （Prunus webbii, Prunus armeniaca, Prunus avium, Prunus eerasus, Prunus dulcis, Prunus mume, Prunus persiea, Prunus salicina, Antirrhinum hispanieum, Petunia integrifolia subsp, inflata） was per- formed by DNAMAN. Phylogenetic analysis between the deduced amino acid sequences and ten proteins of other plant species were processed by MEGA6. The main functions of the deduced amino acid sequenc- es were predicted by ProtFun 2.2 Server. [Results]Two full-length sequences of S-RNase （PtS16-RNase and

关 键 词：野扁桃 自交不亲和性 S-RNASE基因 生物信息学 RACE 

分 类 号：S662.9[农业科学—果树学]