新疆主栽杏品种自交不亲和SFB基因的检测

Identification of Self-incompatibility SFB Genes for Apricot Cultivars in Xinjiang Area

为系统鉴定新疆主栽杏品种自交不亲和SFB基因型,以新疆25个主栽杏品种为试材,利用蔷薇科保守序列设计引物对叶片基因组DNA进行SFB基因特异PCR扩增,筛选出有效的扩增引物;对成功扩增的杏品种的特异条带克隆测序,在GenBank上进行BLASTN比对,确定各品种的SFB基因型。结果显示:引物组合Ⅱ-1、Ⅳ-2,1-Ⅰ、1-Ⅱ对新疆杏品种的扩增效果最好,成功地在18个品种上扩增出了5种大小不同的条带,14种不同的基因型,其中两个基因(SFB-36,SFB-42)与GenBank上已经登录的SFB-13(the Accession No.EF062342)为同一基因,其余12个为新基因,暂命名为:SFB-30、SFB-31、SFB-32、SFB-33、SFB-34、SFB-35、SFB-37、SFB-38、SFB-39、SFB-40、SFB-41和SFB-43。对扩增出的SFB氨基酸序列进行比较,发现3个保守区和两个高变区;将试验所得基因与李属果树62个SFB基因构建系统进化树,显示李属果树SFB基因的种内相似性并不高于种间相似性。

In order to identify SFB gene associated with self-incompatibility of apricot cultivars in Xinjiang area systematically,five pairs of primers designed according to the conserved sequences of Rosaceae SFB were used for specific PCR amplification of 25 mainly apricot cultivars in Xinjiang.The specific fragments amplified successfully were cloned and sequenced.The results showed：The primer combinationⅡ-1、Ⅳ-2,1-Ⅰ、1-Ⅱ was the best optional;Five different size fragments and 14 different genes were cloned from 18 apricot cultivars.Searches for homology with our sequences publicly available DNA databases revealed 2 SFB gene fragments（SFB-36,SFB-42）had been deposited in the GenBank （Accession No：EF062342）as SFB-13,and the other 12 SFB gene fragments were novel SFB genes,temporarily named as：SFB-30,SFB-31,SFB-32,SFB-33,SFB-34,SFB-35,SFB-37,SFB-38,SFB-39,SFB-40,SFB-41 and SFB-43.Comparison of amino acid sequences of this 14 SFB gene fragments in apricot revealed three conserved regions and two high variability regions.Sixty-two SFB genes in Rosaceae Prunus genus were used to form their phyligenetic tree.The evolutionary relationships showed SFB genes did not form a distinct cluster within species.Intra-species similarity was not higher than inter-species one.