11个中国杏品种S-RNase基因的检测与序列分析

以11个未知基因型的中国杏品种为试材,根据李属植物S-RNase基因保守区设计2对引物组合检测各品种S-RNase基因,共获得22条等位扩增片段,电泳检测表明所有品种的扩增条带集中在300~1100bp的范围内,且表现出一定的长度多态性。序列分析进一步确定22个S-RNase基因为10个不同的等位基因,其中6个为首次发现,根据Gen-Bank中已登陆的杏S-RNase基因的顺序,分别命名为S19、S20、S23、S24、S25、S26,序列登陆号为:EF185300、EF185301、EU037262、EU037263、EU037264、EU037265。推导氨基酸序列的同源分析表明,杏的S-RNase与李属植物的S-RNase表现较高的同源性,为59.3%~100%;与苹果和梨的S-RNase同源性较低,为19.6%~31.6%。试验确定11个中国杏品种资源的自交不亲和基因型分别为:‘大果杏’S19/S20,‘张公园’S24/S25,‘二红’S9/S11,‘黄口外’S11/S26,‘植丸子’S11/S17,‘宇宙红’、‘大丰’S17/S23,‘超仁’、‘虹桥’S8/S11,‘冀光’、‘中华大杏梅’S8/S9;部分品种的田间杂交授粉座果与花粉管荧光显微镜观察证实了所鉴定基因型的可靠性。

Detection and sequence analyses of S-RNase gene in eleven Chinese apricot(Prunus armeniaca)cultivars

Eleven native apricot cultivars with unknown genotype were used in the experiments,and 22 specific fragments were amplified with two group primers designed from conserved sequence of S-RNase in Prunus.Amplified fragments were ranged from 300 bp to 1 100 bp in size and showed length polymorphism by electrophoresis.Blast analyses of sequences showed that 22 S-RNase represented 10 different alleles,of which six were reported firstly.We submitted six new S-RNase genes to the GenBank and denoted them as S19,S20,S23,S24,S25 and S26,the accession number being EF185300,EF185301,EU037262,EU037263,EU037264 and EU037265 in sequences.Based on the deduced amino acid sequences alignment,S-RNase of apricot showed high identities with S-RNase of others'in Prunus,whose values were 59.3%-100%,but showed low identities with S-RNase of apple and pear,whose values were 19.6%-31.6%.S-genotypes of eleven Chinese apricot cultivars were putative as follows:S19/S20 for Daguoxing,S24/S25 for Zhanggongyuan,S9/S11 for Erhong,S11/S26 for Huangkouwai,S11/S17 for Zhiwanzi,S17/S23 for Yuzhouhong and Dafeng,S8/S11 for Chaoren and Hongqiao,S8/S9 for Jiguang and Zhonghuadaxingmei.The results were confirmed by fruit setting and fluorescent microscopic observation of pollen tube growth after cross-pollination.