自交亲和苹果品种S基因型的鉴定

Identification of S-gene in Self-compatible Apple Cultivar

为了探讨苹果自交亲和的发生机理,根据保守氨基酸序列"FTQQYQ"和"anti-1/WⅠPNV"设计了苹果花柱S基因的通用引物:P1:5'-TTTACGCAGCAATATCAG-3';P2:5'-ACGTTCGGCCAAATA/CATT-3',根据GeneBank登陆的花粉SLF1(DQ422810)、SLF9(AB270792)基因设计了花粉S基因的引物分别为:P SLF1(上游5'-3':CTGGTGGTGTTCTTTCCTGTGTAT;下游5'-3':ACTTAACTCTTCCCTCAACTCA)和P SLF9(上游5'-3'CAGGTGCGTGAAAGTGAAA;下游5'-3':TGAGCCAAGCATAA-GAACCT),以自交亲和的苹果品种早红香为试验材料,利用PCR-RFLP、克隆、测序等方法研究早红香苹果发生自交亲和的分子性状。结果表明:自交亲和的早红香苹果2条花柱S基因没有发生变异。经BLAST比较,由花粉S基因引物PSLF1和PSLF9扩增得到的花粉S基因的DNA序列,与MdSLF2和MdSLF9基因DNA序列的相似性分别为94%和96%,推导氨基酸序列相似性分别为94%和97%。由P SLF1扩增得到的花粉S基因SLF1?与MdSLF1的DNA序列相似性仅为83%,且二者不一致的碱基分布于整个DNA序列中,自交亲和苹果品种早红香花粉SLF1?基因可能变异为了MdSLF2。

In order to discuss the mechanism of apple self-compatibility,the primer P1：5 -TTTACGCAGCAATATCAG-3 and P2：5 -ACGTTCGGCCAAATA/CATT-3 of apple style S gene were designed according to the conservative amino acid sequence,the primer P SLF1（upriver 5 -3 ：CTGGTGGTGTTCTTTCCTGTGTAT;backward position 5 -3 ：ACTTAACTCTTCCCTCAACTCA）and P SLF9（upriver 5 3 CAGGTGCGTGAAAGTGAAA;backward position 5 -3 ：TGAGCCAAGCATAAGAACCT）of apple pollen S gene were designed according to the SLF1（DQ422810）and SLF9（AB270792）in GeneBank, , the molecular character of self-compatible apple Zaohongxiang was studied in the method of PCR-RFLP, clone and sequence. The result showed that：tow style S gene of self- compatible apple Zaohongxiang did not variation. DNA sequence of pollen S gene which was amplified through pollen S gene primer P SLF1 and P SLF9 compared with the sequence of MdSLF2 and MdSLF9 had a high similarity, which arrived at 94 % and 96 %, amino acid simlarity was 94% and 97% .The similarity of Pollen S gene SLFI？ which was amplified through P SLF1 and DNA sequence of MdSLF1 was 83 %, and the differ bases were distributed in the whole DNA sequence, so that pollen S gene SLF？ could changed into MdSLF2.