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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 investigate the mechanism of apple self-compatibility, a universal primer was designed based on the conserved amino acid sequence “FTQQYQ” and “anti-1 / WⅠPNV ”: P1: 5’-TTTACGCAGCAATATCAG-3 ’ According to the pollen SLF1 (DQ422810) and SLF9 (AB270792) genes of GeneBank, the primers of the pollen S gene were designed as follows: P SLF1 (upstream 5’-3 ’: CTGGTGGTGTTCTTTCCTGTGTAT, downstream of P2: 5’-ACGTTCGGCCAAATA / CATT- The results of PCR-RFLP analysis showed that there was no significant difference between the two cultivars (P <0.05) RFLP, cloning, sequencing and other methods to study the self-compatible molecular characters of early Hongxiang apple. The results showed that there was no variation in the S gene of two styles of self-compatible early Hongxiang apple. The BLAST results showed that the DNA sequence of pollen S gene amplified by pollen S gene primers PSLF1 and PSLF9 was 94% and 96% similar to that of MdSLF2 and MdSLF9, respectively. The deduced amino acid sequence identities were 94% And 97%. The DNA sequence similarity between SLF1? And MdSLF1 amplified by P SLF1 was only 83%, and the inconsistent bases were distributed in the entire DNA sequence. The self-compatible apple variety SLA1? The gene may mutate for MdSLF2.