自交亲和苹果品种S基因型的鉴定

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＇:TGAGCCAAGCATAAGAACCT),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P SLF1 和P SLF9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.

Identification of S-gene in Self-compatible Apple Cultivar

In order to discuss the mechanism of apple self-compatibility, the primer PI:5'-TTTACGCAGC AATATCAG-3' and P2:5'-ACGTTCGGCCAAATA/CATT-3' of apple style S gene were designed according to the conser-vative amino acid sequence,the primer P SLFKupriver 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 com-pared with the sequence of MdSLF2 and MdSLF9 had a high similarity, which arrived at 94% and 96% , amino acid sim-ilarity was 94% and 97% .The similarity of Pollen S gene SLF1? which was amplified through P SLF1 and DNA se-quence of MdSLFl was 83 %, and the differ bases were distributed in the whole DNA sequence, so that pollen S gene SLF? could changed into MdSLF2 .