樱桃自交不亲和s9-单元型特异的基因克隆及其表达分析.pdfVIP

2006,39(5):976-983 Scientia Agricultura Sinica 100094 Cloning and Expression Analysis of the Self-Incompatibility S9-Haplotype Specific F-box Genes in Cherry (Prunus avium) CHENG Jian-hong, BAI Song-ling, HAN Zhen-hai, XU Xue-feng, LI Tian-zhong (Institute of Horticultural Plants, China Agricultural University, Beijing 100094) Abstract: ObjectiveThe purpose of this study is to isolate the pollen S-determiant genes involving self-incompatibility in cherry and settle some basic theories for the researches of self-incompatibility mechanism. MethodA pair of consensus primers were designed according to the cDNA sequences of 16 S-locus F-box genes published in GenBank. With the RT-PCR and RACE methods, two full length genes coding a kind of polypeptides containing 376 amino acids were cloned in pollen cDNA of cherry cultivar Hongdeng. ResultGenBank blast showed that one of the genes coded the same amino acid sequence with the self-incompatibility S3-haplotype-specific SFB (S-locus F-box) gene, SFB3 (AB096857). The other gene coded a novel F-box protein showed no identical sequence in NCBI database and shared 76%-82% amino acid identity with PaSFB1-6. The experimental results showed that the novel gene expressed specifically in pollen and showed the S9-haplotype-specific linkage signal. Conclusion These characters suggested that the novel gene was a new member of pollen S-determinant gene family, PaSFB, in cherry, and was named PaSFB9 (GenBank accession number: DQ422809). The S-genotype of Hongdeng was confirmed as S3S9. Key words: Cherry (Prunus avium); Self-incompatibility; Pollen; F-box gene; Cloning SI