Cloning And Identification Of Pollen SFB Genes In Chinese Cherry (Prunus. Psedocerasus L.)

For isolating the pollen S-determinant genes in Chinese cherry (Prunus. psedocerasus L.), which is a self-compatibility tetroploid species, an Chinese cherry cultivar Taixiaohongying was used as metierial. Using degenerate primers designed based on the conserved regions of the reported SLF/SFB genes, several different cDNA clone were isolated by RT-PCR, The results are as follow:1. One middle segment cDNA clone was isolated from pollen by cDNA RT-PCR., which chosely homologous to those from amond. 3'RACE in combination with RT-PCR allowed isolation of one 3'segment, which chosely homologous to those from amond. 5'RACE combination with RT-PCR allowed isolation of three 5'segments, which chosely homologous to those from amond and Japanese apricot.2. The middle segment, 3'segment and one of the three 5'segment composed one full-length cDNA , length 1171bp. The deduced ORFs encode a F-box protein, length 375, which chosely homologous to those from amond, named SFB. We conclude that SFBc (EU267943) is the pollen SFB genes in Chinese cherry.3. The other two 5'segments, length 656bp, whose corresponding gene SFBa and SBFb were specifically expressed in pollen in an S-haplotype specific manner. The deduced ORFs of the the genes encode twe divergent F-box protein. We conclude that SFBa (GeneBank accession number: EU267941) and SFBb (EU267942) are the pollen SFB genes in Chinese cherry.4. The phylogenetic trees for SFB proteins reported in Solanaceae, Scrophulariaceae and Rosaceae were constructed. It was found that SFB from Chinese cherry were more closely related to those from Prunus (genus of Rosaceae) than those from Malus (genus of Rosaceae), Petunia (a genus of Solanaceae) and Antirrhinum (genus of Scrophulariaceae).