Cloning of Two Haplotypes S-locus Receptor Kinase Extracellular Regoin(eSRK) and S-locus Cysteine Rich Protein(SCR) Genes from Brassica oleracea L. and Detection of Their Allele-specific Interaction
Abstract：S-locus receptor kinase(SRK) and S-locus cysteine rich protein(SCR) are determinant factors of male and female in self-incompatibility, they are receptor-ligand relation and act with each other in allele-specific manner. To establish a technology system suitable for analysis of the allele-specific recognition and interaction mechanism between SRK and SCR, Brassica oleracea L. inbred lines E and F were used as plant materials. Firstly, the compatible index were analyzed by pollination assay, the result showed that E and F were all self-incompatible lines, they belonged to different haplotypes and their cross-pollination were compatible. Secondly, eSRKE, eSRKF, SCRE and SCRF were cloned from genomic DNA(gDNA) of E and F, nucleotide Blast revealed that E was S28 haplotype, and F was S7 haplotype. Thirdly, the interactions among the SCRE, SCRF, eSRKE and eSRKF were detected by MatchmakerTM Gold yeast two-hybrid system. The results indicated that SCRE could specific interact with eSRKE, SCRF could specific interact with eSRKF, whereas SCRF could not interact with eSRKE and SCRE could not interact with eSRKF, which were in entirely agreement with the result of pollination assay. The analysis of the interaction SCRE, SCRF, eSRKE and eSRKF provides a technology system for further study on recognition and interaction mechanism between SRK and SCR.
