Molecular Colning And Expression Analysis Of B-class MADS-box Genes From Ribes(Ribes Diacanthum Pall)

Ribes diacanthum Pall., pertains to Ribesiaceae, A kind of deciduous shrub, separate flower, which widespread. To explain the molecular mechanism of floral organ development, Four full length cDNA sequences of Ribes diacanthum MADS-box genes from Ribes diacanthum as RdMAP3,RdFAP3,RdMPI and RdFPI was isolated and characterized by using RACE. In addition, investigation was also made to analyze sequence characteristics and phylogenetic relationships based on bioinformatics methods. We analyzed the expression pattern of these genes in different organs of Ribes diacanthum according to the semi-quantitative RT-PCR and real-time fluorescence quantitative PCR. The results of this study laid a solid foundation for further interpreting the action mechanism of Ribes diacanthum MADS-box genes and molecular mechanism of floral development.(1)In this study we used Ribes diacanthum as test materials, obtained four full length cDNA sequences contained open reading frames by the method of RACE and named RdMAP3,RdFAP3,RdMPI and RdFPI.(2)In order to interpretate the homology of the four genes to established MADS-box genes, We analyzed sequence characteristics and phylogenetic relationships of Ribes diacanthum acid sequences corresponding to the genes using softwares of bioinformatics. The result of sequence characteristics analysis showed that RdMAP3,RdFAP3,RdMPI and RdFPI encoded deduced proteins with233,233,216and216amino acids residues respectively. all of the amino acid sequences have a typical MIKC-type structural domain, Phylogenetic analysis indicated that RbFAP3andRbMAP3belongs to the euAP3/DEF-lineage of B-function gene family. RbMPI and RbFPI belongs to the, PI/GLO-lineage of B-function gene family.(3) The expression pattern signal of the four Ribes diacanthum MADS-box genes were detected during flowering period. Using semi-quantitative RT-PCR and real-time PCR along with the development, however in the specific period. The high level signal of RdMAP3and RdFAP3were detected. While,the expression of RdMPI and RdFPI genes continued increasing.