Cloning And Preliminary Function Analysis Of B Class Genes, In Bambusa Oldhamii

MADS-box family genes play important roles in the determination of floral meristem identity andthe development of floral organ identity. As a member of MADS-box family genes, AP3regulate thedevelopment of carpel and stamen. Here we reported a full length AP3-like gene obtained from theflower bud of Bambusa oldhamii by RACE techniques.It was named as BoAP3based on sequencesimilarity.The relative expression level of gene was analyzed by Quantitative real time PCR；Theplant expression vectors of BoAP3was constucted. BoAP3and different truncated fragments BoPIwere cloned respectively into pGBKT7and pGADT7. Using yeast two-hybrid system, thePGADT7-BoAP3and PGBKT7-BoPI recombinant vector were constructed and transformed intoyeast Y187and AH109separately.Our results showed that:(1)998bp cDNA sequences of BoAP3was obtained, containing an open reading frame of654bpcoding a polypeptide of218amino acids.The protein showed a typical MADS-box gene structurecontaining MADS domain、K domain、I region and C terminus. Based on bioinformatic analysis, itwas deduced that BoAP3was the homologous gene of AP3, which belonged to B function gene.Theplant expression vectors of BoAP3was constucted.(2) Quantitative real-time PCR analysis revealed that the expression level of BoAP3in flowerbuds was8.1times higher than in vegetative shoots, suggested that BoAP3may be involved in floralorgan development.(3) Auto activation deteetion of recombinant pGBKT7and pGBKT7Plasmid showed that bothof these plasmids did not activate the repoter gene.Yeast two-hybrid result showed that BoAP3andBoPI had no interaction when MADS domain is deleted, while still had interaction when C terminusis deleted，demostrated that MADS domain played a important role in interaction between BoAP3andBoPI.