Cloning And Expression Of Flowering Related Genes In Bambusa Oldhamii

Bambusa oldhamii is one of the best bamboo species with delicious shoots distributing in south of China(including Taiwan).Bambusa oldhamii usually dies in mass after flowering,large scale flowering of bamboo forest not only causes the destruction of environment,but also results in the decrease of bamboo product.The research of flowering gene in Bambusa oldhami is important.Three ESTs named as BoF2,BoF4,BoF6 were cloned from Bambusa oldhami through cDNA library screening.In this study,the full length cDNA sequences of three genes were obtained by Rapid Amplification of cDNA Ends technique.The relative expression level of three genes was analyzed by FQ-PCR.The plant expression vectors of BoF2,BoF4,BoF6 were constucted and the BoF6 was transferred to Arabidopsis thaliana for function analysis.Besides,the homologous genes of FCA and LFY,named as BoFCA and BoLFY,were cloned from Bambusa oldhami.The expression level of BoFCA and BoLFY were analyzed through FQ-PCR.The results were as follows:1)1213bp cDNA sequences of BoF2 were obtained,containing an open reading frame of 792bp coding a polypeptide of 263 amino acids.1059bp cDNA sequences of BoF4 were got,including an open reading frame of 741bp coding a polypeptide of 246 amino acids.1034bp cDNA sequences of BoF6 were acquireded,consist of an open reading frame of 630bp coding a polypeptide of 209 amino acids.The proteins of BoF2,BoF4 and BoF6 had a typical structure of MADS-box gene including MADS domain,I region,K domain and C terminus.It was deduced that BoF2,BoF4 and BoF6 were the homologous genes of AG,SEP3 and PI,which belonged to C,E and B function genes by bioinformatic analysis.2)The plant expression vectors of BoF2,BoF4 and BoF6 were constucted.BoF6 had been transferred to Arabidopsis thaliana.The sepal of BoF6 transgenic Arabidopsis thaliana of F1 developed into petals-like organs.It showed that BoF6 belonged to B function genes.3)502bp genome sequences of BoFCA were obtained,composed of 314bp cDNA sequences, which coded a polypeptide of 103 amino acids.And two genome sequences of BoLFY with 613bp and 201bp were acquired.Bioinformatic analysis showed that BoFCA and BoLFY were highly homologous to FCA and LFY.4)The expression levels of BoF2,BoF4 and BoF6 in flower bud were higher than leaf bud.The expression level of BoFCA had no significant difference between flower bud and leaf bud.It showed that BoF2,BoF4 and BoF6 might be involved in flowering of Bambusa oldhami.