Molecular Cloning And Functional Analysis Of14-3-3Genes In Phyllostachys Violascens

14-3-3proteins are a family of highly conserved regulatory molecules expressed in all eukaryoticcells.14-3-3proteins regulate a diverse set of biological responses in plants.Recent study indicated14-3-3proteins regulate the flowering time by interaction with CO or FT. The identification ofPvGF14c is important to understand flowering mechanismn in bamboo. In this study, four14-3-3geneswere cloned from Phyllostachys violascens and analyzed the expression level by RT-PCR and theirbiological function by transferring them into Arabidopsis and analyzed the intracellular localization ofPVGF14c, confirmed the interaction between Hd3a and GF14c by yeast two-hybrid screen. The resultsare as following:1. Four14-3-3homolog genes have been cloned from Phyllosta chysviolascens, named PvGF14b、PvGF14c、PvGF14e、and PvGF14f respectively. The full-length genome DNA of PvGF14b is2160bpand the full-length cDNA is958bp bp，encoding an262amino acid. The structure of PvGF14b iscomprised of5exons and4introns.The full-length genome DNA of PvGF14c is1935bp and thefull-length cDNA is1086bp，encoding an256amino acid. The structure of PvGF14c is comprised of5exons and4introns. The full-length genome DNA of PvGF14e is2130bp and the full-length cDNA is882bp，encoding an262amino acid. The structure of PvGF14e is comprised of5exons and4introns.The full-length genome DNA of PvGF14f is2030bp and the full-length cDNA is1045bp，encoding an261amino acid. The structure of PvGF14f is comprised of5exons and4introns.2.Analysis of the expression in spatial and temporal showed that the expression of the four genesalmost were stable during flowering stage.The expression of PvGF14b and PvGF14c existed no diurnalrhythm. PvGF14b was expressed in all tissues with high expression in root,stem,and panicle, next inthe mature leaf and low expression in roots.PvGF14cwas detected high expression in leaf, next in stemand no expression in root and panicle, PvGF14e was detected low expression in the root. PvGF14fwas detected with high expression in the stem and next in mature leaf and panicle and low expression inimmature leaf and no expression in root.3. PvGF14c localized in the cytoplasm.4.Yeast two-hybrid assays indicated that PvGF14c cought interact with PvFT.5. We have observed that PvGF14c-overexpressing delayed flowering in T1generation Arabidopsis.31lines PvGF14b transgenic Arabadopsis,17lines PvGF14e transgenic Arabadopsis and15linesPvGF14f transgenic Arabadopsis were harvested.