雷竹PvJ3基因克隆及功能分析

Cloning and functional analysis of the PvJ3 gene from Phyllostachys violascens

拟南芥Arabidopsis thaliana中J3基因参与花期的调节,调控开花时间。为了探究竹类植物中J3基因的功能,根据植物中J同源基因的高度保守性,采用同源克隆技术从雷竹Phyllostachys vilascens中克隆出1个J3基因,命名为PvJ3。该基因的开放阅读框(ORF)区长为1 260 bp,编码419个氨基酸。通过同源比对和系统进化树分析,可知雷竹PvJ3蛋白和其他物种中J3同源蛋白的氨基酸序列同源性很高。通过实时定量聚合酶链式反应(qRT-PCR),发现PvJ3基因在同一竹鞭的笋、花芽、花,开花与不开花雷竹的茎秆、成叶、幼叶中都有表达,且开花雷竹茎秆中表达量最高,表明PvJ3基因在雷竹各组织部位都有表达。亚细胞定位分析结果显示,PvJ3蛋白定位在细胞质和细胞核。通过转化拟南芥对其功能进行了初步分析,表型统计结果显示,PvJ3转基因拟南芥与野生型相比,开花时间明显提早,而且出现多主茎现象,表明PvJ3可以促使植物提前开花并且参与植物茎秆的发育。

The J3 gene mediates the integration of flowering signals in Arabidopsis thaliana.To explore the functions of the J3 gene in bamboo plants,a J3 homolog named PvJ3 was isolated from Phyllostachys violascens,and a preliminary analysis of its function was conducted with Ph.violascens.Analyses included a sequencing alignment and phylogenetic analysis,real-time quantitative PCR,and protein subcellular localization.Results showed that the phenotype of overexpression of PvJ3 in Ph.violascens predated flowering and had more main stems.The full-length ORF of the PvJ3 in Ph.violascens was 1 260 bp which encoded 419 amino acids.The sequencing alignment and phylogenetic analysis of PvJ3 showed a high homology with other species.Real-time quantitative PCR results showed that the PvJ3 was expressed in all organs both in flowering and non-flowering bamboo plants and was highest in the stem of flowering bamboo plants.Protein subcellular localization showed that PvJ3 was located in the cytoplasm and cell nucleus.These results suggest that PvJ3 may promote flowering,may participate in stem development,and may provide a scientific basis for the regulation of bamboo flowering.