甘蓝转录因子BoLH27的克隆与转基因甘蓝的表型分析

Cloning of BoLH27 Gene from Cabbage and Phenotype Analysis of Transgenic Cabbage

bHLH转录因子对植物生长发育、形态调控有重要作用,为了研究BoLH27基因在甘蓝叶片发育及形态建成中的调控功能,本文以甘蓝品种519为材料,克隆出转录因子BoLH27基因。序列分析表明,该基因含有一个795 bp的开放阅读框,编码264个氨基酸,具有一个保守的典型bHLH结构域。以农杆菌介导的遗传转化方法将正义BoLH27基因导入甘蓝品种519中以增强表达,通过PCR筛选出9株T0代转基因单株,结合T2代单株的q RT-PCR分析表明,筛选的转基因植株内BoLH27基因的表达积累量明显高于野生型。试验基地隔离网内种植的转基因甘蓝表型明显,主要表现为莲座期茎叶间距拉长、叶柄伸长以及植株茎和叶柄显示紫色,植株叶片平展,无向上向内卷曲趋势,表明BoLH27基因可能对甘蓝叶片发育有重要的调控作用。

bHLH transcription factor plays an important role in plant growth,development and morphological control.In order to explore BoLH27 gene regulatory function for leaf development and morphologic formation,the BoLH27 gene was cloned from cabbage(Brassica oleracea L.)variety 519.Sequence analysis indicated that the length of BoLH27 gene was 795 bp,which encoded 264 amino acids.The BoLH27 protein contained conservative structure domains of the bHLH family.The sense BoLH27 gene was transformed into cabbage variety 519 by Agrobacterium mediated method,PCR analysis exhibited that BoLH27 was genetically transformed into nine individual plants,qRT-PCR analysis revealed that BoLH27 had higher expression level in T2 transgenic cabbage than in wild type.The phenotype at rosette stage of transgenic cabbage grown in the test base was obvious,showing elongated the stems and leaves,elongated petiole,purple stems and petiole,and flat leaves without upward inward curling trend.It suggested that BoLH27 may play important roles in the control of leaves development.