拟南芥AtPAP1基因植物表达载体构建及在烟草中遗传转化分析

Construction of plant expression vector and genetic transformation analysis of Arabidopsis thaliana AtPAP1 gene in Nicotiana tabacum

为探究花青素调控基因AtPAP1的生物学功能,采用PCR方法从拟南芥花序中克隆出拟南芥MYB家族AtPAP1基因。构建植物表达载体pROKⅡ-AtPAP1,并通过农杆菌介导的叶盘法将外源基因转入野生型烟草。检测结果表明,AtPAP1已成功整合入烟草基因组中,并在mRNA水平表达。形态观察显示AtPAP1基因异源过量表达能显著增强转基因烟草植株花青素的积累,致使转基因烟草的叶片、茎段和花器官等颜色发生变化,呈现出不同程度的紫红色。

In order to analyze the roles ofArabidopsis thalianaAtPAP1 gene, a cDNA sequence of homologous gene AtPAP1 from Arabidopsis inflorescences was cloned by PCR. Plant overexpression vector pROKII-At-PAP1 was constructed and transformed into wild-type tobacco by Agrobacterium-mediated leaf disc transformation to obtain transgenic tobacco overexpressing AtPAP1. The transgenic plants were confirmed by PCR, and the result showed AtPAP1 gene was integrated into tobacco genome. The results of qRT-PCR detection of T2 generation plants showed the mRNA of AtPAP1 was transcribed in transgenic tobaccos. Morphological observation indicated that overexpression ofAtPAP1 gene could enhance the accumulation of anthocyanins in transgenic plantlets leading to color variation of transgenic leaves, stems and flower organs in different degrees of purple red.