《植物生理学报》 2017, 53(7): 1199-1207

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

刘轶¹, 郑唐春^1,2, 代丽娟¹, 刘彩霞¹, 王庆娜¹, 曲冠证^1,*

¹东北林业大学林木遗传育种国家重点实验室, 哈尔滨150040; ²花卉种质创新与分子育种北京市重点实验室, 国家花卉工程技术研究中心, 城乡生态环境北京实验室, 林木花卉遗传育种教育部重点实验室, 园林学院, 北京林业大学, 北京, 100083

通信作者：曲冠证；E-mail: quguanzheng@yahoo.com

摘　要：

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。

关键词：拟南芥; AtPAP1; 花青素; 烟草

收稿：2016-10-24   修定：2017-06-15

资助：教育部新世纪优秀人才支持计划项目(NCET-12-0808)和国家自然科学基金(31370661)。

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

LIU Yi¹, ZHENG Tang-Chun^1,2, DAI Li-Juan¹, LIU Cai-Xia¹, WANG Qing-Na¹, QU Guan-Zheng^1,*

¹State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin 150040, China; ²Beijing Key Laboratory of Ornamental Plants Germplasm Innovation & Molecular Breeding, National Engineering Research Center for Floriculture, Beijing Laboratory of Urban and Rural Ecological Environment, Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants of Ministry of Education, School of Landscape Architecture, Beijing Forestry University, Beijing 100083, China

Corresponding author: QU Guan-Zheng; E-mail: quguanzheng@yahoo.com

Abstract:

In order to analyze the roles of Arabidopsis thaliana AtPAP1 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 of AtPAP1 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.

Key words: Arabidopsis; AtPAP1; anthocyanins; tobacco (Nicotiana tabacum)