毛白杨糖基转移酶基因PtGT3的克隆与遗传转化

于惠敏1, 王艳文2, 任天莹2, 侯丙凯2,*
1齐鲁师范学院生物系, 济南250013; 2山东大学植物细胞工程与种质创新教育部重点实验室, 济南250100

通信作者:侯丙凯;E-mail: bkhou@sdu.edu.cn;Tel: 0531-88364726

摘 要:

通过RT-PCR从毛白杨克隆了一个木质素单体糖基转移酶候选基因PtGT3。该基因编码由465个氨基酸组成的蛋白质, 在其C-端含有一个保守PSPG框, 并与拟南芥木质素单体糖基转移酶具有较高的序列相似性, 推测此酶在杨树中负责木质素单体的糖基化修饰。利用组成型CaMV 35S启动子, 构建了PtGT3基因的植物表达载体。建立毛白杨高效组织培养再生体系, 发现叶柄分化再生率最高, 其次是茎段和叶片。采用农杆菌介导法进行遗传转化, 获得了高表达PtGT3基因的转基因杨树。获得的糖基转移酶转基因杨树为在木本植物中研究木质素单体的糖基化修饰与木质素合成的关系奠定了基础。

关键词:毛白杨; 糖基转移酶; 糖基化修饰; 木质素; 遗传转化

收稿:2013-04-02   修定:2013-06-06

资助:山东大学植物细胞工程与种质创新教育部重点实验室开放课题(2011-03)。

Cloning and Genetic Transformation of Glycosyltransferase Gene (PtGT3) of Poplar (Populus tomentosa Carr.)

YU Hui-Min1, WANG Yan-Wen2, REN Tian-Ying2, HOU Bing-Kai2,*
1Department of Biology, Qilu Normal University, Jinan 250013, China; 2The Key Laboratory of Plant Cell Engineering and Ger-mplasm Innovation, Ministry of Education, Shandong University, Jinan 250100, China

Corresponding author: HOU Bing-Kai; E-mail: bkhou@sdu.edu.cn; Tel: 0531-88364726

Abstract:

In this study, a glycosyltransferase gene, PtGT3, was cloned from poplar (Populus tomentosa Carr.) by RT-PCR. A protein of 465 amino acids, derived from the nucleotide sequences of PtGT3 gene, has a con-served PSPG box in its C-terminal and high sequence similarity with Arabidopsis monolignol glycosyltrans-ferase, suggesting a possible role of PtGT3 in the monolignol glycosylation of poplar. A plant expression vector of PtGT3 driven by CaMV 35S promoter was constructed. The efficient regeneration system of poplar through tissue culture was established and it was found that the petioles had the highest regeneration rate, followed by immature stem segments and leaves. Genetic transformation of poplar plants was conducted using the Agrobac-terium-mediated method and RT-PCR demonstrated that the transgenic poplar plants overexpressing PtGT3 were obtained. These transgenic poplar plants provide useful materials for further study on the relationship be-tween monolignol glycosylation and lignin biosynthesis in wooden plants.

Key words: poplar (Populus tomentosa Carr.); glycosyltransferase; glycosylation modification; lignin; genetic transformation

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