*通信作者:刘选明,博士,教授,主要从植物分子生物学方向研究。E-mail:xml05@hnu.edu.cn

拟南芥GH3.9基因的过表达及其表型分析

周苹,唐冬英,郭明,谭振华,赵小英,刘选明*

(湖南大学 生物学院,植物功能基因组学与发育调控湖南省重点实验室,长沙 410082)

摘要:为研究GH3.9基因在植物生长发育过程中的作用,利用RT-PCR成功克隆到GH3.9基因,该基因全长为1 750 bp。通过构建pEGAD-GH3.9过表达载体转化拟南芥,获得过表达GH3.9基因纯系转基因株系GH3.9ox-3和GH3.9ox-7。对拟南芥野生型(WT)和转基因株系(GH3.9ox-3和GH3.9ox-7)幼苗用不同光强和光质进行处理,结果显示:在蓝光、红光、远红光等不同光照强度下培养,过表达株系幼苗下胚轴的生长均明显受到抑制,且较野生型明显;采用不同光周期处理拟南芥幼苗,过表达幼苗下胚轴的伸长明显低于野生型;对成年植株表型进行观察,发现过表达株系植株矮小、雄蕊变短、果荚短小。研究表明:GH3.9基因参与了拟南芥生长发育调控,过表达GH3.9基因对拟南芥生长有抑制作用。

关键词:拟南芥;GH3.9基因;过表达;表型分析

收稿日期:2014-10-15;修改稿收到日期:2015-01-18

基金项目:湖南省自然科学基金(11JJA002);湖南省生物发育工程及新产品研发协同创新中心(20134486)

作者简介:周苹(1972-),女,硕士,实验师,主要从事生物化学与分子生物学研究。E-mail:76615803@qq.com

中图分类号:Q785;Q789 文献标志码:A

Overexpression and Phenotype Analysis ofGH3.9

Gene inArabidopsisthaliana

ZHOU Ping,TANGdongying,GUO Ming,TAN Zhenhua,ZHAO Xiaoying,LIU Xuanming*

(Hunan Province Key Laboratory of Plant Functional Genomics anddevelopmental Regulation,College of Biology,Hunan University,Changsha 410082,China)

Abstract:To investigate the role of GH3.9 in plant growth anddevelopment,we cloned the 1 750 bp full-length gene with RT-PCR method and obtained GH3.9ox-3 and GH3.9ox-9 pure lines of Arabidopsis thaliana by constructing pEGAD-GH3.9 overexpression vector and transforming A.thaliana.After treating the sowed wild-type and mutant seeds (GH3.9ox-3 and GH3.9ox-9) with blue,red and far-red light atdifferent intensity,we found that:the growth of hypocotyls in the mutants was more sensitive to the strong light suppression comparing with wild-type.Moreover,the hypocotyls of mutants appeared to grow slower when treateddifferent forms of white light.The mutants plant sizes and silique sizes appeared smaller.The results indicate that GH3.9 gene might play an important role in the growth anddevelopment of plants.

Key words:Arabidopsisthaliana;GH3.9 gene;overexpression;phenotype analysis

植物生长素能够诱导一些基因快速高表达,这些基因被称为生长素早期响应基因(primary-response genes)[1]。一般在生长素处理后2～3 min就能检测到基因的表达,进而调控植物相关基因的后续表达,影响植物的生长发育[2]。植物的GH3基因是一种典型的植物生长素早期响应基因[3]。……