擬南芥GH3.9基因的過表達(dá)及其表型分析

*通信作者:劉選明,博士,教授,主要從植物分子生物學(xué)方向研究。E-mail:xml05@hnu.edu.cn

擬南芥GH3.9基因的過表達(dá)及其表型分析

周蘋,唐冬英,郭明,譚振華,趙小英,劉選明*

(湖南大學(xué) 生物學(xué)院,植物功能基因組學(xué)與發(fā)育調(diào)控湖南省重點實驗室,長沙 410082)

摘要:為研究GH3.9基因在植物生長發(fā)育過程中的作用,利用RT-PCR成功克隆到GH3.9基因,該基因全長為1 750 bp。通過構(gòu)建pEGAD-GH3.9過表達(dá)載體轉(zhuǎn)化擬南芥,獲得過表達(dá)GH3.9基因純系轉(zhuǎn)基因株系GH3.9ox-3和GH3.9ox-7。對擬南芥野生型(WT)和轉(zhuǎn)基因株系(GH3.9ox-3和GH3.9ox-7)幼苗用不同光強(qiáng)和光質(zhì)進(jìn)行處理,結(jié)果顯示:在藍(lán)光、紅光、遠(yuǎn)紅光等不同光照強(qiáng)度下培養(yǎng),過表達(dá)株系幼苗下胚軸的生長均明顯受到抑制,且較野生型明顯;采用不同光周期處理擬南芥幼苗,過表達(dá)幼苗下胚軸的伸長明顯低于野生型;對成年植株表型進(jìn)行觀察,發(fā)現(xiàn)過表達(dá)株系植株矮小、雄蕊變短、果莢短小。研究表明:GH3.9基因參與了擬南芥生長發(fā)育調(diào)控,過表達(dá)GH3.9基因?qū)M南芥生長有抑制作用。

關(guān)鍵詞:擬南芥;GH3.9基因;過表達(dá);表型分析

收稿日期:2014-10-15;修改稿收到日期:2015-01-18

基金項目:湖南省自然科學(xué)基金(11JJA002);湖南省生物發(fā)育工程及新產(chǎn)品研發(fā)協(xié)同創(chuàng)新中心(20134486)

作者簡介:周蘋(1972-),女,碩士,實驗師,主要從事生物化學(xué)與分子生物學(xué)研究。E-mail:76615803@qq.com

中圖分類號:Q785;Q789 文獻(xiàn)標(biāo)志碼: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

植物生長素能夠誘導(dǎo)一些基因快速高表達(dá),這些基因被稱為生長素早期響應(yīng)基因(primary-response genes)[1]。一般在生長素處理后2～3 min就能檢測到基因的表達(dá),進(jìn)而調(diào)控植物相關(guān)基因的后續(xù)表達(dá),影響植物的生長發(fā)育[2]。植物的GH3基因是一種典型的植物生長素早期響應(yīng)基因[3]?！?br>