摘要
【目的】本研究旨在探索棉花GhWRN基因在生长发育中的功能。【方法】用生物信息学方法分析GhWRN基因的结构特征、进化关系及上游1500 bp的启动子片段并对其功能进行初步分析,利用实时荧光定量-聚合酶链式反应(Quantitative real-time polymerase chain reaction,qRT-PCR)技术分析该基因在不同熟性棉花品种、不同组织及不同激素处理下的表达特性。构建过表达载体转化拟南芥,观察转基因拟南芥的表型。利用qRT-PCR分析拟南芥开花信号途径中标记基因的表达变化。【结果】以陆地棉cDNA为模板克隆得到GhWRN基因,生物信息学分析表明该基因编码产物含有1个保守的WRN-exo结构域,无内含子,不具有跨膜结构,为非分泌蛋白。qRT-PCR表明该基因在早熟棉花品种的二叶期开始上调表达,在植株各组织中均有表达,在雄蕊和苞片表达量较高;外源激素ABA、IAA处理后0.5 h该基因极显著上调表达。与野生型相比转基因拟南芥开花时间提前,莲座叶数量减少,qRT-PCR分析表明花发育相关关键基因AtSOC1、AtFT和AtFUL上调表达。【结论】GhWRN基因与促进植物开花相关,为进一步探究棉花的开花机制提供新线索。
[Objective]The purpose of this study was to explore the function of GhWRN in the growth and development of cotton.[Methods]GhWRN was analyzed in aspects of gene structure,evolutionary relationship,and gene promoter features through bioinformatic methods.Its expression patterns among different cultivars,tissues,and hormone conditions were gained via quantitative real-time polymerase chain reaction(qRT-PCR).The phenotype of Arabidopsis thaliana with overexpressed GhWRN was observed.The expression pattern of marker genes involved in A.thaliana flowering signal pathway was detected by qRT-PCR.[Results]The GhWRN gene was successfully cloned according to the reference genome of upland cotton.Bioinformatic analysis showed this gene contained a conserved WRN-exo domain,without intron or transmembrane structure,and was not a secretory protein.qRT-PCR showed GhWRN was up-regulated at the second leaf stage in early-maturing cotton cultivars and was expressed in all tissues,especially in stamens and bracts.It was significantly up-regulated at 0.5 h after treatment with exogenous ABA and IAA.The transgenic A.thaliana lines showed early flowering time and decreasing rosettes number compared with the wild-type.The qRT-PCR analysis in A.thaliana indicated that GhWRN could up-regulate the expression of key genes related to flower development,i.e.,AtSOC1,AtFT,and AtFUL.[Conclusion]GhWRN could promote plant flowering,which sheds light on further exploration of flowering mechanism in cotton.
作者
薛羽君
魏恒玲
王寒涛
马亮
程帅帅
郝蓬勃
顾丽姣
付小康
芦建华
喻树迅
Xue Yujun;Wei Hengling;Wang Hantao;Ma Liang;Cheng Shuaishuai;Hao Pengbo;Gu Lijiao;Fu Xiaokang;Lu Jianhua;Yu Shuxun(State Key Laboratory of Cotton Biology/Institute of Cotton Research,Chinese Academy of Agricultural Sciences,Anyang,Henan 455000,China)
出处
《棉花学报》
CSCD
北大核心
2021年第3期189-199,共11页
Cotton Science
基金
国家现代农业产业技术体系——棉花产业技术体系(CARS-15-06)。
