陆地棉早花基因GhPRR9的功能分析及验证  

Functional analysis and validation of early flowering gene GhPRR9 in Gossypium hirsutum

作　　者：张雅楠 徐婷婷 许好标 喻树迅 ZHANG Ya’nan;XU Tingting;XU Haobiao;YU Shuxun(College of Advanced Agricultural Sciences,Zhejiang A&F University,Hangzhou 311300,Zhejiang,China;College of Forestry and Biotechnology,Zhejiang A&F University,Hangzhou 311300,Zhejiang,China)

机构地区：[1]浙江农林大学现代农学院,浙江杭州311300 [2]浙江农林大学林业与生物技术学院,浙江杭州311300

出　　处：《浙江农林大学学报》2025年第1期74-85,共12页Journal of Zhejiang A&F University

基　　金：棉花生物育种与综合利用全国重点实验室开放课题(CB2023A09)。

摘　　要：【目的】研究早花基因GhPRR9在陆地棉Gossypium hirsutum生长发育过程中的功能,为培养早熟棉品种提供理论依据。【方法】从全基因组筛选鉴定陆地棉GhPRR亚家族成员并研究其结构和表达特征;使用实时荧光定量PCR(RT-qPCR)分析GhPRR9在不同品种、不同组织及1 d内的表达差异;在烟草Nicotiana tabacum叶片中采用瞬时转化法进行亚细胞定位;分段克隆GhPRR9基因启动子并构建β-葡萄糖苷酶基因(GUS)植株进行染色实验;构建拟南芥Arabidopsis thaliana过表达株系,统计开花表型及相关基因表达量;进行病毒诱导沉默(VIGS)实验并观察开花时间差异。【结果】生物信息学分析显示:陆地棉GhPRR亚家族14个成员多数定位于细胞核,结构较为保守且主要在开花前的茎叶中集中表达。RT-qPCR分析显示:陆地棉GhPRR9表达量与品种早熟性呈正向相关,在花器官中更高,且在1d内呈周期变化。烟草瞬时转化表明:陆地棉GhPRR9分布在细胞核。GUS染色结果显示:启动子上游500~2 000 bp内可能存在关键调控元件。拟南芥过表达株系可观察到早花。VIGS实验显示:基因沉默导致开花时间延迟。【结论】基因GhPRR9对陆地棉早花性状有正向调控作用,在一定程度上能促进陆地棉生长。[Objective]The objective is to study the function of early flowering gene GhPRR9 in the growth and development of Gossypium hirsutum(cotton),so as to provide theoretical basis for cultivating early maturing cotton cultivars.[Method]Members of GhPRR subfamily were identified by genome-wide screening and the structure and expression characteristics were studied.Real-time quantitative PCR(RT-qPCR)was used to analyze the expression of GhPRR9 among different varieties,tissues and within a single day.Subcellular localization was performed in Nicotiana tabacum(tobacco)leaves by transient transformation method.The promoter of the gene was cloned andβ-D-glucosidase(GUS)plants were constructed for staining experiment.The overexpression lines of Arabidopsis thaliana were constructed to analyze the flowering phenotype data and related gene expression.Virus induced silencing(VIGS)experiment was performed to observe the difference in flowering time.[Result]Bioinformatics analysis showed that most of the 14 members of the GhPRR subfamily were located in the nucleus,with a relatively conserved structure and mainly expressed in stems and leaves before flowering.RT-qPCR analysis indicated that the expression level of GhPRR9 was positively correlated with early maturity of cultivars,higher in flower organs,and exhibited a periodic change within one day.Transient transformation of tobacco revealed that GhPRR9 was distributed in the nucleus.GUS staining suggested that there might be key regulatory elements in the promoter within 500-2000 bp upstream.Early flowering was observed in A.thaliana overexpressed strains.VIGS experiment revealed that gene silencing led to delayed flowering.[Conclusion]Gene GhPRR9 has a positive regulatory effect on the early flowering of upland cotton and can promote its growth and development.

关 键 词：陆地棉 家族分析 GhPRR9 功能验证 早花 

分 类 号：S330[农业科学—作物遗传育种]