森林草莓FvSPL2基因克隆、过表达载体构建及表达分析
Gene Cloning,Over-expression Vector Construction and Expression Analy-sis of FvSPL2 Gene in Fragaria vesca
任子政 1邱安然 1常琳 1侯佳聪 1王丽娟 1龙鸿1
作者信息
- 1. 天津农学院园艺园林学院,天津,300392
- 折叠
摘要
SPL(SQUAMOSA promoter-binding protein-like)转录因子是植物特有的一类基因家族,广泛存在于高等植物中,在植物营养生长和生殖发育中起着重要的调控作用.为了探究SPL基因对森林草莓(Fragaria vesca)花发育的影响,本研究以森林草莓'Hawaii4'为试验材料,基于森林草莓转录组数据分析,克隆得到SPL转录因子基因FvSPL2,对其进行生物信息学和表达分析,并构建了 35S∷FvSPL2过表达载体.结果表明:FvSPL2基因编码区全长639 bp,编码212个氨基酸;蛋白质理化性质和结构分析表明FvSPL2蛋白为不稳定亲水性蛋白,其含有高度保守的SBP结构域和一个核定位信号NLS.RT-qPCR分析结果显示FvSPL2基因在花中表达量最高,在叶柄和叶片中次之.此外,该基因在不同花期表达先上升后下降,在半开花中表达量最高,且与花苞和全开花差异显著.这些结果说明该基因主要参与开花调控过程.研究结果为后续研究FvSPL2基因在森林草莓中的生物学功能提供数据依据.
Abstract
SPL(SQUAMOSA promoter-binding protein-like)transcription factors are plant-specific gene family that are widely found in higher plants and play an important regulatory role in plant vegetative growth and repro-ductive development.This study investigates the effect of SPL genes on flower development in woodland strawberry.In this experiment,the SPL transcription factor gene FvSPL2 was cloned from woodland strawberry(Fragaria vesca)'Hawaii4',and the 35S::FvSPL2 over-expression vector was constructed based on the analysis of woodland straw-berry transcriptome data.The results showed that the coding region of FvSPL2 gene was 639 bp,encoding 212 amino acids.The physicochemical and structural analyses indicated that FvSPL2 protein was an unstable hy-drophobic protein,which contained a highly conserved SBP structural domain and a nuclear localization signal NLS.The highest expression of FvSPL2 gene was detected by RT-qPCR in woodland strawberry flowers,and the lower expression in petiole and leaf.Moreover,the expression level of FvSPL2 at different flowering stages showed an upward trend first and then downward,through period of flower buds(the lowest level),half-blooming flowers(the highest level)and full-blooming flowers,and the difference between the half flowering stage and other two periods was significant.These results indicated that this gene is mainly involved in the flowering regulation pro-cess.The results of the study provide data for the subsequent study of the biological functions of FvSPL2 gene in woodland strawberry.
关键词
森林草莓/FvSPL2/基因克隆/荧光定量RT-PCRKey words
Fragaria vesca/FvSPL2/Gene cloning/Quantitative RT-PCR引用本文复制引用
基金项目
天津市科委种业重大专项(17ZXZYNC00080)
出版年
2024
NETL
NSTL
万方数据