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2025年4月3日 星期四
农业生物技术学报  2021, Vol. 29 Issue (10): 1936-1948    DOI: 10.3969/j.issn.1674-7968.2021.10.008
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
高羊茅FaGI基因克隆、表达及启动子分析
舒健虹1, 罗维2, 路雪萍4, 牟琼1, 吴佳海2,3,*, 王小利1,*
1贵州省农业科学院 草业研究所,贵阳 550006;
2贵州大学 动物科学学院/高原山地动物遗传育种与繁殖教育部重点实验室,贵阳 550025;
3贵州省农业科学院 果树科学研究所,贵阳 550006;
4贵州大学 生命科学学院,贵阳 550006
Cloning, Expression and Promoter Analysis of FaGI Gene from Festuca arundinacea
SHU Jian-Hong1, LUO Wei2, LU Xue-Ping4, MU Qiong1, WU Jia-Hai2,3,*, WANG Xiao-Li1,*
1 Institute of Prataculture, Guizhou Academy of Agricultural Sciences, Guiyang 550006, China;
2 School of Animal Science/Key Laboratory of Animal Genetics, Breeding and Reproduction in the Plateau Mountainous Region, Ministry of Education, Guizhou University, Guiyang 550025, China;
3 Institute of Fruit Research, Guizhou Academy of Agricultural Sciences, Guiyang 550006, China;
4 College of Science,Guizhou University,Guiyang 550006, China
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摘要 GI (GIGANTEA)是节律相关基因,在植物控制节律输出和开花调控中具有重要作用。为探索GI基因在高羊茅(Festuca arundinacea)中的生物学功能,本研究运用cDNA末端快速扩增法(rapid amplification of cDNA ends, RACE)克隆和染色体步移法扩增得到该基因序列和启动子序列,并对其进行生物信息学分析。序列分析表明,FaGI基因(GenBank No. MZ540915)序列全长3 869 bp,其开放阅读框为3 447 bp,编码1 149个氨基酸。启动子序列2 371 bp,有CURECORECR、GT1CONSENSUS、LTRE1HVBLT49和MYB识别位点多个顺式作用元件。系统进化树表明,FaGI基因编码的蛋白与草甸羊茅(Festuca pratensis)、黑麦草(Lolium perenne)、黑麦(Secale cereale)和小麦(Triticum aestivum)等禾本科植物同源蛋白的进化关系比较近。亚细胞定位显示其定位在细胞核,说明该蛋白可能在细胞核中发挥作用。荧光定量分析不同光照处理和不同发育阶段下的表达模式。结果显示,FaGI基因的表达受光周期和生物钟的调控,不同发育时期其表达量不同,苗期的表达量最高,进入生殖生长阶段其表达量下降。构建p1300-FaGI过表达载体转化拟南芥(Arabidopsis thaliana),结果显示,FaGI过表达拟南芥能上调AtCCA1 (CIRCADIAN CLOCK ASSOCIATED 1)和AtTOC1 (TIMING OF CAB EXPRESSION 1)表达,下调AtFT (FLOWERING LOCUS T)和AtCO (CONSTANS)表达。该研究结果为后续验证高羊茅FaGI基因功能提供理论依据。
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舒健虹
罗维
路雪萍
牟琼
吴佳海
王小利
关键词 高羊茅高羊茅GIGANTEA (FaGI)基因启动子克隆表达分析    
Abstract:GIGANTEA (GI) is a rhythm related gene, which plays an important role controlling of rhythm output and flowering regulation in plants.To explore the biological function of GIGANTEA (GI) gene in tall fescue (Festuca arundinacea), in this study, tall fescue was used as the experimental material, and the gene sequence and promoter sequence were amplified by using RACE (rapid amplification of cDNA ends) cloning and chromosome walking method, and analyzed by bioinformatics. Sequence analysis showed that the total length of FaGI gene (GenBank No. MZ540915) was 3 869 bp, and its ORF was 3 447 bp, encoding 1 149 amino acids. The promoter sequence was 2 371 bp and had multiple cis-acting elements such as CURECORECR, GT1CONSENSUS, LTRE1HVBLT49 and MYB recognition site.The phylogenetic tree showed that the protein encoded by the FaGI gene had a close evolutionary relationship with the homologous proteins of gramineous plants such as Festuca pratensis, Lolium perenne, Secale cereale, Triticum aestivum and other grasses.The subcellular location showed that it was located in the nucleus, indicated that the protein might play a role in the nucleus. Fluorescence quantitative analysis of expression patterns under different light treatments and different developmental stages showed that the expression of FaGI gene was regulated by photoperiod and circadian clock. The expression level of FaGI gene was different in different developmental stages. The expression level was the highest in the seedling stage, and the expression level decreases in reproductive growth stage. The p1300-FaGI overexpression vector was constructed to transform Arabidopsis thaliana, and it was found that the Arabidopsis overexpression FaGI could up-regulate the expression of AtCCA1 (CIRCADIAN CLOCK ASSOCIATED 1), AtTOC1 (TIMING OF CAB EXPRESSION 1), and down-regulate the expression of AtFT (FLOWERING LOCUS T) and AtCO (CONSTANS). This study provides a theoretical basis for the follow-up verification of the FaGI gene function of tall fescue.
Key wordsFestuca arundinacea    F. arundinacea GIGANTEA (FaGI) gene    Promoter cloning    Expression analysis
收稿日期: 2021-03-12     
ZTFLH:  S688.4  
基金资助:国家自然科学基金(31860674); 贵州省高层次创新型人才持续培养项目[黔科合人才〔2016〕4024]; 贵州省高层次创新人才培养项目[黔科合平台人才(2018)5634]; 贵州省科技计划项目[黔科合平台人才(2020)5005]
通讯作者: * lhcbqk@163.com; wangxiaolizhenyuan@126.com   
引用本文:   
舒健虹, 罗维, 路雪萍, 牟琼, 吴佳海, 王小利. 高羊茅FaGI基因克隆、表达及启动子分析[J]. 农业生物技术学报, 2021, 29(10): 1936-1948.
SHU Jian-Hong, LUO Wei, LU Xue-Ping, MU Qiong, WU Jia-Hai, WANG Xiao-Li. Cloning, Expression and Promoter Analysis of FaGI Gene from Festuca arundinacea. 农业生物技术学报, 2021, 29(10): 1936-1948.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2021.10.008     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2021/V29/I10/1936
 
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