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
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.
SHU Jian-Hong,LUO Wei,LU Xue-Ping, et al. Cloning, Expression and Promoter Analysis of FaGI Gene from Festuca arundinacea[J]. 农业生物技术学报, 2021, 29(10): 1936-1948.
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