首页 > 分享 > 重瓣山茶花‘金盘荔枝’C功能基因CjAGL6的全长克隆与表达分析

重瓣山茶花‘金盘荔枝’C功能基因CjAGL6的全长克隆与表达分析

花匠小妙招
2024-10-12 20:27

摘要： MADS-box基因家族在调控植物的花器官发育中发挥了重要作用。为研究AG类基因对于山茶花重瓣花形成的作用，采用同源克隆的方法，从山茶花重瓣花品种‘金盘荔枝’（Camellia japonica ‘Jinpanlizhi’）的发育早期花芽中分离到了888bp的山茶花AG基因完整全长序列，命名为CjAGL6，GenBank登录号JX657333。其中开放阅读框长度为747 bp，5′非编码区长37 bp，3′非编码区长141 bp。氨基酸序列分析显示，CjAGL6基因编码的蛋白质含有248个氨基酸，与猕猴桃（Actinidia chinensis）、黑柿（Diospyros digyna）等植物的蛋白同源性都在76%以上。同时，CjAGL6基因编码的蛋白含有重瓣花特有的保守区起始氨基酸序列及显著差异的单个氨基酸。相对荧光定量PCR结果表明，该基因在‘金盘荔枝’花芽不同发育时期的表达量为：花芽发育后期>花芽发育中期>花芽发育早期>现蕾期，表达总体趋势为先渐次升高而后急剧降至最低；不同花器官中CjAGL6基因在花柱中的表达量最高，其次为瓣化雄蕊、雄蕊上部和子房，在萼片和瓣化萼片中的表达量最低，总的趋势是在内轮花器官表达量高，在外轮花器官中的表达量低。重瓣花形成是多基因协同作用的结果，CjAGL6基因可能在调控山茶花重瓣花形成中发挥一定的功能。

关键词: &lsquo, 金盘荔枝&rsquo, CjAGL6基因, 序列分析, 相对荧光定量, 表达

Abstract: MADS-box gene family has played important roles in the floral organs development. In order to investigate the function of A-class genes for double flower development in Camellia japonica, a 888 bp full-length cDNA sequence of AG homologous gene was cloned from early flower bud of C.japonica ‘Jinpanlizhi’, named CjAGL6(GenBank accession JX657333). It contains an opening reading frame of 747 bp, a 5′-untranslated region (UTR) of 37 bp, a 3′-UTR of 141 bp. Amino acids sequence alignment showed that CjAGL6 gene encodes the protein with 248 amino acids, and the homology is more than 76% with Actinidia chinensis and Diospyros digyna. Real-time PCR analysis showed that the relative expression level of this gene was as follows: later flower bud>medium flower bud>early flower bud>squaring stage, it increased slowly at the initial stage and then sharply decreased to the lowest. In different floral organs of ‘Jinpanlizhi’, the highest expression was in style, next in petalody stamens, upper stamens and ovary, the lowest in sepals and petalody sepals. The tendency was the high expression in inter floral organs and the low in outer floral organs. The forming of double flower is the synergistic result of multiple genes. CjAGL6 gene may have some functions in this process.

Key words: Camellia japonica, CjAGL6 gene, sequence analysis, real-time PCR, expression

中图分类号:

S718.46