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本研究利用反转录PCR和RACE方法,从中国珍稀濒危植物金花茶(Camellia nitidissima)花瓣中获得了黄烷酮3-羟化酶(flavanone 3-hydroxylase,F3H)基因的c DNA全长,命名为Cn F3H,Gen Bank登录号为HQ290517。碱基序列分析表明,该Cn F3H基因c DNA序列全长为1 360 bp,5’非翻译区(untranslated regions,UTR)长54 bp,3’UTR长202 bp,开放阅读框长为1 104 bp编码367个氨基酸。氨基酸序列比对分析表明,Cn F3H与茶(C.sinensis)F3H同源性高达99%。二级结构预测表明,无规则卷曲在Cn F3H蛋白结构中所占比例最大,其次为α-螺旋和延伸链。相对荧光定量PCR分析表明,Cn F3H基因的表达量在幼蕾期、初蕾期和膨大期的表达量较高,之后逐渐降低;Cn F3H基因在花器官不同部位中均有表达,其表达量高低顺序为雄蕊、花瓣、萼片、雌蕊和苞片。本研究为全面深入地研究金花茶花色形成的分子调控机理提供了充实的理论依据。
In this study, the full length cDNA of flavanone 3-hydroxylase (F3H) was obtained from the petal of Camellia nitidissima, a rare and endangered plant in China by reverse transcription PCR and RACE. For Cn F3H, Gen Bank accession number is HQ290517. The nucleotide sequence of the Cn F3H gene was 1 360 bp in length, 54 bp in 5 ’untranslated region (UTR), 202 bp in 3’ UTR and 1 104 bp in open reading frame Encoding 367 amino acids. Amino acid sequence alignment analysis showed that the homology between Cn F3H and C. sinensis F3H was as high as 99%. Secondary structure prediction shows that the random coil has the largest proportion of Cn F3H protein structure, followed by α-helix and extension chain. Relative fluorescence quantitative PCR analysis showed that the expression level of Cn F3H gene was higher in young buds, early buds and infancy, and then decreased gradually. Cn F3H gene was expressed in different parts of floral organs and its expression level The order of the stamens, petals, sepals, pistils and bracts. This study provides a solid theoretical basis for a comprehensive and in-depth study on the molecular regulation mechanism of Camellia flower color formation.