为研究A功能基因在山茶花重瓣形成过程中所发挥的功能,利用同源克隆和RACE扩增方法,从重瓣山茶花品种‘金盘荔枝’发育早期的花芽中获得了山茶花AP基因全长cDNA,命名为CjAPL1,GenBank登录号JX657332.该基因全长1149 bp,其中,开放阅读框(ORF)长度为741 bp,5'非编码区长度206 bp,3'非编码区长度202 bp.经氨基酸序列分析表明:CjAPL1基因编码一条含有246个氨基酸的蛋白质,与猕猴桃、八仙花等植物的Ap1蛋白同源性均在75％以上.Rea-time PCR结果显示,CjAPL1基因在‘金盘荔枝’不同发育时期花芽中的表达量为:花芽发育早Ⅲ期＞花芽发育早Ⅱ期＞花芽发育早Ⅰ期＞现蕾期,表达趋势表现为先逐渐升高而后急剧降低；花器官不同部位中的表达量为花柱最高,其次为子房和萼片,在雄蕊下部和外轮花上部中的表达量最低.这些差异表明,CjAPL1基因可能在‘金盘荔枝’重瓣花形成中发挥作用.%In order to study the role of A-class genes in the process of forming plena flower for Camellia japonica,a full-length eDNA sequence of APETALA1 (AP1) gene was cloned from early flower bud of C.japonica ‘ Jinpan Liz-hi' using RT-PCR and RACE,named CjAPL1 (GenBank accession # JX657332).The results of nucleotide sequence analysis show that the length of CjAPL1 is 1149 bp,containing an Opening Reading Frame (ORF) of 741bp,a 5 '-Untranslated Region (5 '-UTR) of 206 bp,and a 3'-UTR of 202 bp.The CjAPL1 gene encodes a protein of 246 amino acids,and has more than 75％ homology with A-class genes from Actinidia chinensis and Hydrangea macrophylla.Quantitative real-time PCR analysis shows that the gene expression pattern is that in early flower bud of Ⅲ stage ＞ early flower bud of Ⅱ stage ＞ early flower bud of Ⅰ stage ＞ squaring stage,and the tendency shows increasing slowly at the early stage and then decreasing sharply.In different floral organs,the highest expression was observed in style,next in ovary and sepals,and the lowest in bottom stamens and upper outer flowers.The difference of gene expression pattern indicates that CjAPL1 may play a role in the morphogenesis of pleiopetalous flower of Camellia.
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