重瓣山茶花器官发育相关基因CjAPL1的全长克隆与表达分析  

cDNA Cloning and Expression Analysis of CjAPL1 Gene from Camellia japonica

作　　者：孙迎坤[1,2] 李纪元[1] 殷恒福[1] 范正琪[1] 周兴文[1] 

机构地区：[1]中国林业科学研究院亚热带林业研究所,浙江富阳311400 [2]青岛农业大学园林与林学院,山东青岛266109

出　　处：《林业科学研究》2013年第4期473-479,共7页Forest Research

基　　金：国家国际科技合作计划项目(2011DFA30490);国家"十二五"科技支撑计划课题(2012BAD01B07);中央级公益性科研院所基本科研业务费专项资金项目(RISF6141)

摘　　要：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1 149 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 cDNA sequence of APETALA1（AP1） gene was cloned from early flower bud of C.japonica‘Jinpan Lizhi’using RT-PCR and RACE,named CjAPL1（GenBank accession # JX657332）.The results of nucleotide sequence analysis show that the length of CjAPL1 is 1 149 bp,containing an Opening Reading Frame（ORF） of 741 bp,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.

关 键 词：山茶花 CjAPL1基因 序列分析 相对荧光定量PCR 表达 

分 类 号：S781.46[农业科学—木材科学与技术]