%0 Journal Article
%T 石榴肉桂酰辅酶A还原酶基因的克隆与表达分析
%A 董丽丽
%A 龚凌燕
%A 陈磊
%A 涂佳丽
%A 张水明
%J 南京农业大学学报
%D 2016
%R 10.7685/jnau.201602011
%X [目的] 肉桂酰辅酶A还原酶（CCR）是木质素合成的关键酶。本文旨在通过克隆和鉴定石榴CCR基因（PgCCR）来探究石榴木质素的合成机制以及培育软籽石榴新品种。[方法] 以石榴品种‘红玉石籽’（Punica granatum ‘Hongyushizi’）籽粒为材料，利用RACE和RT-PCR的方法，获得PgCCR基因的全长序列。通过实时荧光定量PCR分析PgCCR基因在不同品种、不同组织、籽粒不同发育时期的表达水平。[结果] PgCCR基因cDNA全长序列1 017 bp，编码338个氨基酸，包含被认为是CCR蛋白催化位点的保守序列KNWYCYGK。系统进化树分析表明，PgCCR与其他物种起源相同，而与葡萄VvCCR的亲缘关系最近。PgCCR基因在不同品种的石榴籽粒中均有表达，其中‘红玉石籽’中表达较高，‘会理软籽’和‘突尼斯软籽’中表达量较低。PgCCR在叶、花、茎中均有表达，茎中的表达量最高，而叶片中的表达量最低。PgCCR在‘红玉石籽’籽粒的不同时期均有表达，20~80 d相对表达量逐步上升，并在80 d时达到峰值，80 d以后表达量逐渐下降。[结论] 克隆得到石榴CCR基因，其表达水平与木质素的合成及含量有一定相关性。</br>[Objectives] Cinnamoyl coenzyme A reductase(CCR) is a key enzyme in lignin biosynthesis. The aim of this study is to investigate the synthesis mechanism of pomegranate lignin and cultivate new varieties of soft-seed pomegranate by cloning and characterization of pomegranate CCR gene. [Methods] The full length of PgCCR was obtained from the seeds of pomegranate variety ‘Hongyushizi’ by the method of RACE and RT-PCR. The expression levels of PgCCR in different cultivars,different tissues and seeds of different developmental stages were analyzed by qRT-PCR. [Results] The full-length cDNA sequence was 1 017 bp,encoded a deduced protein of 338 amino acids,and contained the conserved sequence KNWYCYGK,which was considered to be the catalytic site of CCR protein. Phylogenetic tree analysis showed that PgCCR shared the same origin with other species,and had the closest relationship with Vitis vinifera VvCCR. PgCCR was detected in three cultivars of pomegranate,and the expression in ‘Hongyushizi’ was higher,while lower in ‘Huiliruanzi’ and ‘Tunisiruanzi’. PgCCR expression was detected in leaves,flowers and stems,and the expression in stems was highest,while the lowest in leaves. PgCCR expression was detected in ‘Hongyushizi’ seeds of different periods. The relative expression of PgCCR gradually increased from 20-80 d,reached a peak at 80 d,and then declined from 80 d. [Conclusions] The pomegranate CCR gene was cloned and its expression level was correlated with the synthesis and content of lignin
%K 石榴
%K 肉桂酰辅酶A还原酶基因(CCR)
%K 基因克隆
%K 表达分析&lt
%K /br&gt
%K pomegranate(Punica granatum L.)
%K cinnamoyl coenzyme A raductase gene(CCR)
%K gene cloning
%K expression analysis
%U http://nauxb.njau.edu.cn/oa/darticle.aspx?type=view&id=201605006