牡丹NCED基因的克隆和表达分析  

Isolation and Expression of 9-cis Epoxycarotenoid Dioxygenase Gene in Tree Peony

作　　者：王晓庆[1] 张超[1] 王彦杰[1] 董丽[1] 

机构地区：[1]北京林业大学园林学院,国家花卉工程技术研究中心,北京100083

出　　处：《园艺学报》2012年第10期2033-2044,共12页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(30972030)

摘　　要：通过RT-PCR和RACE技术克隆得到了牡丹(Paeonia suffruticosa Andr.)中编码9–顺式–环氧类胡萝卜素加双氧酶蛋白(NCED)的一条cDNA基因全长(暂命名为Ps-NCED1)。进化树分析显示Ps-NCED1的氨基酸序列与葡萄、马铃薯、胡萝卜等植物中的NCED一致性均达到了70%以上。通过与拟南芥中CCD(类胡萝卜素分解加双氧酶)家族氨基酸序列比对后发现,Ps-NCED1与调控ABA合成的At-NCED3一致性最高。对花朵中内源ABA的研究发现,ABA在花朵蕾期及衰老期含量较高,盛开期时含量较低,表明ABA对于植物的开放衰老具有促进作用。相对荧光定量PCR分析发现:在花朵完全开放的牡丹植株中,Ps-NCED1在根、茎、叶、花托、萼片、花瓣、心皮、雄蕊中的表达量以在根和雄蕊中较高,在叶和萼片中最低。在外源ABA处理的牡丹花瓣中,Ps-NCED1表达量变化与内源ABA含量变化具有相同的趋势,推测该基因是调控内源ABA含量的主要基因。A full-length cDNA（temporarily name it Ps-NCED1）encoding 9-cis-epoxycarotenoid （NCED）was isolated with the techniques of RT-PCR and RACE from petals of tree peony flower and characterized. Phylogenetic tree analysis revealed that amino acids sequence of Ps-NCED1 shared more than 70% identity with the sequence of NCED from plants such as Vitis vinifera,Solanum tuberosum, Daucus carota and so on. Ps-NCED1 also had the highest amino acids sequence identity with At-NCED3 of CCD（carotenoid cleavage dioxygenase）family in Arabidopsis. Cut flowers during full open stage had the lowest endogenous ABA levels,to the contrary,flowers during primary and senescent stages had higher ABA levels. The relative real-time PCR analysis was used to identify the expression of Ps-NCED1 in different parts of tree peony at blooming stage. The expression in root and stamen presented the highest,while that in leaf and sepal the lowest. The expression of Ps-NCED1 in the petals from ABA treated flowers during vasing increased with cut flowers opening and senescence,coincided with the ABA content increase,resulted in the assumption that it might be the main member in regulation the ABA biosynthesis in tree peony responding to the postharvest stress during opening and senescence process.

关 键 词：牡丹 ABA NCED 基因 实时定量PCR 

分 类 号：S685.11[农业科学—观赏园艺]