牡丹隐花色素基因PsCRY2的克隆与表达分析  

Molecular Cloning and Expression Analysis of Cryptochrome Gene PsCRY2 in Tree Peony

作　　者：任秀霞[1] 王顺利[1] 薛璟祺[1] 朱富勇 刘传娇 张秀新[1] 

机构地区：[1]中国农业科学院蔬菜花卉研究所,农业部园艺作物生物学与种质创制重点实验室,北京100081

出　　处：《园艺学报》2015年第11期2229-2236,共8页Acta Horticulturae Sinica

基　　金：中国农业科学院院所基金项目(1610092015002-01);中国农业科学院科技创新工程项目(CAAS-ASTIP-IVFCAAS);国家‘863’计划项目(2011AA10020703);国家公益性行业(农业)科研专项(201203071)

摘　　要：以‘秋发1号'和‘洛阳红'牡丹(Paeonia suffruticosa)为试验材料,采用RT-PCR的方法从花芽中克隆得到1个隐花色素基因Cryptochrome 2,其ORF全长为1902 bp,编码633个氨基酸,基因登录号为KP982893。序列比对和结构域分析表明,此蛋白包含1个PHR和1个CCT结构域,与拟南芥中AtCRY2最为相似,将其命名为PsCRY2。系统进化树分析表明,PsCRY2与甜橙(Citrus sinensis)CsCRY2亲缘关系最近。实时荧光定量PCR表明,PsCRY2在牡丹的不同组织器官中均有表达。其中,成年植株的花芽以及种子胚的表达量最高,幼苗根、茎、叶次之。在整个花芽分化不同时期,PsCRY2的表达呈现较高的表达水平;在花蕾发育不同时期,PsCRY2的表达呈现先降低再升高而后有降低的趋势,大风铃期表达量最高,推测PsCRY2在花芽分化和花蕾发育的过程中均起到重要作用。在‘洛阳红'和‘秋发1号'牡丹春季花芽发育过程中,PsCRY2的表达均呈升高的趋势,‘秋发1号'显著高于‘洛阳红'。不同光周期条件下,PsCRY2表达稍有不同。与长日照条件相比,短日照条件下PsCRY2在现蕾期和小风铃期的表达量均下降。Cryptochrome 2（CRY2） was isolated from Paeonia sujfruticosa by RT-PCR.The full open reading fragment（ORF） length of PsCRY2 encoding 633 amino acids was 1 902 bp,and the GenBank No.was KP982893.Sequence alignment and motif analysis showed that the deduced amino acids contained a PHR domain near the amino terminus and a CCT domain near the carboxy terminus.It was designated as PsCRY2,according to its high identity with AtCRY2 in Arabidopsis.Phylogenetic analysis showed that it had close relationship with Citrus sinensis.Gene expression analysis revealed that the highest expression levels of PsCRY2 were appeared in the bud and seed embryo of P.sujfruticosa,followed by root,stem and leaves.The expression of PsCRY2 in the whole process of bud differentiation was high.While the expression of PsCRY2 was low in the early stage of bud development and quite high in the middle and late stage.The expression of PsCRY2 reached a peak in big bell-like flower-bud.The results suggested that PsCRY2 play an important role both in the process of bud differentiation and bud development.The expression patterns of PsCRY2 in the bud of ＇Luoyanghong＇ and ＇Qiufa 1＇ were similar,while the expression level of PsCRY2 in the bud of ＇ Qiufa 1＇ was significantly higher than that in the bud of＇Luoyanghong＇.The expression patterns of PsCRY2 in the buds with different photoperiod treatments were different.The expression of PsCRY2 decreased in bud sprouting and small bell-like flower bud under short day photoperiod,compared with that in long day photoperiod.

关 键 词：牡丹 光周期途径 隐花色素 PsCRY2 实时荧光定量PCR 

分 类 号：S685.11[农业科学—观赏园艺]