怀黄菊CmPAN基因的克隆与表达分析  

Cloning and Expression Analysis of CmPAN Gene in Chrysanthemum morifolium cv. Huaihuang

作　　者：赵喜亭[1,2,3] 刘克 朱玉婷[1] 马孟丹[1] 吕萌 宋灵雨 蒋丽微 王苗[1] 李明军[1,2,3] ZHAO Xi-Ting;LIU Ke;ZHU Yu-Ting;MA Meng-Dan;LV Meng;SONG Ling-Yu;JIANG Li-Wei;WANG Miao;LI Ming-Jun(College of Life Science, Henan Normal University, Xinxiang 453007, China;Engineering Laboratory of Biotechnology for Green MedicinalPlant of Henan Province, Xinxiang 453007, China;Engineering Technology Research Center of Nursing and Utilization of Genuine Chinese Crude Drugs in Universities of Henan Province, Xinxiang 453007, China)

机构地区：[1]河南师范大学生命科学学院,新乡453007 [2]绿色药材生物技术河南省工程实验室,新乡453007 [3]河南省高校道地中药材保育及利用工程技术研究中心,新乡453007

出　　处：《农业生物技术学报》2019年第4期615-623,共9页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金-河南联合基金(No.U1704120);国家自然科学基金(No.31372105);中国博士后科学基金(No.2011M500457);中央高校基本科研业务费专项资金(No.2011JS076);河南省创新型科技人才队伍建设工程项目(No.C20130037)

摘　　要：怀菊花(Chrysanthemum morifolium)是著名的四大怀药之一,具有很高的观赏、食用及药用价值,但其花器官发育缺乏相关研究。为探究PERIANTHIA (PAN)基因在怀菊花花发育中的功能,首先利用同源克隆与cDNA末端快速扩增(rapid amplification of cDNA ends, RACE)技术成功克隆获得怀菊花优良种质怀黄菊(Ch. morifolium cv. Huaihuang)中CmPAN的cDNA序列,全长为1 484 bp,其中CDS区为1 314bp,5'UTR和3'UTR分别为64和106 bp。预测其编码437个氨基酸,与其他植物的PAN序列具有较高同源性,且C末端具有碱性亮氨酸拉链(basic leucine zipper motif, bZIP)家族D亚族的特征区和2个谷氨酰胺丰富区,与拟南芥(Arabidopsis thaliana) AtPAN的亲缘关系最近,故命名为CmPAN (GenBank No.KX380854)。生物信息学分析显示,CmPAN蛋白质的预测分子量和等电点分别为48.265 k D和8.82;CmPAN为非分泌性蛋白,并具有核定位序列,无序化程度小于AtPAN。qRT-PCR检测结果表明,CmPAN在花与茎中的表达量高于根与叶;在花发育的现蕾期至透色期表达量较高,之后呈下降趋势;在花器官中表达水平从高至低依次为管状花瓣、管状花雄蕊、舌状花瓣、舌状花心皮、花托。上述结果提示,CmPAN可能在花发育初期发挥重要作用,并且与管状花的花瓣和雄蕊的发育密切相关。本研究为进一步探究CmPAN基因的功能提供了参考依据。Chrysanthemum morifolium originated from the ancient Huaiqingfu is one of the Four Famous Huai Medicines, which has high ornamental, edible and medicinal value. But it is rare reported about the research on its flower organ development. To study the function of PERIANTHIA(PAN) gene in flower development of Ch. morifolium, a 1 484 bp full-length cDNA sequence of homologous PAN gene was cloned by employing homology gene cloning and Rapid amplification of cDNA ends(RACE) from Ch. morifolium cv.Huaihuang. The cDNA sequence contained 64 bp 5’ UTR, 106 bp 3’ UTR and 1 314 bp CDS which encoded437 amino acids. The protein of CmPAN had very high similarity with PANs from other plant species. Cterminal amino acids sequence of CmPAN included group D structural features with a well characterized family of plant bZIPs(basic leucine zipper motif) and 2 glutamine-rich regions. CmPAN was most close to AtPAN, so the cloned sequence was named CmPAN(GenBank No. KX380854). Putative protein molecular weight was 48.265 kD, the theoretical isoelectric point was 8.82, and it was non-secretory protein with nuclear localization sequence, its disordered degree was smaller than that of AtPAN. The qRT-PCR detection showed that CmPAN was higher expressed in flowers and stems than that in roots and leaves, and the CmPAN expession in floral development increased from bud emergence stage to visible color stage, then decreased at late phase.The CmPAN expession in floral organ ranked from high to low was petal of tubular flower, stamen of tubular flower, petal of ray floret, carpel of ray floret and receptacle. As a result, CmPAN might have important roles in early phase of floral development, and be closely related to the development of petals and stamens of tubular flower. The study provides some reference for further exploring the function of CmPAN gene.

关 键 词：怀黄菊 PERIANTHIA (PAN) 基因克隆 生物信息学分析 表达分析 

分 类 号：S682.11[农业科学—观赏园艺] Q945.45[农业科学—园艺学]