http://www.ahs.ac.cn/images/0513-353X/images/top-banner1.jpg|#|苹果
http://www.ahs.ac.cn/images/0513-353X/images/top-banner2.jpg|#|甘蓝
http://www.ahs.ac.cn/images/0513-353X/images/top-banner3.jpg|#|菊花
http://www.ahs.ac.cn/images/0513-353X/images/top-banner4.jpg|#|灵芝
http://www.ahs.ac.cn/images/0513-353X/images/top-banner5.jpg|#|桃
http://www.ahs.ac.cn/images/0513-353X/images/top-banner6.jpg|#|黄瓜
http://www.ahs.ac.cn/images/0513-353X/images/top-banner7.jpg|#|蝴蝶兰
http://www.ahs.ac.cn/images/0513-353X/images/top-banner8.jpg|#|樱桃
http://www.ahs.ac.cn/images/0513-353X/images/top-banner9.jpg|#|观赏荷花
http://www.ahs.ac.cn/images/0513-353X/images/top-banner10.jpg|#|菊花
http://www.ahs.ac.cn/images/0513-353X/images/top-banner11.jpg|#|月季
http://www.ahs.ac.cn/images/0513-353X/images/top-banner12.jpg|#|菊花

园艺学报 ›› 2020, Vol. 47 ›› Issue (1): 161-168.doi: 10.16420/j.issn.0513-353x.2019-0135

• 研究报告 • 上一篇    下一篇

菊花胚发育相关基因Cm2S的克隆与表达分析

徐素娟1,2,吴 泽1,2,赵静雅1,2,侯慧中1,2,陈发棣1,滕年军1,2,*   

  1. 1南京农业大学园艺学院,农业农村部景观设计重点实验室,南京 210095
  • 出版日期:2020-01-25 发布日期:2020-01-25
  • 基金资助:

    国家自然科学基金项目(31672182);中央高校基本业务费专项基金项目(KYZ201602);江苏省自然科学基金项目(BK20161449)

Cloning and Expression Analysis of Cm2S,a Gene Related to Embryo Development in Chrysanthemum

XU Sujuan1,2,WU Ze1,2,ZHAO Jingya1,2,HOU Huizhong1,2,CHEN Fadi1,and TENG Nianjun1,2,*   

  1. 1Key Laboratory of Landscape Design,Ministry of Agriculture and Rural Affairs,College of Horticulture,Nanjing Agricultural University,Nanjing 210095,China;2Jiangsu Graduate Workstation Jointed by Nanjing Agricultural University and Nanjing Lidao Modern Agriculture Development Co.,Ltd,Nanjing 210043,China
  • Online:2020-01-25 Published:2020-01-25

摘要:

通过对菊花(Chrysanthemum morifolium)品种‘雨花落英’与四倍体菊花脑(C. nankingense)杂交后胚的转录组和蛋白组数据分析,筛选到1个菊花胚不同发育时期差异表达的基因Cm2S。采用高保真PCR技术,克隆得到Cm2S的开放阅读框(ORF)。序列分析表明,Cm2S开放阅读框为852 bp,编码283个氨基酸。利用SMART在线网站预测Cm2S蛋白的功能结构域,结果显示,Cm2S蛋白属于AAI_LTSS家族,具有两个保守的AAI(Alpha-Amylase Inhibitors)结构域。对其进行同源性比对及系统进化树分析发现,Cm2S与青蒿(Artemisia annua)种子贮藏蛋白AaSSP6亲缘关系最近。亚细胞定位结果显示,Cm2S蛋白在细胞质和细胞核中均有分布。荧光实时定量PCR分析表明,Cm2S在菊花的胚中特异性表达,且在授粉后18 d正常胚中的表达量显著高于授粉后12 d的胚,是其315.11倍,同时也显著高于授粉后18 d败育的胚,是其1.98倍。

关键词: 菊花, 菊花脑, 胚胎发育, Cm2S, 基因克隆, 亚细胞定位, 组织定量

Abstract:

Based on the analysis of transcriptome and proteome data of embryos in the hybridization of Chrysanthemum morifolium var.‘Yuhualuoying’× tetraploid C. nankingense,a gene named Cm2S was identified that could express differentially in different stages of embryo development. Cm2S was isolated from normal embryos of‘Yuhualuoying’by a high-fidelity PCR,and the Open Reading Frame(ORF)length of Cm2S was 852 bp,encoding a protein of 283 amino acids. The SMART online website was used to predict the functional structural domains of Cm2S protein,the results showed that Cm2S protein belongs to the AAI_LTSS family and it has two conservative AAI(alpha-amylase Inhibitors)structural domains. Homology comparison and phylogenetic tree analysis showed that Cm2S had the highest genetic relationship with AaSSP6 in Artemisia annua. The result of subcellular localization showed that Cm2S was located in cytoplasm and nucleus. Quantitative real-time PCR(qRT-PCR)showed that Cm2S was specifically expressed in embryos,and the expression level in normal embryos at 18 days after pollination was significantly higher than that in the other two samples. It was 315.11 times of that in normal embryos at 12 days after pollination while 1.98 times of that in abortion embryos at 18 days,respectively.

Key words: Chrysanthemum morifolium, C. nankingense, embryo development, Cm2S, gene cloning, subcellular localization, tissue quantification

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