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 (4): 686-698.doi: 10.16420/j.issn.0513-353x.2019-0246

• 研究论文 • 上一篇    下一篇

月季花瓣特异表达启动子的筛选和鉴定

王 焕*,郑日如*,曹声海,张 通,史若明,王彩云,罗 靖**   

  1. 华中农业大学园艺林学学院,园艺植物生物学教育部重点实验室,武汉 430070
  • 出版日期:2020-04-25 发布日期:2020-04-25
  • 基金资助:
    国家自然科学基金项目(31401911);湖北省自然科学基金项目(2014CFB944)

Selection and Identification of Petal-specific Promoter in Rose

WANG Huan*,ZHENG Riru*,CAO Shenghai,ZHANG Tong,SHI Ruoming,WANG Caiyun,and LUO Jing**   

  1. Key Laboratory for Biology of Horticultural Plants,Ministry of Education,College of Horticulture and Forestry Sciences,Huazhong Agricultural University,Wuhan 430070,China
  • Online:2020-04-25 Published:2020-04-25

摘要: 以月季品种‘萨曼莎’(Rosa hybrida‘Samantha’)为试验材料,选取了13个花色、花香相关的基因,研究其在根、茎、叶和花等器官中的表达规律及其启动子活性。结果表明,RhOOMT2(Rosa hybrida orcinol o-methyltransferase 2)和RhCCD4(Rosa hybrida carotenoid cleavage dioxygenase 4)在花瓣中具有相对较高的表达量,RhNUDX1(Rosa hybrida nudix hydrolase 1)在根部表达量最高,花瓣中其次,在茎和叶中几乎没有表达,另外10个类黄酮代谢途径相关基因在各器官中表达差异较小。使用FPNI-PCR的方法克隆到ATG上游1 601 bp的RhOOMT2启动子、1 539 bp的RhCCD4启动子和1 433 bp的RhNUDX1启动子。将携带RhOOMT2、RhCCD4和RhNUDX1启动子的PBI121载体通过农杆菌的介导,在月季‘Samantha’、洋桔梗(Eustoma grandiflorum‘Green Pelleted’)和百合(Lillium Oriental hybrida‘Siberia’)花瓣中进行瞬时表达,GUS染色结果表明,与阳性对照35S启动子相比,RhOOMT2启动子在月季、洋桔梗和百合花瓣中具有更强的活性,RhCCD4启动子在百合花瓣中具有较弱的活性,而在月季和洋桔梗中活性很弱,RhNUDX1启动子则在3种花中均无活性。进一步在4个月季品种‘萨曼莎’、‘戴安娜’、‘香槟月季’和‘雪山’中比较了RhOOMT2和35S的启动子活性,证实了在花瓣中RhOOMT2启动子具有比35S更高的活性。

关键词: 月季, RhOOMT2, 启动子, 花瓣特异性表达

Abstract: Thirteen color/scent-related genes in Rosa hybrida‘Samantha’were selected,and their expression in root,stem,leaf and petal showed that RhOOMT2(orcinol o-methyltransferase 2)and RhCCD4(carotenoid cleavage dioxygenase 4)had relatively high expression in petals. RhNUDX1 had high expression in root,moderate in petal,and weak in stem and leaf. The other ten anthocyanin synthesis-related genes did not show much difference in root,stem,leaf,and petal. The promoter sequences of RhOOMT2,RhCCD4 and RhNUDX1 were got by using FPNI-PCR method,whose length was 1 601,1 539 and 1 433 bp,separately. Agrobacterium tumefaciens EHA105 carrying 35S::GUS,proRhOOMT2::GUS,proRhCCD4::GUS,or proRhNUDX1::GUS was transferred into petals of rose (R. hybrida‘Samantha’),eustoma(Eustoma grandiflorum‘Green Pelleted’),and lily(Lillium Oriental hybrida‘Siberia’)by vacuum aspiration. The GUS staining result showed that the promoter of RhOOMT2 had stronger activity than 35S in rose,eustoma and lily petals,the promoter of RhCCD4 had relatively weak activity in lily petals,and very weak activity in rose and eustoma,and the promoter of RhNUDX1 almost had no activity in three kinds of flowers. The promoter activity of RhOOMT2 and 35S was also compared in four cultivars of rose,‘Samantha’,‘Diana’,‘Champagne’,and‘Snow Mountain’,the result confirmed that the promoter of RhOOMT2 had higher activity than 35S in petals.

Key words: rose;RhOOMT2, promoter, petal-specific expression

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