https://www.ahs.ac.cn/images/0513-353X/images/top-banner1.jpg|#|苹果
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https://www.ahs.ac.cn/images/0513-353X/images/top-banner12.jpg|#|菊花

园艺学报 ›› 2017, Vol. 44 ›› Issue (7): 1388-1396.doi: 10.16420/j.issn.0513-353x.2016-0884

• 研究报告 • 上一篇    下一篇

‘云香’水仙ACC 氧化酶基因克隆及遗传转化

孙申申,温秀萍,杨菲颖,申艳红,陈晓静*   

  1. 福建农林大学园艺学院,福建农林大学园艺植物遗传育种研究所,福州 350002
  • 出版日期:2017-07-25 发布日期:2017-07-25

Cloning of ACC Oxidase Gene from Narcissus tazetta var.‘Yunxiang’and Its Transformation

SUN Shenshen,WEN Xiuping,YANG Feiying,SHEN Yanhong,and CHEN Xiaojing*   

  1. College of Horticulture,Fujian Agriculture and Forestry University;Institute of Genetics & Breeding in Horticultural
    Plants,Fujian Agriculture and Forestry University,Fuzhou 350002,China
  • Online:2017-07-25 Published:2017-07-25

摘要:

以‘云香’水仙花瓣为试验材料,采用RT-PCR 技术克隆出乙烯生物合成途径中的关键
酶ACC 氧化酶(ACO)基因的全长序列1 162 bp,命名为NtACO1(GenBank 登录号为KX082935)。
其开放阅读框(ORF)长度为939 bp,编码312 个氨基酸。NtACO1 蛋白包括ACO 家族保守的DIOX_N
和20G-FeⅡ_Oxy 结构域。分子进化树分析表明,NtACO1 蛋白与‘金盏银台’水仙亲缘关系最近。
通过qRT-PCR 检测NtACO1 在‘云香’水仙花朵开放过程中的表达模式,结果表明,NtACO1 在花瓣
和副冠中的表达量随着花朵发育衰老而逐渐上升,并且各个时期花瓣中的表达量都高于副冠,推测
NtACO1 可能参与了‘云香’花的发育和衰老。构建了NtACO1 的正、反义植物表达载体,并以烟草
叶片为外植体,通过根癌农杆菌介导的遗传转化体系,获得NtACO1 过表达转基因植株。转基因烟草
与野生型相比缩短了营养生长期,开花较早,叶片较少。该研究为进一步探究水仙花ACO 基因功能,
以及延长水仙花花期的分子育种奠定了试验基础。

关键词: 水仙, 基因克隆, 基因工程, 延长花期, ACC 氧化酶

Abstract:

ACC oxidase(ACO)is a key enzyme in the biosynthesis pathway of ethylene. In order to
explore the characteristics and function of the ACO gene in narcissus,the following work has been done in
this study. Petals of Narcissus tazetta var.‘Yunxiang’were used as experimental materials,a full-length
1 162 bp sequence was obtained by RT-PCR and named as NtACO1(GenBank KX082935). The open
reading frame(ORF)length of NtACO1 is 939 bp,which encodes 312 amino acids. Amino acid domain
analysis showed that this gene has DIOX_N and 20G-FeⅡ_Oxy domains and was identified as a member
of the ACO protein family. Molecular phylogenetic tree analysis showed that ACO gene of‘Yunxiang’was
closely related to that of Narcissus tazetta var. chinensis. qRT-PCR results showed that the expression of
NtACO1 increased gradually with Narcissus flower development and senescence,and the expression level
in the petals were higher than that in contemporaneous coronas,indicating that the NtACO1 gene may be involved in the flower development and senescence of‘Yunxiang’. We obtained several NtACO1
overexpression transgenic tobacco plants using the Agrobacterium mediated. Compared with wild type,the
vegetative growth stage of transgenic tobaccos was reduced,as well as the transgenic tobaccos showed
early flowering and less leaves. This study lays an experimental foundation for further research on the
function of ACO gene and provide a theoretical basis for the molecular breeding in prolonging plant
florescence of Narcissus.

Key words: Narcissus, gene clone, genetic engineering, prolonged flowering, ACC oxidase