园艺学报 ›› 2018, Vol. 45 ›› Issue (12): 2371-2382.doi: 10.16420/j.issn.0513-353x.2018-0433

• 研究论文 • 上一篇    下一篇


陈义挺,冯 新*,赖瑞联,高敏霞,陈文光,吴如健   

  1. 福建省农业科学院果树研究所,福州 350013
  • 出版日期:2018-12-25 发布日期:2018-12-25
  • 基金资助:

Cloning and Expression Regulation of the AdCCS1 from Actinidia chinensis var. deliciosa

CHEN Yiting,FENG Xin*,LAI Ruilian,GAO Minxia,CHEN Wenguang,and WU Rujian   

  1. Fruit Research Institute,Fujian Academy of Agricultural Sciences,Fuzhou 350013,China
  • Online:2018-12-25 Published:2018-12-25

摘要: 采用RT-PCR法从‘米良1号’猕猴桃中分离到1 066 bp的铜锌超氧化物歧化酶分子伴侣蛋白基因AdCCS1,登录号为KY471361。AdCCS1的开放阅读框为984 bp,编码327个氨基酸,包含3个典型结构域(N端结构域、中间结构域和C端结构域)和2个保守的金属结合位点(MXCXXC和CXC)。基因结构分析显示AdCCS1由6个外显子和5个内含子组成。系统进化分析表明AdCCS1聚在双子叶植物分支中,与茶树CsCCS的亲缘关系最近。此外,AdCCS1的5′端调控区存在多种光响应、胁迫响应和激素响应应答元件。定量PCR分析显示,AdCCS1在猕猴桃叶片中的表达量最高,其次是成熟果、花、幼果和茎,在根中的表达量最低。猕猴桃果实中AdCCS1在4 ℃低温贮藏过程中的表达量均比25 ℃贮藏中同期的低,说明低温抑制AdCCS1的表达。脱落酸和赤霉素处理后AdCCS1的表达下调,但二者的应答模式不同。

关键词: 猕猴桃, AdCCS1, 贮藏温度, ABA, GA3, 表达分析

Abstract: Copper/zinc superoxide dismutases(Cu/Zn SOD),acting as an essential antioxidant enzyme,play important roles in plant growth and development,stress response and fruit senescence,while their activities depended on the copper chaperone for Cu/Zn SOD(CCS)to delivering copper to them. Therefore,CCS is also involved in the plant growth and development and stress response. In this study,a 1 066 bp gene of copper chaperone for Cu/Zn SOD(AdCCS1)was isolated from Actinidia chinensis var. deliciosa‘Miliang 1’by using reverse transcription(RT-PCR),and its accession number was KY471361. The AdCCS1 gene had an open reading frame of 984 bp and encoded 327 amino acids,which contained three typical domains(N-terminal domain,central domain and C-terminal domain)and two conserved metal binding sites(MXCXXC and CXC). Gene structure analysis showed that AdCCS1 gene consisted of six extrons and five introns. Phylogenetic analysis revealed that AdCCS1 was clustered in the dicotyledonous branch with close relation to tea CsCCS. In addition,a variety of cis-acting elements associated with light response,stress response and hormone response were present in the 5′ terminal regulatory region of the AdCCS1 gene. Quantitative PCR analysis showed that AdCCS1 was expressed strongly in kiwifruit leaves,followed by ripe fruits,flowers,young fruits and stems,but weakly in roots. The expression level of AdCCS1 in kiwifruits stored at 4 ℃ was lower than that of the same time stored at 25 ℃,which suggested that low temperature inhibit the expression of AdCCS1. The expression of AdCCS1 in fruits was down-regulated after being treated with abscisic acid and gibberellin,while their expression patterns were different.

Key words: Actinidia chinensis, AdCCS1, storage temperature, ABA, GA3, expression analysis