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园艺学报 ›› 2011, Vol. 38 ›› Issue (6): 1081-1088.

• 果树 • 上一篇    下一篇

番木瓜果实成熟相关基因的cDNA-AFLP分析及克隆

申艳红,陈晓静*,卢秉国,何玮毅   

  1. (福建农林大学园艺学院,园艺植物遗传育种研究所,福州 350002)   
  • 收稿日期:2010-01-25 修回日期:2011-05-16 出版日期:2011-06-25 发布日期:2011-06-25
  • 通讯作者: 陈晓静

cDNA-AFLP Analysis and Cloning of Fruit Ripening-related Genes from Papaya

SHEN Yan-hong,CHEN Xiao-jing*,LU Bing-guo,and HE Wei-yi   

  1. (College of Horticulture,Institute of Genetics & Breeding in Horticultural Plants,Fujian Agriculture and Forestry University,Fuzhou 350002,China)   
  • Received:2010-01-25 Revised:2011-05-16 Online:2011-06-25 Published:2011-06-25
  • Contact: CHEN Xiao-jing

摘要: 为了探讨番木瓜果实成熟机理,利用cDNA-AFLP技术对破色期和半黄期的番木瓜果实进行基因差异表达分析,获得了50个与果实成熟相关的差异基因片段,其中28个与GenBank数据库中的功能基因同源,分别参与了果实成熟过程中基因表达调控,DNA与蛋白质合成及运输,蛋白质降解,能量代谢,营养物质代谢和逆境胁迫反应等过程。在差异片段序列基础上设计特异引物,采用cDNA末端快速扩增(RACE)技术成功克隆了4条差异基因cDNA全长,分别命名为CpGMP、CpPAA1、CpPOD和CpMYB。半定量RT-PCR分析结果表明,随着果实的成熟衰老,4个基因的表达量相应变化,说明其可能参与果实的成熟过程。

关键词: 番木瓜, 果实, 成熟, cDNA-AFLP, 差异表达, RACE

Abstract: In order to explore molecular mechanism of fruit ripening and the key softening-related genes,cDNA-AFLP technique was used to identify differentially expressed genes during papaya fruit ripening. Fifty transcript-derived fragments(TDFs)were obtained from the analysis of semi-yellow and color-break stage of papaya fruit. Bioinformatics analysis showed that 28 TDFs were homologous to the known functional genes,which could be divided into different functional groups including gene expression regulation,DNA and protein synthesis and transport,protein degradation,energy metabolism,nutrient metabolism and stress responding. Using specific primers designed from the sequences of TDFs,four differential expression genes were cloned by rapid amplification of cDNA ends(RACE). Semi-quantitative RT-PCR analysis showed that the expression of four genes changed accordingly during fruit ripening,indicating the possible role of these genes in the ripening of papaya.  

Key words: Carica papaya, fruit, ripening, cDNA-AFLP, differential expression, RACE

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