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园艺学报 ›› 2011, Vol. 38 ›› Issue (6): 1052-1062.

• 果树 • 上一篇    下一篇

H2O2与葡萄VvIPK2基因表达及其低温胁迫响应的关系

李希东1,侯丽霞1,刘 新1,*,卢 江2   

  1. (1青岛农业大学生命科学学院,山东青岛 266109;2中国农业大学食品科学与营养工程学院,北京 100093)   
  • 收稿日期:2011-02-17 修回日期:2011-04-27 出版日期:2011-06-25 发布日期:2011-06-25
  • 通讯作者: 刘 新

Involvement of H2O2 in Regulating the Expression of VvIPK2 in Response to Low Temperature Stress in Leaves of Vitis

LI Xi-dong1,HOU Li-xia1,LIU Xin1,*,and LU Jiang2   

  1. (1College of Life Science,Qingdao Agriculture University,Qingdao,Shandong 266109,China;2College of Food Science and Nutritional Engineering,China Agriculture University,Beijing 100093,China)  
  • Received:2011-02-17 Revised:2011-04-27 Online:2011-06-25 Published:2011-06-25
  • Contact: LIU Xin

摘要: 以葡萄砧木品种‘F-242’试管苗为材料,克隆了葡萄(Vitis L.)的多磷酸肌醇激酶(inositol polyphosphate kinase,IPK2)基因VvIPK2,并对其进行生物信息学分析;利用实时定量PCR和植物生理学技术,结合药理学实验探究了H2O2在葡萄响应低温胁迫中的作用及其与VvIPK2的关系。结果表明,克隆的VvIPK2其推导的翻译产物(VvIPK2)与数据库中推测的氨基酸序列一致;与栽培亚麻和拟南芥多磷酸肌醇激酶高度同源。在低温胁迫下,叶片VvIPK2表达与H2O2水平均明显升高;外源H2O2能提高VvIPK2表达水平,而H2O2清除剂抗坏血酸(ascorbic acid,AsA)降低VvIPK2表达水平。外施一定浓度的H2O2可提高超氧化物歧化酶(superoxide dismutase,SOD)活性,上调Cu/Zn SOD基因表达,降低丙二醛(malondialdehyde,MDA)含量和细胞膜相对透性;但IPK2抑制剂LY-294002对内源H2O2却无明显影响;LY-294002和AsA均能降低SOD活性,下调Cu/Zn SOD基因表达,提高MDA含量和细胞膜相对透性。推测H2O2调节VvIPK2的表达,参与葡萄响应低温胁迫过程。   

关键词: 葡萄, 过氧化氢, VvIPK2, 低温

Abstract: An inositol polyphosphate kinase gene named VvIPK2 from Vitis vinifera was cloned and its biomatics was analyzed,the role of H2O2 and the interaction between H2O2 and VvIPK2 in response to cold stress of‘F-242’leaves were investigated by real-time PCR,combined with physiological and pharmacological methods. The results showed that the deduced translation product(VvIPK2)was completely the same with the sequence in database which shares 64% and 62% identity with the inositol polyphosphate kinase(LuIPK2)in Linum usitatissimum and that in Arabidopsis(AtIPK2). Low temperature induced H2O2 accumulation and enhanced VvIPK2 transcription level. Exogenous H2O2 could enhance expression of VvIPK2 and H2O2 cleaner(ascorbic acid,AsA)could reduce its expression. At the same time,exogenous H2O2 could enhance the activities of superoxide dismutase(SOD)and Cu/Zn SOD expression,reduce the content of malondialdehyde(MDA)and the relative permeability of cell membrane under cold stress. But LY-294002 had no effect on cold-induced H2O2 production. Moreover,LY-294002 and AsA reversed the changes of SOD,MDA and the relative permeability of cell membrane,implying that VvIPK2 was assumed to be the downstream of H2O2 in grape response to low temperature stress.  

Key words: Vitis vinifera, hydrogen peroxide, VvIPK2, low temperature stress

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