H_2O_2与葡萄VvIPK2基因表达及其低温胁迫响应的关系  

Involvement of H_2O_2 in Regulating the Expression of VvIPK2 in Response to Low Temperature Stress in Leaves of Vitis

作　　者：李希东[1] 侯丽霞[1] 刘新[1] 卢江[2] 

机构地区：[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

出　　处：《园艺学报》2011年第6期1052-1062,共11页Acta Horticulturae Sinica

基　　金：农业部‘948’项目(2006-G26)

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。An inositol polyphosphate kinase gene named VvIPK2 from Vitis vinifera was cloned and its biomatics was analyzed,the role of H_2O_2 and the interaction between H_2O_2 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 H_2O_2 accumulation and enhanced VvIPK2 transcription level.Exogenous H_2O_2 could enhance expression of VvIPK2 and H_2O_2 cleaner（ascorbic acid,AsA）could reduce its expression.At the same time,exogenous H_2O_2 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 H_2O_2 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 H_2O_2 in grape response to low temperature stress.

关 键 词：葡萄 过氧化氢 VvIPK2 低温 

分 类 号：S663.1[农业科学—果树学]