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园艺学报 ›› 2012, Vol. 39 ›› Issue (10): 1893-1902.

• 果树 • 上一篇    下一篇

苹果酰基辅酶A结合蛋白2 编码基因MdACBP2的克隆和表达分析

柏素花, 祝军, 戴洪义   

  1. (1 湖南农业大学园艺园林学院,长沙 410128;2 青岛农业大学园林园艺学院,山东青岛 266109;3 青岛农业大学生命科学学院,山东省高校植物生物技术重点实验室,山东青岛 266109)
  • 出版日期:2012-10-25 发布日期:2012-10-25

Cloning and Expression Analysis of a New Acyl-CoA-binding Protein 2 (MdACBP2)Identified from Malus domestica

 BAI  Su-Hua, ZHU  Jun, DAI  Hong-Yi   

  1. (1College of Horticulture and Landscaping,Hunan Agricultural University,Changsha 410128,China;2College of Landscaping and Horticulture,Qingdao Agricultural University,Qingdao,Shandong 266109,China;3Key Lab of Plant Biotechnology of Shandong Province,College of Life Sciences,Qingdao Agricultural University,Qingdao,Shandong 266109,China)
  • Online:2012-10-25 Published:2012-10-25

摘要: 酰基辅酶A结合蛋白(ACBP)是一个保守的蛋白家族,在酰基酯辅酶A的转运过程中发挥重要作用。在水稻和拟南芥中鉴定的ACBP除了参与长链酰基辅酶A的转运以外,还参与了植物对生物和非生物胁迫的反应。为了探讨在苹果树中是否也存在参与植物对逆境反应的ACBP,并可用于改良苹果品种,以提高抗逆能力,从苹果品种‘鸡冠’中克隆、鉴定了一个ACBP基因,命名为MdACBP2MdACBP2包含一个378 bp的开放阅读框,编码包含125个氨基酸残基的蛋白质;推定的氨基酸序列中包含有一个ACB结构域,无信号肽序列。序列和结构特征表明该基因是ACBP家族ClassⅠ亚族的成员。利用荧光定量PCR技术检测MdACBP2的表达,发现在苹果的根、叶、花、幼果、枝皮、芽等组织中均有表达,在叶中的表达量最高,在幼果中的表达量最低,这一表达模式暗示MdACBP2可能参与多种生理途径。不同胁迫处理的时序表达分析表明,MdACBP2的表达能被轮纹病病原菌Botryosphaeria dothidea侵染和10% PEG渗透胁迫所抑制,低温胁迫则能诱导MdACBP2的表达,1 mmol·L-1 Pb(NO3)2处理对MdACBP2的表达没有显著影响。推测MdACBP2可能参与了苹果对低温胁迫的防御反应。

关键词: 苹果, 酰基辅酶A 结合蛋白, 轮纹病菌, 低温胁迫, 渗透胁迫

Abstract: To investigate the function of acyl-CoA-binding protein in Malus domestica,we isolated a new ACBP gene(designated as MdACBP2) from leaf cDNA using RACE technique. The full-length cDNA has an open reading frame(ORF) of 378 bp encoding a protein of 125 amino acids. The putative amino acid sequence contains an ACB domain consisting of 85 amino acid residues as predicted with  SMART program. Sequence analysis and structural feature indicated that MdACBP2 belongs to Class Ι of ACBP family. Quantitative real-time PCR experiment revealed that MdACBP2 is expressed in root,leaf,flower,young fruit,bark and bud. The highest mRNA expression was found in leaf,and the lowest in young fruit. The responses of MdACBP2 expression to biotic or abiotic stress were also investigated using quantitative real-time PCR. The results showed that MdACBP2 expression was inhibited by osmotic stress and Botryosphaeria dothidea infection,but was enhanced by cold stress. There was no significant changes among the apple seedlings treated with 1 mmol · L-1 Pb(NO3)2. It was speculated that MdACBP2 gene might be involved in the defense of apple seedling against cold stress.

Key words: Malus domestica, acyl-CoA-binding protein, Botryosphaeria dothidea, cold stress, osmotic stress

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