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2025年8月14日 星期四
  2015, Vol. 23 Issue (2): 170-180    
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
甘蔗抗坏血酸过氧化物酶基因(ScAPX)的克隆及表达分析
王竹青1,陈云1,杨玉婷1,苏亚春2,陈珊珊1,吴期滨3,3,许莉萍1
1. 福建农林大学国家甘蔗产业研究中心
2. 国家甘蔗产业技术研发中心
3.
Cloning and Expression Analysis of Ascorbate Peroxidase Gene (ScAPX) in Sugarcane (Saccharum officinarum)
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摘要 抗坏血酸过氧化物酶(ascorbate peroxidase, APX)是清除活性氧(reactive oxygen species, ROS)的重要酶类。本研究对针刺接种黑粉菌(Sporisorium scitamineum)后的抗、感基因型甘蔗(Saccharum officinarum)进行APX活性测定。结果表明,甘蔗接种黑穗病菌48 h内,抗病品种(崖城05-179) APX活性显著高于感病品种(柳城03-182) (P<0.05)。借助电子克隆技术,结合RT-PCR方法,从甘蔗中分离到一个APX基因,并命名为ScAPX(GenBank登录号: KJ7565501)。生物信息学分析显示,ScAPX基因全长1 171 bp,包含一个1 038 bp的完整开放阅读框,编码345个氨基酸。ScAPX编码的蛋白不含信号肽,推测其为非分泌蛋白,定位于线粒体基质和叶绿体基质中的概率分别为91.1%和88.7%。实时荧光定量PCR检测结果显示,ScAPX在甘蔗根、芽、叶、蔗皮和蔗髓中均有表达,为组成型表达基因,表达量在蔗皮中最高,叶中最低,前者是后者的19.7倍;在外源因子处理后0~48 h,ScAPX基因的表达受水杨酸(salicylic acid, SA)、茉莉酸甲酯(methyl jasmonate, MeJA)、过氧化氢(hydrogen peroxide, H2O2)、脱落酸(abscisic acid, ABA)、氯化钠(sodium chloride, NaCl)和聚乙二醇(polyethylene glycol, PEG)诱导,SA、MeJA和H2O2处理后的ScAPX转录本积累量峰值较外源激素(ABA)和环境(NaCl和PEG)胁迫早。前者的ScAPX转录本变化呈现出胁迫初期积累量增加,达到峰值后又逐步下降的特点;在同样的测试时间(处理后24 h)内,ABA、NaCl和PEG处理后ScAPX转录本在达到峰值后未见下降。虽然在外源胁迫下,ScAPX表达量变化存在明显差异,但其表达均表现出正响应上述外源因子对甘蔗的胁迫。本研究为后续深入鉴定该基因的功能与进一步应用提供基础资料。
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王竹青
陈云
杨玉婷
苏亚春
陈珊珊
吴期滨
许莉萍
关键词 甘蔗甘蔗黑穗病菌抗坏血酸过氧化物酶基因(ScAPX)非生物胁迫qRT-PCR    
Abstract:Ascorbate peroxidase (APX) is one of the important enzymes which can remove the reactive oxygen species (ROS) in the plant. In this study, APX activity in sugarcane (Saccharum officinarum) smut resistant variety (Yacheng05-179) was significantly higher (P<0.05) than that in the susceptible one (Liucheng03-182) after inoculated with Sporisorium scitamineum within 48 h. ScAPX (GenBank accession No. KJ7565501) was separated from sugarcane with cloning and RT-PCR technologies. The bioinformatic analysis showed that the total length of ScAPX gene was 1 171 bp, which contained a complete open reading frame (1 038 bp) and encoded 345 amino acids. The ScAPX contained no signal peptide and belonged to nonsecretory protein which was likely located in the matrix of mitochondrial (91.1%) and chloroplast (88.7%). The results of tissue specificity analysis showed that the highest expression level of ScAPX was in stem skin which was 19.7 times of that in leaves. The transcript of ScAPX increased under the stresses of salicylic acid (SA), methyl jasmonate (MeJA), hydrogen peroxide (H2O2), abscisic acid (ABA), sodium chloride (NaCl) and polyethylene glycol (PEG). The peak of the ScAPX transcript under the stresses of SA, MeJA and H2O2 was accumulated during the initial period, which was earlierly found than that under ABA, NaCl and PEG stresses, and then it declined gradually. The expression pattern of ScAPX in response to ABA, NaCl and PEG treatments was not clear after 24 h, while the transcripts were kept in the peak at 24 h. Though the gene expression under the exogenous stresses were different, it was undoubted that ScAPX was positive response to the external stress. This study provides the basis for further functional analysis and application of this gene in the future.
Key wordsSugarcane    Sporisorium scitamineum    Ascorbate peroxidase gene(ScAPX)    Abiotic stresses    qRT-PCR
收稿日期: 2014-06-03      出版日期: 2015-01-13
基金资助:现代农业产业技术体系建设专项资金
通讯作者: 许莉萍     E-mail: xlpmail@126.com
引用本文:   
王竹青 陈云 杨玉婷 苏亚春 陈珊珊 吴期滨 许莉萍. 甘蔗抗坏血酸过氧化物酶基因(ScAPX)的克隆及表达分析[J]. , 2015, 23(2): 170-180.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2015/V23/I2/170
 
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