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作物学报 ›› 2009, Vol. 35 ›› Issue (6): 1161-1166.doi: 10.3724/SP.J.1006.2009.01161

• 研究简报 • 上一篇    下一篇

甘蔗中一个NBS-LRR类基因的全长克隆与表达分析

阙友雄，许莉萍*，张木清，张积森，陈如凯   

  1. 福建农林大学/农业部甘蔗遗传改良重点开放实验室，福建福州350002
  • 收稿日期:2008-09-13 修回日期:2008-12-13 出版日期:2009-06-12 网络出版日期:2009-04-16
  • 通讯作者: 许莉萍，E-mail:xlpmail@yahoo.com.cn
  • 基金资助:

    本研究由国家高技术研究发展计划（863计划）项目（2007AA100701），农业部引进国际先进农业科学技术计划（948计划）项目（2006-G37），国家自然科学基金项目（30170639），福建省科技厅国家科技项目备案（F2007AA100701）资助。

Cloning and Expression Analysis of an NBS-LRR Type Gene from Sugarcane

QUE You-Xiong,XU Li-Ping*,ZHANG Mu-Qing,ZAHNG Ji-Sen,CHEN Ru-Kai   

  1. Key Laboratory of Sugarcane Genetic Improvement,Ministry of Agriculture,Fujian Agriculture and forestry University,Fuzhou 350002,China
  • Received:2008-09-13 Revised:2008-12-13 Published:2009-06-12 Published online:2009-04-16
  • Contact: XU Li-Ping，E-mail:xlpmail@yahoo.com.cn

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2162

被引次数

16

摘要：

采用RACE技术, 从甘蔗高抗黑穗病品种NCo376中克隆了一个NBS-LRR类基因的cDNA全长序列, 命名为SNLR。生物信息学分析显示, 甘蔗SNLR基因的cDNA全长为2 985 bp (Accession No. EF155654), 包括一个2 661 bp的完整开放读码框以及一个典型的29 bp poly-A。同时, 还具有NBS-LRR类抗病基因的所有保守结构域, 包括4个NBS区域保守结构域和6个潜在LRR结构域；蛋白疏水性分析和二级、三级结构分析表明, 该基因编码的蛋白质为弱碱性蛋白, pI为7.76, 无明显的疏水结构域, 以卷曲结构和螺旋结构为骨架, 三级结构未见明显的跨膜信号蛋白区。定量PCR分析表明, 甘蔗SNLR基因的表达受到黑穗病菌、水杨酸和过氧化氢的影响, 分别表现出“抑-扬”、“全程抑制”和“扬-抑”的表达模式, 也具有抗病基因组成型和组织特异性表达的特点。推测甘蔗SNLR基因可能为抗病相关基因。

关键词: 甘蔗, 分子克隆, 抗病基因, 定量PCR

Abstract:

In the present study, a non-TIR-NBS-LRR type disease-related gene was cloned by rapid amplification of cDNA ends (RACE) using the high-resistant sugarcane variety NCo376.This gene was termed as SNLR, with the GenBank accession No. of EF155648. The full-length cDNA sequence of SNLR is 2 985 bp, including an open reading frame (ORF) of 2661 bp and the typical 29 bp poly-A. The SNLR gene contained all the four typical conserved motifs of the NBS: P-loop (GMGGVGGKTT), Kinase-2 (LIVLDD), Kinase3a (GSR/KILVIIR) and hydrophobic region (GLPLAL), plus six putative LRR regions. It can be deduced from the hydrophobic character, secondary structure and 3D model analysis of the corresponding coding protein that the SNLR protein was alkalescent, with pI of 7.76 and without any obvious hydrophobic domain; coil and helices were the framework of secondary structure; no transmembrane region was found in its protein 3D model. The gene expression profile under the treatment ofU. scitaminea, SA and H2O2 wereinvestigated by Real-time qPCR. Results showed that expression ofsugarcane SNLR gene was influenced by the fungus, SA and H2O2, with the expression patterns of “down-up”, “down in the whole process” and “up-down”, respectively. It was inferred that expression of SNLR gene occurs both via an H2O2- and SA-dependent pathway. At the same time, SNLR gene was found to be expressed highly in leaves, mildly in stalks and slightly in roots, which indicated its relation to resistance in this aspect.

Key words: Sugarcane, Molecular cloning, Disease resistance gene, Real-time quantity PCR

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