海岛棉抗枯萎病相关GbNPR1基因的克隆及其表达分析

Cloning and Expression Analysis of Fusarium Oxysporum Resistance-related GbNPR1 Genes in Gossypium barbadense

目的:克隆海岛棉GbNPR1基因并研究该基因在抗病及抗病信号传导中的功能。方法:根据已报道的棉花NPR1基因序列设计1对特异引物,利用RT-PCR从新海21号克隆GbNPR1基因并对其序列进行生物信息学分析;通过实时定量PCR技术检测海岛棉经枯萎病菌侵染、水杨酸和乙烯诱导后该基因的表达特性。结果:GbNPR1基因编码587个氨基酸,与可可树中NPR1基因编码的氨基酸同源性最高(89.61%),其蛋白含有保守的BTB/POZ、ANK和NPR1_like_C结构域;枯萎病菌侵染,水杨酸和乙烯诱导均可使GbNPR1明显上调表达,枯萎病菌侵染3h,水杨酸和乙烯诱导2h表达量最高。结论:GbNPR1基因在棉花抵御枯萎病菌侵染及植物抗病信号转导过程中起一定作用。

Objective： To clone the GbNPR1 gene from Gossypium barbadense and study its function of disease resistance as well as resist- ance signal transduction. Method：A specific primer were designed according to the reported sequence of cotton NPR1 gene. Using the method of RT- PCR, GbNPR1 gene was cloned from Xinhai 21 which was induced by Fusarium oxysporum f. sp. Vasinfectum（Fov）. , Then bioinformatics analysis on the gene sequence was conducted. We detected the expression characterristics of GbNPRI after induced by FOV, alicylic acid and ethylene using qPCR. Result： GbNPR1 was encoded 587 amino acids,which had a highest homology of 89. 61% with amino acids encoded by NPR1 gene of cacao tree, the protein of GbNPR1 contained three conserved structure domains, incluingBTB/POZ, ANK and NPR1 like C. The expression of GbNPR1 gene was obviously up - regulated by the induction of Fov, salicylic acid and ethylene. There was a highest expression level at three hours after fusarium infection and two hours after induced by salicylic acid and ethylene. Conclusion：GbNPR1 gene played a certain role in resistance to fusarium infection as well as the process of disease resistance sig- nal transduction.