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拟南芥突变体npr1-2的鉴定及水杨酸感受性分析

Identification and Salicylic Acid Sensitivity Analysis of Arabidopsis Mutant npr1-2
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摘要 【目的】拟南芥的NPR1 (nonexpresser pathogenesis-related gene 1)是植物系统获得抗性(systemic acquired resistance,SAR)的关键调节基因,SAR是一种有效的先天免疫反应,提供广谱抗病性。大量的研究表明:水杨酸为诱导SAR的细胞外信号,而NPR1蛋白作为水杨酸的受体串联并启动细胞内的反应。拟南芥突变体npr1-2是研究拟南芥水杨酸信号转导途径的材料之一。为了筛选出纯合的npr1-2突变体,测定纯合突变体中NPR1基因的表达水平,鉴定突变幼苗对水杨酸的敏感性。【方法】采用PCR扩增、DNA测序、定量RT-PCR以及水杨酸敏感性鉴定等方法。【结果】鉴定并筛选出了npr1-2纯合突变体,发现突变体NPR1基因表达量下调为野生型的30%左右,突变体幼苗对水杨酸的敏感性降低。【结论】npr1-2纯合突变体为进一步的研究奠定了材料基础。 [Purpose]NPR1(nonexpresser of pathogenesis-related gene 1)is a key regulator of SAR(systemic acquired resistance)in Arabidopsis thaliana.SAR is an effective innate immune response that provides protection against a broad range of pathogens.Numerous studies have revealed that NPR1 as a salicylic acid(SA)perception chains together extracellular SA signaling and intracellular resistant reaction.Mutant npr1-2 is one of the materials for studying SA signal transduction pathway in Arabidopsis.The objective of this research is to screen out homozygous npr1-2 mutants,detection of NPR1 gene expressional level in homozygous mutants and identification of the sensitivity of mutant seedlings to SA.[Method]PCR amplification,DNA sequencing,quantitative RT-PCR detection and SA sensitivity analysis approaches were used in this study.[Result]The homozygous npr1-2 mutant was obtained.And the dramatic reduction in expression of NPR1 in the homozygous npr1-2 mutant,to about 30%of wild type plants has been found by qRT-PCR detection.Compared with the wild type,the sensitivity of homozygous npr1-2 seedling roots to high concentration SA was decreased.[Conclusion]The homozygous mutant of npr1-2 can provide the material for further research.
作者 任杰 吴嘉 张照利 乔菊香 王云月 杨红玉 REN Jie;WU Jia;ZHANG Zhaoli;QIAO Juxiang;WANG Yunyue;YANG Hongyu(Ministry of Education Key Laboratory for Agrobiodiversity and Pest Management,Yunnan Agricultural University,Kunming 650201,China;Department of Life Science and Technology,Kunming University,Kunming 650214,China;College of Forestry,Southwest Forestry University,Kunming 650224,China)
出处 《云南农业大学学报(自然科学版)》 CSCD 北大核心 2018年第5期964-968,共5页 Journal of Yunnan Agricultural University:Natural Science
基金 国家自然科学基金项目(31760078,31260062)。
关键词 NPR1 npr1-2 纯合体 水杨酸感受性 NPR1 npr1-2 homozygote salicylic acid sensitivity
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