| 西洋参根腐病抗性相关基因PqDELLA1的克隆及表达分析 |
| 投稿时间：2021-05-07 点此下载全文
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| 引用本文：杨姗姗,王仪,刘紫祺,邵慧慧,高微微.西洋参根腐病抗性相关基因PqDELLA1的克隆及表达分析[J].中国现代中药,2022,24(5):776-783 |
| DOI：10.13313/j.issn.1673-4890.20210507001 |
| 摘要点击次数: 453 |
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| 作者中文名 | 作者英文名 | 单位中文名 | 单位英文名 | E-Mail |
| 杨姗姗 |
YANG Shan-shan |
中国医学科学院 北京协和医学院 药用植物研究所，北京 100193 |
Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China |
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| 王仪 |
WANG Yi |
中国医学科学院 北京协和医学院 药用植物研究所，北京 100193 |
Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China |
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| 刘紫祺 |
LIU Zi-qi |
中国医学科学院 北京协和医学院 药用植物研究所，北京 100193 |
Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China |
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| 邵慧慧 |
SHAO Hui-hui |
中国医学科学院 北京协和医学院 药用植物研究所，北京 100193 |
Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China |
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| 高微微* |
GAO Wei-wei |
中国医学科学院 北京协和医学院 药用植物研究所，北京 100193 |
Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China |
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| 基金项目:中国医学科学院医学与健康科技创新工程项目（2016-I2M-1-012）；山东省重点研发计划项目（重大科技创新工程）（2019JZZY020905） |
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| 中文摘要:目的 克隆西洋参PqDELLA1基因，进行生物信息学分析，并探讨其在西洋参抗根腐病中的表达模式。方法 根据已获得的西洋参转录组数据，设计PqDELLA1基因全长扩增引物，利用无缝克隆的方法获得全长互补脱氧核糖核酸（cDNA）；采用国家生物技术信息中心（National Center for Biotechnology Information，NCBI）在线Blastp、DNAMAN和MEGA 6.0等软件对序列进行生物信息学分析；通过实时荧光定量多聚核苷酸链式反应（real-time quantitative polymerase chain reaction，qPCR）检测PqDELLA1基因的组织表达及诱导表达模式。结果 扩增得到西洋参PqDELLA1基因，其cDNA序列为1743 bp，编码580个氨基酸；PqDELLA1蛋白相对分子质量为63 910，理论等电点为5.01，不含信号肽，为非跨膜蛋白，定位于细胞核，与番茄SlDELLA蛋白氨基酸相似度最高。qPCR结果显示PqDELLA1基因在叶中表达量最高，其次为茎，根中最少；该基因的表达受根腐病病原菌茄病镰刀菌（Fusarium solani）的诱导，接种后5 d内持续升高后降低。结论 首次克隆出西洋参PqDELLA1基因并进行了生物信息学和表达特性分析，发现该基因可响应F. solani的侵染，为后续解析其参与西洋参抗病反应的分子机制提供参考。 |
| 中文关键词:西洋参 根腐病 PqDELLA1 生物信息学 表达分析 |
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| Cloning and Expression Analysis of PqDELLA1 Gene in Root Rot Resistance of Panax quinquefolius L. |
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| Abstract:Objective To clone PqDELLA1 gene of Panax quinquefolius L. and explore the expression patterns of PqDELLA1 gene in root rot resistance of P. quinquefolius L. with bioinformatics analysis.Methods The full-length amplification primer of PqDELLA1 gene was designed according to the obtained transcriptome data of P. quinquefolius L., and the full-length complementary deoxyribonucleic acid (cDNA) was obtained by in-fusion cloning. The National Center for Biotechnology Information （NCBI) online Blastp, DNAMAN, and MEGA 6.0 were used for bioinformatics analysis of the sequence. The tissue expression and induced expression patterns of PqDELLA1 gene were detected by real-time quantitative polymerase chain reaction (qPCR).Results The PqDELLA1 gene of P. quinquefolius L. was amplified. The cDNA sequence of P. quinquefolius L. was 1743 bp, encoding 580 amino acids. The protein molecular weight of PqDELLA1 gene was 63 910, and theoretical isoelectric point was 5.01. The PqDELLA1 gene, as a non-transmembrane protein without signal peptide, was located in the nucleus and was most similar to the S1DELLA protein amino acid of tomato. The results of real-time qPCR showed that the expression level of PqDELLA1 gene was highest in leaves, followed by stems and roots. The expression of PqDELLA1 gene was induced by Fusarium solani, the pathogen of root rot disease, and continued to increase within 5 days and then decreased after F. solani vaccination.Conclusion The cloning of PqDELLA1 gene for the first time with the bioinformatics analysis and expression characteristics analysis found that PqDELLA1 gene responded to the infestation of F. solani. The findings of this study provided references for the subsequent analysis of the molecular mechanism of PqDELLA1 gene in the disease resistance of P. quinquefolius L. |
| keywords:Panax quinquefolius L. root rot PqDELLA1 gene bioinformatics expression analysis |
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