| 橡胶树红根病病原菌LAMP检测方法的建立及应用 |
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| 引用本文： | 贺春萍,董文敏,吴伟怀,梁艳琼,李锐,谢立,黄兴,易克贤.橡胶树红根病病原菌LAMP检测方法的建立及应用[J].热带作物学报,2021,42(1):182-187. |
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| 作者姓名： | 贺春萍 董文敏 吴伟怀 梁艳琼 李锐 谢立 黄兴 易克贤 |
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| 作者单位： | 1.中国热带农业科学院环境与植物保护研究所/农业农村部热带作物有害生物综合治理重点实验室/海南省热带农业有害生物监测与控制重点实验室,海南海口 5711012.南京农业大学植物保护学院,江苏南京 2100953.海南大学林学院,海南海口 570228 |
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| 基金项目： | 海南省自然科学基金项目(320RC692);海南省院士创新平台科研专项(SQ2020PTZ0011);国家天然橡胶产业技术体系建设专项资金项目(CARS-33-BC1) |
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| 摘 要： | 由灵芝菌(Ganoderma pseudoferreum)引起的红根病是橡胶上危害面积最广、影响最大的世界性根部传染性病害.本研究利用环介导等温扩增技术(LAMP),以G.pseudoferreum线粒体Large rDNA特异片段为靶标序列,设计出G.pseudoferreum特异性LAMP引物,以SYBR Gree...
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| 关 键 词： | 灵芝菌 环介导等温扩增 分子检测 |
| 收稿时间： | 2020-01-08 |
Establishment and Application of a Loop-mediated Isothermal Amplification Assay for Detection of Ganoderma pseudoferreum on Hevea |
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| HE Chunping,DONG Wenmin,WU Weihuai,LIANG Yanqiong,LI Rui,XIE Li,HUANG Xing,YI Kexian.Establishment and Application of a Loop-mediated Isothermal Amplification Assay for Detection of Ganoderma pseudoferreum on Hevea[J].Chinese Journal of Tropical Crops,2021,42(1):182-187. |
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| Authors: | HE Chunping DONG Wenmin WU Weihuai LIANG Yanqiong LI Rui XIE Li HUANG Xing YI Kexian |
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| Institution: | 1. Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences / Key Laboratory of Integrated Pest Management on Tropical Crops, Ministry of Agriculture and Rural Affairs / Hainan Key Laboratory for Monitoring and Control of Tropical Agricultural Pests, Haikou, Hainan 571101, China2. College of Plant Protection, Nanjing Agricultural University, Nanjing, Jiangsu 210095, China3. Institute of Forestry, Hainan University, Haikou, Hainan 570228, China |
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| Abstract: | Rubber tree red root disease caused by Ganoderma pseudoferreum is proved to be a serious disease, which is the most widespread and harmful to rubber tree (Hevea brasiliensis) production. This study reports the development of a loop-mediated isothernal amplification (LAMP) assay targeting the mitochondria large rDNA for visual detection of G. pseudoferreum. The mitochondria large rDNA-LAMP assay efficiently amplified the target gene within 80 min at 64 ℃ (in 1 hour) and was evaluated for specificity and sensitivity. A positive color or (green) was only observed in the presence of G. pseudoferreum by SYBR Green I as an indicator reaction after amplification; however none of other pathogens changed color (still orange). The lower limit of detection of the LAMP assay was about 1×10 -5 ng/μL of genomic DNA, which was 1000 times more sensitive than the PCR method. Twenty strains of suspected samples in the field were tested by the LAMP system with the pathogen detection rate was 85%. This method would provide a new technology for the rapid identification of G. pseudoferreum. |
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| Keywords: | Ganoderma pseudoferreum loop-mediated isothermal amplification molecular detection |
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