基于环介导等温扩增技术快速诊断由Fusarium andiyazi引起的水稻恶苗病
Rapid diagnosis of rice bakanae caused by Fusarium andiyazi using a loop-mediated isothermal amplification assay
查看参考文献20篇
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文摘
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水稻恶苗病是水稻上的重要病害,在我国各水稻主要种植区均有发生,造成水稻产量的严重损失.Fusarium andiyazi是近年来国外报道的水稻恶苗病的病原菌之一,本研究基于环介导等温扩增技术(loop-mediated isothermal amplification, LAMP),以F. andiyazi的TAT(trichothecene 3-O-acetyltransferase)基因为靶标设计并筛选出一套灵敏、特异的LAMP引物,建立了可快速诊断该病菌所引起的水稻恶苗病的LAMP检测技术.在等温条件下(64℃)只需进行核酸扩增反应80 min,反应前向体系中加入了金属离子指示剂HNB(羟基萘酚蓝),反应后即可肉眼观察反应产物颜色变化判断检测结果,阳性反应呈天蓝色,阴性呈紫色.该TAT-Fan-LAMP技术的最低检测灵敏度为100 pg·μL~(-1).应用该技术成功地对南京江宁和镇江句容田间采集的由F. andiyazi引起的水稻恶苗病进行快速诊断.该LAMP检测技术的建立为F. andiyazi引起的水稻恶苗病的诊断提供了简便快速的新技术. |
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其他语种文摘
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Bakanae is an important disease of rice across all major rice-growing regions in China leading to serious rice yield losses. Here we describe a loop-mediated isothermal amplification (LAMP)assay on direct detection of F. andiyazi in the diseased rice tissues,for rapid diagnoses of rice bakanae caused by this pathogen. Based on the sequence of trichothecene 3-O-acetyltransferase (TAT)gene in F. andiyazi,a set of species-specific primers was designed and screened to generate a positive color (sky blue)in the presence of F. andiyazi with the addition of hydroxynaphthol blue (HNB)indicator prior to amplification,whereas no color change (purple)in the negative reaction tubes of other species isolates under isothermal condition at 64℃ for 80 min. Using the TAT-Fan-LAMP assay with the detection limit at 100 pg·μL~(-1) genomic DNA for F.andiyazi,we successfully and rapidly diagnosed the suspected diseased rice plant samples collected from Jiangning,Nanjing and Jurong,Zhenjiang in Jiangsu Province. This method can be used as a new way of diagnosing rice bakanae caused by F. andiyazi. |
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来源
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植物病理学报
,2018,48(2):256-262 【核心库】
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DOI
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10.13926/j.cnki.apps.000128
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关键词
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水稻恶苗病
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Fusarium andiyazi
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环介导等温扩增技术
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TAT
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地址
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1.
南京农业大学植物保护学院卵菌与真菌分子生物学实验室, 南京, 210095
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镇江市农科院, 镇江, 212400
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语种
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中文 |
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文献类型
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研究性论文 |
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ISSN
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0412-0914 |
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学科
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植物保护 |
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基金
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国家863计划
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镇江市重点研发项目-现代农业
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文献收藏号
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CSCD:6220430
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