| 张海青，杜春梅，董瑞芳，姚 强.青稞禾生指葡孢霉（Dactylobotrys gramincola）LAMP快速检测方法的建立及应用[J].麦类作物学报,2025,(7):993 |
| 青稞禾生指葡孢霉（Dactylobotrys gramincola）LAMP快速检测方法的建立及应用 |
| Establishment and Application of Loop-Mediated Isothermal Application Assay for Detecting Dactylobotrys gamincola in Highland Barley |
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| DOI： |
| 中文关键词: 青稞 禾生指葡孢霉 环介导等温扩增 田间快速检测 |
| 英文关键词:Highland barley Dactylobotrys gramincola Loop-mediated isothermal application Rapid detection in the field |
| 基金项目:青海省科技计划-杰出青年基金项目（2023-ZJ-944J） |
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| 中文摘要: |
| 青稞鞘腐病是由禾生指葡孢霉（Dactylobotrys gramincola）引起的穗部病害，严重影响青稞的产量和品质。本研究基于环介导等温扩增技术（loop-mediated isothermal amplification，LAMP），以D.gramincola的ITS序列为基础，设计并筛选出1组特异性引物，通过优化反应温度，建立了灵敏度高且可快速检测青稞D.gramincola的LAMP检测方法。进一步对该方法的特异性和灵敏度进行评估，并对田间疑似发病植株的茎、旗叶、穗组织进行检测。结果表明，该LAMP检测体系在64 ℃下反应70 min即可从6种青稞病原菌中有效检测D.graminicola，检测灵敏度达13.07 fg·μL-1，比常规PCR技术灵敏度高100倍。对田间疑似发病植株组织进行检测，穗部检出10份阳性样本，旗叶检出8份阳性样本，茎秆部位未检出，检出率为50%。该方法能够特异性检测青稞发病组织中的D.graminicola，为青稞鞘腐病的田间快速诊断及病原菌检测提供了新方法。 |
| 英文摘要: |
| Highland barley sheath rot is a panicle disease caused by Dactylobotrys gramincola, which seriously affects the yield and quality of highland barley. In this study, a set of specific primers were designed and screened based on the ITS sequence of D.gramincola based on loop-mediated isothermal amplification(LAMP). By optimizing the reaction temperature, a LAMP detection method with high sensitivity and rapid detection of D. gramincola was established. The specificity and sensitivity of the method were further evaluated, and the stem, flag leaf, and panicle tissues of the suspected diseased plants in the field were detected. The results showed that the LAMP detection system can effectively detect D. graminicola from six highland barley pathogens at 64 ℃ for 70 min. The detection sensitivity was 13.07 fg·μL-1, which was 100 times higher than that of the conventional PCR. The tissues of the suspected diseased plants in the field were detected. Ten positive samples were detected in the panicle; eight positive samples were detected in the flag leaf; and no positive samples were detected in the stem. The detection rate was 50%. |
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