重组酶聚合酶扩增-横向流动试纸快速检测水稻黄单胞菌。米中的米

IF 2.5 2区农林科学 Q1 AGRONOMY

Crop Protection Pub Date : 2025-02-06 DOI:10.1016/j.cropro.2025.107153

Jieling Wang , Agen Li , Chengxin Mao , Xinyue Li , Chuanqing Zhang

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引用次数: 0

摘要

细菌性叶枯病（BLB），主要由米黄单胞菌引起。oryzae （Xoo）是全球水稻生产的主要威胁。种子携带的Xoo是BLB暴发期间一次接种的主要来源之一。因此，准确、快速地检测Xoo对于采取有效的疾病管理策略至关重要。在这项研究中，我们设计并评估了一套基于糖核苷酸外聚酶/脱水酶编码基因的特异性引物和探针，以建立Xoo的重组酶聚合酶扩增（RPA）点对点（POC）检测。该试验对Xoo具有高度特异性，不能检测到其他水稻病原体。优化的RPA条件（在50 μL反应体系中，36°C, 10 min）可检测Xoo， DNA限幅为100 fg/μL。比较了RPA -横向流动检测法（LFD）和PCR检测方法，证实了Xoo在水稻种子中的存在。结果表明，该方法可快速、灵敏、特异地检测水稻种子中的Xoo。

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Recombinase polymerase amplification-lateral flow dipstick for rapid detection of Xanthomonas oryzae pv. oryzae in rice

Bacterial leaf blight (BLB), primarily caused by Xanthomonas oryzae pv. oryzae (Xoo), is a major threat to global rice production. Seed borne Xoo is one of the key sources of primary inocula during BLB outbreaks. Therefore, accurate and rapid detection of Xoo is crucial for adopting effective disease management strategies. In this study, we designed and evaluated a set of specific primers and probes based on a sugar nucleotide epimerase/dehydratase coding gene to develop the recombinase polymerase amplification (RPA) point-of-care (POC) detection of Xoo. This assay is highly specific to Xoo and does not detect other rice pathogens. Optimized RPA conditions (36 °C for 10 min in a 50 μL reaction system) enabled Xoo detection with a DNA limit of 100 fg/μL. This RPA -lateral flow dipstick (LFD) and PCR detection methods were compared to confirm Xoo's presence in rice seeds. Our results demonstrated the rapid, sensitive, and specific detection of Xoo in rice seeds using this RPA-LFD method.

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来源期刊

Crop Protection 农林科学-农艺学

CiteScore

6.10

自引率

3.60%

发文量

200

审稿时长

29 days

期刊介绍： The Editors of Crop Protection especially welcome papers describing an interdisciplinary approach showing how different control strategies can be integrated into practical pest management programs, covering high and low input agricultural systems worldwide. Crop Protection particularly emphasizes the practical aspects of control in the field and for protected crops, and includes work which may lead in the near future to more effective control. The journal does not duplicate the many existing excellent biological science journals, which deal mainly with the more fundamental aspects of plant pathology, applied zoology and weed science. Crop Protection covers all practical aspects of pest, disease and weed control, including the following topics: -Abiotic damage- Agronomic control methods- Assessment of pest and disease damage- Molecular methods for the detection and assessment of pests and diseases- Biological control- Biorational pesticides- Control of animal pests of world crops- Control of diseases of crop plants caused by microorganisms- Control of weeds and integrated management- Economic considerations- Effects of plant growth regulators- Environmental benefits of reduced pesticide use- Environmental effects of pesticides- Epidemiology of pests and diseases in relation to control- GM Crops, and genetic engineering applications- Importance and control of postharvest crop losses- Integrated control- Interrelationships and compatibility among different control strategies- Invasive species as they relate to implications for crop protection- Pesticide application methods- Pest management- Phytobiomes for pest and disease control- Resistance management- Sampling and monitoring schemes for diseases, nematodes, pests and weeds.