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将孢子捕捉技术与环介导等温扩增检测法相结合用于水稻稻曲病的监测与可持续管理。

Integrating spore trapping technology with loop-mediated isothermal amplification assay for surveillance and sustainable management of rice false smut disease.

作者信息

Arumugam Gopalakrishnan Meena, Chellappan Gopalakrishnan, Ayyanar Kamalakannan, Ramasamy Jagadeeswaran, Santhosh Ganapati Patil, Nagaranai Karuppasamy Sathyamoorthy

机构信息

Department of Plant Pathology, Tamil Nadu Agricultural University, Coimbatore, India.

Department of RS and GIS, Tamil Nadu Agricultural University, Coimbatore, India.

出版信息

Front Microbiol. 2024 Dec 4;15:1485275. doi: 10.3389/fmicb.2024.1485275. eCollection 2024.

Abstract

Rice ( L.) is a vital crop feeding more than half of the world's population, with production occurring predominantly in Asian countries. However, rice cultivation faces challenges from various fronts, including biotic stresses intensified by climate change. False smut, caused by , has emerged as a significant threat to rice production globally. The application of curative fungicides after symptom appearance has limited scope in managing this disease since the infection process usually starts during the early flowering stage of rice crops. This study investigates the utilization of spore-trapping technology coupled with Loop-Mediated Isothermal Amplification (LAMP) assay for monitoring airborne inocula in rice fields. For early detection and quantification of , sampling rods coated with silicone grease were deployed to collect airborne spores, and DNA extraction was performed using a modified method. Both PCR and LAMP assays were employed for detection, with LAMP offering advantages of rapidity, sensitivity, and simplicity. The study demonstrated the superior sensitivity of LAMP compared to PCR, detecting DNA at concentrations as low as 100 femtograms. Continuous monitoring of inoculum using spore trapping revealed the spatio-temporal dynamics of dispersal, providing valuable insights for disease management. Implementing a fungicidal application schedule based on airborne inoculum detection led to significant reductions in both false smut incidence and severity and improved crop yield. The meteorological parameters including minimum temperature, relative humidity in the morning and evening, sunshine, and solar radiation were found to be correlated with disease incidence. Multi-operator validation confirmed the robustness and specificity of the LAMP assay. Overall, this integrated approach offers a proactive strategy for monitoring and managing false smut disease, enhancing sustainable rice production and food security.

摘要

水稻(Oryza sativa L.)是养活全球一半以上人口的重要作物,其生产主要集中在亚洲国家。然而,水稻种植面临着来自各个方面的挑战,包括气候变化加剧的生物胁迫。由Ustilaginoidea virens引起的稻曲病已成为全球水稻生产的重大威胁。由于感染过程通常在水稻作物的始花期开始,因此在症状出现后施用治疗性杀菌剂在控制这种病害方面的作用有限。本研究调查了利用孢子捕捉技术结合环介导等温扩增(LAMP)分析法监测稻田空气中的接种体。为了早期检测和定量U. virens,部署了涂有硅脂的采样棒来收集空气中的孢子,并使用改良方法进行DNA提取。采用PCR和LAMP分析法进行检测,LAMP具有快速、灵敏和简便的优点。研究表明,LAMP比PCR具有更高的灵敏度,能检测低至100飞克浓度的U. virens DNA。利用孢子捕捉技术对U. virens接种体进行连续监测,揭示了其传播的时空动态,为病害管理提供了有价值的见解。根据空气中接种体的检测结果实施杀菌剂施用计划,可显著降低稻曲病的发病率和病情严重程度,并提高作物产量。发现包括最低温度、早晚相对湿度、日照和太阳辐射在内的气象参数与病害发病率相关。多操作员验证证实了LAMP分析法的稳健性和特异性。总体而言,这种综合方法为监测和管理稻曲病提供了一种积极主动的策略,有助于加强可持续水稻生产和粮食安全。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e424/11652660/9d6896157d53/fmicb-15-1485275-g001.jpg

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