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Phytophthora capsici, the causal agent of Phytophthora blight, is a prominent and economically damaging oomycete pathogen in South Georgia. P. capsici causes crown, root, leaf, stem, and fruit infections on a wide range of vegetable crops. Oomycete pathogens such as P. capsici are dispersed in water, as their zoospores are flagellated and can move through runoff. Irrigation ponds are often reservoirs for different pathogens, and reusing the captured runoff is increasing in popularity to decrease irrigation costs. This combination allows for unintended outbreaks of diseases by pumping the contaminated runoff onto susceptible crops. Detection and identification of these pathogens are crucial steps in disease management, and rapid detection can ensure timely application of disease control measures. In this study, 42 irrigation ponds in nine counties from South Georgia were surveyed for the presence of P. capsici using a novel filtration method in conjunction with a LAMP assay specific for P. capsici. Ten ponds in five counties were found to have P. capsici as detected from the assay, suggesting that testing of irrigation ponds for P. capsici and other pathogens should be conducted to assist in preventing disease outbreaks.

Phytophthora capsici is a devastating oomycete pathogen that affects many important solanaceous and cucurbit crops causing significant economic losses in vegetable production annually. Phytophthora capsici is soil-borne and a persistent problem in vegetable fields due to its long-lived survival structures (oospores and chlamydospores) that resist weathering and degradation. The main method of dispersal is through the production of zoospores, which are single-celled, flagellated spores that can swim through thin films of water present on surfaces or in water-filled soil pores and can accumulate in puddles and ponds. Therefore, irrigation ponds can be a source of the pathogen and initial points of disease outbreaks. Detection of P. capsici in irrigation water is difficult using traditional culture-based methods because other microorganisms present in the environment, such as Pythium spp., usually overgrow P. capsici making it undetectable. To determine the presence of P. capsici spores in water sources (irrigation water, runoff, etc.), we developed a hand pump-based filter paper (8-10 µm) method that captures the pathogen's spores (zoospores) and is later used to amplify the pathogen's DNA through a novel loop-mediated isothermal amplification (LAMP) assay designed for the specific amplification of P. capsici. This method can amplify and detect DNA from a concentration as low as 1.2 x 102 zoospores/mL, which is 40 times more sensitive than conventional PCR. No cross-amplification was obtained when testing closely related species. LAMP was also performed using a colorimetric LAMP master mix dye, displaying results that could be read with the naked eye for on-site rapid detection. This protocol could be adapted to other pathogens that reside, accumulate, or are dispersed via contaminated irrigation systems.