Antifungal defenses of seagrasses from the Indian River Lagoon,Florida

We investigated the antifungal chemical defenses and physiological responses of five seagrasses collected from nearshore seagrass beds from the Indian River Lagoon, Florida, against a panel of co-occurring marine fungi isolated from nearby coastal communities. Whole plant tissues from Thalassia testudinum, Halodule wrightii and Syringodium filiforme prevented overgrowth by three of the seven fungi used in this study. Organic extracts from four of the five seagrasses inhibited the growth of at least one fungal strain. The extract from Ruppia maritima exhibited the highest antifungal activity, inhibiting the growth of three fungi including the pathogen Lindra thalassiae. Among the fungal panel, Fusarium sp. 2 was the most susceptible to seagrass extracts, whereas none of the extracts disrupted the growth of Dendryphiella salina and Fusarium sp. 3. Under laboratory conditions fungal inoculation elicited hydrogen peroxide production in all specimens within 25 min post-inoculation as measured with a redox sensitive dichlorodihydrofluorescein diacetate (DCFH-DA) assay. The concentration of H₂O₂ released into the immediate vicinity of infected seagrasses varied between 0.10 and 0.85 μmol g⁻¹ FW min⁻¹ depending on seagrass species and pathogen combination. Longer term incubation (days) of T. testudinum with homogenates of D. salina or L. thallasiae resulted in the induction of caspase activity, a known proteolytic activator of apoptotic and inflammatory activities. The application of micromolar concentrations of H₂O₂ to blades of T. testudinum induced caspase activity suggesting that fungal detection, H₂O₂ production, and caspase activation occur in a consecutive order. The seagrasses examined in this study appear to use a combined strategy to combat fungal infection, including microbial chemical defenses and signaling pathways observed in terrestrial plants.