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Lygodium microphyllum is an invasive exotic plant species taking over many sites in freshwater and moist habitats in Florida. Managing it has been a significant challenge for land resource managers and researchers due to its extensive rapid invasion. To assess the effects of soil pH on growth, nutrient uptake, and mycorrhizal colonization in the roots of L. microphyllum, we conducted a 60-day greenhouse experiment by growing it in pots filled with pH-adjusted soils to a range from 4.5 to 8.0. L. microphyllum was able to survive and grow at all soil pH levels; however, final biomass, relative growth rate, photosynthesis, and specific leaf area were all greater in soil pH 5.5–6.5 compared to the other treatments. Correspondingly, nitrogen concentration was also related to these four plant parameters. Root colonization by mycorrhizal fungi was higher in soil pH 5.5–7.5 and lowest for plants growing in 4.5 or 8.0 and was correlated with plant growth parameters as well as elemental concentration in the leaves. Soil pH 8.0 was not strong enough for a pronounced growth decline, thus further increasing soil pH could provide a desired outcome and merit further investigation, although its potential negative impact on native flora (both plants and microorganisms) would need to be assessed.