Loblolly pine decline, characterized by deteriorating root systems leading to shortening and thinning of foliage, has been observed throughout portions of the south-eastern United States. Several root-inhabiting ophiostomatoid fungi, including Leptographium procerum, Leptographium terebrantis, Leptographium serpens, and Grosmannia huntii are associated with lateral root damage on declining loblolly pine. Trees of various ages were inoculated in primary lateral roots during fall (2006 and 2007) and spring (2007 and 2008). All fungi caused a darkened, resin-filled lesion on the surface of the phloem, extending into the xylem that was larger than that of controls. Only lesions associated with G. huntii infection were significantly larger in the spring season, compared with the fall. Grosmannia huntii was found to be the most virulent fungus, causing lesions that were longer, deeper and larger than all other fungal species during the spring and larger than L. terebrantis and L. procerum in the fall. Leptographium serpens was the second most virulent fungal species, causing lesions larger than L. procerum and L. terebrantis (with the exception of lesion depth) during both seasons. These tests indicate that G. huntii and L. serpens are significant root pathogens, capable of causing considerable damage, while L. terebrantis and L. procerum may be less virulent. Depending on the actions of their vectors, G. huntii and L. serpens may be responsible for significant root deterioration and tree disease.