Use of TEM in Plant Disease Diagnosis: A Xylem-Limited Bacterium Associated with Diseased ‘Toronto’ Creeping Bentgrass

Plant disease diagnosis is most often accomplished by examination of symptoms and observation or isolation of causal organisms. Occasionally, diseases of unknown etiology occur and are difficult or impossible to accurately diagnose by the usual means. In 1980, such a disease was observed on Agrostis palustris Huds. c.v. Toronto (creeping bentgrass) putting greens at the Butler National Golf Course in Oak Brook, IL.The wilting symptoms of the disease and the irregular nature of its spread through affected areas suggested that an infectious agent was involved. However, normal isolation procedures did not yield any organism known to infect turf grass. TEM was employed in order to aid in the possible diagnosis of the disease.Crown, root and leaf tissue of both infected and symptomless plants were fixed in cold 5% glutaraldehyde in 0.1 M phosphate buffer, post-fixed in buffered 1% osmium tetroxide, dehydrated in ethanol and embedded in a 1:1 mixture of Spurrs and epon-araldite epoxy resins.