Agrobacterium tumefaciens has been associated with the occurrence of bronze wilt of cotton, a disease first recognized in 1995. The disease is characterized by the sudden wilting and bronzing or reddening of the foliage and occurs primarily on a number of short-season determinate cultivars. One of the complicating factors in demonstrating the bacterium’s role in bronze wilt is the report that Agrobacterium tumefaciens is endophytic and ubiquitous in cotton. A collaborative research project was initiated to examine the frequency of occurrence of Agrobacterium spp. in seedlots of cotton. Cultivars used were DP 50, Sure-Grow 125, Sure-Grow 125RR, Sure-Grow 125BR, PM 1218 BR, PM 1220 RR, PM 1220 BR, and Stoneville 373. Two methods were used to assess to occurrence of Agrobacterium spp. with cottonseed. The first method, described by Bell (1999), involves planting cottonseed, harvesting seedlings 2 weeks after planting, removing a 1-cm proximal section of roots, grinding the root sections in sterile distilled water, and plating the homogenate on microbiological media. Media used to isolate bacteria included TSA, D-1, and EMB. Method 2 involves shaking seed in broths of media selective for the growth of Agrobacterium spp. (D-1 or D-5) for 48 hrs, homogenizing the seed and broth, and dilution plating the homogenate on TSA, D-1 and EMB. Bacteria were commonly detected on/in seed using either method. Biolog or FAME analysis indicated that none of the isolates from seed of any cultivar by the seed assay method (Method 2) were Agrobacterium spp. Bacteria from a number of genera commonly associated with plants were recovered, including Enterobacter agglomerans, Pantoea agglomerans, Stenotrophomonas maltophila, Pseudomonas spp. Paenibacillus spp., and Bacillus spp. Artificially infesting seed with isolates of Agrobacterium and incubating in D-1 broth increased populations approximately two log units when seed were assayed 48 hrs after inoculation. By Method 1, Agrobacterium tumefaciens/ radiobacter was isolated from PM 1218 BR and PM 1220 RR at very low frequencies in the Clemson and Arkansas laboratories, respectively. This preliminary information suggests that Agrobacterium spp. may be present at very low frequencies in cottonseed of some cultivars. The apparent latent infection of cotton seedlings by Agrobacterium spp. warrants further investigation. Agrobacterium spp. isolated from cotton are currently being characterized using PCR with primers specific for virulence genes of Agrobacterium tumefaciens.