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With future global temperatures predicted to increase, the relationship between a host, pathogen, and environment, becomes less predictable and epidemics may pose a greater risk to forests worldwide. Resistance breeding is an important disease management tool, but because tree species require long breeding times, it is necessary to develop techniques for testing current pathogen isolates against their hosts. Pitch canker disease of pines, caused by the pathogen Fusarium circinatum, is no exception and represents a threat to pine forests and commercial plantations worldwide, as it thrives at warm temperatures and high humidity. We tested growth of 15 F. circinatum isolates in culture at three temperatures: 25, 27, and 31 degrees C. We also evaluated the sporulation and pathogenicity of eight of the isolates on two susceptible Pinus elliotti (slash pine) open-pollinated families and one tolerant open-pollinated Pinus taeda (loblolly pine) family. Our results showed significant differences among isolates in the temperature and pathogenicity tests. All isolates showed a significant decrease in growth at 31 degrees C, although some showed similar growth at 25 and 27 degrees C. Several of the new isolates tested were more pathogenic than the isolates that the USDA Forest Service Resistance Screening Center (RSC) had been using. The new isolates have now been incorporated into their operational screening program.