In the Cerrado-Amazonian ecotone in the State of Mato Grosso, intensely altered by anthropic action, the knowledge and processes of energy conversion and energy balance are still incipient, making the monitoring and modeling of diffuse radiation essential for several applications. The objective of this study was to evaluate the seasonality of the diurnal evolution and estimate the hourly diffuse radiation (Hdh) and incident radiation in the horizontal plane between June 2011 and October 2016. The instantaneous measurements (5 min) of diffuse radiation underwent geometric, astronomical, and anisotropic corrections, with subsequent hourly integrations. The seasonality of diffuse radiation and its radiometric fractions was evaluated. The estimates were made considering total and seasonal data groupings (water stations in the region) and in different cloudiness classes (atmospheric transmissivity index—KTh). The diurnal behavior of diffuse radiation (Hdh) was similar to that of global radiation and at the top of the atmosphere, with maximum values at solar noon. The correlations between Kdh and KTh showed third-order polynomial behavior, with maximum observed values of Kdh ranging from 0.8 to 0.9, for KTh less than 0.2. Estimation equations based on radiometric fractions underestimated the values of diffuse radiation, with a better performance presented by models adjusted in annual data groupings. Among the parameterized models for estimating diffuse radiation obtained in the literature, those calibrated regionally in this study, together with those developed for tropical regions, presented better statistical performances.