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Reference evapotranspiration (ET0) plays a critical role in irrigation planning and is also important for hydrological cycle, environmental, and other studies. Thus, this research examined the trends in ET0 on seasonal and annual timescales in Hubei Province, China. ET0 was estimated using the Penman Monteith method (P-M) at 16 meteorological stations located in different areas of Hubei Province during the period 1960-2014. The trends in seasonal and annual ET0 were investigated using the Mann-Kendall test and Sen's slope estimator. The periodicities of ET0 in different regions were investigated using wavelet analysis. The major meteorological factors affecting ET0 were investigated using partial correlation analysis and the contribution rate method. The results showed, on a seasonal timescale, that in spring, ET0 increased in all geographic zones. In summer, ET0 decreased in all geographic zones. In autumn and winter, ET0 displayed no significant changes in any of the geographic zones. On an annual timescale, ET0 decreased in all geographic zones, and the magnitudes of the negative trend in annual ET0 were 2.58-10.04 mm 10a(-1). In the five geographic zones, the periodic characteristics of ET0 were identical; the significant wavelet power spectra of ET0 had 3-7, 13-17, and 24-32-year modulations in variation. Among the meteorological factors, sunshine hours were the major climate element that influenced the variability in ET0. The results will provide important references for scientific planning for agriculture, water resource allocation, and water-saving irrigation.