The watershed ecological compensation mechanism plays an important role in the creation of ecological civilization, and improving the watershed ecological compensation mechanism is an important policy arrangement to implement the reform of China's ecological civilization system. Upstream residents seldomly receive the ecological compensation funds in practice, so the universal benefit of ecological compensation in the river basin needs to be improved. The universal benefit of ecological compensation should not only increase the sense of gain of upstream residents, but also enable them to obtain ecological compensation funds matching the contribution of ecological conservation by accurately understanding the upstream residents' willingness and preference. This paper takes the ecological compensation project of the Xin'an River Basin as an example, which is the first trans-boundary pilot of watershed ecological compensation project at the provincial level in China, then estimates the upstream residents' willingness to accept (WTA) based on the contingent valuation method and explores its determinants. It is shown that:(1) the annually ecological compensation fund demand for upstream area is 16.560 billion Yuan based on the upstream residents' WTA estimates. Specifically, the annually ecological compensation fund demand of Chun'an County and Huangshan City are 4.432 billion Yuan and 12.128 billion Yuan, respectively. (2) The WTA estimates of the upstream area become larger as the respondent's age or her/his family number increases, while the rural residents need be compensated more than the urban residents, and the income elasticity of the allocation of compensation fund is approximately 0.04. (3) The perceived intensity of environmental regulation of Chun'an is significantly higher than that of Huangshan, and the willingness to accept of Chun'an is also significantly higher. The frequency of the upstream residents' voluntary participation in environmental protection is positively correlated with the WTA. (4) The upstream residents' perceived burden of government's expenditure of environmental protection does not match reality. Therefore, the government needs to increase the publicity on ecological protection to realize the value of government investment in environmental protection and the effectiveness of governance. (5) The allopatric employment policy can serve as a beneficial supplement to alleviate the monetization payment pressure of ecological compensation and employment pressure in the local area. While the allopatric employment policy including the employment subsidy policy, free education and training policy can be implemented, the sustainability needs to be explored by more empirical analysis.