The extensive and intensive use of herbicides has resulted in the spread of herbicide-resistant weeds in many crop production systems; therefore, it is imperative to devise new organic weed control methods. Recently, the application of spent coffee grounds (SCG) in agricultural fields has been found to inhibit plant growth and germination and is thus considered a potentially effective weed control measure. This study aimed to evaluate the effects of different amounts and methods of SCG application on weed growth through field experiments. The field experiments were conducted in an upland field converted from a paddy in western Japan. The results show that the plow-in application of over 10 kg m−2 of SCG and mulching application of 20 kg m−2 decreased the weed dry weight compared with the control. In addition, the growth of weed species of families other than Gramineae, such as wingleaf primrose-willow and horseweed, was not significantly affected by SCG application. Weed species of families other than Gramineae are dominant in some upland fields. Hence, the inhibitory effect of SCG on weeds may be lower in original upland fields than in the upland field converted from paddy field that was investigated in the present study. Overall, this study demonstrated that the plow-in application of 10 kg m−2 of SCG every 4 mo was effective for weed control in an upland field converted from a paddy field. Because SCG worked against grass weeds under the specific conditions in this study, it would be valuable to explore other potential applications of this novel means of weed control.