The pronounced gender gap in Italian agriculture is reflected by lower levels of female labor force participation, labor supply and managerial positions in the farm sector, coupled with their higher incidence of part-time work. The objective of the paper is to investigate the drivers of farm holders’ labor supply decisions while controlling for gender differences. Using micro-data from the Italian agricultural business survey (REA), this study employs a random effects ordered probit over the period 2002-2009. The results highlight significant gender differences in labor market responses. In particular, farm size and livestock systems are found to increase the on-farm labor supply of male farm holders, reflecting the role of men in the farm and gender differences in ownership, control and decision making over productive resources. The diverse impact of farm subsidies on labor supply may suggest the presence of credit constraints in female-operated households, preventing the capitalization of subsidies into fixed assets.