The exploitation of the existing genetic variability for the physiological traits related to the yield and quality of maize forage can assist in the development of superior inbred lines and hybrids. The objective of this work was to evaluate the general and the specific combining ability and the nature of gene effects of physiological and chemical traits of forage maize genotypes. Two groups of genotypes and 16 hybrids resulting from a 4x4 partial diallel scheme were evaluated. Group I consisted of two commercial hybrids (AG8025, P30B39) and two elite inbred lines (LEM2 and LEM3). Group II consisted of four experimental inbred lines originated from different populations of forage maize breeding program. In total, 24 treatments were evaluated, formed by the genotypes of both groups and the respective crossings. The traits evaluated were: CO2 assimilation (A), stomatal conductance (gs), internal CO2 concentration (Ci), transpiration (E), calculated activity of Rubisco (A/Ci) and efficiency of water use (A/E). Forage acid detergent fiber (ADF), neutral detergent fiber (NDF) and digestibility in situ were obtained. There was a predominance of non-additive gene effects for most of the chemical and physiological traits. Crossings LEM2 x 203-218.3, LEM3 x 201-107.2, LEM2 x 101-7.2 and LEM3 x 101-7.2 stood out regarding CO2 assimilation, and are indicated for future research considering the physiological traits. Inbred lines 101-7.2 and 203-218.3 presented high concentration of favorable alleles to increase carboxylation efficiency, in which inbred line 101-7.2 stood out for NDF, ADF and DIG. Inbred line 201-107.2 has a high concentration of favorable alleles for efficiency of water use. Physiological parameters can assist the selection of inbred lines and hybrids in maize breeding for forage purpose.