Salinity is the second most important abiotic stress in the world, especially in Iran. The research dedicated to reveal the responses of the some physiological and biochemical parameters (proline content, carbohydrates, total protein and peroxides enzymes) of Chrysopogon zizanioides to soil salinity in vitro. Prior to salinity application, the plant seedlings placed in greenhouse for adaptation purpose. To apply soil salinity stress on Chrysopogon zizanioides, sodium chloride solution was applied in 6 different concentrations in the period of 4 days. All the measurements, including leaf proline content, soluble and non-soluble sugar, protein content, and peroxidase enzyme, were measured after 2 month of salinity treatment application. The results indicated that salinity stress could have a significant increasing effects on proline content, soluble sugar and non-soluble one of leaves (p≤0.05). Leaf total protein content and peroxidase enzyme are also significantly affected by salinity stress, showing an increasing trend up to 32 ds/m following by a decreasing trend at 44 ds/m. According to physiologic and biochemical parameters, the results confirms high tolerance of Vetiver grass against 32 ds/m salinity in short term.