Background and Objective: Vitamins are natural substances that are important in tiny quantities to sustain the normal growth and development of the plant. Thus this study was carried out to study the physiological effect of pyridoxine on increasing yield quantity and quality of Egyptian clover plant. Materials and Methods: In sandy soil at the Research and Production Station, National Research Centre, Nubaria Province, Behaira Governorate, two field experiments were carried out during two consecutive winter seasons to study the effects of pyridoxine treatment on the yield and nutritional value of Egyptian clover. Results: Spraying Egyptian clover with pyridoxine significantly increased yield attributes, crude protein, Ash contents, however, crude fibre content decreased by 50 or 75 mg LG 1 . Pyridoxine levels significantly decreased nitrogen, free extract contents. Pyridoxine with 100 mg LG 1 significantly increased gross, digestible, metabolizable and net energy, total digestible nutrients, digestible crude protein, neutral detergent fibre, acid detergent fibre, acid detergent lignin and cellulose contents in comparison with control and the other treatments. Cut stages affect the significantly chemical composition and their contents of both energetic and nutritive values and cell wall constituents. Moreover, there were significant interactions between pyridoxine supplementation and berseem cut stages on chemical composition and their contents of energetic and nutritive values and cell wall constituents. Also, photosynthetic pigments, carbohydrates constituents, indole acetic acid, phenolic and flavonoids contents were increased gradually with increasing pyridoxine concentrations. Conclusion: In conclusion, different pyridoxine treatments have a promotive effect on increasing the forage yield of the berseem plant in addition to its nutritive values.