The aim of the present work was to determine the carbohydrate content in plantlets of Clematis pitcheri cultured in vitro depending on the temperature (15°C, 20°C and 25°C) and sucrose (10 and 30 g·dm⁻³) and nitrogen (100% and 50% of normal MS strength) level in the medium. 100% N meant 1.90 g KNO₃ and 1.65 g NH₄NO₃, and 50% N meant 0.950 g KNO₃ and 0.825 g NH₄NO₃ in 1 dm⁻³ of MS medium. The highest accumulation of starch in shoots took place at 30 g·dm⁻³ of sucrose and lower level of nitrogen compounds in the medium. Also, at 30 g·dm⁻³ of sucrose in the medium the content of fructose in shoots was higher and independent from the level of nitrogen. The temperature of 25°C stimulated glucose accumulation in shoots at the highest degree at 30 g·dm⁻³ of sucrose and lower level of nitrogen. Regardless of the treatment, no sucrose was found in plantlets. Generally, the higher concentration of sucrose in the medium the higher content of sum of carbohydrates in plantlets. A lower nitrogen compound level (50% N) led to increase of glucose and starch content in shoots, and had a smaller effect on the level of fructose. Shoots cultured at the temperature of 20°C and 25°C accumulated more carbohydrates than at 15°C.