Phalaenopsis ‘KS Little Gem’ is a new cultivar with superior ornamental qualities. Newly produced cultivars are often studied to determine the optimum growing conditions, as production management practices vary within species. We focused on the effects of temperature and fertilizer application on the growth and carbohydrate content of ‘KS Little Gem’ plantlets. Two-month-old plantlets were subjected to different temperatures (20, 25, and 30°C) and fertilizer rates (0.33, 0.5, and 1 g・L-1) inside a growth chamber. Data were collected from leaves and roots regarding their growth, fresh weight, sugar content, and starch content. The results revealed that temperature significantly affected leaves and roots parameters, fresh weight, and carbohydrate content of the plantlets. However, fertilizer rates alone showed no significant effects on the growth and carbohydrate content of plantlets. The highest leaf and root growth, root fresh weight, and leaf carbohydrate content were observed in plantlets subjected to 25°C. In addition, ‘KS Little Gem’ plants grown under 30°C had significantly inhibited growth and reduced carbohydrate content in both the leaves and roots.