Polysaccharide is the main active component of Dendrobium huoshanense, and it is also an important indicator to evaluate its quality. In this study, the polysaccharides and invertase components in the stems and leaves of D. huoshanense were studied to provide a theoretical basis for the full exploitation and utilization of its resources. Taking the D. huoshanense grown in the wild, indoors and outdoors as a test material and choosing stems and roots of D. huoshanense for 1 year, 2 years and 3 years of growth, the polysaccharide content of different samples was determined, and the relative expression activity of sucrose metabolism-related genes in different samples was detected by qRT-PCR. Western blot was used to detect the changes of sucrose metabolism-related proteins in different samples. The results showed that the content of polysaccharide in the wild D. huoshanense was the highest, the indoor growth was the second, the growth in the outdoor was the lowest, and the content of polysaccharide in the stem was significantly higher than that in the root. The expression of D. huoshanense neutral/alkaline invertase (DhNI), D. huoshanense sucrose synthase (DHSS) and D. huoshanense sucrose phosphate synthase (DHSPS) was the highest in wild D. huoshanense, followed by indoor growth, and the expression in stem was much higher than in root. Western blot results also indicated that DhNI, DHSS and DHSPS protein expression was highest in wild, followed by indoors. The stem is much higher than the root and is grown for 2 years in the same growing environment for more than 1 year and 3 years. The above results show that the nutritional value of D. huoshanense is related to the decomposition of sucrose and the accumulation of polysaccharides in plants. The experimental results provide a theoretical basis for better exploitation and utilization of D. huoshanense resources.