Abstract: In April 2017, 1-year Tetrastigma hemsleyanum cuttings of different provenances such as Zhejiang Wenzhou, Zhejiang Jinhua, Zhejiang Quzhou and Zhejiang Lishui were planted under Camellia oleifera standin Suichang, Yunhe and Yongkang of Zhejiang province with 4 cultivation patterns like field cultivation, field cultivation with non-woven fabrics at the bottom of ditch, cultivationin non-woven bag and cultivation in root control container, and with 4 substrates like humus soil under C. oleifera stand, 40% special matrix+60% yellow soil, 30% sawdust+60% yellow soil+10% gravel, vegetable soil+10% ash. Orthogonal design was used for the experiment. In December 2019, root tubers of random 10 individuals of each treatment (provenance of Jiangxi Shangrao as CK) were harvested to determine their fresh weight and total flavonoids content. The results showed that the soil substrate had the strongest effect on the individual fresh weight of root tubers in Suichang and Yunhe, while the cultivation pattern had the strongest effect on the fresh weight of individual root tubers in Yongkang. The provenance had the greatest influence on the total flavonoids content of root tubers in three experimental sites. Seedlings of Zhejiang Jinhua provenance cultivated by root control container or non-woven bag with 40% special substrate+60% yellow soil had good effect in the three sites. Seedlings of Zhejiang Lishui provenance, field cultivation by non-woven bag, humus soil+10% ash had the greatest total flavonoid content under C. oleifera.