CH257 is a three-pistil material bred after three backcrosses between "three-grain wheat" and common wheat "Shiyou 20", which can greatly increase the number of grains per ear. In order to analyze the inheritance of three pistil traits in wheat, dig out its control genes and develop related molecular markers, this study used CH257 and "China Spring" to formulate a combination, and obtained its F1, F2 and BC1F1 populations for genetic analysis; using homozygous normal and For homozygous three-pistil F2 lines, bulked segrgant analysis (BSA) was used to construct single-pistil and three-pistil mixed pools for 90K gene chip scanning, and molecular markers were developed in the corresponding segments based on the chip results. The results show that the development of the three pistils in CH257 is controlled by a pair of dominant single gene Pis-CH257; using 90K gene chip scanning to locate Pis-CH257 on the 2DL chromosome, develop markers in the corresponding segment of 2DL, and obtain a total of 5 and Pis- The sequence of CH257-linked co-dominant SSR markers is 2DL07, 2DL17, 2DL22, Pis-CH257, 2DL25, 2DL38, the linkage markers 2DL22 and 2DL25 on both sides of Pis-CH257 and their genetic distances are 1.1cM and 2.5cM respectively. This study provides a basis for cloning the genes that control the three pistil traits of wheat.