Background: Mungbean yellow mosaic virus disease is the most devastating disease on Mungbean production. The virus is transmitted by whitefly and can cause yield losses from 75 to 100 per cent. The development of mungbean cultivars resistant to both virus and its vector is considered as one of the most desirable means of managing the disease as it is environmentally safe and highly efficient. The selection of resistant genotypes in conventional methods is complex and time consuming. Hence, the use of molecular markers linked with resistance genes is powerful as it hastens the breeding programmes. The current study was aimed to develop mapping population and to validate molecular markers associated with Mungbean yellow mosaic virus (MYMV). Methods: The present investigation was carried out with 260 F2 individuals that were derived from crossing DGGV-2 and IPM 2-14 during Kharif-2017 at Main Agril Research Station, UAS, Dharwad. Hybrid seeds of this cross were harvested individually and sown during rabi 2017 along with the two parents, as checks for distinguishing the true hybrids. Hybridity of F1s was confirmed through molecular marker analysis and the true F1s were selfed to raise the F2 generation. Result: Of the 24 previously reported simple sequence repeat markers used for detecting the polymorphism, two markers viz., CEDG305 and CEDG115 were found to be polymorphic between DGGV-2 and IPM-2-14. Two hundred and sixty F2 plants segregated in the ratio of 3 S:1 R (202 susceptible: 58 resistant) as phenotypic and 1: 2 :1 as genotypic ratio implying that single recessive gene controlled resistance. Single marker analysis revealed that the molecular markers CEDG305 and CEDG115 were associated with MYMV resistance with a phenotypic variance of 24.5 and 10.3 per cent respectively.