NAGESHA, N.U. M., ANKITH,2023-01-162023-01-162020-12-30https://krishikosh.egranth.ac.in/handle/1/5810192392Ridge gourd is one of the important vegetable crops of cucurbit species. The recent emergence of Ridge gourd yellow mosaic disease (RgYMD) caused by ToLCNDV is becoming the major constraints in ridge gourd production. In present investigation, the preliminary work has been carried out in the process of development of transgenic ridge gourd to confer resistance against ToLCNDV using coat protein mediated resistance (CPMR) strategy. The survey was conducted in southern parts of Karnataka to assess the economic importance of ToLCNDV infection. The results of survey revealed the ToLCNDV is predominant in all surveyed areas and all varieties were found susceptible to virus. The total DNA from infected leaves of ridge gourd and coat protein gene was amplified using gene specific primers. The purified CP gene was successfully cloned topTZ57R/T vector. Further, CP gene was moved into plant expression vector pBI121 with the help of the pET32a vector. Subsequently, the vector pBI121 having CP gene construct was transformed into Agrobacterium tumefaciens. For bacterial expression studies, the CP gene was cloned into pET28a and expressed in E. coli BL21 cells. The presence of distinct protein band of size ~34 kDa in the recombinant cells confirms the over-expression of protein. The efforts were made to standardize the growth media for in vitro regeneration of ridge guard. The best organogenic callus was obtained in 2.5 mg/L of 2, 4- D + 0.5 mg/L of BAP. These research findings will be helpful for thedevelopment of transgenic ridge gourd.EnglishThesis