Inter-varietal hybridization is a powerful tool for genetic improvement and production of new genotypes for a trait of interest. Four parents of faba beans (Vicia faba L.) were hybridized using agromorphological and molecular characterization to obtain genotypes resistant to the chocolate spot disease. The study was done at the Nubaria Research Station, Giza, Egypt. Eight traits including resistance to chocolate spot, days to flowering, plant height (cm), number of branches/plant, number of pods/plant, number of seeds/plant, 100-seed weight (g), and seed yield/plant were estimated during the three growth seasons of 2016/2017, 2017/2018, and 2018/2019. Genetic parameters revealed by RAPD and ISSR markers assessed the genetic variation of genotypes with their generations. Crosses 1 (P1 “Nubaria-1” x P2 “Sakha-1”), 2 (P1 “Nubaria-1” x P3 “T.W”), and 3 (P1 “Nubaria-1” x P4 “Camolina”) revealed high resistance to disease with high yield. Markers patterned specific loci of resistant parents at a length of 360, 470, 450, 660, and 140 bp in RAPD and 1100, 810, 650, 700, 480 bp in ISSR. Inter-varietal hybridization between the resistant and susceptible genotypes is considered one of the most promising methods to obtain germplasm with resistance and high yield.