Beans one of the essential plant protein sources for human and animal diets. Conventional breeding methods have been used to develop the cultivars of beans with high quality and high yield. However, conventional methods of plant breeding are time-consuming and laborious. Biotechnological methods can accelerate the breeding process in conventional plant breeding. However, the beans are thought to be a recalcitrant crop plant for applying biotechnological methods since plant regeneration under in vitro conditions in beans is not successful. Developing an appropriate method for in vitro bean regeneration remains a significant problem. The objective of this study was to develop a protocol for the culture of unfertilized ovaries of beans. Culture media and genotype are effective on the success of in vitro cultivation. For this reason, 12 genotypes of beans and some nutrient media such as MS and B5 with various 2,4-D/kinetin combinations were tested to obtain callus from unfertilized ovaries. The highest callus induction was obtained with a medium containing 2,4-D (0.5 mg L−1) and Kinetin (2.5 mg L−1). A literature review on beans indicates that no ovary culture has been carried out on tested varieties in this study to date.