Plant breeding has significantly increased the yield, quality, and disease resistance of crops. Advances in genomic resources have revolutionized this field, allowing for the precise identification of genes and the application of marker-assisted selection (MAS) through molecular breeding techniques. These advancements are supported by various powerful tools, including genomic sequences, molecular markers, genetic transformation, and genome editing methods, all utilized to improve crops such as tomatoes. Tomatoes represent a major global agriculture commodity, with an estimated value of USD 1.4 billion. In North Carolina alone, tomato cultivation contributes approximately USD 35 million annually, spanning 3,200 acres of cultivated land. This presentation will focus on the genomic and biotechnological resources employed in crop improvement, using North Carolina (NC), USA, as a representative case study. The NC State Tomato Breeding Program enhances tomato varieties by improving their disease resistance and fruit quality. Substantial progress has been made by integrating conventional and molecular breeding approaches. However, new challenges, particularly from evolving pests and diseases, require ongoing research and development efforts. Fruit quality is a multifaceted trait assessed based on various parameters such as shape, size, smoothness, lycopene content, and flavor. The NC State Tomato Breeding Program strives to optimize these attributes to meet market demands and consumer preferences. The program's efforts include the development of new tomato varieties that exhibit superior qualities in these areas. This presentation will outline the ongoing breeding initiatives aimed at addressing the challenges that are faced by tomato cultivation. It will delve into the methodologies employed, encompassing conventional breeding techniques and cutting-edge molecular approaches. Through this comprehensive overview, the audience will gain a deeper understanding of the critical role that genomic and biotechnological resources play in modern crop improvement efforts.