Fruit crops are an important component of horticultural biodiversity, contributing significantly to nutritional security, farm income, and sustainability of agricultural systems. However, their genetic base is increasingly threatened by genetic erosion, climate change, emerging pests and diseases, and intensive cultivation of a limited number of commercial cultivars. Germplasm resources, comprising landraces, traditional varieties, wild relatives, and advanced breeding lines, serve as the primary reservoir of genetic variability for fruit crop improvement. Systematic germplasm activities such as exploration, collection, characterization, evaluation, documentation, and conservation are essential for the effective management and utilization of these genetic resources. Conservation of fruit crop germplasm is particularly challenging due to the perennial nature of crops, recalcitrant seeds, vegetative propagation, and high maintenance costs, necessitating the use of in situ, ex situ, in vitro, and cryopreservation approaches. Pre-breeding plays a critical role in bridging the gap between conserved germplasm and varietal development by identifying and introgression valuable genes from unadopted and wild germplasm into usable breeding materials. Integration of conventional pre-breeding approaches with modern biotechnological tools such as molecular markers, genomics, tissue culture, and gene editing has significantly enhanced the efficiency of germplasm utilization in fruit crops. Strengthening germplasm conservation and pre-breeding programs is vital for developing climate-resilient, high-yielding, and quality fruit cultivars to meet future challenges in horticultural production.