The secret art of silk culture along with mulberry seeds is presumed to have spread from China to other parts of the world including India through the famous Silk Road. In this study, we investigated a set of 36 important mulberry genotypes (designated as ‘breeders’ collection’) of historical importance that have been frequently used in Indian crop improvement programmes over the last five decades. This study is the first to employ a large number of microsatellite markers (140 screened; 70 used for profiling) to elucidate the diversity, structure and breeding history of mulberry. The cluster and STRUCTURE analysis corroborated with the known genetic relationships and origin. The groupings by STRUCTURE (k= 4) confirm parallel breeding efforts undertaken in the eastern, southern and northern regions of the country. The cultivar ‘Mysore Local’ shares a common parent with ‘Berhampore Local’, and this supports the historical records of its introduction from eastern India to the Kingdom of Mysore by the erstwhile ruler Tippu Sultan. The popular variety ‘Kanva-2’ shared a common male parent with the Japanese variety ‘Kousen’, supporting the contribution of exotic progenitors in Indian cultivars. The findings of this study will be useful in formulating new strategies for mulberry improvement and reveals the historical events of the introduction and spread of cultivated mulberry in the Indian subcontinent.