Tomato germplasm is the raw material, the source of new generic variability, used by plant breeders to develop new cultivars of tomatoes. Regeneration of the national collection of tomato germplasm, now stored at the Australian Tropical Field Crops Genetic Resource Centre, Biloela was required many accessions had small quantities of seed. A number of experiments were conducted to develop a procedure for germinating “fresh” seed. The best method involved scarification of seeds by nicking them with a scalpel under sterile conditions, germination rates increased from 0% to 90%. This procedure for germinating and growing tomato seedlings was used to regenerate the national tomato germplasm collection which currently consists of 15 species and a total of 1216 accessions.