Hematopoietic stem cells (HSCs) replenish all types of blood cells. It is debating whether HSCs in adults solely originate from the aorta-gonad-mesonephros (AGM) region, more specifically, the dorsal aorta, during embryogenesis. Here we report that somite hematopoiesis, a previously unwitnessed hematopoiesis, can generate definitive HSCs (dHSCs) in zebrafish. We noticed that in several transgenic lines, in which GFP is expressed in somites during embryonic development, embryos and adults carry hematopoietic cells in the circulation. We established Tg(foxc1b:EOS) transgenic line, which expresses the photoconvertible fluorescence reporter EOS in somites. By photoactivating EOS in specific somite areas, we found that a subset of cells within the forming somites emigrate ventrally and mix with lateral plate mesoderm-derived primitive hematopoietic cells before the blood circulation starts. These somite-derived hematopoietic precursors and stem cells (sHPSCs) subsequently enter the circulation and colonize the kidney of larvae and adults. RNA seq analysis reveals that sHPSCs express hematopoietic genes with sustained expression of many muscle/skeletal genes, which is in contrast to absent or low expression of muscle/skeletal genes in gata1a-positive primitive hematopoietic precursors. Embryonic sHPSCs transplanted into wild-type embryos expand during growth and survive for life time with differentiation into various hematopoietic lineages, indicating self-renewal and multipotency features. In contrast, transplanted gata1a-positive primitive hematopoietic precursors cannot survive over 9 days in the recipients. Therefore, we propose that dHSCs can also originate from embryonic somites.