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It is well accepted that the HSPCs are emerged directly from the aortic endothelium of the dorsal aorta(DA)in AGM region via endothelial haematopoietic transition(EHT)in zebrafish.However,through the living image of Tg(flk1:eGFP),we observed that the aortic endothelial cells of the DA in the CHT region could also bend to the HSPCs directly,just succeeding the termination of HSPCs formation in AGM at around 60 hpf,suggesting a second wave of HSPCs formation via EHT existed.