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The grass goldfish appeared early in the evolutionary history of goldfish, and shows heritable stability in the development of the caudal fin. The twin-tail phenotype is extremely rare, however, some twin-tail individuals were produced in the process of breeding for ornamental value. From mutations in the twin-tail goldfish genome, we identified two kinds of  Tgf2  transposons. One type was completely sequenced  Tgf2  and the other type was  ΔTgf2 , which had 858 bp missing. We speculate that the bifurcation of the axial skeletal system in goldfish may be caused by an endogenous  ΔTgf2  insertion mutation in  Chordin A , as  ΔTgf2  has no transposition activity and blocks the expression of  Chordin A . The twin-tail showed doubled caudal fin and accumulation of red blood cells in the tail. In addition,  in situ  hybridization revealed that ventral embryonic tissue markers ( eve1 ,  sizzled , and  bmp4 ) were more widely and strongly expressed in the twin-tail than in the wild-type embryos during the gastrula stage, and  bmp4  showed bifurcated expression patterns in the posterior region of the twin-tail embryos. These results provide new insights into the artificial breeding of genetically stable twin-tail grass goldfish families.

Insertional mutagenesis in ChordinA induced by endogenous ΔTgf2 transposon leads to bifurcation of axial skeletal systems in grass goldfish