In the present study, the transient expression of maize C1-R regulatory genes of anthocyanin biosynthesis in maize calli, wheat immature embryos, rice calli and tobacco leaves were described. C1-R regulatory genes could activate anthocyanin biosynthesis in vivo and the pigmented cells were visible. For major crops, including maize, wheat and rice, which were transformed mainly by biolistic method, C1-R genes were novel visible markers, and the expression was easier to detect than GUS gene. And these genes also worked well in transient expression in biolistic transformation system of tobacco leaves.