LjCYC genes constitute floral dorsoventral asymmetry in Lotus japonicus

Previous study shows that LjCYC2, a CYC-like TCP (TB1, CYC and PCFs) gene in the model legume, Lotus japonicus, is involved in dorsal petal development, which together with the other two homologous genes, LjCYC1 and LjCYC3, belongs to an LjCYC gene cluster. In this report, we modified the transformation system in L. japonicus, and constructed different RNAi transgenes to target different LjCYC genes. The expression of three endogenous LjCYC genes was specifically suppressed by different specific RNAi transgenes, and a chimerical RNAi transgene that contains the specific sequences from LjCYC1 and LjCYC2 was found to downregulate the expression of both endogenous genes simultaneously. Effects of silencing three LjCYC genes were mainly restricted on either dorsal or lateral petals, demonstrating their dorsalizing and lateralizing activities during the development of zygomorphic flower. Furthermore, abolishing the expression of three LjCYC genes could give rise to complete loss of dorsoventral (DV) differentiation in the flower whose petals all resembled the ventral one in the wild type and displayed intact organ internal (IN) asymmetry. Our data demonstrate that during zygomorphic flower development, the DV asymmetry is constituted by the LjCYC genes, while the floral organ IN asymmetry is independently determined by other genetic factors.