Characterization of the leaf color mutant hy and identification of the mutated gene in Chinese cabbage

Abstract

Leaf color mutants play an important role in our understanding of chlorophyll biosynthesis and catabolism. In this study, we obtained a yellow-green leaf mutant hy in an ethyl methanesulfonate mutagenized population of chinese cabbage (Brassica rapa ssp. pekinensis). The hy phenotype was controlled by a recessive allele at a single locus. The intrinsic photochemical activity of photosystem II (PSII) is impaired in hy, suggesting that absorbed light energy is not efficiently transferred from the light-harvesting complexes antenna to the PSII reaction centers and dissipated as heat or fluorescence. We measured chlorophyll content and chlorophyll precursors and analyzed the expression of key genes in the chlorophyll synthetic pathway in hy and wild type. The mutation phenotype was consistent with inhibited expression of chlorophyll a oxygenase (CAO) gene in the chlorophyll synthetic pathway. In mutant hy, CAO cDNA was cloned so that a C to T mutation at 1099 bp caused a conserved proline (Pro) to serine (Ser) mutation at the 367th amino acid in C-domain, which changed the secondary structure of CAO protein. We speculate that the mutation amino acid changed in the C-domain may affect the catalytic function in mutant CAO.