Overexpression of CONSTANS-like 16 enhances chlorophyll accumulation in petunia corollas

We have previously found that a gene closely related to Arabidopsis CONSTANS-like 16 (COL16) was coordinately expressed with chlorophyll content in chrysanthemum petals and leaves. Here, to elucidate whether COL16 is involved in the regulation of chlorophyll biosynthesis and accumulation, we analyzed the function of COL16 in petunia (Petunia hybrida). We identified three petunia COL16 homologs: PhCOL16a, PhCOL16b, and PhCOL16c. Expression patterns of all three homologs were associated with chlorophyll content, with lower levels in white corollas than in pale green corollas, and relatively high levels in leaves. The result suggests that PhCOL16 homologs are involved in chlorophyll accumulation. We introduced a PhCOL16a overexpression construct into petunia. The transgenic plants had pale green corollas with a higher chlorophyll content than wild-type plants. Expression of genes encoding key enzymes of chlorophyll biosynthesis was significantly higher in the transgenic plants than in the wild-type plants. The results indicate that PhCOL16 positively regulates chlorophyll biosynthesis.