Anthocyanin mutants of Japanese and common morning glories exhibit normal proanthocyanidin accumulation in seed coats

Abstract

Anthocyanin and proanthocyanidin biosynthesis pathways are believed to overlap. This study examined proanthocyanidin accumulation in seed coats of morning glories (Ipomoea nil and I. purpurea) carrying mutations in CHS-D, CHI, and ANS genes encoding chalcone synthase, chalcone isomerase, and anthocyanidin synthase, respectively. Chemical staining revealed that mutants accumulate proanthocyanidin normally. Thus, the tested genes are not essential to proanthocyanidin biosynthesis, but are essential to anthocyanin biosynthesis in flowers and stems. Based on the results and the I. nil draft genome sequence, the genes involved in proanthocyanidin biosynthesis, including a new copy of the flavanone 3-hydroxylase gene could be predicted. Moreover, the genome has no homologs for known enzymes involved in producing flavan-3-ols, the starter and extension units of proanthocyanidin. These results suggested that I. nil produces flavan-3-ols through an undiscovered biosynthesis pathway. To characterize proanthocyanidin pigmentation further, we conducted mutant screening using a large I. nil population. We discovered that the brown mutant lines (exhibiting brown seeds and normal anthocyanin pigmentation) do not accumulate proanthocyanidin in their seed coats. Thus, the brown mutation should be useful for further investigations into the various mechanisms controlling anthocyanin and proanthocyanidin pathways.