Two distinct spontaneous mutations involved in white flower development in Lilium speciosum

Abstract

Lilium speciosum is one of the most important wild species used for breeding Oriental hybrid lily cultivars. Typical L. speciosum flowers possess red tepals and dark red anthers because of anthocyanin accumulation. However, white-flowered L. speciosum with two distinct phenotypes, one with white tepals and dark red anthers and the other with white tepals and yellow anthers, also exists. To elucidate the molecular mechanisms underlying white colouration, we isolated two regulatory genes and eight biosynthetic genes responsible for anthocyanin pigmentation in L. speciosum and compared these in red- and white-flowered lines. In the white tepal line with dark red anthers, we detected an amino acid substitution in the R2 repeat of LsMYB12, the R2R3-MYB that regulates tepal-specific anthocyanin biosynthesis, which led to reduced anthocyanin biosynthetic gene transcription and no tepal pigmentation. This LsMYB12 sequence was identical to that of an LhMYB12 in the white-flowered Oriental hybrid lily cultivar ‘Rialto’, indicating that this LsMYB12 sequence is present in lines that have been used for breeding. In addition, a nonsense mutation was detected in the dihydroflavonol 4-reductase (DFR) sequence of the white tepal line with yellow anthers, which resulted in a lack of anthocyanin accumulation in tepals and anthers. This mutated DFR has not been used for lily breeding because white tepal Oriental hybrid lily cultivars usually have dark red anthers. These results suggest that several mechanisms generate white flowers in lilies, and provide a substantial foundation for the production of Oriental hybrid lily cultivars with novel flower colour combinations.