Floral organ- and temperature-dependent regulation of anthocyanin biosynthesis in Cymbidium hybrid flowers.

Anthocyanins are responsible for red, purple, and pink pigmentation of flowers in Cymbidium hybrids. Although anthocyanin content in all floral organs increases with flower development, they increase markedly in the tepals compared with the labella or columns. Using next-generation sequencing technology, we identified three anthocyanin biosynthesis regulatory genes, CyMYB1, CybHLH1, and CybHLH2, from Cymbidium 'Mystique'. Yeast two-hybrid analysis showed that the CyMYB1 protein can form a heterodimer with either CybHLH1 or CybHLH2. In the tepals, the expression level of CyMYB1 increased as the flower developed, whereas the high expression level of CyMYB1 was detected at the early flower developmental stages in the labella and columns, remaining constant until increasing at the late developmental stage. These expression profiles of CyMYB1 positively correlated with the profiles of anthocyanin accumulation in the tepals. When Cymbidium Sazanami 'Champion' was grown at 30 °C/25 °C, reduced anthocyanin levels were observed, specifically in the tepals, compared with those in flowers grown at 20 °C/15 °C. The transcription of CyMYB1 in the tepals was suppressed at high temperatures, and the expressions of CyDFR and CyANS were also synchronously suppressed. This study revealed that CyMYB1 activates the transcription of CyDFR and CyANS and regulates the temporal- and temperature-dependent anthocyanin accumulation in Cymbidium tepals.