Flowering time control in ornamental gloxinia (Sinningia speciosa) by manipulation of miR159 expression

Gloxinia (Sinningia speciosa) is a popular commercial plant for its attractive and colourful flowers. However, the genetic mechanism of flowering time regulation in gloxinia is largely unknown. Recent studies on model plants have elucidated that miR159 acts as a negative regulator of floral transition in short-day photoperiods. The aim of this study was to investigate whether genetic modification of miR159 expression can offer an effective approach for regulation of flowering characteristics in gloxinia. Transgenic gloxinia plants were generated that over-express or suppress miR159 by means an efficient Agrobacterium-mediated transformation system in order to study the effect of miR159 on flowering time. In addition, the full-length cDNA of gloxinia GAMYB (SsGAMYB) was also cloned, and was verified to be a target of miR159 by modified RNA ligase-mediated 5 rapid amplification of cDNA ends. Transgenic gloxinia plants that over-express or suppress miR159 exhibited significantly late or early flowering, respectively. During flower development, the expression level of miR159 was negatively correlated with SsGAMYB in gloxinia. MiR159-mediated SsGAMYB expression affected the expression levels of SsLEAFY (SsLFY) and three MADS-box genes (SsAP1, SsAP3 and SsAG), which regulated floral transition downstream of GAMYB. In addition, suppression of miR159 caused a conversion of petals and sepals in a few transgenic plants. miR159-regulated GAMYB expression is an effective pathway of flowering time control in gloxinia. Transgenic manipulation of miR159 can be used as an applicable strategy to regulate flowering time in commercial ornamental plants.