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Petunia  line Mitchell [MP,  Petunia axillaris  × ( P. axillaris  ×  P. hybrida )] and  Eustoma grandiflorum  (lisianthus) plants were produced containing a transgene for over-expression of the R2R3-MYB transcription factor [TF; ROSEA1 (ROS1)] that up-regulates flavonoid biosynthesis in  Antirrhinum majus . The petunia lines were also crossed with previously produced MP lines containing a  Zea mays  flavonoid-related basic helix-loop-helix TF transgene (LEAF COLOR, LC), which induces strong vegetative pigmentation when these  35S:LC  plants are exposed to high-light levels.  35S:ROS1  lisianthus transgenics had limited changes in anthocyanin pigmentation, specifically, precocious pigmentation of flower petals and increased pigmentation of sepals. RNA transcript levels for two anthocyanin biosynthetic genes,  chalcone synthase  and  anthocyanidin synthase , were increased in the  35S:ROS1  lisianthus petals compared to those of control lines. With MP, the  35S:ROS1  calli showed novel red pigmentation in culture, but this was generally not seen in tissue culture plantlets regenerated from the calli or young plants transferred to soil in the greenhouse. Anthocyanin pigmentation was enhanced in the stems of mature  35S:ROS1  MP plants, but the MP white-flower phenotype was not complemented. Progeny from a  35S:ROS1  ×  35S:LC  cross had novel pigmentation phenotypes that were not present in either parental line or MP. In particular, there was increased pigment in the petal throat region, and the anthers changed from yellow to purple pigmentation. An outdoor field trial was conducted with the  35S:ROS1 ,  35S:LC ,  35S:ROS1  ×  35S:LC  and control MP lines. Field conditions rapidly induced intense foliage pigmentation in  35S:LC  plants, a phenotype not observed in control MP or equivalent  35S:LC  plants maintained in a greenhouse. No difference in plant stature, seed germination, or plant survival was observed between transgenic and control plants.

frontiers in plant science

MYB and bHLH transcription factor transgenes increase anthocyanin pigmentation in petunia and lisianthus plants, and the petunia phenotypes are strongly enhanced under field conditions