The procedures involving binary vectors for rose transformation
were also described previously (Fukuchi-Mizutani et al. 2003).
In brief, pBin19 and pBinPLUS (van Engelen et al. 1995), which
contain neomycin phosphotransferase II (NPTII) as the selection
marker, were used to construct a binary vector, and the enhanced
cauliflower mosaic virus (CaMV) 35S promoter derived from
pE2113 (Mitsuhara et al. 1996) was used to regulate the transgenes.
Torenia anthocyanin 5-acyltransferase cDNA (database accession
No. AB332099) was obtained from its petal cDNA library (Suzuki
2000) as described previously (Yonekura-Sakakibara et al. 2000).
Irishollandica DFR cDNA (AB332098) was cloned from its
petal cDNA (Imayama et al. 2004) as described before (Tanaka
et al. 1995). Viola spp. contains two F3050H genes (Brugliera et al.
2004), and one of them (clone BP40) was used in this study
(AB332097).