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A. thaliana accession Columbia-0 (Lehle Se … A. thaliana accession Columbia-0 (Lehle Seeds), or accession Nossen (Fedoroff and Smith, 1993)] were used as wild-types in this study. The mutant brt1-1 (ugt84a2-2) was described previously (Sinlapadech et al., 2007). The Arabidopsis transposon-tagged lines Ds53-4592-1 and Ds54-1263-1 for UGT79B1 (ugt79b1-1 and ugt79b1-2, respectively), and Ds11-5836-1 for UGT84A2 (ugt84a2-1) were obtained from RIKEN Bioresource Center. The T-DNA-insertion mutant GABI_765F10 for UGT84A1 (ugt84a1-1) was obtained from the Arabidopsis Biological Resource Center. Homozygous knockout lines were screened by PCR using specific primers for UGT79B1, UGT84A1, UGT84A2, Ds transposon and T-DNA: UGT79B1f, UGT79B1r, UGT84A1f, UGT84A1r, UGT84A2f, UGT84A2r, Ds5-2a, Ds3-2a, o8409, o3144. PCR products were sequenced to determine the exact insertion points.<br />
For analyses of anthocyanin accumulation, plants were cultured on one-half-strength MS-agar medium containing 1% sucrose (Valvekens et al., 1988) in a growth chamber at 22°C with 16 h/8 h light and dark cycles for 14 days with a light intensity of 40 μmol of photons m−2 s−1, then transferred on one-half-strength MS-agar medium containing 12% sucrose for 3 days with a light intensity of 80 μmol of photons m−2 s−1. Plants were harvested, immediately frozen with liquid nitrogen, and stored at −80°C until use. At least three biological replicates were used for anthocyanin analysis. icates were used for anthocyanin analysis.
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