| S Preparation
|
Arabidopsis thaliana (accession Columbia-0 … Arabidopsis thaliana (accession Columbia-0; Lehle Seeds) was used as the wild type in this study. The mutants ugt75c1, ugt78d1, ugt78d2, ugt89c1, rol1-1, rol1-2, and rhm2-1 were described previously (Jones et al., 2003; Usadel et al., 2004; Tohge et al., 2005; Diet et al., 2006; Yonekura-Sakakibara et al., 2007). The T-DNA–inserted knockout mutants of omt1 (CS25167) and tt7 (CS6509) were obtained from the ABRC. The tt4 (C1) mutant was obtained from tt mutant lines induced by ion beam irradiation of Arabidopsis (Shikazono et al., 2003). The T-DNA–inserted mutant of Arabidopsis, lines SALK_114099 for UGT78D3 and SALK_085051 (rhm2-3, a homozygous knockout line) for RHM2, were obtained from the Salk Institute. T-DNA insertion lines were screened by PCR using specific primers for UGT78D3, RHM2 ,and T-DNA: UGT78D3f, UGT78D3r, RHM2f, RHM2r, LBa1, and RBa1 (see Supplemental Table 1 online). PCR products were sequenced to determine the exact insertion points.<br />
<br />
Plants were cultured on Murashige and Skoog–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 18 d or in a greenhouse at 22°C with 16 h/ 8 h light/dark for 4 weeks. Plants were harvested, immediately frozen with liquid nitrogen, and stored at −30°C until use. Five biological replicates were used for flavonoid profiling. licates were used for flavonoid profiling.
|
