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For the metabolite measurements, qRT–PCR, … For the metabolite measurements, qRT–PCR, and microarray analysis, the plant material was sterilized, grown, and harvested according to the following procedure. Surface-sterilized T2 seeds of the rice FOX Arabidopsis lines and the Arabidopsis At-LBD37/ASL39-overexpressor lines were inoculated on solid Murashige and Skoog (MS) medium containing 1% sucrose and 20 mg l−1 hygromycin. Seeds of the plants of the activation-tagged lines of the AS2 and ASL1 genes (Chalfun-Junior et al., 2005; Iwakawa et al., 2002; Nakazawa et al., 2003), and the LBD38/ASL40 gene originating from the same mutant collection, as well as seeds of the LBD39/ASL41-overexpressing Arabidopsis FOX line F15720, were inoculated under the same growth conditions using the above-mentioned procedure.
SALK lines (Alonso et al., 2003) were obtained from the Arabidopsis Biological Resource Center (ABRC). The homozygous SALK lines 057939 and 058867, which harbor a T-DNA insertion site in the ORF and the 5′ UTR of LBD37/ASL39, respectively, were grown on MS medium supplemented with 50 mg l−1 kanamycin.
The plants were grown under long-day (16 h light) and low-light conditions (50 μmol s−1) at 22°C for 18 d in growth chambers (SANYO; MLR-350h/MLR-350 HT) until sampling. The plates were prepared in duplicate using 60 seeds of six independent lines; each plate contained five seeds of a particular line and five seeds of the empty-vector control line ‘BIG’. The plates were numbered, placed in a growth chamber in a defined position, and rotated twice a week to ensure equal light reception. At 18 DAS, the seedlings were harvested without roots, weighed, immediately deep-frozen in liquid nitrogen, and stored at –80°C until further analysis.
For assays using homozygous T3 lines, the seeds were treated using the above-mentioned procedure and inoculated on solid hygromycin-free MS medium. Plant growth and harvesting were performed using the above-mentioned protocol.
The soil-grown plants in the greenhouse were grown under the same conditions as those used for the plants grown in growth chambers. d for the plants grown in growth chambers.
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