Fig. 3. Suppression of FLC-dependent late flowering by esd1 mutations. (A) Photograph illustrating the flowering phenotype of double mutant esd1 fve and esd1 fca plants grown under LD. (B) Flowering phenotype of lines where an active allele of FRI is combined with esd1 grown under LD. (C) Analysis of the expression of FLC in the late-flowering genotypes FRI, fve and fca combined with esd1. RNA blot hybridizations were performed using total mRNA from 9-day-old Col, esd1-3, FRI, esd1-3 FRI, Ler, esd1-2, fve-1, esd1-2 fve-1, fca-1 and esd1-2 fca-1 plants grown under LD. (D) Analysis of the expression of FT and SOC1 genes in esd1 mutants. RT-PCR assays comparing FT and SOC1 expression in 9 day-old Col and esd1-3 plants. The samples were taken at the time of the day with the maximum expression; for FT expression analysis, before dusk, and for SOC1 analysis, 8 hours after dawn. (E) Flowering phenotype of esd1 flc double mutant plants grown under LD. (F) Analysis of the expression of MAF genes in esd1 mutant plants. Total RNA was extracted from pools of 50 9-day-old seedlings grown under LD conditions. Expression was monitored by RT-PCR over 32 cycles for MAF1, 28 cycles for MAF2, and 35 cycles for MAF3, MAF4 and MAF5. For the UBQ10 control, we amplified during 22 cycles. RT-PCR products were blotted and hybridized with specific probes for each MAF gene.