R ecently, we reported on a gene, FLOWERING LOCUS F (FLF), from Arabidopsis whose activity, as determined by steady-state mRNA levels, is correlated quantitatively with the period taken to initiate flowering (1). The FLF gene encodes a putative transcription factor of the MADS-box class, which, we concluded, acts as a repressor of floral induction. Genetic analyses of Arabidopsis ecotypes previously identified FLOW-ERING LOCUS C (FLC) as a semidominant repressor of floral induction (2-4). Because FLF has similar properties to FLC and maps to the same chromosomal region, the possibility arose that FLF and FLC are one and the same gene; this hypothesis has been confirmed by Michaels and Amasino (5). We suggest that the gene be referred to as FLC and the mutants flf-1 to flf-9 now be referred to as flc-11 to flc-19.In many plant species, flowering is promoted by a period of exposure to low temperature through a process known as vernalization. A number of the late-flowering mutants and ecotypes of Arabidopsis have a vernalization response. We found that the vernalization-responsive late-flowering mutants had a higher level of FLC transcript than was present in the wild types and, importantly, that the non-vernalization-responsive lateflowering mutants did not have increased FLC transcript levels. The vernalization-responsive ecotype C24, which is somewhat late-flowering, also has a high level of FLC transcript that is reduced by the low-temperature treatment (1). These observations suggested to us that FLC must be directly involved in the vernalization response.We have shown that reduction in the level of genomic methylation largely substitutes for the low-temperature treatment in promoting flowering (6-8), implicating a reduction in DNA methylation level as a component in the vernalization response. Early-flowering METHYLTRANSFERASE1 antisense plants, with a decreased level of genomic methylation, have a reduced level of FLC activity as compared with the nontransformed control (1). The correlation between FLC transcription and methylation levels implies that FLC is regulated by methylation status, but we are not able to say whether this is through an alteration of the methylation status of the FLC gene itself, or whether it involves other genes that regulate FLC expression.In this paper, we demonstrate that the FLC gene is involved directly in controlling the response to vernalization and more generally in the timing of the transition to the reproductive phase of development. We find that a reduction in FLC activity is a key component of the vernalization response in six late-flowering vernalization-responsive mutants and in the C24 and Pitztal ecotypes. The late-flowering mutants are mutated in genes that regulate the activity of the FLC gene. We further show that the quantitative responses in the promotion of flowering to various periods of exposure to low temperature are paralleled by the levels of FLC transcript, and that the changes observed at the transcriptional level are reflected in the level of the FLC pr...