Contribution à l'analyse fonctionnelle des gènes FLOWERING LOCUS C (FLC) et CONSTANS (CO) impliqués dans la floraison de Sinapis alba

Onset of flowering is a major transition in the plant life cycle and is controlled by environmental factors including photoperiod, light quality and temperature. Prevalence of controlling factors depends on species, hence physiological models were selected for their strong requirement for one or another environmental cue. Among Brassicaceae, Sinapis alba was intensively studied for its strong response to photoperiod while molecular-genetic analyses of Arabidopsis thaliana disclosed complex interactions between pathways inducing flowering in response to photoperiod and other environmental cues, such as vernalization. We were therefore interested in studying the vernalization process in S. alba and its interactions with the previously characterized floral response to long days (LDs). Two-week old seedlings grown in non-inductive short days (SDs) were vernalized at 7°C for increasing durations and a strong promotive effect of vernalization was observed. In contrast to the common view of vernalization as a preliminary step bringing the competence to flower, we observed that vernalization had a direct inductive effect on flowering: floral buds were initiated during cold-exposure. Floral integrator genes SaMADSA (homologous to SUPPRESSOR OF OVEREXPRESSION OF CO 1) and SaLEAFY were up-regulated in the shoot apex after 3-4 weeks of vernalization. To monitor the vernalization process at the molecular level, we isolated SaFLC which, based on sequence analysis, expression patterns and complementation test, appeared as orthologous to FLOWERING LOCUS C. Down-regulation of SaFLC by vernalization was fast since transcript level was already very low after one week of vernalization, but stability of the repression required longer exposure to cold. To test the physiological significance of these observations, we studied the floral response to 16-h LDs after unstable and stable repression of SaFLC. We observed that one week of vernalization – which was sufficient for SaFLC repression but not for maintenance of the silenced state - increased the flowering response of S. alba to LDs when the LDs just followed the cold treatment. This effect was lost after two weeks post-vernalization. In contrast, the promotive effect of longer vernalization on flowering response to LDs was maintained post-vernalization. These results suggested that vernalization not only works when plants experience long exposure to cold in winter: short cold periods might stimulate flowering of LD-plants if occurring when photoperiod is increasing, i.e. in spring.