概要: This study aim to understand how the long and short day flowering pathways are integrated and the mechanism of photoperiod perception in rice. Trascriptome at different time points under LD and SD conditions reveal that photoperiodism in rice is controlled by the evening complex. Mutants in LUX ARRYTHMO (LUX) and EARLY FLOWERING3 (ELF3) orthologs abolish flowering. We show that light causes a rapid and sustained degradation of ELF3-1, and this response is dependent on phyB. ChIP-seq of ELF3 and LUX reveal that EC controls both LD and SD flowering pathways by directly binding and suppressing the expression of key floral repressors, including PRR7 orthologs and Ghd7.
项目整体设计: Transcriptome in WT, phyb-1 and elf3-1 elf3-2 lines at multiple points in the day under SD or LD conditions.
For the measurement of flowering-time, wild-type, lux, elf3-1, elf3-2, and elf3-1 elf3-2 rice seeds were germinated in water-soaked paper at 28 ºC for 3 days. Germinated seedlings were planted in containers with soil (soil:peat:vermiculite in 2:2:1 volume ratio) and placed in controlled conditions at 28 ºC under 12h/12h light/dark photoperiod (ND conditions) at 70 % humidity.
处理方案:
Plants were grown under 14h/10h (LD conditions) and 10h/14h (SD conditions) photoperiod
提取方案:
Homogenizing samples to powder using a freeze cold mortar and pestle followed by total RNA using the RNeasy plant kit (Qiagen), followed by DNAse I treatment (Ambion)
建库方案:
Prepare high-throughput mRNA sequencing libraries using NEBNext Ultra II Directional RNA Library Prep Kit (New England Biolabs)
