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OsLHY regulates photoperiodic flowering through the unique pathways under long-day conditions in rice (Oryza sativa L.)
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  • Chao Li,
  • Xue-Jiao Liu,
  • Yan Yan,
  • Alam Mohammad Shah,
  • Zhen Liu,
  • Ruo-Fu Tao,
  • Er-Kui Yue,
  • Ming-Hua Duan,
  • Jianhong Xu
Chao Li
Zhejiang University
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Xue-Jiao Liu
Zhejiang University
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Yan Yan
Zhejiang University
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Alam Mohammad Shah
Zhejiang University
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Zhen Liu
Zhejiang University
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Ruo-Fu Tao
Zhejiang University
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Er-Kui Yue
Zhejiang University
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Ming-Hua Duan
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Jianhong Xu
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Abstract

Flowering time (or heading date in crops) is a critical agronomic trait for rice reproduction and adaptation. The circadian clock is an endogenous oscillator that is involved in controlling photoperiodic flowering. The rice LATE ELONGATED HYPOCOTYL (OsLHY), the core oscillator component of circadian clock, is a homolog of the LHY/CCA1 in Arabidopsis. Here we showed that CRISPR/Cas9-engineered mutations in OsLHY caused late flowering in rice only under long-day (LD) conditions. In the oslhy mutant, the diurnal expression of circadian clock-related genes was seriously affected under both LD and short-day (SD) conditions. Furthermore, the expression of the flowering activators Ehd1, Hd3a and RFT1 was down-regulated and flowering repressors Hd1 and Ghd7 was up-regulated in the oslhy mutant under LD conditions. While the transcripts of flowering-related genes were not dramatically influenced under SD conditions. Dual-luciferase assays showed that OsLHY repressed the transcription of OsGI, Hd1, Ghd7, Hd3a, RFT1 and OsELF3, and activated the transcription of Ehd1. Moreover, the yeast one hybrid assay confirmed that OsLHY directly repressed OsGI, RFT1 and OsELF3 by binding to their promoters, which is consistent with that in Arabidopsis. These results suggested that the OsLHY can promote rice flowering mainly through regulating Hd1 and Ehd1 under LD conditions.