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Chen W.,Zhejiang Agriculture And forestry University | Qin Q.,Zhejiang Agriculture And forestry University | Zhang C.,Zhejiang Agriculture And forestry University | Zheng Y.,Zhejiang Senhe Seed Co. | And 5 more authors.
Plant Cell Reports | Year: 2015

Key message: DhEFL2, 3and4regulate the flowering ofDoritaenopsis. These genes could rescueelf4-1phenotype in Arabidopsis while its overexpression delayed flowering.Abstract: Phalaenopsis are popular floral plants, and studies on orchid flowering genes could help develop off-season cultivars. EARLY FLOWERING 4 (ELF4) of A. thaliana has been shown to be involved in photoperiod perception and circadian regulation. We isolated two members of the ELF4 family from Doritaenopsis hybrid (Doritaenopsis ‘Tinny Tender’ (Doritaenopsis Happy Smile × Happy Valentine)), namely, DhEFL2 and DhEFL3 (DhEFL4 has been previously cloned). Multiple alignment analysis of the deduced amino acid sequences of the three DhEFL homologs showed that DhEFL4 and DhEFL2 are similar with 72 % identical amino acids, whereas DhEFL3 is divergent with 72 % similarity with DhEFL2 and 68 % similarity with DhEFL4. DhEFL3 forms a separate phylogenetic subgroup and is far away from DhEFL2 and DhEFL4. The diurnal expression patterns of DhEFL2, 3, and 4 are similar in the long-day photoperiod conditions; however, in the short-day conditions, DhEFL3 is different from DhEFL2 and 4. For the DhEFL2, 3, and 4 genes, the strongest audience expression organs are the stem, petal and bud, respectively. The ectopic expression of DhEFL2, 3, or 4 in transgenic A. thaliana plants (Ws-2 ecotype) showed novel phenotypes by late flowering and more rosette leaves. The ectopic expression of DhEFL2, 3, or 4 could complement the elf4-1 flowering time and hypocotyl length defects in transgenic A. thaliana elf4-1 mutant plants. These results strongly suggest that DhEFL2, 3, and 4 may be involved in regulation of flower formation and floral induction in Doritaenopsis. © 2015 Springer-Verlag Berlin Heidelberg

Chen W.,Zhejiang Agriculture And forestry University | Qin Q.,Zhejiang Agriculture And forestry University | Zheng Y.,Zhjiang Senhe Seed Co. | Zheng Y.,Oceanwide International Center | And 6 more authors.
Plant Molecular Biology Reporter | Year: 2015

The EARLY FLOWERING 4 (ELF4) gene has been reported as a requirement for photoperiod perception and circadian function. In this study, an ELF4 homolog gene EARLY FLOWERING 4-like4 (EFL4) was isolated from Doritaenopsis “Tinny Tender” (Doritaenopsis Happy Smile × Happy Valentine) and designated as DhEFL4. The acquired DhEFL4 cDNA was 593 bp with a 345 bp open reading frame encoding 114 amino acids. The putative DhEFL4 protein contained a conserved region, DUF1313, which is shared by the ELF4 family. Amino acid sequence alignment showed that DhEFL4 shared high similarity to ELF4 and ELF4-like homologs from other species. The DhEFL4 promoter was also obtained, and 241 cis-elements were found. The diurnal expression of DhEFL4, which may be affected by the EVENINGAT (EE) and circadian motif was analyzed by quantitative PCR under short- and long-day conditions. Reverse-transcription PCR analysis results showed that DhEFL4 is an evening-expressed circadian gene. DhEFL4 was expressed in both vegetative and reproductive organs, and its transcripts reached higher levels in the buds. To further study the function of the DhEFL4 gene, DhEFL4 was overexpressed in Arabidopsis thaliana by Agrobacterium-mediated transformation. The flowering of these transgenic plants was more delayed than that of wild-type Arabidopsis by 12−14 days, and flower stems increased. Results indicated that DhEFL4 may perform an important function in the regulation of flowering. © 2015 Springer Science+Business Media New York

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