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DhEFL2, 3 and 4, the three EARLY FLOWERING4-like genes in a Doritaenopsis hybrid regulate floral transition

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Abstract

Key message

DhEFL2, 3 and 4 regulate the flowering of Doritaenopsis . These genes could rescue elf4 - 1 phenotype 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.

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Abbreviations

ELF4 :

EARLY FLOWERING 4

EFL2 :

EARLY FLOWERING 4-like2

EFL3 :

EARLY FLOWERING 4-like3

EFL4 :

EARLY FLOWERING 4-like4

SD:

Short day

LD:

Long day

GI :

GIGANTEA

CCA1:

CIRCADIAN CLOCK ASSOCIATED 1

LHY:

LATE ELONGATED HYPOCOTYL

TOC1 :

TIMING OF CAB EXPRESSION 1

ELF3:

EARLY FLOWERING 3

LUX:

LUX ARRHYTHMO

CaMV:

Cauliflower mosaic virus

EST:

Expressed sequence tag

CO :

CONSTANS

RACE:

Rapid amplification of cDNA ends

ZT:

Zeitgeber time

qRT-PCR:

Quantitative real-time PCR

PCR:

Polymerase chain reaction

MS:

Murashige and Skoog

ORF:

Open reading frame

RT-PCR:

Reverse transcription-polymerase chain reaction

WT:

Wild type

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Acknowledgments

We thank Dr. Zhi John Lu, MOE Key of Bioinformatics, School of Life Science, Tsinghua University, for the useful suggestions on improving the manuscript, and Mr. B. B. Shi, School of Medicine, Tsinghua University, for the useful suggestions on drafting figures.

Author information

Authors and Affiliations

  1. The Nurturing Station for the State Key Laboratory of Subtropical Silviculture, Zhejiang Agriculture and Forestry University, Lin’an, Hangzhou, 311300, Zhejiang, People’s Republic of China

    Weiwei Chen & Mingbing Zhou

  2. School of Agriculture and Food Science, Zhejiang Agriculture and Forestry University, 88 Huancheng Street, Lin’an, Hangzhou, 311300, Zhejiang, People’s Republic of China

    Weiwei Chen, Qiaoping Qin, Chi Zhang & Yongyi Cui

  3. Zhejiang Senhe Seed Co., Ltd.,, Hangzhou, 310012, Zhejiang, People’s Republic of China

    Yongping Zheng & Chun Wang

  4. Oceanwide International Center, 19th Floor, Building A, No. 2 Xiangzhang Road, Qianjiang CBD, Hangzhou, 310007, Zhejiang, People’s Republic of China

    Yongping Zheng & Chun Wang

Authors
  1. Weiwei Chen
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  2. Qiaoping Qin
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  3. Chi Zhang
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  4. Yongping Zheng
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  5. Chun Wang
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  7. Yongyi Cui
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Corresponding author

Correspondence to Yongyi Cui.

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Conflict of interest

No conflicts of interest declared.

Funding

This work was supported by National Natural Science Foundation of China (Grant Nos. 30771762, 31170658, and 31301811) and the Science & Technology Key Project of Zhejiang Province (Grant No. 2012C12909-11).

Additional information

Communicated by P. Lakshmanan.

The nucleotide sequence reported in this paper has been submitted to GenBank with accession numbers KP728997 (DhEFL2 mRNA), KP728998 (DhEFL3 mRNA).

Electronic supplementary material

Below is the link to the electronic supplementary material.

299_2015_1848_MOESM1_ESM.tif

Supplementary material 1 (TIFF 14597 kb) Online Resource 1 Nucleotide and deduced amino acid sequences of DhEFL2 and 3 cDNAs. The DUF1313 superfamily region is indicated with the parenthesis in bold

299_2015_1848_MOESM2_ESM.docx

Supplementary material 2 (DOCX 72 kb) Online Resource 2 Sequence information on alignment of the DhEFL2, 3, 4 amino acid sequences with ELF4 and ELF4-like from different plant species.

299_2015_1848_MOESM3_ESM.docx

Supplementary material 3 (DOCX 16 kb) Online Resource 3 Sequence information on phylogenetic analysis of the DhEFL2, 3, and 4 encoded amino acid sequences from different plant species

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Chen, W., Qin, Q., Zhang, C. et al. DhEFL2, 3 and 4, the three EARLY FLOWERING4-like genes in a Doritaenopsis hybrid regulate floral transition. Plant Cell Rep 34, 2027–2041 (2015). https://doi.org/10.1007/s00299-015-1848-z

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  • DOI: https://doi.org/10.1007/s00299-015-1848-z

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