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Characterization of TEMINAL FLOWER1 homologs CmTFL1c gene from Chrysanthemum morifolium

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The CmTFL1c gene of Chrysanthemum morifolium inhibits flowering, regulates inflorescence architecture and floral development.

Abstract

The timing of flowering is an important ornamental trait of chrysanthemum. The gene TERMINAL FLOWER1 (TFL1) has been shown to be involved in the regulation of meristem fate and flowering time regulation. Here, a TFL1 gene named as CmTFL1c, was cloned from Chrysanthemum morifolium and further characterized. The open reading frame of CmTFL1c comprises 522 bp, which encodes a polypeptide of 173 amino acids. Phylogenetic analysis revealed that CmTFL1c belongs to the CEN/TFL1 clade. CmTFL1c protein localizes to the nucleus as well as to plasma membrane, which suggests that CmTFL1c may be a transcription factor. The CmTFL1c gene was most highly expressed in vegetative stems, and weakly expressed in leaves and flower buds; both shoot apices and stems had sensitivity to photoperiod. Overexpression of CmTFL1c in wild Arabidopsis and tfl1-13 mutant led to late flowering and altered architecture, including increased secondary branching, and abnormal inflorescences and flowers. The CmTFL1c gene negatively regulated flowering by inhibiting the up-regulation of the AtFT, AtLFY and AtAP1. The biological function of CmTFL1c was further characterized in C. morifolium via Agrobacterium-mediated transformation, which showed that CmTFL1c not only delayed flowering and promoted axillary bud formation, but also played an important role in inflorescence formation of chrysanthemum. These results showed that the CmTFL1c affects flowering time and regulates inflorescence architecture.

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Acknowledgements

We acknowledge Chunxin Lin professor from Zhejiang Agriculture and Forestry University for the Arabidopsis tfl1-13 mutant. We thank the China Ministry of Science and Technology for providing funding (Grant No. 2013AA102706).

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YG and YG conceived and designed research. YG and XB conducted experiments. MF and ZW analyzed data. YG and MF wrote the manuscript. QZ guided the writing of the manuscript. All authors read and approved the manuscript.

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Correspondence to Yike Gao.

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The authors declare that they have no conflict of interests.

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11103_2019_838_MOESM1_ESM.docx

Supplementary Figure 1 DNA sequence of CmTFL1c (Grey highlighting indicate exon sequences and yellow highlighting indicate shear sites) (DOCX 23 KB)

11103_2019_838_MOESM2_ESM.docx

Supplementary Figure 2 PCR identification of partially CmTFL1c transgenic Arabidopsis. (M.DL2000 Maker, 1. Positive control, 2.WT, 3.ddH2O, 6-13. The CmTFL1c transgenic plants). (DOCX 622 KB)

11103_2019_838_MOESM3_ESM.docx

Supplementary Figure 3 Generation of CmTFL1c transgenic chrysanthemum seedlings. Initial stage of resistance screening; b. Resistance callus; c. Resistant buds; d. Resistant plant (DOCX 243 KB)

11103_2019_838_MOESM4_ESM.docx

Supplementary Figure 4 Detection of CmTFL1c transgenic chrysanthemum. a. PCR detection, M was DL2000 Maker, 1 indicated positive control, 2 indicated WT, 3 indicated ddH2O, 4-13 indicated resistant plants; b. Chlorophenol red staining identification (DOCX 201 KB)

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Gao, Y., Gao, Y., Wu, Z. et al. Characterization of TEMINAL FLOWER1 homologs CmTFL1c gene from Chrysanthemum morifolium. Plant Mol Biol 99, 587–601 (2019). https://doi.org/10.1007/s11103-019-00838-6

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  • DOI: https://doi.org/10.1007/s11103-019-00838-6

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