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ClE2F1 Overexpression Enhances Plant Growth in Chrysanthemum lavandulifolium (Fisch. ex Trautv.) Makino

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Abstract

E2F transcription factors play key roles in plant growth and development by regulating the cell cycle. In this study, ClE2F1 was isolated from Chrysanthemum lavandulifolium. This gene encodes a 452-amino-acid protein that contains a DNA-binding domain, marked box domain, pRBR-binding domain, and a nuclear localization signal. Phylogenetic analysis showed that ClE2F1 was most closely related to Arabidopsis thaliana AtE2Fa, AtE2Fb, and AtE2Fc. Transient expression experiments demonstrated that the ClE2F1 protein localizes to the nucleus. Yeast one-hybrid assays showed that ClE2F1 promotes transcriptional activation. The morphology of the OX-6, OX-7, and OX-10 transgenic and wild-type lines was investigated. Plant height, leaf length and width, inflorescence diameter, disc flower diameter, the number of tabulate florets, and ligulate floret length were greater in the OX-6, OX-7, and OX-10 lines than in wild-type plants. Microscopic observation revealed a higher number of cells per unit area but a smaller cell area in the OX-6, OX-7, and OX-10 lines than those in wild-type plants. In addition, the relative expression of cell cycle-associated genes in the OX-6, OX-10, and wild-type plants revealed that the ClPCNA3, ClRNR, ClASF, ClDPa, ClCDKA, and ClCYCD transcript levels were up-regulated in the OX-6 and OX-10 plants, whereas ClRBR and ClKRP5 expression was down-regulated.

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References

  • Apostolova MD, Ivanova IA, Dagnino C, D’Souza SJ, Dagnino L (2002) Active nuclear import and export pathways regulate E2F-5 subcellular localization. J Biol Chem 277:34471–34479

    Article  PubMed  CAS  Google Scholar 

  • Attwooll C, Lazzerini Denchi E, Helin K (2004) The E2F family: specific functions and overlapping interests. EMBO J 23(24):4709–4716

  • Bracken AP, Ciro M, Cocito A, Helin K (2004) E2F target genes: unraveling the biology. Trends Biochem Sci 29:409–417

    Article  PubMed  CAS  Google Scholar 

  • Cam H, Dynlacht BD (2003) Emerging roles for E2F: beyond the G1/S transition and DNA replication. Cancer Cell 3:311–316

    Article  PubMed  CAS  Google Scholar 

  • Cheng Y, Cao L, Wang S, Li Y, Shi X, Liu H, Li L, Zhang Z, Fowke LC, Wang H, Zhou Y (2013) Down-regulation of multiple CDK inhibitor ICK/KRP genes up-regulates E2F pathway and increases cell proliferation, organ and seed sizes in Arabidopsis. Plant J 75:642–655

    Article  PubMed  CAS  Google Scholar 

  • Cockcroft CE, den Boer BG, Healy JM, Murray JA (2000) Cyclin D control of growth rate in plants. Nature 405:575–579

    Article  PubMed  CAS  Google Scholar 

  • Czerednik A, Busscher M, Angenent GC, de Maagd RA (2015) The cell size distribution of tomato fruit can be changed by overexpression of CDKA1. Plant Biotechnol J 13:259–268

    Article  PubMed  CAS  Google Scholar 

  • De Veylder L et al (2001) Functional analysis of cyclin-dependent kinase inhibitors of Arabidopsis. Plant Cell 13:1653–1668

    Article  PubMed  PubMed Central  Google Scholar 

  • De Veylder L et al (2002) Control of proliferation, endoreduplication and differentiation by the Arabidopsis E2Fa-DPa transcription factor. EMBO J 21:1360–1368

    Article  PubMed  PubMed Central  Google Scholar 

  • DeGregori J, Johnson DG (2006) Distinct and overlapping roles for E2F family members in transcription, proliferation and apoptosis. Curr Mol Med 6:739–748

    PubMed  CAS  Google Scholar 

  • Di Stefano L, Jensen MR, Helin K (2003) E2F7, a novel E2F featuring DP-independent repression of a subset of E2F-regulated genes. EMBO J 22(23):6289–6298

  • Dimova DK, Dyson NJ (2005) The E2F transcriptional network: old acquaintances with new faces. Oncogene 24:2810–2826

    Article  PubMed  CAS  Google Scholar 

  • Dyson N (1998) The regulation of E2F by pRB-family proteins. Genes Dev 12:2245–2262

    Article  PubMed  CAS  Google Scholar 

  • Egelkrout EM, Mariconti L, Settlage SB, Cella R, Robertson D, Hanley-Bowdoin L (2002) Two E2F elements regulate the proliferating cell nuclear antigen promoter differently during leaf development. Plant Cell 14:3225–3236

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Francis D (2011) A commentary on the G(2)/M transition of the plant cell cycle. Ann Bot 107:1065–1070

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Gao R, Wang H, Dong B, Yang X, Chen S, Jiang J, Zhang Z, Liu C, Zhao N, Chen F (2016) Morphological, genome and gene expression changes in newly induced autopolyploid Chrysanthemum lavandulifolium (Fisch. Ex Trautv.) Makino. Int J Mol Sci 17:1690–1702

    Article  PubMed Central  CAS  Google Scholar 

  • Humbert PO, Rogers C, Ganiatsas S, Landsberg RL, Trimarchi JM, Dandapani S, Brugnara C, Erdman S, Schrenzel M, Bronson RT, Lees JA (2000) E2F4 is essential for normal erythrocyte maturation and neonatal viability. Mol Cell 6:281–291

    Article  PubMed  CAS  Google Scholar 

  • Imajuku Y, Ohashi Y, Aoyama T, Goto K, Oka A (2001) An upstream region of the Arabidopsis thaliana CDKA;1 (CDC2aAt) gene directs transcription during trichome development. Plant Mol Biol 46:205–213

    Article  PubMed  CAS  Google Scholar 

  • Inze D (2005) Green light for the cell cycle. EMBO J 24:657–662

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • de Jager SM, Menges M, Bauer UM, Murray JAH (2001) Arabidopsis E2F1 binds a sequence present in the promoter of S-phase-regulated gene AtCDC6 and is a member of a multigene family with differential activities. Plant Mol Biol 47:555–568

    Article  PubMed  Google Scholar 

  • Judah D, Chang WY, Dagnino L (2010) EBP1 is a novel E2F target gene regulated by transforming growth factor-β. PLoS One 5:pone.0013941

    Article  CAS  Google Scholar 

  • Kong LJ, Orozco BM, Roe JL, Nagar S, Ou S, Feiler HS, Durfee T, Miller AB, Gruissem W, Robertson D, Hanley-Bowdoin L (2000) A geminivirus replication protein interacts with the retinoblastoma protein through a novel domain to determine symptoms and tissue specificity of infection in plants. EMBO J 19:3485–3495

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Kosugi S, Ohashi Y (2002a) E2Ls, E2F-like repressors of Arabidopsis that bind to E2F sites in a monomeric form. J Biol Chem 277:16553–16558

    Article  PubMed  CAS  Google Scholar 

  • Kosugi S, Ohashi Y (2002b) Interaction of the Arabidopsis E2F and DP proteins confers their concomitant nuclear translocation and transactivation. Plant Physiol 128:833–843

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Kosugi S, Ohashi Y (2003) Constitutive E2F expression in tobacco plants exhibits altered cell cycle control and morphological change in a cell type-specific manner. Plant Physiol 132:2012–2022

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • del Pozo JC, Diaz-Trivino S, Cisneros N, Gutierrez C (2006) The balance between cell division and endoreplication depends on E2FC-DPB, transcription factors regulated by the ubiquitin-SCFSKP2A pathway in Arabidopsis. Plant Cell 18:2224–2235

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Lario LD, Ramirez-Parra E, Gutierrez C, Spampinato CP, Casati P (2013) Anti-silencing function1 proteins are involved in ultraviolet-induced DNA damage repair and are cell cycle regulated by E2F transcription factors in Arabidopsis. Plant Physiol 162:1164–1177

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Li P, Song A, Gao C, Jiang J, Chen S, Fang W, Zhang F, Chen F (2015) The over-expression of a chrysanthemum WRKY transcription factor enhances aphid resistance. Plant Physiol Biochem 95:26–34

    Article  PubMed  CAS  Google Scholar 

  • Lindeman GJ, Aubatz S, Livingston D, Insberg D (1997) The subcellular localization of E2F-4 is cell-cycle dependent. Proc Natl Acad Sci 94:5095–5100

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Luisa M, Barbara P, Rita C (2002) The E2F family of transcription factors from Arabidopsis thaliana. Novel and conserved components of the retinoblastoma/E2F pathway in plants. J Biol Chem 277:9911–9919

    Article  CAS  Google Scholar 

  • Macaluso M, Cinti C, Russo G, Russo A, Giordano A (2003) pRb2/p130-E2F4/5-HDAC1-SUV39H1-p300 and pRb2/p130-E2F4/5-HDAC1-SUV39H1-DNMT1 multimolecular complexes mediate the transcription of estrogen receptor-alpha in breast cancer. Oncogene 22:3511–3517

    Article  PubMed  CAS  Google Scholar 

  • Magyar Z, de Veylder L, Atanassova A, Bakó L, Inzé D, Bögre L (2005) The role of the Arabidopsis E2FB transcription factor in regulating auxin-dependent cell division. Plant Cell 17:2527–2541

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Magyar Z, Horváth B, Khan S, Mohammed B, Henriques R, de Veylder L, Bakó L, Scheres B, Bögre L (2012) Arabidopsis E2FA stimulates proliferation and endocycle separately through RBR-bound and RBR-free complexes. EMBO J 31:1480–1493

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Mariconti L, Pellegrini B, Cantoni R, Stevens R, Bergounioux C, Cella R, Albani D (2002) The E2F family of transcription factors from Arabidopsis thaliana. Novel and conserved components of the retinoblastoma/E2F pathway in plants. J Biol Chem 277:9911–9919

    Article  PubMed  CAS  Google Scholar 

  • Perrot-Rechenmann C (2010) Cellular responses to auxin: division versus expansion. Cold Spring Harb Perspect Biol 2:a001446

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29(e45):45e–445e

  • Polyn S, Willems A, Veylder LD (2015) Cell cycle entry, maintenance, and exit during plant developmental cell. Curr Opin Plant Biol 23:1–7

    Article  PubMed  CAS  Google Scholar 

  • Ramirez-Parra E, Xie Q, Boniotti MB, Gutierrez C (1999) The cloning of plant E2F, a retinoblastoma-binding protein, reveals unique and conserved features with animal G(1)/S regulators. Nucleic Acids Res 27:3527–3533

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Ramirez-Parra E, Frundt C, Gutierrez C (2003) A genome-wide identification of E2F-regulated genes in Arabidopsis. Plant J 33:801–811

    Article  PubMed  CAS  Google Scholar 

  • Ramirez-Parra E, Lopez-Matas MA, Frundt C, Gutierrez C (2004) Role of an atypical E2F transcription factor in the control of Arabidopsis cell growth and differentiation. Plant Cell 16:2350–2236

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Raynaud C, Sozzani R, Glab N, Domenichini S, Perennes C, Cella R, Kondorosi E, Bergounioux C (2006) Two cell-cycle regulated SET-domain proteins interact with proliferating cell nuclear antigen (PCNA) in Arabidopsis. Plant J 47:395–407

    Article  PubMed  CAS  Google Scholar 

  • Rempel RE, Saenz-Robles MT, Storms R, Morham S, Ishida S, Engel A, Jakoi L, Melhem MF, Pipas JM, Smith C, Nevins JR (2000) Loss of E2F4 activity leads to abnormal development of multiple cellular lineages. Mol Cell 6:293–306

    Article  PubMed  CAS  Google Scholar 

  • Roa H, Lang J, Culligan KM, Keller M, Holec S, Cognat V, Montane MH, Houlne G, Chaboute ME (2009) Ribonucleotide reductase regulation in response to genotoxic stress in Arabidopsis. Plant Physiol 151:461–471

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Rossignol P, Stevens R, Perennes C, Jasinski S, Cella R, Tremousaygue D, Bergounioux C (2002) AtE2F-a and AtDP-a, members of the E2F family of transcription factors, induce Arabidopsis leaf cells to re-enter S-phase. Mol Gen Genomics 266:995–1003

    Article  CAS  Google Scholar 

  • Schnittger A, Schobinger U, Stierhof YD, Hulskamp M (2002) Ectopic B-type cyclin expression induces mitotic cycles in endoreduplicating Arabidopsis trichomes. Curr Biol 12:415–420

    Article  PubMed  CAS  Google Scholar 

  • Shen WH (2002) The plant E2F-Rb pathway and epigenetic control. Trends Plant Sci 7:505–511

    Article  PubMed  CAS  Google Scholar 

  • Sozzani R, Maggio C, Varotto S, Canova S, Bergounioux C, Albani D, Cella R (2006) Interplay between Arabidopsis activating factors E2Fb and E2Fa in cell cycle progression and development. Plant Physiol 140:1355–1366

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Storre J, Elsässer HP, Fuchs M, Ullmann D, Livingston DM, Gaubatz S (2002) Homeotic transformations of the axial skeleton that accompany a targeted deletion of E2f6. EMBO Rep 3:695–700

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Sung YH, Jin Y, Kang Y, Devkota S, Lee J, Roh JI, Lee HW (2013) Ei24, a novel E2F target gene, affects p53-independent cell death upon ultraviolet C irradiation. J Biol Chem 288:31261–31267

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Tracy K, Dibling BC, Spike BT, Knabb JR, Schumacker P, Macleod KF (2007) BNIP3 is an RB/E2F target gene required for hypoxia-induced autophagy. Mol Cell Biol 27:6229–6242

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Trimarchi JM, Lees JA (2002) Sibling rivalry in the E2F family. Nat Rev Mol Cell Biol 3:11–20

    Article  PubMed  CAS  Google Scholar 

  • Trimarchi JM, Fairchild B, Wen J, Lees JA (2001) The E2F6 transcription factor is a component of the mammalian bmi1-containing polycomb complex. Proc Natl Acad Sci 98:1519–1524

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Vandepoele K, Vlieghe K, Florquin K, Hennig L, Beemster GT, Gruissem W, van de Peer Y, Inzé D, de Veylder L (2005) Genome-wide identification of potential plant E2F target genes. Plant Physiol 139:316–328

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Vidal M, Braun P, Chen E, Boeke JD, Harlow E (1996) Genetic characterization of a mammalian protein-protein interaction domain by using a yeast reverse two-hybrid system. Proc Natl Acad Sci 93:10321–10326

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Wang H, Jiang J, Chen S, Qi X, Peng H, Li P, Song A, Guan Z, Fang W, Liao Y, Chen F (2013) Next-generation sequencing of the Chrysanthemum nankingense (Asteraceae) transcriptome permits large-scale unigene assembly and SSR marker discovery. PLoS One 8:e62293

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Weinmann AS, Bartley SM, Zhang T, Zhang MQ, Farnham PJ (2001) Use of chromatin immunoprecipitation to clone novel E2F target promoters. Mol Cell Biol 21:6820–6832

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Weinmann AS, Yan PS, Oberley MJ, Huang TH, Farnham PJ (2002) Isolating human transcription factor targets by coupling chromatin immunoprecipitation and CpG island microarray analysis. Genes Dev 16:235–244

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Wu LZ, Timmers C, Maiti B, Saavedra HI, Sang L, Chong GT, Nuckolls F, Giangrande P, Wright FA, Field SJ, Greenberg ME, Orkin S, Nevins JR, Robinson ML, Leone G (2001) The E2F1-3 transcription factors are essential for cellular proliferation. Nature 414:457–462

    Article  PubMed  CAS  Google Scholar 

  • Yang WW, Wang ZH, Zhu Y, Yang HT (2007) E2F6 negatively regulates ultraviolet-induced apoptosis via modulation of BRCA1. Cell Death Differ 14:807–817

    Article  PubMed  CAS  Google Scholar 

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Funding

This work was supported by the National Natural Science Foundation of China (31425022, 31372092), a Project of the “333 project” in Jiangsu Province (BRA2017382), the Natural Science Fund of Jiangsu Province (BK20150661, BK20151429), the Fundamental Research Funds for the Central Universities (KYZ201606, KJQN201652, KYZ201507), the Fund for Independent Innovation of Agricultural Sciences in Jiangsu Province [CX (16)1025], and the new project in Jiangsu Province (SXGC [2016] 318).

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  1. College of Horticulture, Key Laboratory of Landscaping, Ministry of Agriculture, Nanjing Agricultural University, Nanjing, 210095, China

    Ri Gao, Yajun Yan, Xiaodong Yang, Yinjie Wang, Weimin Fang, Sumei Chen, Jiafu Jiang, Haibin Wang & Fadi Chen

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  1. Ri Gao
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  2. Yajun Yan
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  3. Xiaodong Yang
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Correspondence to Fadi Chen.

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Fig. S1

Relative expression level of E2F (a) and ClE2F1 in the different organs of chrysanthemums (b) (JPG 81 kb)

Fig. S2

Relative expression levels of ClORC3, ClORC1, ClCDC6b, ClCDC4, and ClCDC45 in wild-type and transgenic plants. Letters indicate significant differences at p = 0.05 by Duncan’s test. (JPG 321 kb)

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Gao, R., Yan, Y., Yang, X. et al. ClE2F1 Overexpression Enhances Plant Growth in Chrysanthemum lavandulifolium (Fisch. ex Trautv.) Makino. Plant Mol Biol Rep 36, 341–349 (2018). https://doi.org/10.1007/s11105-018-1084-0

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