RcSPL1–RcTAF15b regulates the flowering time of rose (Rosa chinensis)

Abstract

Rose ( Rosa chinensis), which is an economically valuable floral species worldwide, has three types, namely once-flowering (OF), occasional or re-blooming (OR), and recurrent or continuous flowering (CF). However, the mechanism underlying the effect of the age pathway on the duration of the CF or OF juvenile phase is largely unknown. In this study, we observed that the RcSPL1 transcript levels were substantially upregulated during the floral development period in CF and OF plants. Additionally, accumulation of RcSPL1 protein was controlled by rch-miR156. The ectopic expression of RcSPL1 in Arabidopsis thaliana accelerated the vegetative phase transition and flowering. Furthermore, the transient overexpression of RcSPL1 in rose plants accelerated flowering, whereas silencing of RcSPL1 had the opposite phenotype. Accordingly, the transcription levels of floral meristem identity genes ( APETALA1, FRUITFULL, and LEAFY) were significantly affected by the changes in RcSPL1 expression. RcTAF15b protein, which is an autonomous pathway protein, was revealed to interact with RcSPL1. The silencing and overexpression of RcTAF15b in rose plants led to delayed and accelerated flowering, respectively. Collectively, the study findings imply that RcSPL1–RcTAF15b modulates the flowering time of rose plants.

Acknowledgements

This work was funded by Guest Investigator Grant of the State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Science (SKLPG2016A-29). We thank Professor Nan Ma (China Agricultural University) for providing the TRV1, TRV2, pSuper1300-GFP, and pSuper: NF-YA4-mCherry plasmids, as well as cDNA library of Y2H. We also thank Professor Jian Ye (Institute of Microbiology, Chinese Academy of Sciences) for kindly providing the psTRV1 and psTRV2 vectors. We thank Liwen Bianji (Edanz) (www.liwenbianji.cn) for editing the English text of a draft of this manuscript.

Author contributions

R.Y. designed and performed the experiments, analyzed the data, and wrote the manuscript. Z.Y.X. performed gene silencing and protein interaction assays; and X.H.Z. performed the overexpression assays; P.P.F. performed the RT–qPCR analyses and morphology observations; Z.Y.H. revised the manuscript; R.X.F. and Y.M.Z. guided the research and revised the manuscript; and Q.L.L. designed the experiments, conceived the project, and revised the manuscript. All authors read and approved the manuscript.

Data availability

The sequencing data that support the findings of this study are available in the genome database of ‘Old Blush’ (https://lipm-browsers.toulouse.inra.fr/pub/RchiOBHm-V2/) and genome sequence archive (https://ngdc.cncb.ac.cn/search/?dbId=gwh&q=GWHAAAF00000000&page=1).

Conflict of interest

The authors declare no competing interest.

References

[1] Hibrand Saint-Oyant L, Ruttink T, Hamama L et al. A high-quality genome sequence of Rosa chinensis to elucidate ornamental traits . Nat Plants. 2018; 4: 473-84.

[2] Iwata H, Gaston A, Remay A et al. The TFL1 homologue KSN is a regulator of continuous flowering in rose and strawberry . Plant J. 2012; 69: 116-25.

[3] Wang L, Liu Y, Zhang Y et al. The expression level of Rosa Terminal flower 1 (RTFL1) is related with recurrent flowering in roses . Mol Biol Rep. 2012; 39: 3737-46.

[4] Randoux M, Davière JM, Jeauffre J et al. RoKSN, a floral repressor, forms protein complexes with RoFD and RoFT to regulate vegetative and reproductive development in rose. New Phytol. 2014; 202: 161-73.

[5] Bai M, Liu J, Fan C et al. KSN heterozygosity is associated with continuous flowering of Rosa rugosa Purple branch . Hortic Res. 2021; 8: 26.

[6] Hamama L, Naouar A, Gala R et al. Overexpression of RoDELLA impacts the height, branching, and flowering behaviour of Pelargonium × domesticum transgenic plants . Plant Cell Rep. 2012; 31: 2015-29.

[7] Randoux M, Jeauffre J, Thouroude T et al. Gibberellins regulate the transcription of the continuous flowering regulator, RoKSN, a rose TFL1 homologue . J Exp Bot. 2012; 63: 6543-54.

[8] Kawamura K, Ueda Y, Matsumoto S et al. The identification of the Rosa S-locus provides new insights into the breeding and wild origins of continuous-flowering roses . Hortic Res. 2022; 9: uhac155.

[9] Foucher F, Chevalier M, Corre C et al. New resources for studying the rose flowering process. Genome. 2008; 51: 827-37.

[10] Zlesak DC . Rosa. Rosa × Hybrida . In: Anderson NO, ed. Flower Breeding and Genetics: Issues, Challenges and Opportunities for the 21st Century. Springer: Dordrecht, 2006, 695-738.

[11] Putterill J, Laurie R, Macknight R . It’s time to flower: the genetic control of flowering time. BioEssays. 2004; 26: 363-73.

[12] Amasino RM, Michaels SD . The timing of flowering. Plant Physiol. 2010; 154: 516-20.

[13] Srikanth A, Schmid M . Regulation of flowering time: all roads lead to Rome. Cell Mol Life Sci. 2011; 68: 2013-37.

[14] Lee J, Lee I . Regulation and function of SOC1, a flowering pathway integrator. J Exp Bot. 2010; 61: 2247-54.

[15] Wang JW . Regulation of flowering time by the miR156-mediated age pathway. J Exp Bot. 2014; 65: 4723-30.

[16] Teotia S, Tang G . To bloom or not to bloom: role of microRNAs in plant flowering. Mol Plant. 2015; 8: 359-77.

[17] Bartel DP . MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 2004; 116: 281-97.

[18] Zhang B, Pan X, Cobb GP et al. Plant microRNA: a small regulatory molecule with big impact. Dev Biol. 2006; 289: 3-16.

[19] Rhoades MW, Reinhart BJ, Lim LP et al. Prediction of plant microRNA targets. Cell. 2002; 110: 513-20.

[20] Klein J, Saedler H, Huijser P . A new family of DNA binding proteins includes putative transcriptional regulators of the Antirrhinum majus floral meristem identity gene SQUAMOSA . Mol Gen Genet. 1996; 250: 7-16.

[21] Yamasaki K, Kigawa T, Inoue M et al. A novel zinc-binding motif revealed by solution structures of DNA-binding domains of Arabidopsis SBP-family transcription factors . J Mol Biol. 2004; 337: 49-63.

[22] Hultquist JF, Dorweiler JE . Feminized tassels of maize mop1 and ts1 mutants exhibit altered levels of miR156 and specific SBP-box genes . Planta. 2008; 229: 99-113.

[23] Yang Z, Wang X, Gu S et al. Comparative study of SBP-box gene family in Arabidopsis and rice . Gene. 2008; 407: 1-11.

[24] Salinas M, Xing S, Höhmann S et al. Genomic organization, phylogenetic comparison and differential expression of the SBP-box family of transcription factors in tomato. Planta. 2012; 235: 1171-84.

[25] Hou H, Li J, Gao M et al. Genomic organization, phylogenetic comparison and differential expression of the SBP-box family genes in grape. PLoS One. 2013; 8: e59358.

[26] Xu M, Hu T, Zhao J et al. Developmental functions of miR156-regulated SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) genes in Arabidopsis thaliana . PLoS Genet. 2016; 12: e1006263.

[27] Wang JW, Czech B, Weigel D . miR156-regulated SPL transcription factors define an endogenous flowering pathway in Arabidopsis thaliana . Cell. 2009; 138: 738-49.

[28] Zhang H, Zhang L, Han J et al. The nuclear localization signal is required for the function of squamosa promoter binding protein-like gene 9 to promote vegetative phase change in Arabidopsis . Plant Mol Biol. 2019; 100: 571-8.

[29] Zhang Y, Schwarz S, Saedler H et al. SPL8, a local regulator in a subset of gibberellin-mediated developmental processes in Arabidopsis . Plant Mol Biol. 2007; 63: 429-39.

[30] Yu S, Galvão VC, Zhang YC et al. Gibberellin regulates the Arabidopsis floral transition through miR156-targeted SQUAMOSA PROMOTER BINDING-LIKE transcription factors . Plant Cell. 2012; 24: 3320-32.

[31] Gou J, Fu C, Liu S et al. The miR156- SPL4 module predominantly regulates aerial axillary bud formation and controls shoot architecture . New Phytol. 2017; 216: 829-40.

[32] Long J, Liu CY, Feng MQ et al. miR156- SPL modules regulate induction of somatic embryogenesis in citrus callus . J Exp Bot. 2018; 69: 2979-93.

[33] Eom H, Park SJ, Kim MK et al. TAF15b, involved in the autonomous pathway for flowering, represses transcription of FLOWERING LOCUS C . Plant J. 2018; 93: 79-91.

[34] Choi K, Kim J, Hwang HJ et al. The FRIGIDA complex activates transcription of FLC, a strong flowering repressor in Arabidopsis, by recruiting chromatin modification factors . Plant Cell. 2011; 23: 289-303.

[35] Lago C, Clerici E, Mizzi L et al. TBP-associated factors in Arabidopsis . Gene. 2004; 342: 231-41.

[36] Lago C, Clerici E, Dreni L et al. The Arabidopsis TFIID factor AtTAF6 controls pollen tube growth . Dev Biol. 2005; 285: 91-100.

[37] Waterworth WM, Drury GE, Blundell-Hunter G et al. Arabidopsis TAF1 is an MRE11-interacting protein required for resistance to genotoxic stress and viability of the male gametophyte . Plant J. 2015; 84: 545-57.

[38] Chimonidou D. Flower development and abscission zone. In: Roberts AV, ed. Encyclopedia of Rose Science, 1st edn, Vol. 2. Morphology and Anatomy. Elsevier Academic: Amesterdam, 2003, 504-12.

[39] Zhang S, Feng M, Chen W et al. In rose, transcription factor PTM balances growth and drought survival via PIP2;1 aquaporin. Nat Plants. 2019; 5: 290-9.

[40] Siddika T, Heinemann IU . Bringing microRNAs to light: methods for microRNA quantification and visualization in live cells. Front Bioeng Biotechnol. 2021; 8: 619583.

[41] Wu G, Park MY, Conway SR et al. The sequential action of miR156 and miR172 regulates developmental timing in Arabidopsis . Cell. 2009; 138: 750-9.

[42] Tang HB, Wang J, Wang L et al. Anisotropic cell growth at the leaf base promotes age-related changes in leaf shape in Arabidopsis thaliana . Plant Cell. 2023; 35: 1386-407.

[43] Qu J, Ye J, Geng YF et al. Dissecting functions of KATANIN and WRINKLED1 in cotton fiber development by virus-induced gene silencing . Plant Physiol. 2012; 160: 738-48.

[44] Wu G, Poethig RS . Temporal regulation of shoot development in Arabidopsis thaliana by miR156 and its target SPL3 . Development. 2006; 133: 3539-47.

[45] Wang JW, Park MY, Wang LJ et al. miRNA control of vegetative phase change in trees. PLoS Genet. 2011; 7: e1002012.

[46] Zheng C, Ye M, Sang M et al. A regulatory network for miR156-SPL module in Arabidopsis thaliana . Int J Mol Sci. 2019; 20: 6166.

[47] Blázquez M, Ferrándiz C, Madueo F et al. How floral meristems are built. Plant Mol Biol. 2006; 60: 855-70.

[48] Moyroud E, Kusters E, Monniaux M et al. LEAFY blossoms. Trends Plant Sci. 2010; 15: 346-52.

[49] Birkenbihl R, Jach G, Saedler H et al. Functional dissection of the plant-specific SBP-domain: overlap of the DNA-binding and nuclear localization domains. J Mol Biol. 2005; 352: 585-96.

[50] Jung JH, Ju Y, Seo PJ et al. The SOC1-SPL module integrates photoperiod and gibberellic acid signals to control flowering time in Arabidopsis . Plant J. 2012; 69: 577-88.

[51] Yamaguchi N, Winter CM, Wu MF et al. Gibberellin acts positively then negatively to control onset of flower formation in Arabidopsis . Science. 2014; 344: 638-41.

[52] Wang R, Albani MC, Vincent C et al. Aa TFL1 confers an age-dependent response to vernalization in perennial Arabis alpina . Plant Cell. 2011; 23: 1307-21.

[53] Zhou CM, Zhang TQ, Wang X et al. Molecular basis of age-dependent vernalization in Cardamine flexuosa . Science. 2013; 340: 1097-100.

[54] Liu J, Fu X, Dong Y et al. MIKC ^c-type MADS-box genes in Rosa chinensis: the remarkable expansion of ABCDE model genes and their roles in floral organogenesis . Hortic Res. 2018; 5: 25.

[55] Kumar S, Stecher G, Li M et al. MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol. 2018; 35: 1547-9.

[56] Sui M, Hui-Jun Y, Zhen-Zhen W et al. Identification of microRNA associated with flower organ development in Rosa chinensis ’Viridiflora’ . Plant Sci J. 2019; 37: 37-46.

[57] Clough SJ, Bent AF . Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana . Plant J. 1998; 16: 735-43.

[58] Chen J, Li Y, Li Y et al. AUXIN RESPONSE FACTOR 18-HISTONE DEACETYLASE 6 module regulates floral organ identity in rose (Rosa hybrida) . Plant Physiol. 2021; 186: 1074-87.