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Cytokinin oxidase/dehydrogenase inhibitors stimulate 2iP to induce direct somatic embryogenesis in Coffea arabica

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Abstract

Somatic embryogenesis in Coffea spp. is important for mass production and genetic engineering. Coffee is an exceptional case, as somatic embryogenesis can be induced by applying a cytokinin such as 2iP as the only plant growth regulator. 1-(2-(2-Hydroxyethyl)phenyl)-3-(3-(trifluoromethoxy)phenyl)urea (3TFM-2HE) and 1-(3-bromo-5-(trifluoromethoxy)phenyl)-3-(2-(2-hydroxyethyl)phenyl)urea (3TFM,5Br-2HE) are two newly designed cytokinin oxidase/dehydrogenase inhibitors (CKX) from the diphenylurea group. We used a Coffea arabica leaf disks bioassay to demonstrate the indirect somatic embryo induction potential of these compounds. The leaf disks were incubated on modified Murashige and Skoog (MS) semi-solid medium in which 3TFM-2HE or 3TFM,5Br-2HE were combined with N6-(2-isopentenyl)adenine (2iP). Although these compounds do not possess intrinsic cytokinin activity, they enhanced the activity of 2iP, resulting in direct somatic embryogenesis after seven weeks. The best results were obtained with 1 µM 3TFM-2HE and 5 or 10 µM 2iP. Maturation of somatic embryos into fully developed plants took place on medium supplemented with 0.5 µM kinetin and the somatic embryos developed true leaves and a root system.

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All data is available on request to corresponding author.

References

  • Almeida JAS, Silvarolla MB (2009) Induction of somatic embryos of Coffea arabica genotypes by 6-benzyladenine. Int J Plant Dev Biol 3(1):5–9

    Google Scholar 

  • Campos NA, Panis B, Carpentier SC (2017) Somatic embryogenesis in coffee: the evolution of biotechnology and the integration of omics technologies offer great opportunities. Front Plant Sci 8:1460

    Article  PubMed  PubMed Central  Google Scholar 

  • Chatfield JM, Armstrong DJ (1986) Regulation of cytokinin oxidase activity in callus tissues of Phaseolus vulgaris L. cv Great Northern. Plant Physiol 80(2):493–499

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chone RMS, Rocha DI, Monte-Bello CC, Pinheiro HP, Dornelas MC, Haddad CRB, Almeida JAS (2018) Brassinosteroid increases the cytokinin efficiency to induce direct somatic embryogenesis in leaf explants of Coffea arabica L. (Rubiaceae). Plant Cell Tissue Organ Cult (PCTOC) 135(1):63–71

    Article  CAS  Google Scholar 

  • Etienne H (2005) Somatic embryogenesis protocol: coffee (Coffea arabica L. and C. canephora P.). In: Jain SM, Gupta PK (eds) Protocol for somatic embryogenesis in woody plants. Forestry sciences, vol 77. Springer, Dordrecht

    Google Scholar 

  • Etienne H, Bertrand B (2003) Somaclonal variation in Coffea arabica: effects of genotype and embryogenic cell suspension age on frequency and phenotype of variants. Tree Physiol 23(6):419–426

    Article  CAS  PubMed  Google Scholar 

  • Gatica-Arias AM, Arrieta G, Espinoza AM (2008) Direct somatic embryogenesis in Coffea arabica L. cvs. Caturra and Catuaí: effect of tricontanol, light condition, and medium consistency. Agron Costarric. 32(1):139–147

    Google Scholar 

  • Giridhar P, Indu EP, Ravishankar GA, Chandrasekar A (2004) Influence of triacontanol on somatic embryogenesis in Coffea arabica L. and Coffea canephora P. ex Fr. In Vitro Cell Dev Biol Plant 40(2):200–203

    Article  Google Scholar 

  • Hare PD, Van Staden J (1994) Inhibitory effect of thidiazuron on the activity of cytokinin oxidase isolated from soybean callus. Plant Cell Physiol 35(8):1121–1125

    Article  CAS  Google Scholar 

  • Hatanaka T, Arakawa O, Yasuda T, Uchida N, Yamaguchi T (1991) Effect of plant growth regulators on somatic embryogenesis in leaf cultures of Coffea canephora. Plant Cell Rep 10(4):179–182

    Article  CAS  PubMed  Google Scholar 

  • Huetteman CA, Preece JE (1993) Thidiazuron: a potent cytokinin for woody plant tissue culture. Plant Cell, Tissue Organ Cult 33(2):105–119

    Article  CAS  Google Scholar 

  • Kahia J, Kirika M, Lubabali H, Mantell S (2016) High-frequency direct somatic embryogenesis and plantlet regeneration from leaves derived from in vitro-germinated seedlings of a Coffea arabica hybrid cultivar. HortScience 51(9):1148–1152

    Article  CAS  Google Scholar 

  • Kopečný D, Briozzo P, Popelková H et al (2010) Phenyl- and benzylurea cytokinins as competitive inhibitors of cytokinin oxidase/dehydrogenase: a structural study. Biochimie 92:1052–1062

    Article  PubMed  Google Scholar 

  • Leroux O, Van Der Kinderen G, Viane RLL (2007) A sandwich-embedding method for oriented sectioning. J Microsc 227(1):79–82

    Article  CAS  PubMed  Google Scholar 

  • Los Santos-Briones D, Hernández-Sotomayor SM (2006) Coffee biotechnology. Braz J Plant Physiol 18(1):217–227

    Article  Google Scholar 

  • Menéndez-Yuffá A, Barry-Etienne D, Bertrand B, Georget F, Etienne H (2010) A comparative analysis of the development and quality of nursery plants derived from somatic embryogenesis and from seedlings for large-scale propagation of coffee (Coffea arabica L.). Plant Cell. Tissue Organ Cult (PCTOC) 102(3):297–307

    Article  Google Scholar 

  • Muniswamy B, Sreenath HL, Samuel SD, Jayarama J (2015) In vitro multiplication of Coffea arabica F1 hybrid (S. 2800) and its performance in the field. Short Sci Rep J Plant Crops 43:225–230

    Google Scholar 

  • Nisler J, Kopečný D, Končitíková R, Zatloukal M, Bazgier V, Berka K, Spíchal L (2016) Novel thidiazuron-derived inhibitors of cytokinin oxidase/dehydrogenase. Plant Mol Biol 92(1–2):235–248

    Article  CAS  PubMed  Google Scholar 

  • Nisler J, Kopečný D, Pěkná Z, Končitíková R, Koprna R, Murvanidze N, Werbrouck SPO, Havlíček L, De Diego N, Kopečná M, Wimmer Z, Briozzo P, Moréra S, Zalabák D, Spíchal L, Strnad M (2020) Diphenylurea-derived cytokinin oxidase/dehydrogenase inhibitors for biotechnology and agriculture. J Exp Bot. https://doi.org/10.1093/jxb/eraa437

    Article  Google Scholar 

  • Schindelin J, Arganda-Carreras I, Frise E (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9(7):676–682

    Article  CAS  Google Scholar 

  • Schmülling T, Werner T, Riefler M, Krupková E, y Manns IB, (2003) Structure and function of cytokinin oxidase/dehydrogenase genes of maize, rice, Arabidopsis and other species. J Plant Res 116(3):241–252

    Article  PubMed  Google Scholar 

  • Söndahl MR, Sharp WR (1977) High frequency induction of somatic embryos in cultured leaf expiants of Coffea arabica L. Z Pflanzenphysiol 81(5):395–408

    Article  Google Scholar 

  • Staritsky G (1970) Embryoid formation in callus tissues of coffee. Acta Bot Neerland 19(4):509–514

    Article  Google Scholar 

  • Yasuda T, Fujii Y, Yamaguchi T (1985) Embryogenic callus induction from Coffea arabica leaf explants by benzyladenine. Plant Cell Physiol 26(3):595–597

    Article  CAS  Google Scholar 

  • Zatloukal M, Gemrotová M, Doležal K, Havlíček L, Spíchal L, Strnad M (2008) Novel potent inhibitors of A. thaliana cytokinin oxidase/dehydrogenase. Bioorg Med Chem 16(20):9268–9275

    Article  CAS  PubMed  Google Scholar 

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Acknowledgement

The authors acknowledge Dr. Ir. Stephen Andersen for English proofreading.

Author information

Authors and Affiliations

  1. Department of Plants and Crops, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium

    Nino Murvanidze & Stefaan P. O. Werbrouck

  2. Laboratory of Growth Regulators, Institute of Experimental Botany of the Czech Academy of Sciences &, Palacký University, Olomouc, Czech Republic

    Jaroslav Nisler

  3. Department of Biology, Faculty of Sciences, Ghent University, Ghent, Belgium

    Olivier Leroux

Authors
  1. Nino Murvanidze
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  2. Jaroslav Nisler
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  3. Olivier Leroux
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  4. Stefaan P. O. Werbrouck
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Contributions

NM performed the experiments, SW and NM designed the experiment and wrote the manuscript, JN synthesized the cytokinin oxidase/dehydrogenase inhibitors, OL advised for histology.

Corresponding author

Correspondence to Stefaan P. O. Werbrouck.

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Conflict of interest

JN has filed a Czech patent application (PV 2020-144), which claims the protection of CKX inhibitors described in this paper and their use as plant growth regulators.

Additional information

Communicated by J. Celenza.

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Murvanidze, N., Nisler, J., Leroux, O. et al. Cytokinin oxidase/dehydrogenase inhibitors stimulate 2iP to induce direct somatic embryogenesis in Coffea arabica. Plant Growth Regul 94, 195–200 (2021). https://doi.org/10.1007/s10725-021-00708-6

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  • DOI: https://doi.org/10.1007/s10725-021-00708-6

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