Skip to main content
Log in

Influences of factors affecting the induction of high protoplast yield and callus induction in two different carnations

  • Original Article
  • Published:
Plant Biotechnology Reports Aims and scope Submit manuscript

A Correction to this article was published on 17 May 2024

This article has been updated

Abstract

This study aims to establish an efficient protocol for protoplast isolation, cell division, and callus induction in two carnation cultivars, Chabaud and Giant Chabaud, by examining the influence of different types of plant tissue, enzyme concentrations, incubation times, cotyledon ages, and medium compositions. Our results indicate that protoplast yield varies significantly between different plant tissues, with true leaves offering the highest yield and viability, especially under a 0.1% driselase concentration and a 6-h incubation time. We observed that increasing the driselase concentration to 1.0% significantly reduced protoplast yields in all tissues tested. In addition, the age of the cotyledons notably affected protoplast yield, with younger cotyledons providing higher yields. The Murashige and Skoog medium supplemented with 1 mg/L zeatin and 1 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) proved to be the most effective for promoting cell division and colony formation from protoplasts derived from cotyledons and true leaves. The study also found that plant growth regulators (PGRs) significantly influence callus proliferation, with differences observed between protoplast sources.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Change history

References

  • Adedeji OS, Naing AH, Kim CK (2020) Protoplast isolation and shoot regeneration from protoplast-derived calli of Chrysanthemum cv. White ND. Plant Cell Tissue Organ Cult (PCTOC) 141:571–581

    Article  Google Scholar 

  • Arai M, Sugawara Y, Matsushima H, Takeuchi M (1989) Enhancement of colony formation of carnation (Dianthus caryophyllus) mesophyll protoplasts by abscisic acid pretreatment. Plant Tissue Culture Letters 6(2):80–84

    Article  CAS  Google Scholar 

  • Cui J, Mackenzie KK, Eeckhaut T, Müller R, Lütken H (2019) Protoplast isolation and culture from Kalanchoë species: optimization of plant growth regulator concentration for efficient callus production. Plant Cell Tissue Organ Cult (PCTOC) 138(2):287–297

    Article  CAS  Google Scholar 

  • Duquenne B, Eeckhaut T, Werbrouck S, Van Huylenbroeck J (2007) Effect of enzyme concentrations on protoplast isolation and protoplast culture of Spathiphyllum and Anthurium. Plant Cell Tissue Organ Cult 91(2):165–173

    Article  CAS  Google Scholar 

  • Grzebelus E, Szklarczyk M, Baranski R (2012) An improved protocol for plant regeneration from leaf-and hypocotyl-derived protoplasts of carrot. Plant Cell. Tissue Organ Cult (PCTOC) 109:101–109

    Article  Google Scholar 

  • Jawaharlal M, Ganga M, Padmadevi K, Jegadeeswari V, Karthikeyan S (2009) A technical guide on carnation. A technical guide on carnation. Tamil Nadu Agricultural University, Coimbatore, pp 1–56

    Google Scholar 

  • Jomori H, Takahata Y, Kaizuma N (1994) Plant regeneration from leaf-derived calli of gentians and their protoplast culture. Genetic Improv Hortic Crops Biotechnol 392:81–86

    Google Scholar 

  • Kang HH, Naing AH, Kim CK (2020) Protoplast isolation and shoot regeneration from protoplast-derived callus of Petunia hybrida Cv. Mirage Rose. Biology 9(8):228

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kiełkowska A, Adamus A (2012) An alginate-layer technique for culture of Brassica oleracea L. protoplasts. In Vitro Cell Dev-Pl 48:265–273

    Article  Google Scholar 

  • Kim JC, Lee EA (1996) Plant regeneration from mesophyll protoplasts of Dianthus superbus. Plant Cell Rep 16(1):18–21

    Article  CAS  PubMed  Google Scholar 

  • Meyer L, Serek M, Winkelmann T (2009) Protoplast isolation and plant regeneration of different genotypes of Petunia and Calibrachoa. Plant Cell, Tissue and Organ Cult (PCTOC) 99(1):27–34

    Article  Google Scholar 

  • Naing AH, Adedeji OS, Kim CK (2021) Protoplast technology in ornamental plants: current progress and potential applications on genetic improvement. Sci Hortic-Amst 283:110043

    Article  CAS  Google Scholar 

  • Nakano M, Mii M (1992) Protoplast culture and plant regeneration of several species in the genus Dianthus. Plant Cell Rep 11(5–6):225–228

    CAS  PubMed  Google Scholar 

  • Nakano M, Mii M (1995) Plant regeneration from protoplasts in Dianthus: comparison of cultural behavior of different donor tissues. Plant Tissue Culture Lett 12(1):62–67

    Article  Google Scholar 

  • Nakano M, Mii M, Genetics A (1993) Somatic hybridization between Dianthus chinensis and D. barbatus through protoplast fusion. Theoret Appl Genet 86(1):1–5

    Article  CAS  Google Scholar 

  • Nassour M, Chasseriaux G, Dorion N (2003) Optimization of protoplast-to-plant system for Pelargonium× hortorum ‘Alain’ and genetic stability of the regenerated plants. Plant Sci 165(1):121–128

    Article  CAS  Google Scholar 

  • Nhut DT, Teixeira Da Silva JA, Aswath CR (2003) The importance of the explant on regeneration in thin cell layer technology. In Vitro Cell Dev Biol Plant 39(3):266–276. https://doi.org/10.1079/IVP2002408

    Article  Google Scholar 

  • Nirmal Babu K, Samsudeen K, Divakaran M, Pillai GS, Sumathi V, Praveen K, Ravindran P, Peter K (2016) Protocols for in vitro propagation, conservation, synthetic seed production, embryo rescue, microrhizome production, molecular profiling, and genetic transformation in ginger (Zingiber officinale Roscoe.). In: Protocols for in vitro cultures and secondary metabolite analysis of aromatic and medicinal plants, Second Edition, pp 403–426

  • Nishihara M, Higuchi A, Watanabe A, Tasaki K (2018) Application of the CRISPR/Cas9 system for modification of flower color in Torenia fournieri. BMC Plant Biol 18(1):1–9

    Article  Google Scholar 

  • Pongchawee K, Na-Nakorn U, Lamseejan S, Poompuang S, Phansiri S (2006) Factors affecting the protoplast isolation and culture of Anubias nana Engler. To Bot 2:193–200

    Google Scholar 

  • Rahmani M-S, Pijut PM, Shabanian N (2016) Protoplast isolation and genetically true-to-type plant regeneration from leaf-and callus-derived protoplasts of Albizia julibrissin. Plant Cell Tissue Organ Culture (PCTOC) 127(2):475–488

    Article  CAS  Google Scholar 

  • Reed KM, Bargmann BOR (2021) Protoplast regeneration and its use in new plant breeding technologies. Front Genome Edit 3:734951

    Article  Google Scholar 

  • Shiba T, Mii M (2005) Plant regeneration from mesophyll-and cell suspension-derived protoplasts of Dianthus acicularis and characterization of regenerated plants. In Vitro Cell Dev Biol-Plant 41:794–800

    Article  CAS  Google Scholar 

  • Xu J, Kang BC, Naing AH, Bae SJ, Kim JS, Kim H, Kim CK (2020) CRISPR/Cas9-mediated editing of 1-aminocyclopropane-1-carboxylate oxidase1 enhances Petunia flower longevity. Plant Biotechnol J 18(1):287–297

    Article  CAS  PubMed  Google Scholar 

  • Xu J, Naing AH, Bunch H, Jeong J, Kim H, Kim CK (2021) Enhancement of the flower longevity of petunia by CRISPR/Cas9-mediated targeted editing of ethylene biosynthesis genes. Postharvest Biol Technol 174:111460

    Article  CAS  Google Scholar 

  • Xu J, Naing AH, Kang H, Lee SY, Li W, Chung MY, Kim CK (2023) CRISPR/Cas9-mediated editing of PhMLO1 confers powdery mildew resistance in petunia. Plant Biotechnol Rep 17(5):767–775

    Article  CAS  Google Scholar 

Download references

Funding

This work was supported by the National Research Foundation (NRF) grant funded by the Korean government (MSIT) (No. 2021R1A2C2008951).

Author information

Authors and Affiliations

Authors

Contributions

AHN designed the study. AOS conducted the experiments. JRC and KIP assisted with the experiments. AHN and AOS wrote the manuscript. CKK supervised the project. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Aung Htay Naing or Chang Kil Kim.

Ethics declarations

Conflict of interest

The authors report no conflicts of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

The original online version of this article was revised due to co-corresponding author information was missing and included in this version.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Adedeji, O.S., Naing, A.H., Campol, J.R. et al. Influences of factors affecting the induction of high protoplast yield and callus induction in two different carnations. Plant Biotechnol Rep 18, 375–383 (2024). https://doi.org/10.1007/s11816-024-00903-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11816-024-00903-7

Keywords

Navigation