Protoplast Fusion as a Breeding Tool for Berries: A Review of Prospects and Current Limitations
DOI:
https://doi.org/10.65888/icraft.3.1.14Keywords:
Plant biotechnology, Protoplast fusion, Somatic hybridization, Breeding, Berry cropsAbstract
Berries of all different species, such as (Vitis, Fragaria, Vaccinium), are known for their abundance and diversity of health benefits and economic importance. However, their traditional breeding and production are hindered by long generation cycles and limited gene pools, which slow and limit the growth rate of new varieties that have been developed. The fusion technique allows the transfer and enhancement of important plant traits, including increased disease resistance, environmental stress tolerance, and improved fruit characteristics. Although research on berry crops is still in its infancy, proven research on grapes, strawberries, and others indicates the usefulness of this technique for enhancing genetic diversity. Nevertheless, major challenges continue, particularly in protoplast differentiation, protoplast fusion, regeneration efficiency, and selection of stable mixtures. The integration of emerging tools such as genome editing, and analysis is expected to increase the effectiveness of protoplast fusion. In short, this technology will achieve significant advances in berry genetics, poised to become a cornerstone of both basic research and applied breeding programs.
References
1. Compounds, B. (2015). Antioxidant Activity in Different Types of Berries/S. Skrovankova, D. Sumczynski,J. Mlcek, T. Jurikova, J. Sochor. International Journal of Molecular Sciences, 16(10), 24673-24706.
2. Basu, A., Rhone, M., & Lyons, T. J. (2010). Berries: emerging impact on cardiovascular health. Nutrition reviews, 68(3), 168-177.
3. Miller, M. G., & Shukitt-Hale, B. (2012). Berry fruit enhances beneficial signaling in the brain. Journal of agricultural and food chemistry, 60(23), 5709-5715.
4. Montes Ninaquispe, J. C., Arbulú Ballesteros, M. A., Cruz Salinas, L. E., García Juárez, H. D., Farfán Chilicaus, G. C., Martel Acosta, R., ... & Coronel Estela, C. V. (2024). A strategy for the sustainability of Peru’s blueberry exports: diversification and competitiveness. Sustainability, 16(15), 6606.
5. Singh, R. P., Singh, P. K., Gupta, R., & Singh, R. L. (2018). Biotechnological tools to enhance sustainable production. In Biotechnology for sustainable agriculture (pp. 19-66). Woodhead Publishing.
6. Akhtar, S., Rao, E., Uike, A., & Saatu, M. (2023). Plant breeding strategies: traditional and modern approaches. Genetic revolution in agriculture: unleashing the power of plant genetics Elite Publishing House, New Delhi.
7. Ranaware, A. S., Kunchge, N. S., Lele, S. S., & Ochatt, S. J. (2023). Protoplast technology and somatic hybridisation in the family Apiaceae. Plants, 12(5), 1060.
8. Soriano, L., Omar, A. A., & Martinelli, A. P. (2022). Citrus protoplast isolation and plant regeneration through somatic embryogenesis. In Somatic Embryogenesis: Methods and Protocols (pp. 111-126). New York, NY: Springer US.
9. Pasternak, T., Lystvan, K., Betekhtin, A., & Hasterok, R. (2020). From single cell to plants: mesophyll protoplasts as a versatile system for investigating plant cell reprogramming. International Journal of Molecular Sciences, 21(12), 4195.
10. Zong, Y., Wang, Y., Li, C., Zhang, R., Chen, K., Ran, Y., ... & Gao, C. (2017). Precise base editing in rice, wheat and maize with a Cas9-cytidine deaminase fusion. Nature biotechnology, 35(5), 438-440.
11. Zhao, X., Song, H., Liu, J., Feng, K., Wu, Q., Arif, T., ... & Zhang, L. (2025). Efficient Protoplast Isolation and PEG-mediated Transformation protocols for blueberry Vaccinium corymbosum. Scientia Horticulturae, 340, 113916.
12. Yang, W., Ren, J., Liu, W., Liu, D., Xie, K., Zhang, F., ... & Wu, X. (2023). An efficient transient gene expression system for protein subcellular localization assay and genome editing in citrus protoplasts. Horticultural Plant Journal, 9(3), 425-436.
13. Najafi, S., Bertini, E., D’Incà, E., Fasoli, M., & Zenoni, S. (2023). DNA-free genome editing in grapevine using CRISPR/Cas9 ribonucleoprotein complexes followed by protoplast regeneration. Horticulture Research, 10(1), uhac240.
14. Tian, S., Jiang, L., Gao, Q., Zhang, J., Zong, M., Zhang, H., ... & Xu, Y. (2017). Efficient CRISPR/Cas9-based gene knockout in watermelon. Plant Cell Reports, 36(3), 399-406.
15. Naing, A. H., Adedeji, O. S., & Kim, C. K. (2021). Protoplast technology in ornamental plants: current progress and potential applications on genetic improvement. Scientia horticulturae, 283, 110043.
16. Kafkas1a, N. E., Ates, S., Nasır, N., & Mezzetti, B. Protoplast Technology and Somatic Hybridization in
Strawberry.
17. Mezzetti, B., Giampieri, F., Zhang, Y. T., & Zhong, C. F. (2018). Status of strawberry breeding programs and cultivation systems in Europe and the rest of the world. Journal of Berry Research, 8(3), 205-221.
18. Vondracek, K., Altpeter, F., Liu, T., & Lee, S. (2024). Advances in genomics and genome editing for improving strawberry (Fragaria× ananassa). Frontiers in Genetics, 15, 1382445.
19. Migicovsky, Z., & Myles, S. (2017). Exploiting wild relatives for genomics-assisted breeding of perennial crops. Frontiers in Plant Science, 8, 460.
20. Ipek, A., Ates, D., & Tangu, N. A. (2021). Molecular breeding in berries. In Genome Engineering for Crop Improvement (pp. 202-225). Springer, Cham.
21. [21] Sakin, M., et al. (2021). Türk Bilimsel Derlemeler Dergisi, 14(1), 47-55.
22. Clark, J. R., & Finn, C. E. (2011). In Fruit, Vegetable and Cereal Science and Biotechnology 5(1), 27-43.
23. Qu, Y., Li, M., & Mao, J. (2021). Advances in the study of self-incompatibility in fruit trees. Journal of Fruit Science, 38(2), 252-265.
24. Hancock, J. F. (Ed.). (2008). Temperate fruit crop breeding: germplasm to genomics. Springer Science & Business Media.
25. Bassi, R., & Dall'Osto, L. (2021). Dissipation of light energy absorbed in excess: the molecular mechanisms. Annual review of plant biology, 72(1), 47-76.
26. Liu, Y., & Liu, Y. (2021). Somatic hybridization for plant breeding. In Plant Breeding Reviews (Vol. 44, pp. 323–372). Wiley.
27. Wang, J., Wang, Y., Lü, T., Yang, X., Liu, J., Dong, Y., & Wang, Y. (2022). An efficient and universal protoplast isolation protocol suitable for transient gene expression analysis and single-cell RNA sequencing. International Journal of Molecular Sciences, 23(7), 3419.
28. Eeckhaut, T., Lakshmanan, P. S., Deryckere, D., Van Bockstaele, E., & Van Huylenbroeck, J. (2013). Progress in plant protoplast research. Planta, *238*(6), 991–1003.
29. Tu, L., Subburaj, S., Lee, K., Jeon, Y., Yan, F., Yao, J., ... & Lee, G. J. (2023). Optimized regeneration of petunia protoplast and its association with tissue identity regulators. Horticulturae, 9(2), 216.
30. Zaranek, M., Pérez-Pérez, R., Milewska-Hendel, A., Betekhtin, A., & Grzebelus, E. (2023). Promotive effect of phytosulfokine-peptide growth factor-on protoplast cultures development in Fagopyrum tataricum (L.) Gaertn. BMC Plant Biology, 23(1), 385.
31. Kaur, S., Ijaz, S., & Nasir, B. (2024). Tissue Culture and Somatic Fusion in Plants. Trends in Plant Biotechnology, 1-21.
32. Moemi, B., Masanga, J., & Runo, S. (2023). An optimized protocol for in vitro regeneration of tropical maize inbred lines through cell suspension and semi-protoplast cultures.
33. Dutt, M., Mahmoud, L. M., Chamusco, K., Stanton, D., Chase, C. D., Nielsen, E., ... & Grosser, J. W. (2021). Utilization of somatic fusion techniques for the development of HLB tolerant breeding resources employing the Australian finger lime (Citrus australasica). PLoS One, 16(8), e0255842.
34. Lin, J., & Wong, K. C. (2018). Off-target predictions in CRISPR-Cas9 gene editing using deep learning. Bioinformatics, 34(17), i656-i663.
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