Abstract
Protoplasts are an attractive explant source for biotechnological tools widely used on citrus genetic improvement, such as somatic hybridization and direct genetic transformation. These delicate and responsive materials are subjected to cell proliferation induction and differentiation of somatic embryos which further regenerate into entire plants. The isolation of viable protoplasts followed by regeneration of plants through somatic embryogenesis is an important methodology for breeding applications. The methods presented here can also be used as a reference for protoplast work in other species, followed by protocol optimization for different species/genotypes.
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References
Johnson AAT, Veilleux RE (2001) Somatic hybridization and application in plant breeding. In: Janick J (ed) Plant breeding rev 20. John Wiley & Sons, New York, pp 167–225
Davey M, Anthony P, Patel D, Power J (2010) Plant protoplasts: isolation, culture and plant regeneration. In: Davey M, Anthony P (eds) Plant cell culture: essential methods. John Wiley & Sons, New York, pp 2153–2173
Omar AA, Song WY, Grosser JW (2007) Introduction of the Xa21, Xanthomonas resistance gene from rice, into ‘Hamlin’ sweet orange [citrus Sinensis (L.) Osbeck] using protoplast/GFP co-transformation and single plasmid transformation. J Hortic Sci Biotechnol 82(6):914–923
Grosser JW, Gmitter Junior FG (2011) Protoplast fusion for production of tetraploids and triploids: applications for scion and rootstock breeding in citrus. Plant Cell Tissue Organ Cult 104:343–357
Liu Y, Xue Y, Tang J, Chen J, Chen M (2019) Efficient mesophyll protoplast isolation and development of a transient expression system for castor-oil plant (Ricinus communis L.). Biol Futur 70(1):8–15
Grosser JW, Gmitter FG Jr (1990) Protoplast fusion and citrus improvement. Plant Breed Rev 8:339–374
Soriano L, Mourão Filho FAA, Camargo LEA, Cristofani-Yaly M, Latado RR, Pacheco CA et al (2012) Regeneration and characterization of somatic hybrids combining sweet orange and mandarin/mandarin hybrid cultivars for citrus scion improvement. Plant Cell Tissue Organ Cult 111:385–392
Kim MJ, Yun SH, Park SM, Jin SB, Song K (2020) Characterization of allotetraploids derived from protoplast fusion between navel orange and kumquat. In Vitro Cell Dev Biol Plant 56:634–643
Ruiz M, Pensabene-Bellavia G, Quiñones A, García-Lor A, Morillon R, Ollitrault P et al (2018) Molecular characterization and stress tolerance evaluation of new allotetraploid somatic hybrids between Carrizo Citrange and Citrus macrophylla W. rootstocks. Front Plant Sci 9:901 https://doi.org/10.3389/fpls.2018.00901.
Eeckhaut T, Lakshmanan P, Deryckere D, Van Huylenbroeck J, Van Bockstaele E (2013) Progress in plant protoplast research. Planta 238:991–1003
Hassanein A, Hamama L, Loridon K, Dorion N (2009) Direct gene transfer study and transgenic plant regeneration after electroporation into mesophyll protoplasts of pelargonium x hortorum, ‘panache Sud’. Plant Cell Rep 28:1521–1530
Masani MY, Noll GA, Parveez GK, Sambanthamurthi R, Prüfer D (2014) Efficient transformation of oil palm protoplasts by PEG-mediated transfection and DNA microinjection. PLoS One 9(5):e96831
Lin CS, Hsu CT, Yang LH, Lee LY, Fu JY, Cheng QW et al (2018) Application of protoplast technology to CRISPR/Cas9 mutagenesis: from single-cell mutation detection to mutant plant regeneration. Plant Biotechnol J 16(7):1295–1310
Salonia F, Ciacciulli A, Poles L, Pappalardo HD, La Malfa S, Licciardello C (2020) New plant breeding techniques in citrus for the improvement of important agronomic traits - a review. Front Plant Sci 11:1234
Murashige T, Tuker DPH (1969) Growth factor requirement of citrus tissue culture. In: International citrus symposium. University of California, Riverside, pp 1155–1169
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Soriano, L., Omar, A.A., Martinelli, A.P. (2022). Citrus Protoplast Isolation and Plant Regeneration Through Somatic Embryogenesis. In: Ramírez-Mosqueda, M.A. (eds) Somatic Embryogenesis. Methods in Molecular Biology, vol 2527. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2485-2_9
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DOI: https://doi.org/10.1007/978-1-0716-2485-2_9
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