Fertile Plant Regeneration from Protoplasts of the Korean Lettuce Cultivar “Cheongchima” (Lactuca sativa L.)
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초록
Protoplasts were isolated from the primary leaves of lettuce (Lactuca sativa L.) seedlings 10 days after in vitro germination. The leaves were stripped and incubated in an enzyme mixture consisting of 1.2% Cellulase R-10 and 0.3% Macerozyme R-10 in cell and protoplast washing solution (CPW) overnight. The average protoplast yield was 8.25 x 106 protoplasts per g of fresh leaf tissue. When protoplasts were cultured at a density of 3.0 × 105 protoplasts/mL in agarose solid KM8P/KM8 medium, first and second divisions were observed in the protoplasts within a week. Protoplast-derived microcolonies formed after 4 weeks of culture, and visible colonies were present after 3 months of culture. Protoplast-derived microcalli were transferred to Murashige and Skoog medium supplemented with 2.0 mg/L kinetin and 0.1 mg/L NAA and incubated in the light for 3 weeks. They grew into callus, which then regenerated into plants after 7 weeks of culture. The regenerated plants grew as apparently normal flowering fertile plants.
Abstract
Protoplasts were isolated from the primary leaves of lettuce (Lactuca sativa L.) seedlings 10 days after in vitro germination. The leaves were stripped and incubated in an enzyme mixture consisting of 1.2% Cellulase R-10 and 0.3% Macerozyme R-10 in cell and protoplast washing solution (CPW) overnight. The average protoplast yield was 8.25 x 106 protoplasts per g of fresh leaf tissue. When protoplasts were cultured at a density of 3.0 × 105 protoplasts/mL in agarose solid KM8P/KM8 medium, first and second divisions were observed in the protoplasts within a week. Protoplast-derived microcolonies formed after 4 weeks of culture, and visible colonies were present after 3 months of culture. Protoplast-derived microcalli were transferred to Murashige and Skoog medium supplemented with 2.0 mg/L kinetin and 0.1 mg/L NAA and incubated in the light for 3 weeks. They grew into callus, which then regenerated into plants after 7 weeks of culture. The regenerated plants grew as apparently normal flowering fertile plants.
참고문헌 (26)
- Brown CJ, "Plant regeneration from protoplasts of a wild lettuce species (Lactuca saligna L.)", Plant Cell Rep, 6, pp.180~182 (1987)
- Chaipakdee W, "Isolation and culture of protoplast from leaves of Lactuca sativa", Songklanakarin J. Sci. Technol, 29, pp.929~935 (2007)
- Seung Hee Choi, "TSA Promotes CRISPR", Cas9 Editing Efficiency and Expression of Cell Division-Related Genes from Plant Protoplasts, 2021, pp.22(15), 7817~[DOI]
- Chupeau MC, "Transgenic plants of lettuce(Lactuca sativa)obtained through electroporation of protoplasts", Bio, 1989, pp.7, 503~508
- Engler DE, "Isolation, culture and regeneration of lettuce leaf mesophyll protoplasts", Plant Sci. Lett, 28, pp.223~229 (1983)
- Gupta HS, "Plant regeneration from mesophyll protoplasts of rice (Oryza sativa L.)", Bio, 1993, pp.11, 90~94
- Jeon JM, "Efficient transient expression and transformation of PEG-mediated gene uptake into mesophyll protoplasts of pepper (Capsicum annuum L.)", Plant Cell Tiss. Organ Cult, 88, pp.225~232 (2007)
- Kang BC, "Chloroplast and mitochondrial DNA editing in plants", Nature Plants, 7, pp.899~905 (2021)
- Kao KN, "Nutritional requirements for growth of Vicia hajastana cells and protoplasts at a very low density in liquid media", Planta, 126, pp.105~110 (1975)
- Lin CS, "Application of protoplast technology to CRISPR", Cas9 mutagenesis: From single-cell mutation detection to mutant plant regeneration, 2018, pp.16, 1295~1310
- Matsumoto E, "Interspecific somatic hybridization between lettuce (Lactuca sativa) and wild species L. virosa", Plant Cell Rep, 9, pp.531~534 (1991)
- Mazier M, "Interest of protoplasts for lettuce breeding", Proceedings of the Eucarpia Meeting on Leafy Vegetables Genetic and Breeding, pp.239~244 (1999)
- Mohebodini M, "Effects of genotype, explant age and growth regulators on callus induction and direct shoot regeneration of lettuce (Lactuca sativa L.)", AJCS, 5, pp.92~95 (2011)
- Mou B, "Handbook of plant breeding. vol. 1", pp.75~116 (2008), Springer
- Murashige T, "A reversed medium for rapid growth and bioassays with tobacco tissue cultures", Physiol. Plantarum, 15, pp.473~497 (1962)
- Nassour M, "Plant regeneration from protoplasts of micropropagated Pelargonium x hortorum 'Alain' : Effect of some environmental and medium factors on protoplats system efficiency", Plant Sci, 163, pp.169~176 (2002)
- Nishio T, "Simple and efficient protoplast culture procedure of lettuce", Japan. J. Breed, 38, pp.165~171 (1988)
- Power JB, "Plant cell culture", pp.37~66 (1985), IRL Press
- Pupilli F, "Plant regeneration from callus protoplasts of Onobrychis viciifolia Scop. (Sainfoin)", Plant Sci, 63, pp.87~94 (1989)
- Quattrocchio FM, "Transgenes and protein localization : Myths and legends", Trends in Plant Science, 18, pp.473~476 (2013)
- Jinbo Shen, "Isolation, Culture, and Transient Transformation of Plant Protoplasts", Current Protocols in Cell Biology, 63(1), pp.2.8.1~2.8.27 (2014), Wiley[DOI]
- Siddiqui MR, "Somatic hybridization via protoplasts fusion in Lactuca sativa (lettuce) and its fused product response to culture media", J. Agric. Res, 52, pp.1~9 (2014)
- 손보화, "상추의 자엽 및 제 1엽 절편체들로부터 효율적인 식물체 재분화", 생명과학회지, 17(6), pp.822~827 (2007), 한국생명과학회
- Suzuki H, "Plant regeneration from leaf mesophyll protoplasts of Lactuca sativa L", Plant Tiss. Cult, 3, pp.83~85 (1986)
- Webb CL, "Plant regeneration from mesophyll protoplasts of Lactuca perennis", Plant Cell Tiss. Organ Cult, 38, pp.77~79 (1994)
- Woo JW, "DNA-free genome editing in plants with preassembled CRISPR-Cas9 ribonucleoproteins", Nature Biotechnol, 11, pp.1162~1165 (2015)