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Open AccessArticle

Protoplast Isolation and Shoot Regeneration from Protoplast-Derived Callus of Petunia hybrida Cv. Mirage Rose

Department of Horticultural Science, Kyungpook National University, Daegu 41566, Korea
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Authors to whom correspondence should be addressed.
Authors equally contributed this work.
Biology 2020, 9(8), 228; https://doi.org/10.3390/biology9080228
Received: 1 July 2020 / Revised: 3 August 2020 / Accepted: 13 August 2020 / Published: 16 August 2020
Despite the increasing use of protoplasts in plant biotechnology research, shoot regeneration from protoplasts remains challenging. In this study, we investigated the factors involved in protoplast isolation, callus induction, and shoot regeneration in Petunia hybrida cv. Mirage Rose. The following conditions were found to be most optimal for protoplast yield and viability: 0.6 M mannitol, 2.0% cellulase, and 6 h digestion time. A plating density of 10 × 104 protoplasts/mL under osmoticum condition (0.58 M mannitol) showed high microcolony viability in liquid culture. The Kao and Michayluk medium was found to be appropriate for callus proliferation from microcalli under a 16-h light photoperiod. Calli cultured in Murashige and Skoog medium containing 1.0 mg/L 6-benzylaminopurine and 0.2 mg/L 3-indole butyric acid showed the highest shoot regeneration frequency and number of shoots obtained per explant. Random amplification of polymorphic DNA analysis showed that the protoplast-derived shoots exhibited the same banding patterns as those of donor plants. Collectively, these findings can contribute to solving problems encountered in protoplast isolation and shoot regeneration in other petunia cultivars and related species. As the protocol developed by us is highly reproducible, it can be applied in biotechnology research on P. hybrida cv. Mirage Rose. View Full-Text
Keywords: digestion time; osmoticum conditions; plating density; plant growth regulators; protoplast viability digestion time; osmoticum conditions; plating density; plant growth regulators; protoplast viability
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MDPI and ACS Style

Kang, H.H.; Naing, A.H.; Kim, C.K. Protoplast Isolation and Shoot Regeneration from Protoplast-Derived Callus of Petunia hybrida Cv. Mirage Rose. Biology 2020, 9, 228. https://doi.org/10.3390/biology9080228

AMA Style

Kang HH, Naing AH, Kim CK. Protoplast Isolation and Shoot Regeneration from Protoplast-Derived Callus of Petunia hybrida Cv. Mirage Rose. Biology. 2020; 9(8):228. https://doi.org/10.3390/biology9080228

Chicago/Turabian Style

Kang, Hyun H.; Naing, Aung H.; Kim, Chang K. 2020. "Protoplast Isolation and Shoot Regeneration from Protoplast-Derived Callus of Petunia hybrida Cv. Mirage Rose" Biology 9, no. 8: 228. https://doi.org/10.3390/biology9080228

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