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<p>Protoplast isolation and regeneration remain challenging for many ornamental species, including the hybrid <i>Dianthus</i> cultivar ‘Telstar Scarlet,’ due to low regeneration capacity and genotype-dependent responses. This study aimed to establish an efficient protoplast-to-plant regeneration system by comparing liquid culture with an alginate-embedding method and evaluating the effects of various plant growth regulator (PGR) combinations. Protoplasts were isolated from seven-day-old cotyledons and three-week-old leaves, yielding 2.61 × 10<sup>6</sup> and 3.23 × 10<sup>6</sup> protoplasts g<sup>−1</sup> of fresh weight, respectively. While both liquid- and alginate-embedded cultures supported initial cell division, only alginate embedding enabled sustained development and microcallus formation within six weeks. Among the five PGR-supplemented protoplast culture media (PCM) tested, combining zeatin and 2,4-dichlorophenoxyacetic acid (2,4-D) in PCM 2 resulted in the highest division frequency and plating efficiency. Shoot induction from protoplast-derived calli was influenced by both explant type and age; younger leaf-derived calli responded best to benzyladenine (BA) and naphthaleneacetic acid (NAA), while cotyledon-derived calli tended to promote root formation. The optimized protocols enabled successful plant regeneration from both leaf and cotyledon protoplasts, and flow cytometric analysis confirmed stable ploidy levels in the regenerated plants.</p>

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Efficient plant regeneration from protoplasts of Dianthus “Telstar Scarlet” through alginate embedding and optimized PGR treatment

  • 期刊论文

  • 发表日期：

    2025年9月10日

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  • Oluwaseun Suleimon Adedeji,
  • Aung Htay Naing,
  • Kyeung Il Park,
  • Chang Kil Kim

摘要

Protoplast isolation and regeneration remain challenging for many ornamental species, including the hybrid Dianthus cultivar ‘Telstar Scarlet,’ due to low regeneration capacity and genotype-dependent responses. This study aimed to establish an efficient protoplast-to-plant regeneration system by comparing liquid culture with an alginate-embedding method and evaluating the effects of various plant growth regulator (PGR) combinations. Protoplasts were isolated from seven-day-old cotyledons and three-week-old leaves, yielding 2.61 × 106 and 3.23 × 106 protoplasts g−1 of fresh weight, respectively. While both liquid- and alginate-embedded cultures supported initial cell division, only alginate embedding enabled sustained development and microcallus formation within six weeks. Among the five PGR-supplemented protoplast culture media (PCM) tested, combining zeatin and 2,4-dichlorophenoxyacetic acid (2,4-D) in PCM 2 resulted in the highest division frequency and plating efficiency. Shoot induction from protoplast-derived calli was influenced by both explant type and age; younger leaf-derived calli responded best to benzyladenine (BA) and naphthaleneacetic acid (NAA), while cotyledon-derived calli tended to promote root formation. The optimized protocols enabled successful plant regeneration from both leaf and cotyledon protoplasts, and flow cytometric analysis confirmed stable ploidy levels in the regenerated plants.