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Efficient In Vitro Plantlet Regeneration from Stolon Explants and Genetic Stability Assessment Using ISSR Markers in the Ornamental Fern Hypolepis punctata
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Efficient In Vitro Plantlet Regeneration from Stolon Explants and Genetic Stability Assessment Using ISSR Markers in the Ornamental Fern Hypolepis punctata
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Efficient In Vitro Plantlet Regeneration from Stolon Explants and Genetic Stability Assessment Using ISSR Markers in the Ornamental Fern Hypolepis punctata
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Journal Article
Efficient In Vitro Plantlet Regeneration from Stolon Explants and Genetic Stability Assessment Using ISSR Markers in the Ornamental Fern Hypolepis punctata
2025
Overview
Hypolepis punctata, an aromatic fern with insect-resistant and ornamental potential. Up to date, no studies have reported its micropropagation, particularly using vegetative organs as explants. The optimized stolon sterilization (81.11%) employed 75% ethanol (30 s) and 15% sodium hypochlorite (12 min). The optimal conditions for GGB induction (75.56%) and proliferation (8.46 mm) were achieved using Murashige and Skoog (MS) medium + 2.0 mg/L 6-benzylaminopurine (BA) + 0.2 mg/L 1-naphthaleneacetic acid (NAA). The optimal plant growth regulator (PGR) formula for sporophyte regeneration was 0.5 mg/L BA + 0.1 mg/L NAA + 2 g/L activated charcoal (AC), achieving a 98.89% induction rate and 49.19 buds per explant. The 1/4 MS medium had the greatest promoting effect on biomass accumulation and leaf expansion. Optimal shoot elongation (97.78% success, 4.83 cm) was achieved in 1/4 MS + 0.5 mg/L BA + 0.1 mg/L NAA + 2 g/L AC, and optimized rooting (92.22%) was achieved using 1/4 MS + 0.5 mg/L indole-3-butyric acid (IBA) + 0.1 mg/L NAA + 2 g/L AC, producing 25.27 roots per plantlet. Crucially, ISSR analysis confirmed the genetic stability of all regenerants. This optimized protocol establishes a scalable micropropagation system, enhancing both commercial cultivation and genetic improvement potential in Hypolepis punctata.
