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<p><i>Pterocarpus marsupium</i> Roxb., an important multipurpose threatened forest tree, is a vital species in the Fabaceae family. Its natural propagation is hindered by low seed germination rates and overexploitation, highlighting the urgent requirement for innovative methods to enhance its populations due to its diverse utility. Somatic embryogenesis (SE) faces challenges in meeting demand for elite materials, necessitating a high-throughput approach for optimizing technique by multiplying experimental conditions quickly. A protocol has been developed for the first time to propagate genetically uniform plantlets from immature cotyledons (ICs) of <i>P. marsupium</i> through direct somatic embryogenesis (DSE). The study found that ICs from 8-day-old seedlings had the greatest incidence of SE (83.33%) and produced 24.2 somatic embryos / explant in Murashige and Skoog (MS) medium enriched with 2,4-D (2.0 mg L<sup>− 1</sup>) and Kn (0.5 mg L<sup>− 1</sup>). After 6 weeks of transferring globular shape to the same medium, an ideal inducing rate of 62.3% from globular to mature stage embryos was achieved. The formation and maturation of SE decreased when sugar concentrations exceeded 3%. Mature embryos germinated to yield plantlets on ½ strength MS media fortified with 1.0 mg L<sup>− 1</sup> GA<sub>3</sub>, achieving a germination frequency of 61.2%. The study reveals that SE, as depicted through stereomicroscopic and histological examinations of explants, comprises various developmental stages. The plantlets generated from somatic embryos acclimatized in a greenhouse showed the highest survival rate (82%) after two months. The study used ISSR-PCR analysis to analyze the genetic uniformity of regenerated plantlets, revealing that all banding patterns were uniform and identical to the mother plant, indicating no DNA polymorphism. The development of a direct somatic embryogenesis regeneration system and clonal fidelity studies for <i>P. marsupium</i> could facilitate large-scale propagation, genetic improvements, and germplasm conservation in this plant.</p>

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Clonal fidelity analysis using ISSR markers in Pterocarpus marsupium Roxb. plantlets regenerated through direct somatic embryogenesis

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年8月2日

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  • Radhika Tippani,
  • Mahendar Porika,
  • Christopher Thammidala

摘要

Pterocarpus marsupium Roxb., an important multipurpose threatened forest tree, is a vital species in the Fabaceae family. Its natural propagation is hindered by low seed germination rates and overexploitation, highlighting the urgent requirement for innovative methods to enhance its populations due to its diverse utility. Somatic embryogenesis (SE) faces challenges in meeting demand for elite materials, necessitating a high-throughput approach for optimizing technique by multiplying experimental conditions quickly. A protocol has been developed for the first time to propagate genetically uniform plantlets from immature cotyledons (ICs) of P. marsupium through direct somatic embryogenesis (DSE). The study found that ICs from 8-day-old seedlings had the greatest incidence of SE (83.33%) and produced 24.2 somatic embryos / explant in Murashige and Skoog (MS) medium enriched with 2,4-D (2.0 mg L− 1) and Kn (0.5 mg L− 1). After 6 weeks of transferring globular shape to the same medium, an ideal inducing rate of 62.3% from globular to mature stage embryos was achieved. The formation and maturation of SE decreased when sugar concentrations exceeded 3%. Mature embryos germinated to yield plantlets on ½ strength MS media fortified with 1.0 mg L− 1 GA3, achieving a germination frequency of 61.2%. The study reveals that SE, as depicted through stereomicroscopic and histological examinations of explants, comprises various developmental stages. The plantlets generated from somatic embryos acclimatized in a greenhouse showed the highest survival rate (82%) after two months. The study used ISSR-PCR analysis to analyze the genetic uniformity of regenerated plantlets, revealing that all banding patterns were uniform and identical to the mother plant, indicating no DNA polymorphism. The development of a direct somatic embryogenesis regeneration system and clonal fidelity studies for P. marsupium could facilitate large-scale propagation, genetic improvements, and germplasm conservation in this plant.