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<p><i>Begonia grandis subsp. sinensis</i> (A. DC.) Irmsch (<i>B. grandis</i>) has high ornamental and medicinal value. This study aimed to establish a regeneration protocol for <i>B. grandis</i> by assessing explant responses to specific hormone combinations, providing a system for clonal propagation and conservation. Here, two distinct regeneration pathways, direct somatic embryogenesis (SE) and shoot organogenesis in <i>B. grandis</i> are reported for the first time. Four explant types were evaluated: shoots with one leaf (SL) and bulbils derived from mother plants and aseptic leaf and petiole segment explants derived from in vitro-cultured shoots. Compared with either plant growth regulator (PGR) alone, the auxin-cytokinin combination was more effective at inducing regeneration in all explant types. Single-leaf shoots and petiole segments as explant materials led to plant regeneration via SE. Both explants showed the optimal regeneration response on Murashige and Skoog medium (MS) supplemented with 0.2 mg L<sup>− 1</sup> α-naphthaleneacetic acid (NAA) and 1.0 mg L<sup>− 1</sup> 6-benzyladenine (BA), with SE induction frequencies of 96.3% and 93.5%, averaging 41.6 and 35 somatic embryos per explant, respectively. When bulbils and leaf segments were used as explants, the regeneration process proceeded via dual pathways of both direct and indirect shoot organogenesis, with the most effective response [highest induction rates (organogenesis frequencies of 82.4% and 70.7%, respectively) and maximum shoot production (an average of 18 and 14 adventitious shoots per explant, respectively)] observed on MS medium enriched with 0.2 mg L<sup>− 1</sup> NAA and 1.0 mg L<sup>− 1</sup> thidiazuron (TDZ). Histological examination revealed structures corresponding to various somatic embryos developmental stages, including globular, heart-shaped and torpedo-shaped embryos. Genetic fidelity among the regenerated and mother plants were assessed using inter-simple sequence repeat (ISSR) and start codon targeted (SCoT) markers, both of which exhibited monomorphic banding patterns, indicating the high genetic fidelity of the regenerated plants to the mother plants. Flow cytometry analysis demonstrated stable genome size across regenerants, with no significant ploidy variation. Thus, the developed in vitro propagation protocol is efficient and cost effective and demonstrates strong potential for the large-scale production of this valuable plant.</p>

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A robust system for somatic embryogenesis and shoot organogenesis in Begonia grandis subsp. sinensis (A. DC.) Irmsch and genetic fidelity assessment of the regenerants

  • 期刊论文

  • 发表日期：

    2026年2月27日

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  • Feimin Ma,
  • Zhengli Shan,
  • Haonan Su,
  • Ruijing Han,
  • Wenjing Zhou,
  • Mu Ze,
  • Ming Dong,
  • Qinggui Wu,
  • Lijuan Zou

摘要

Begonia grandis subsp. sinensis (A. DC.) Irmsch (B. grandis) has high ornamental and medicinal value. This study aimed to establish a regeneration protocol for B. grandis by assessing explant responses to specific hormone combinations, providing a system for clonal propagation and conservation. Here, two distinct regeneration pathways, direct somatic embryogenesis (SE) and shoot organogenesis in B. grandis are reported for the first time. Four explant types were evaluated: shoots with one leaf (SL) and bulbils derived from mother plants and aseptic leaf and petiole segment explants derived from in vitro-cultured shoots. Compared with either plant growth regulator (PGR) alone, the auxin-cytokinin combination was more effective at inducing regeneration in all explant types. Single-leaf shoots and petiole segments as explant materials led to plant regeneration via SE. Both explants showed the optimal regeneration response on Murashige and Skoog medium (MS) supplemented with 0.2 mg L− 1 α-naphthaleneacetic acid (NAA) and 1.0 mg L− 1 6-benzyladenine (BA), with SE induction frequencies of 96.3% and 93.5%, averaging 41.6 and 35 somatic embryos per explant, respectively. When bulbils and leaf segments were used as explants, the regeneration process proceeded via dual pathways of both direct and indirect shoot organogenesis, with the most effective response [highest induction rates (organogenesis frequencies of 82.4% and 70.7%, respectively) and maximum shoot production (an average of 18 and 14 adventitious shoots per explant, respectively)] observed on MS medium enriched with 0.2 mg L− 1 NAA and 1.0 mg L− 1 thidiazuron (TDZ). Histological examination revealed structures corresponding to various somatic embryos developmental stages, including globular, heart-shaped and torpedo-shaped embryos. Genetic fidelity among the regenerated and mother plants were assessed using inter-simple sequence repeat (ISSR) and start codon targeted (SCoT) markers, both of which exhibited monomorphic banding patterns, indicating the high genetic fidelity of the regenerated plants to the mother plants. Flow cytometry analysis demonstrated stable genome size across regenerants, with no significant ploidy variation. Thus, the developed in vitro propagation protocol is efficient and cost effective and demonstrates strong potential for the large-scale production of this valuable plant.