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<p>An optimized, multi-step somatic embryogenesis protocol was developed for large-scale propagation of adult <i>Coffea arabica</i> cv. ‘Catuaí Vermelho’. A factorial approach was used to evaluate the influence of key culture parameters at each developmental phase, from callus induction to embryo maturation. The highest induction of embryogenic calluses was achieved with 3% sucrose, casein hydrolysate combined with malt extract, and 10 µM 2iP. For callus multiplication, optimized liquid culture conditions included controlled inoculum density (5–10&#xa0;g L⁻¹), adequate culture volume, and dark incubation, which together enhanced proliferation while maintaining embryogenic potential. Differentiation and maturation were maximized using four to six explants per plate in solid medium for 90 days, followed by transfer to liquid maturation medium at an initial density of 5&#xa0;g L⁻¹, yielding well-formed somatic embryos with improved developmental uniformity. This integrated optimization resulted in embryo yields surpassing those reported in classical protocols, while maintaining morphological uniformity. The methodology provides a robust and scalable platform for high-efficiency clonal propagation, with direct applications in genetic improvement and germplasm conservation programs for high-value coffee cultivars. Further validation in bioreactors and integration of genetic fidelity analyses will be essential to fully realize its potential for commercial propagation and long-term conservation initiatives.</p>

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Integrated optimization of a multi-step somatic embryogenesis protocol enabling large-scale propagation of Coffea arabica L.

  • 期刊论文

  • 发表日期：

    2025年10月31日

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  • Patrícia Monah Cunha Bartos,
  • Hugo Teixeira Gomes,
  • Inaê Mariê de Araújo Silva-Cardoso,
  • João Batista Teixeira,
  • Jonny Everson Scherwinski-Pereira

摘要

An optimized, multi-step somatic embryogenesis protocol was developed for large-scale propagation of adult Coffea arabica cv. ‘Catuaí Vermelho’. A factorial approach was used to evaluate the influence of key culture parameters at each developmental phase, from callus induction to embryo maturation. The highest induction of embryogenic calluses was achieved with 3% sucrose, casein hydrolysate combined with malt extract, and 10 µM 2iP. For callus multiplication, optimized liquid culture conditions included controlled inoculum density (5–10 g L⁻¹), adequate culture volume, and dark incubation, which together enhanced proliferation while maintaining embryogenic potential. Differentiation and maturation were maximized using four to six explants per plate in solid medium for 90 days, followed by transfer to liquid maturation medium at an initial density of 5 g L⁻¹, yielding well-formed somatic embryos with improved developmental uniformity. This integrated optimization resulted in embryo yields surpassing those reported in classical protocols, while maintaining morphological uniformity. The methodology provides a robust and scalable platform for high-efficiency clonal propagation, with direct applications in genetic improvement and germplasm conservation programs for high-value coffee cultivars. Further validation in bioreactors and integration of genetic fidelity analyses will be essential to fully realize its potential for commercial propagation and long-term conservation initiatives.