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<p>Avocado (<i>Persea americana</i> Mill.) represents a crop of strategic importance in Colombia, yet its cultivation faces significant challenges arising from high genetic heterogeneity in propagated plant material and unfavorable phytosanitary conditions. These constraints underscore the need for robust strategies to enable the clonal propagation of elite seedlings. In this study, we developed an optimized protocol for the in vitro micropropagation of Creole rootstocks and the cv. Hass. Apical and nodal explants derived from etiolated mother plants were successfully established under aseptic conditions. Etiolation of donor plants for seven weeks significantly enhanced shoot elongation and reduced tissue lignification, thereby improving explant competence and morphogenic responsiveness. Extended darkness preconditioning (15, 30, and 45 days) markedly reduced phenolic oxidation across genotypes, with “Creole 1” exhibiting the lowest phenolization rate (40%) following 45 days of dark treatment. An effective surface disinfection regime was achieved using 1% sodium hypochlorite (NaClO) for 5&#xa0;min, supplemented with Benlate 50 WP<sup>®</sup> (1&#xa0;g L⁻¹), and the antibiotics vancomycin (250&#xa0;mg L⁻¹) and cefotaxime (500&#xa0;mg L⁻¹). The hormonal combination of 6-benzylaminopurine (BAP, 1.5&#xa0;mg L⁻¹), 1-naphthaleneacetic acid (NAA, 0.4&#xa0;mg L⁻¹), and gibberellic acid (GA₃, 0.4&#xa0;mg L⁻¹) resulted in the highest shoot induction frequency (62%) and elongation (1.35&#xa0;cm). Continuous supplementation with activated charcoal (1&#xa0;g L⁻¹) promoted spontaneous adventitious rooting during the multiplication phase (44–56%), thereby reducing dependence on exogenous auxin application. Overall, this integrated protocol effectively mitigates phenolic oxidation, enhances organogenic potential, and facilitates adventitious root formation, providing a reproducible framework for the clonal propagation of elite avocado rootstocks adapted to Colombian agroecological conditions.</p>

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An efficient protocol for clonal propagation of avocado (Persea americana Mill.) rootstocks via etiolation-enhanced micropropagation

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年3月5日

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  • Catalina Restrepo-Osorio,
  • Alejandro Gil-Correal,
  • Susan Saavedra-Porras,
  • Luisa Cárdenas-Ortiz,
  • Lina Chamorro-Gutiérrez,
  • Diego Villanueva-Mejía

摘要

Avocado (Persea americana Mill.) represents a crop of strategic importance in Colombia, yet its cultivation faces significant challenges arising from high genetic heterogeneity in propagated plant material and unfavorable phytosanitary conditions. These constraints underscore the need for robust strategies to enable the clonal propagation of elite seedlings. In this study, we developed an optimized protocol for the in vitro micropropagation of Creole rootstocks and the cv. Hass. Apical and nodal explants derived from etiolated mother plants were successfully established under aseptic conditions. Etiolation of donor plants for seven weeks significantly enhanced shoot elongation and reduced tissue lignification, thereby improving explant competence and morphogenic responsiveness. Extended darkness preconditioning (15, 30, and 45 days) markedly reduced phenolic oxidation across genotypes, with “Creole 1” exhibiting the lowest phenolization rate (40%) following 45 days of dark treatment. An effective surface disinfection regime was achieved using 1% sodium hypochlorite (NaClO) for 5 min, supplemented with Benlate 50 WP® (1 g L⁻¹), and the antibiotics vancomycin (250 mg L⁻¹) and cefotaxime (500 mg L⁻¹). The hormonal combination of 6-benzylaminopurine (BAP, 1.5 mg L⁻¹), 1-naphthaleneacetic acid (NAA, 0.4 mg L⁻¹), and gibberellic acid (GA₃, 0.4 mg L⁻¹) resulted in the highest shoot induction frequency (62%) and elongation (1.35 cm). Continuous supplementation with activated charcoal (1 g L⁻¹) promoted spontaneous adventitious rooting during the multiplication phase (44–56%), thereby reducing dependence on exogenous auxin application. Overall, this integrated protocol effectively mitigates phenolic oxidation, enhances organogenic potential, and facilitates adventitious root formation, providing a reproducible framework for the clonal propagation of elite avocado rootstocks adapted to Colombian agroecological conditions.