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<p>Somatic embryogenesis in <i>Cannabis sativa</i> is notoriously difficult, standing out as one of the most recalcitrant morphogenetic pathways in plant biotechnology. A major hurdle is how differently each genotype responds to treatment. This study evaluated callus induction, differentiation, embryogenic maturation, and histological characteristics in two Moroccan varieties, Beldia and Khardala. We first initiated callus induction on ½ MS medium using 0.5&#xa0;mg/L 2,4-D alone, 0.5&#xa0;mg/L BAP alone, or a combination of both. For the maturation phase, we transferred the cultures to ECO+ medium supplemented with 0.2&#xa0;mg/L BAP alone, 0.2&#xa0;mg/L 2iP alone, or 0.05&#xa0;mg/L IBA alone. The results highlighted a sharp contrast between the two genotypes. While Beldia showed a strong initial callus formation response on leaf explants—with induction rates reaching 68.75%—it struggled to progress, with embryo maturation stalling below 25%. On the other hand, Khardala followed a more successful embryogenic pathway, particularly on stem explants, where embryogenic callus maturation reached 46.87–50% under BA and 2iP treatments. Our histological analysis further confirmed these differences. In Khardala, we observed well-developed globular, heart-shaped, and early cotyledonary structures characterized by dense meristematic clusters and organized embryogenic domains. However, a significant bottleneck remains: even with successful maturation, germination rates did not exceed 40% for either variety. Ultimately, these findings underscore that somatic regeneration in <i>C. sativa </i>is highly genotype-specific, meaning that a universal protocol remains elusive.</p>

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Somatic embryogenesis and histological characterization of callus development in moroccan varieties of Cannabis sativa L.

  • 期刊论文

  • 发表日期：

    2026年7月10日

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  • Meryem Tahtah,
  • Aicha Nordine,
  • Karima Samir,
  • Toufik Ibrahim

摘要

Somatic embryogenesis in Cannabis sativa is notoriously difficult, standing out as one of the most recalcitrant morphogenetic pathways in plant biotechnology. A major hurdle is how differently each genotype responds to treatment. This study evaluated callus induction, differentiation, embryogenic maturation, and histological characteristics in two Moroccan varieties, Beldia and Khardala. We first initiated callus induction on ½ MS medium using 0.5 mg/L 2,4-D alone, 0.5 mg/L BAP alone, or a combination of both. For the maturation phase, we transferred the cultures to ECO+ medium supplemented with 0.2 mg/L BAP alone, 0.2 mg/L 2iP alone, or 0.05 mg/L IBA alone. The results highlighted a sharp contrast between the two genotypes. While Beldia showed a strong initial callus formation response on leaf explants—with induction rates reaching 68.75%—it struggled to progress, with embryo maturation stalling below 25%. On the other hand, Khardala followed a more successful embryogenic pathway, particularly on stem explants, where embryogenic callus maturation reached 46.87–50% under BA and 2iP treatments. Our histological analysis further confirmed these differences. In Khardala, we observed well-developed globular, heart-shaped, and early cotyledonary structures characterized by dense meristematic clusters and organized embryogenic domains. However, a significant bottleneck remains: even with successful maturation, germination rates did not exceed 40% for either variety. Ultimately, these findings underscore that somatic regeneration in C. sativa is highly genotype-specific, meaning that a universal protocol remains elusive.