Vaccarini Cristian, Mercado María Inés, Ponessa Graciela Inés, McCarthy Andrés, McCarthy Christina, Amado Cattáneo Raúl, Sedan Daniela, Andrinolo Darío
Centro de Investigaciones del Medioambiente (CIM-CONICET-CIC), Universidad Nacional de La Plata, La Plata, Argentina.
Instituto de Morfología Vegetal, Fundación Miguel Lillo, Tucumán, Argentina.
Protoplasma. 2025 Mar 19. doi: 10.1007/s00709-025-02054-0.
This study analyzed the morphoanatomical, histochemical, and molecular characteristics of three Cannabis sativa strains, CAT1, CAT2, and CAT3, acronyms for Argentine therapeutic strains in Spanish (Cepas Argentinas Terapéuticas), using bright light, fluorescence, and scanning electron microscopy. The strains were previously cultivated and chemically characterized at CIM, UNLP, CONICET. Five plants from each strain were collected at the end of the vegetative and flowering phases; part of the material was fixed for anatomical studies, and part was used fresh for histochemical analyses. Anatomical features of roots, stems, leaves, and flowers were analyzed, focusing on stomatal density, trichome types, and the presence of laticifers. Histochemical analyses detected phenolic compounds, cannabinoids, lipophilic compounds, and other metabolites using specific staining techniques. Additionally, short sequence repeat (SSR) molecular markers were employed to characterize and confirm the genomic identity of the strains. Morphoanatomical and histochemical traits enabled differentiation among the strains, revealing significant variations in leaflet dimensions, trichome density, and metabolic profiles. For example, CAT1 exhibited thicker leaves and larger stomata, CAT2 had a higher density of laticifers, and CAT3 showed a greater density of cannabinoid-rich glandular trichomes. Using SSR molecular markers, strain genomic identity was confirmed with a probability greater than 99.99999983%. This integrative approach, combining morphoanatomical, histochemical, and molecular analyses, highlights the unique features of CAT1, CAT2, and CAT3 and underscores the importance of molecular markers in validating strain identity.
本研究利用明场显微镜、荧光显微镜和扫描电子显微镜，分析了三种大麻品种CAT1、CAT2和CAT3（西班牙语中阿根廷治疗用品种的首字母缩写，即Cepas Argentinas Terapéuticas）的形态解剖学、组织化学和分子特征。这些品种此前已在国立拉普拉塔大学化学研究所（CIM）、国立拉普拉塔大学（UNLP）和国家科学技术研究委员会（CONICET）进行过种植和化学特征分析。在营养生长阶段和开花阶段结束时，从每个品种中采集了五株植物；部分材料用于解剖学研究，部分材料新鲜用于组织化学分析。分析了根、茎、叶和花的解剖特征，重点关注气孔密度、腺毛类型和乳汁管的存在情况。组织化学分析使用特定染色技术检测酚类化合物、大麻素、亲脂性化合物和其他代谢产物。此外，还采用短串联重复序列（SSR）分子标记来鉴定和确认这些品种的基因组身份。形态解剖学和组织化学特征能够区分这些品种，揭示出小叶尺寸、腺毛密度和代谢谱的显著差异。例如，CAT1的叶子更厚，气孔更大；CAT2的乳汁管密度更高；CAT3的富含大麻素的腺毛密度更大。使用SSR分子标记，确认品种基因组身份的概率大于99.99999983%。这种结合形态解剖学、组织化学和分子分析的综合方法，突出了CAT1、CAT2和CAT3的独特特征，并强调了分子标记在验证品种身份方面的重要性。
