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Sessão de Pósteres n.º 2 Localização: Átrio ESAC | |
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Micropropagation and slow growth germplasm conservation of Ilex aquifolium L. 1: CBPBI - Centro de Biotecnologia de Plantas da Beira Interior, Portugal; 2: CERNAS-IPCB - Centro de Estudos em Recursos Naturais, Ambiente e Sociedade, Instituto Politécnico de Castelo Branco. Castelo Branco, Portugal; 3: The Navigator Company, Setúbal, Portugal; 4: Viveiros Aliança, Herdade de Espirra, Pegões, Portugal; 5: CIBIO (Research Center in Biodiversity and Genetic Resources) - InBIO (Research Network in Biodiversity and Evolutionary Biology), University of Porto. Campus Agrário de Vairão. Vairão, Portugal; 6: BIOPOLIS Program in Genomics, Biodiversity and Land Planning, CIBIO. Campus de Vairão. Vairão, Portugal.; 7: MHNC-UP - Museu de História Natural e da Ciência da Universidade do Porto – Herbário PO, Universidade do Porto. Porto, Portugal; 8: proMetheus - Research Unit in Materials, Energy and Environment for Sustainability, Instituto Politécnico de Viana do Castelo. Viana do Castelo, Portugal; 9: Departamento de Biologia, Faculdade de Ciências, Universidade do Porto. Porto, Portugal Ilex aquifolium L., commonly known as European Holly, is a dioecious tree species of significant ecological and ethnobotanical value. Due to its critical decline caused by overexploitation for ornamental purposes and habitat loss, I. aquifolium has been legally protected in Portugal since 1989 (Decree-Law No. 423/89), which strictly prohibits the collection, transport, or sale of wild specimens. Given its legal protection and the critical decline of wild populations, the development of efficient biotechnological tools is crucial to ensure its long-term survival and genetic diversity. The aim of this study was a development of an efficient micropropagation protocol and a slow growth conservation assay. The establishment was done from an adult plant tree from Monchique mountains. Nodal segments were cultured on Woody Plant Medium (WPM) supplemented with different cytokinins to evaluate their effects on shoot induction and multiplication. The experimental design evaluated 2-isopentenyladenine (2iP), 6-benzylaminopurine (BAP), and kinetin (KIN), each tested at concentrations of 0.5, 1.0, and 2.0 mg/L. For the rooting assay, indole-3-butyric acid (IBA) was applied at 1, 3, and 5 mg/L. During the multiplication stage, the best results were obtained with 0.5 mg/L BAP, yielding a maximum shoot length of 20 mm and a multiplication rate of 2.7, although the explants exhibited mild hyperhydricity. The highest rooting efficiency was achieved using 5 mg/L IBA, which resulted in 97% and 100% rooting after 7 and 30 days, respectively. At acclimatization phase, under a relative humidity control environment, 100% of survival was achieved. Regarding slow growth conservation, viable explants were successfully retrieved after 12 months of cold storage across different conservation interval times. This study establishes the baseline protocol for micropropagation and conservation of I. aquifolium as essential tools for the preservation of this species. Funding: These studies were supported by the TransForm Mobilisation Agenda (PRR 02/C05-i01/2022), funded by the European Union through the Recovery and Resilience Plan (RRP) and the NextGenerationEU fund, under component C5 – Capitalisation and Business Innovation | |