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Tuyển tập các báo cáo nghiên cứu về lâm nghiệp được đăng trên tạp chí lâm nghiệp quốc tế đề tài: Towards genetic engineering of maritime pine (Pinus pinaster Ait.). | Ann. For. Sci. 59 2002 687-697 INRA EDP Sciences 2002 DOI 10.1051 forest 2002057 687 Original article Towards genetic engineering of maritime pine Pinus pinaster Ait. Jean-Franẹois Trontin Luc Harvengt Elizabeth Garin Manuel Lopez-Vernaza Lydia Arancio Josiane Hoebeke Francis Canlet and Marc Pâques AFOCEL Laboratoire de Biotechnologie biotech@afocel.fr Domaine de I Etan on 77370 Nangis France Received 1 September 2001 accepted 22 January 2002 Abstract - Using our improved protocols for somatic embryogenesis in Pinuspinaster transgenic tissues and plantlets were recovered after microprojectile bombardment biolistic or cocultivation of embryonal-suspensor masses ESM with Agrobacterium tumefaciens. Transformation experiments were carried out with selectable hpt gene hygromycin B resistance and reporter gus gene P-glucuronidase activity . With both methods hygromycin was shown to be an effective selective agent of transformed cells within 4-19 weeks. The mean number of hygromy-cin-resistant lines expressing gus per gram ESM subjected to DNA transfer ranged from 7.0 to 8.5 using biolistic and 0 to 67.3 during Agrobacterium experiments. Mature somatic embryos obtained from some transformed lines were converted into plantlets and grown in the greenhouse. The whole process from transformation to plant acclimatisation could be completed within only 12 months. The transgenic state of ESM somatic embryos and plants was confirmed by histochemical GUS assays and molecular methods. Pinus pinaster somatic embryogenesis biolistic Agrobacterium tumefaciens transgenic plant Résumé - Transformation génétique du pin maritime Pinuspinaster Ait. . En appliquant nos protocoles d embryogenese somatique déve-loppés pour Pinuspinaster des tissus et plantes transgéniques ont été obtenus après bombardement avec des microparticules biolistique ou coculture de masses embryonnaires ESM avec Agrobacterium tumefaciens. Les expériences de transformation ont été conduites à l aide du gene de .
