ABSTRACT
The preservation of embryogenic lines derived from several endangered local grapevine cultivars was studied. Embryogenic calluses were obtained from immature anthers of eight cultivars, sampled on both fruity-cuttings and field grown vines. Anthers at the 'separated flower' stage, derived from fruity-cuttings, resulted in an increased induction of somatic embryogenesis, compared to those derived from the field. Pro-embryogenic calluses were induced on Chée and Pool (1987) basal medium, supplemented with 9 micron M of 2,4-dichlorophenoxyacetic acid (2,4-D) and 11.35 micron M of thidiazuron (TDZ) under dark conditions. Different anther zones (filament, abaxial, adaxial, lateral zones and entire anthers) were involved in somatic embryogenesis induction. The percentages of granular and yellowish pro-embryogenic calluses ranged between 15.6 percent and 34.8 percent in 'Kahli Kerkennah' and 'Muscat Raf-raf' cultivars, respectively. Although, morphological diversifications of pro-embryogenic calluses (several necrosis and spontaneous maturation) were observed on the induction mediumafter 5 subcultures. The reduction of 2,4-D and TDZ levels to 4.52 micron M and 2.89 micron M respectively, induced granular and yellowish embryogenic material. Thus, Chée and Pool (1987) (CP) enriched with 4.52 micron M of 2,4-D and 2.89 micron M of TDZ revealed to be the most appropriate for long-term maintenance. In fact, all the cultivars presented high and regular embryo maturation rates after 12, 24, 36 and 48 months of cultivation on this medium, under light conditions. After 4 years, they still exhibit high germination and regeneration abilities. Germination of somatic embryos was achieved on Murashige and Skoog (1962) basal-medium, with rates ranging from 69 percent to 96 percent. Only 5 percent of somatic embryos were concerned by morphological variations. The regenerated plantlets presented a normal phenotype under controlled greenhouse conditions, compared to mother plants.