Protoplast isolation from cell suspension of two Algerian cultivars of date palm ('Takerbucht' and 'Tegaza')
2013
Yatta, D. | Abed, F. | Amara, B. | Yakhou, M.S. | Bouguedoura, N.
The development of the culture of embryogenic cell suspensions can have a material of choice for production in many plants and for genetic improvement by biotechnological methods. The purpose of this study is the establishment of cell suspensions from date palm in order to have the materiel necessary for the isolation of protoplasts with the aim of obtaining somatic hybrids resistant to bayoud through protoplast fusion. For this embryogenic callus (compact and friable) were used to initiate cell suspensions in liquid medium enriched with 5 mg/L picloram Proliferation was obtained with 3.2 of coefficient of multiplication after two and a half months of subculture. Counting of cells was performed using Bürker cell (Marienfeld, United Kingdom). These suspensions showing cellular structures which were elongated and of rounded shape of varying size and viability were evaluated by fluorescein diacetate (FDA) under inverted microscope. From the first hours of incubation in the enzyme solution, the first protoplasts were released. The isolation of protoplasts from these suspensions allowed the obtaining of a large number of protoplasts containing large spherical nuclei. The viability is observed using fluorescein diacetate (FDA) (widhom, 1972). The best yields are obtained with solutions E1 (Cellulase 0.5%, Pectolyase Y23 0.5%, Hemicellulase 2%), E2 (Cellulase 2%, Pectolyase Y23 0.5% Hemicellulase 2%).
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