Mahdi Ghabouli; Ahmadreza Bahrami; Farajollah Shahriari; Jafar Zolala; Ali Mohammadi
Abstract
Plant tissue culture techniques are used as basic requirement of common plant transformation systems. In most cases of plant transformation, a reproducible regeneration protocol is the limiting step due to long time lasting, specialized facilities and well experienced persons. Furthermore, tissue culture ...
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Plant tissue culture techniques are used as basic requirement of common plant transformation systems. In most cases of plant transformation, a reproducible regeneration protocol is the limiting step due to long time lasting, specialized facilities and well experienced persons. Furthermore, tissue culture procedures induce somaclonal variation among regenerated transgenic plants. Therefore, recently current studies in plant molecular biology prefer plant transformation procedures avoiding tissue culture phase. Various in plant a transformation procedures have been explored, among which the floral dip method is the most reliable in vivo transformation method. In this research, with the aim of evaluating the ability of floral dip method for genetic transformation of some Apiaceae plants, we studied Dill (Anethum graveolens), Fennel (Foeniculum vulgare), Coriander (Coriandrum sativum), Carrot (Daucus carrota), Parsley (Petrocelium sativum) and Celery (Apium graveolens). Arabidopsis thaliana was used as a model plant of experimental procedure. Flowers, in different stages of inflorescence development, were immersed in different suspension of Agrobacterium tumefaciens carrying the plant binary vector pBI121. This vector carries plant reporter gene uidA (gus) and the plant selectable marker gene npt II. Although, producing transgenic Arabidopsis plants with a high transformation rate of 4% verified the accuracy of experimental procedure, floral dip method was not successful for transformation of Apiaceae plants. Only one transformed celery plantlet, carrying nptII gene with no expression of GUS, was obtained bby screening more than 10000 seeds produced by treated plants from all the species. Transgenic Arabidopsis plants expressing gus reporter gene were confirmed through PCR and histochemical assays.
