Thus, an efficient, fast, and easy system of plant regeneration from leaf- and hypocotyl-derived protoplasts is desired and would be valuable, e.g. To our knowledge, other types of carrot green tissue have not been used as donor material for protoplast isolation. In carrot, petiole-derived protoplasts were successfully used for callus development and then plant regeneration by indirect somatic embryogenesis (Dirks et al. Several types of plant material like leaves, cotyledons or hypocotyls are used as protoplast sources in many species. In contrast, leaf tissue can serve as an easy and direct source of protoplasts.
Despite the fact, that suspension cultures are an excellent source of protoplasts due to their high embryogenic ability, the establishment and maintenance of suspension culture is laborious and time consuming, usually requiring several weeks in addition to the time required for callus induction. To date, cell suspension cultures have been usually used as a protoplast donor material in carrot studies, especially in research on intra- and inter-specific as well intergeneric somatic hybridization (Dudits et al. At the same time, both cell division and plant development via direct somatic embryogenesis for protoplasts isolated from carrot cell suspension cultures were described (Grambow et al. Research on protoplast culture in carrot began when successful protoplast isolation from sliced, 50-day-old, field-cultivated roots was reported (Kameya and Uchimiya 1972). Since then, carrot has become a model species for plant tissue culture systems. Early reports indicated the induction of callus tissue from root discs or embryoids formation from free-floating somatic carrot cells (Gautheret 1939 Steward 1958). sativus Hoffm., 2n = 2x = 18) is one of the first plants used for successful growth in vitro.
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