Morfogeneza i regeneracija biljaka šalota (Allium ascalonicum L.) i vlašca (A. schoenoprasum L.) in vitro
In vitro morphogenesis and plant regeneration in shallot (Allium ascalonicum L.) and chive (A. schoenoprasum L.)
Doctoral thesis (Published version)
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Shallot (Allium ascalonicum L.) and chive (A. schoenoprasum L.) belong to the genus Allium, which represents one of the largest genera of the monocotyledonous plants. These economically important plant species have been used worldwide as culinary herbs due to their specific odor, taste and proven potent antioxidative capacity. Crop improvement using modern biotechnological approaches requires an efficient and reliable protocol for in vitro plant regeneration. Since no such protocols for in vitro regeneration of shallot and chive had been available, the main objective of this dissertation was to develop efficient protocols for the induction of bud/somatic embryo regeneration in these plant species.Regeneration of shoots/somatic embryos was induced from root sections of shallot and chive. In both species, only the root-tip sections had the regeneration capacity under the experimental conditions tested, but the regeneration proceeded via different pathways: indirect caulogenesis in shallot and indirect somatic embryogenesis in chive. The strong influence of genotype on regeneration capacity was observed in both species. The regeneration capacity was tested in 30 randomly chosen lines of shallot. A line constitutes the root-tip-derived regenerants originating from a single seed-derived plant. High variability in the frequency of regeneration (0.93-100%) and the mean bud number per explant (0.01-20.67) was observed among these lines. Among them, line 6 exhibited both the highest caulogenic and rizogenic capacity, and the fastest regeneration response. Using the root-tip-derived in vitro regenerated plants of line 6 as an explant source, the regeneration procedure was further optimized. Light intensity, the 2,4-dichlorophenoxyacetic acid (2,4-D)/6-benzyladenine (BA) ratio in callus induction medium and the duration of the callus induction phase significantly affected the caulogenic capacity of this line. Thus, the optimized protocol included a 5-week-long cultivation of the explants on callus induction medium supplemented with 5 μM 2,4-D and 5 μM BA followed by an 8-week-long cultivation on regeneration induction medium containing 5 μM BA, under light with a photosynthetic photon flux density of 100 μmol m-2s-1 during both phases. Using this protocol, a 100% frequency ofregeneration and 18.4 buds per explant were attained in line 6 after 13 weeks of treatment...
Keywords:Adventive buds; Allium ascalonicum; A. schoenoprasum; Antioxidative enzyme; Apical root sections; Basal plate; Cytokinins; Caulogenesis; Growth regulators; Somatic embryogenesis
Source:University of Belgrade, Faculty of Biology, 2016, 1-189