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Kyung Moon Kim,P. Stephen Baenziger,Thummala Chandrasekhar,Hyo Yeon Lee 한국육종학회 2007 한국육종학회지 Vol.39 No.2
The effect of osmotic condition on β-glucuronidase (GUS) transient expression was evaluated in microspore-derived embryos of wheat. Microspore explants were treated on medium containing various mannitol concentrations prior to and post bombardment with plasmid DNA pAHC25 containing uidA gene controlled by maize ubiquitin 1 (UBI1) promoter. GUS expression in the bombarded explants was examined by histochemical and fluorometric assays. Increased GUS expression was observed with mannitol treatment when compared to untreated explants. The histochemical study showed that the number of blue (GUS) foci were the highest in the bombarded explants treated with 0.6 M mannitol medium. The fluorometric assay of bombarded explants also proved 3.5-fold increase in GUS activity with 0.6 M mannitol treatment when compared to without mannitol treatment. These results indicate that 0.6 M mannitol is beneficial for improving transformation efficiency of wheat microspore-derived embryos or embryogenic calli through biolistic transformation.
( Kyung Moon Kim ),( Min Young Kim ),( Pil Yong Yun ),( Thummala Chandrasekhar ),( Hyo Yeon Lee ),( Pill Soon Song ) 한국식물학회 2007 Journal of Plant Biology Vol.50 No.4
High frequency of multiple shoots and plant regeneration has been obtained from the leaf segments of fig tree (Ficus carica L.). Budbreak from dormant buds is highly dependent upon cultivar, so we chose cv. Seungjung Dauphine because it shows an excellent degree of budbreak. Tissue-browning can be an important limiting factor during in vitro culture. This phenomenon could be substantially delayed or reduced by treating the tissues with 0.5 mM phloroglucinol, thus oxidizing the phenolic substances exuded from the segments. Wounded leaf explants cultured on MS medium supplemented with TDZ in combination with IBA produced more multiple shoots than did other combinations of auxin and cytokinin. For example, 2 mg L-1 IBA along with either 0.5 or 1.0 mg L-1 TDZ resulted in 8.1 or 10.8 multiple shoots per explant, respectively. We achieved a frequency of approximately 90% when tissues were first maintained under darkness in the culture medium for one week before being transferred to the light. Regenerated shoots rooted best in a full-strength MS basal medium. In vitro regenerated plantlets were then successfully transferred to greenhouse conditions. Here, we have demonstrated a regeneration protocol that is suitable for use in conservation as well as genetic transformation studies of figs and related species.