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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.
Ganesan, Markkandan,Han, Yun-Jeong,Bae, Tae-Woong,Hwang, Ok-Jin,Chandrasekkhar, Thummala,Shin, Ah-Young,Goh, Chang-Hyo,Nishiguchi, Satoshi,Song, In-Ja,Lee, Hyo-Yeon,Kim, Jeong-Il,Song, Pill-Soon Springer-Verlag 2012 Planta Vol.236 No.4
<P>Phytochrome A (phyA) in higher plants is known to function as a far-red/shade light-sensing photoreceptor in suppressing shade avoidance responses (SARs) to shade stress. In this paper, the Avena PHYA gene was introduced into creeping bentgrass (Agrostis stolonifera L.) and zoysiagrass (Zoysia japonica Steud.) to improve turf quality by suppressing the SARs. In addition to wild-type PHYA, a hyperactive mutant gene (S599A-PHYA), in which a phosphorylation site involved in light-signal attenuation was removed, was also transformed into the turfgrasses. Phenotypic traits of the transgenic plants were compared to assess the suppression of SARs under a simulated shade condition and outdoor field conditions after three growth seasons. Under the shade condition, the S599A-PhyA transgenic creeping bentgrass plants showed shade avoidance-suppressing phenotypes with a 45?% shorter leaf lengths, 24?% shorter internode lengths, and twofold increases in chlorophyll concentrations when compared with control plants. Transgenic zoysiagrass plants overexpressing S599A-PHYA also showed shade-tolerant phenotypes under the shade condition with reductions in leaf length (15?%), internode length (30?%), leaf length/width ratio (19?%) and leaf area (22?%), as well as increases in chlorophyll contents (19?%) and runner lengths (30?%) compared to control plants. The phenotypes of transgenic zoysiagrass were also investigated in dense field habitats, and the transgenic turfgrass exhibited shade-tolerant phenotypes similar to those observed under laboratory shade conditions. Therefore, the present study suggests that the hyperactive phyA is effective for the development of shade-tolerant plants, and that the shade tolerance nature is sustained under field conditions.</P>
( 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.