• Journal of Plant Biotechnology
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• v.31 no.4
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• pp.279-283
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• 2004

To lily (Lilium longflorum cv. Georgia) pollen, impacts by some physical, chemical and biological factors were examined in respects of its growth and transient gene expression via agro-infiltration. Rolling movement in liquid medium or vacuum pressure during Agro-infiltration was regarded as a impact that should be minimized for normal pollen growth. Pollen growth was maintained well in relatively broad range of temperature (19 to 27$^{\circ}C$) or pH (5.0 to 8.0). Chemical factors such as cefotaxime (up to 300mg/L), acetosyringone (up to 800 $\mu$M) and syringealdehyde (up to 800 $\mu$M) did not show any harmful effects but kanamycin severely did even at concentration as low as 25mg/L in some cases. For GUS gene expression, acetosyringone at 200 to 400 $\mu$M slightly improved the efficiency while syringealdehyde did not. Brief agro-infiltration followed by 18 hr of co-incubation of pollen along with Agrobacterium was suggested as a condition basically required for the transient expression system using lily pollen regardless of the presence of acetosyringone.

• Journal of agriculture & life science
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• v.44 no.1
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• pp.9-15
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• 2010

Barley, a monocotyledonous plant, is relatively recalcitrant to the process of Agrobacterium-mediated genetic transformation. In this study, seedlings of six barley cultivars (Keunal-1-Ho, Saessal, Ol, Saechalssal, Seodunchal and Pungsanchalssal) were injured using alkali, oxidizing or reducing agents. They were then transformed using Agrobacterium via vacuum infiltration for the analysis of comparative GUS gene expression. It was determined that chemical injuries causing a slight growth retardation could overall enhance the GUS transformation rate. Hydrogen peroxide was determined to be the most effective.

• KSBB Journal
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• v.21 no.6 s.101
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• pp.489-492
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• 2006

For molecular breeding purpose, genetic transformation of Brassica napus cultivars has been extensively performed using Agrobacterium method. B. napus cv. napus, one of major oil crops, can be transformed via Agrobacterium-based method. We demonstrated that Agrobacterium-mediated transformation via vacuum infiltration slightly worked for the seedlings of B. napus cv. napus according to fluorometric GUS enzyme analysis. In contrast, transformation efficiency was highly enhanced when the seedlings, prior to agroinfiltration, were treated with sodium hydrosulfite solution as a chemical wounding agent. GUS gene expression in transformed seedlings that was confirmed by RT-PCR suggests their usefulness for the development of transient expression system.

• Journal of Life Science
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• v.15 no.6 s.73
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• pp.1034-1036
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• 2005

Chinese cabbage (Brassica campestris ssp. napus var. pekinensis Makino) seeds/seedlings were transformed via vacuum-infiltration with recombinant Agrobacterium tumefaciens LBA4404 cells. The agroinfiltration method was determined to be unsuccessful for Chinese cabbage transformation during the analysis of hepatitis B surface antigen expression by ELISA. However, treatment of sodium hypochlorite solution, prior to agroinfiltration, to pregerminated or germinating 1 day- or 2 days-old seeds was proven effectively to enhance transformation efficiency, suggesting that chemical wounding caused by sodium hypochlorite reaction might facilitate Agrobacterium infection and, therefore, transient gene expression in Chinese cabbage sprouts.

• Journal of Plant Biotechnology
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• v.34 no.4
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• pp.293-298
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• 2007

Robinia pseudoacacia var. umbraculifera, commonly called umbrella black locust were regenerated after co-cultivation of internode segments with Agrobacterium tumefaciens which included yeast cadmium factor 1 (YCF 1) gene. The tolerance to cadmium and lead for plants can be increased by the YCF1 gene expression. Moreover, the recent studies have shown that YCF1 gene transgenic plants increase the accumulation of cadmium and lead into plant vacuoles. The effect of plant growth regulator such as 2,4-dichlorophenoxyacetic acid (2,4-D), ${\alpha}$-naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and thidiazuron (TDZ) were studied to evaluate the propagation of plants through internode explants. The efficient induction of multiple adventitious shoots and callus were observed on a medium supplemented with 0.1 mg/L TDZ + 0.2 mg/L BA. To induce shoot elongation and rooting, regenerated shoots were transferred into basal MS medium without any plant growth regulator. Successful Agrobacterium tumefaciens mediated transformation was obtained by 20 min vacuum-infiltration with $50{\mu}M$ acetosyringone on the optimal multiple shoot induction medium with 30 mg/L hygromycin and 300 mg/L cefotaxime. To confirm the integration and expression of transgene, Polymerase Chain Reaction (PCR) and Reverse Transcriptase PCR (RT-PCR) were performed with specific primers. The frequency of transformation was approximately 18.94%. This study can be used to genetic engineering of phytoremediator.