• Journal of Ginseng Research
• /
• v.15 no.2
• /
• pp.124-130
• /
• 1991

This study was conducted to obtain basic information about the transformation of ginseng tissue, identification of opine compound and protein, and saponin production from ginseng callus transformed with Ti-plasmic of AW$.$obacterium tumefaiens C58. Ginseng crown gall callus induced by pTiC58 could be continuously cultured on the Phytohormone-free medium. The transformation was reconfirmed by the detection and identification of opine compound, from the gall callus. The transformed ginseng callus contained higher amounts of protein than normal callus and the protein pattern of transformed callus was quite different from that of normal callus. The xylose which is not detected in the normal callus and ginseng root was identified in gall callus. The saponin contents of gall callus of ginseng were three times higher than that of normal callus, and ginsenoside composition of the transformed callus was similar to that of the cultivated ginseng root, but quite different from that of normal callus.

• Korean Journal of Plant Tissue Culture
• /
• v.25 no.3
• /
• pp.177-181
• /
• 1998

This study was carried out to investigate activity difference in the superoxide dismutase (SOD) and peroxidase (POD) of tomato callus transformed with Agrobacterium containing the GUS gene. Than those of other two tomato cultivars, the hypocotyl explant of JA101 was shown to have higher POD and SOD specific activity of 23 unit/mg protein and 2,156 unit/mg protein, respectively. Relatively high frequency of callus formation was obtained from the hypocotyl explant on MS medium containing 1 mg/L 2,4-D for 30 days and its POD(47 unit/mg protein) and SOD (95,786 unit/mg protein) specific activities were higher than other 2,4-D concentration. The hypocotyl explant and callus cocultivated with Agrobacterium for 72 hours were transferred to MS medium supplemented with 1 mg/L 2,4-D, 30 mg/L kanamycin, 30 g/L sucrose and 4 g/L Gelrite. The hypocotyl explants transferred to the medium formed callus with 45.5% effeciency after 8 weeks. The transformation efficiency confirmed by GUS assay was 21.6%. POD specific activity of the transformed callus (54 unit/mg protein) were somewhat lower than the non-transformed callus (64 unit/mgg protein) and SOD specific activity of the transformed callus (30,300 unit/mg protein) were also lower than the non-transformed callus (37,077 unit/mg protein). However there was no significant difference in POD and SOD isozyme patterns between the transformed and the non-transformed calluses. From these results, it revealed that there was no difference of antioxidant enzyme activities between the transformed callus and the non-transformed callus in tomato.

• Journal of The Korean Society of Grassland and Forage Science
• /
• v.21 no.3
• /
• pp.145-150
• /
• 2001

This study was conducted to obtain the transformed birdsfoot trefoil (Lotus corniculatus L.) plants with BcHSP17.6 gene using Agrobacterium turnefaciens LBA4404 and we confirmed transformed gene from the regenerated birdsfoot trefoil plants. The expression vector, pBKH4 vector, harboring BcHSP17.6 gene was used for production of transgenic birdsfoot trefoil plants. The callus of birdsfoot trefoil was cocultivated with Agrobacteriurn turnefaciens and transformed calli were selected on kanamycin-containing SH-kc medium to regenerate into plants. The transformed birdsfoot trefoil plants were produced 4 momths after cultivation on BOi2Y medium. The transgenic birdsfoot trefoil plants were analyzed by isolation of genomic DNA and genomic Southern hybridization using a -32P labelled BcHSPl7.6 fragments. (Key words : Birdsfoot trefoil, Transgenic plant. BcHSP17.6 gene, Callus induction, Plant regeneration)

• Plant Biotechnology Reports
• /
• v.4 no.4
• /
• pp.253-260
• /
• 2010

Plant secondary metabolites have always been a focus of study due to their important roles in human medicine and nutrition. We transferred the isoflavone synthase (IFS) gene into soybean [Glycine max (L.) Merr.] using the Agrobacterium-mediated transformation method in an attempt to produce transformed soybean plants which produced increased levels of the secondary metabolite, isoflavone. Although the trial to produce transgenic plant failed due to unestablished hygromycin selection, transformed callus cell lines were obtained. The induction rate and degree of callus were similar among the three cultivars tested, but light illumination positively influenced the frequency of callus formation, resulting in a callus induction rate of 74% for Kwangan, 67% for Sojin, and 73% for Duyou. Following seven to eight subcultures on selection media, the isoflavone content of the transformed callus lines were analyzed by high-performance liquid chromatography. The total amount of isoflavone in the transformed callus cell lines was three- to sixfold higher than that in control callus or seeds. Given the many positive effects of isoflavone on human health, it may be possible to adapt our transformed callus lines for industrialization through an alternative cell culture system to produce high concentrations of isoflavones.

• Journal of the Korean Society of Tobacco Science
• /
• v.13 no.2
• /
• pp.34-41
• /
• 1991

To explore the possibility of introducing Zea mays transposable element Ac(activator) which can be used as a mutagen and gene tag in tobacco plants other than maitre, we tried to introduce a cloned Ac element into tobacco cells by an Agrobacterium tumefaciens binary vector system. Transformation of N. babacum cv. Burley 21 tissues and regeneration to whole plant were carried out. The frequency of the transformed callus induced in shoot induction media was higher than that of transformed callus induced in callus induction media. However, the calli were not grown in the second selection media, and became yellow senescent calli. Regenerated tobacco plantlets with foreign gene were also obtained in shoot induction media containing 100 $\mu\textrm{g}$/ml kanamycin and 100$\mu\textrm{g}$/ml carbenicillin. The leaf tissues of transformant was also resistant to 1000 $\mu\textrm{g}$/ml kanamycin. The chromosomal DNAs of transformant and normal plant of N. tabacum were digested by EcoR I and Hind III but not by Pst I.

• Korean Journal of Breeding Science
• /
• v.42 no.4
• /
• pp.351-356
• /
• 2010

Brachypodium distachyon is rapidly emerged in biological study and has been currently used as a model system for genetics and functional studies for crop improvement and biofuel production. Phosphinothricin (PPT) has been widely used as a selectable agent, which raises ammonium content and induces toxicity in non-transformed plant cells. However PPT selection is not much effective on Brachypodium callus consequently reducing transformation efficiency. In order to identify the efficient conditions of PPT selection, calli obtained from mature seeds of Brachypodium (PI 254867) were cultured on the callus inducing medium (CIM) or regeneration medium (ReM) containing serial dilutions of the PPT (0, 2, 5, 10, and 15 mg/l) in dark or light condition. Callus growth and ammonium content of each treatment were measured 2 weeks after the treatment. Although callus growth and ammonium content did not show much difference in CIM, slow callus growth and increased ammonium accumulation were found in ReM. No significant difference of ammonium accumulation in response to PPT was found between dark and light conditions. In order to identify major factors affecting increased ammonium accumulation, callus was cultured on the media in combined with phytohormones (2,4-D or kinetin) and carbon sources (sucrose or maltose) containing with PPT (5 mg/l). The highest ammonium content in callus was found in the kinetin and maltose media.

• Journal of Plant Biology
• /
• v.36 no.1
• /
• pp.43-49
• /
• 1993

In order to survey possibility to produce saikosaponin from in vitro hairy root culture, culture of callus, adventitous root, and hairy root of Bupleurum falcatum L. were estabilished, and quantitative and qualitative aspects in saikosaponin extracted from these were compared with those of cultivated root. Callus grew well in MS medium containing 0.9 $\mu$M 2, 4-D. In contrast, both of adventitous root and hairy root grew well in hormone-free MS medium. However, hairy root showed more rapid growth with extensive lateral root branches, characteristics of lower content of water and softer than in adventitous root. Among the selected lines of adventitous root and hairy root were observed difference in the growth rate. Mannopine, one of opine synthesized in the transformed tissue with Agrobacterium rhizogenes. A4 were detected in the extract of hairy root lines. Pattern and content of crude saponin from adventitous and hairy root showed no difference, but somewhat difference from those of cultivated root. However, in callus, distinct production-aspect of saponin was not observed.

• Journal of Plant Biotechnology
• /
• v.2 no.3
• /
• pp.135-142
• /
• 2000

The activity of promoter sequences was evaluated in garlic cells using the $\beta$-glucuronidase (GUS) gene as a reporter. Histochemical GUS assay indicated transient GUS activity in leaf, callus and root cells 48 hours after particle bombardment transformation. Quantitative fluorometric assays in extracts of transformed leaves demonstrated that the CsVMV promoter induced the highest level of gene expression, which was, on average, ten fold the level induced by CaMV35S and by the Arabidopsis Act2 promoters and two fold the level expression observed with a construct containing a double CaMV35S plus the untranslated leader sequence from AMV. No activity or very low levels were observed when cells were transformed with plasmids rontaining the typical monocot promoters, Actl, from rice or the Ubi-1, from maize. The green fluorescent protein (GFP) was also tested as a marker gene for garlic transformation. Intense fluorescence was observed in leaf, callus and root cells transformed with a construct containing the gfp gene under control of the CaMV35 Promoter. No fluorescence was detected when the gfp was under control of the Ubi-1 promoter.

• Journal of Plant Biology
• /
• v.39 no.2
• /
• pp.93-98
• /
• 1996

In order to enhance the system of potato transformation and further regeneration, potato was transformed using the Agrobacterium tumefaciens harboring $\beta$-glucuronidase (GUS) gene. We found that a series fo modified medium ttained 100% shoot regeneration within 5 weeks after the preincubated explants on stage I medium were infected with Agrobacterium. Callus appeared at the cut edges of stem segments on stage II medium, mainly at the basal parts. Some explants started to form shoots after two to three weeks on stage III medium containing kanamycin (50 mg/L). When transferred to MS medium containing 200 mg/L kanamycin, 81% of the transformed shoots formed roots at the cut edge of the plantlets. In contrast, untrasformed shoots never rooted and became yellowish after few weeks under the same conditions. Southern and northern analysis indicated in vitro shoot regeneration on the callus derived from the potato explants, which were incubated with Agrobacteria. The regeneration cycle was shortened after the transformatin and finally the transformation efficiency was highly enhanced.

• Journal of Plant Biotechnology
• /
• v.31 no.1
• /
• pp.13-17
• /
• 2004

To improve transformation efficiency, the callus of Lilium lancifolium Thunb. were bombarded by particles coated with pIG 121 Hm which include NPT II and GUS genes, and then cocultivated with Agrobacterium tumefaciens EHA101 which contain pIG121Hm binary vector, carrying neomycin phosphotransferase (NPT II) and $\beta$-Glucuronidase (GUS) genes. Three days after cocultivation with Agrobacterium tumefaciens and particle bombardment, the callus clusters were transferred to MS medium containing 1mg/L 2,4-D, 0.1mg/L BAP, 100mg/L kanamycin and 200mg/L carbenicillin. Four weeks after transfer to the selection medium, GUS expression was detected and PCR analysis revealed the presence of NPT II fragment of the expected size (700 bp) in the transformed callus. The GUS expression from Agrobacterium-mediated transformants after particle bombardment increased to over 3-folds compared with that of callus cocultivated with Agrobacterium tumefaciens without particle bombardment. The callus clusters containing kanamycin resistant gene were transferred to MS medium containing 1mg/L NAA and 1mg/L BAP. Somatic embryos were developed in four weeks and microbulbs expressing GUS were formed.