• Journal of Marine Bioscience and Biotechnology
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• v.1 no.4
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• pp.260-267
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• 2006

The Liquid Culture Assay using a recombinant Agrobacterium tumefaciens strain has been developed as a means for quorum sensing autoinducer screening. However, the low aqueous solubility of marine natural product extracts used as potential autoinducers has been a hindrance in the screening process. Although the addition of organic solvents and/or surfactants could increase aqueous solubility, errors in data interpretation including false positive results could be a serious problem. Therefore, determining the best possible solvent and surfactant at the optimum concentration is crucial. Evaluating methanol, ethanol, 1-propanol, DMSO and DMF at concentration ranges of 0~10% revealed < 2% methanol to be most favorable when tested for ${\beta}$-gal activity and growth inhibition of the recombinant A. tumefaciens strain. On the other hand, among surfactants tested, Triton X-100 was similarly effective in increasing the delivery of autoinducers for activity at less than 0.05% concentration.

• Journal of Microbiology and Biotechnology
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• v.18 no.7
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• pp.1245-1251
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• 2008

Oxalate decarboxylases (OXDCs) (E.C. 4.1.1.2) are enzymes catalyzing the conversion of oxalate to formate and $CO_2$. The OXDCs found in fungi and bacteria belong to a functionally diverse protein superfamily known as the cupins. Fungi-originated OXDCs are secretory enzymes. However, most bacterial OXDCs are localized in the cytosol, and may be involved in energy metabolism. In Agrobacterium tumefaciens C58, a locus for a putative oxalate decarboxylase is present. In the study reported here, an enzyme was overexpressed in Escherichia coli and showed oxalate decarboxylase activity. Computational analysis revealed the A. tumefaciens C58 OXDC contains a signal peptide mediating translocation of the enzyme into the periplasm that was supported by expression of signal-peptideless and full-length versions of the enzyme in A. tumefaciens C58. Further site-directed mutagenesis experiment demonstrated that the A. tumefaciens C58 OXDC is most likely translocated by a twin-arginine translocation (TAT) system.

• Journal of Plant Biology
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• v.33 no.2
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• pp.97-104
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• 1990

In order to investigate the host range of Agrobacterium tumefaciens KU12 containing pTi-12, 28 species of dicotyledonous plants were infected with KU12, A136 without Ti plasmid and A348 containing pTi A6, respectively. KU12 and A348 induced tumor in 20 species and 14 species, respectively. This results showed that KU12 has a wide host range. Therefore, it was confirmed that KU12 and pTi-12 are very useful for developing plant vector system having a broad host range.

• Korean Journal of Microbiology
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• v.32 no.1
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• pp.53-59
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• 1994

Agrobacterium tumefaciens KU12 contains pTiKU12 (240kb) of the octopine type Ti plamsid and pTi12 (45 kb) of the cryptic plasmid. To make the avirulent A. tumefaciens, the octopine type Ti plasmid, pTiKU12, was cured with elevated temperature (37${\circ}C$) and ethidium bromide (EtBr), respectively. Also the cryptic plasmid, pTi12, was cured by the introduction of recombinant plasmid, pYWXP, made by pTi12 replication origin and pUC19. pYWXP was cured by elevated temperature (37${\circ}C$) and EtBr simultaneously.

• Journal of Plant Biology
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• v.39 no.3
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• pp.179-184
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• 1996

The purpose of this study is to obtain the most efficient combination of Agrobacterium tumefaciens strains and Ti plasmids for the construction of dicotyledonous plant vector system. Ti plasmid-curing A. tumefaciens A136 and KU12C3 were transformed with four kinds of Ti plasmids, pTiBo542, pTiA6, pTiKU12 and pTiAch5, respectively. The stems of 28 species of dicotyledonous plants were then inoculated with these transformants and examined for crown gall formation. The different combination of A. tumefaciens strains and Ti plasmids showed quite a difference in terms of the crown gall formation. Agrobacterium strins A136 and KU12C3 have a same plant host range in case that both strains harour the same kind of Ti plasmid, pTiBo542 or pTiAch5. However, the above-mentioned both strains have quite different host range in the event of containing the same Ti plasmid, pTiKU12 or pTiA6. In case that KU12C3 contains pTiA6 or pTiKU12, this strain has a wider plant host range than A136. The plant host range of pTiBo542 is the widest, followed by pTiA6, pTiKU12 and pTiAch5. Twelve plants among 28 tested plants are not transformed by any virulent Agrobacterium strains used in this study. In conclusion, A. tumefaciens KU12C3 and A136 harboring pTiBo542 showed the widest host range for transforming dicotyledonous plants. Also, it was acertained that the host range of Ti plasmids is affected by chromosomal level.

• Korean Journal of Microbiology
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• v.25 no.1
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• pp.17-22
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• 1987

For the purpose of securing of strains which can be usefully utilized to study symbiosis between Rhizobium and legume plant, A. tumefaciens T7 was isolated and characterized and then subgroup biovar was determined. A. tumefaciens T7 induced smooth tumor like nopaline type one and did not grow at $37^{\circ}C$ and in the presence of 2% NaCl on yeast extract mannitol medium. The strain was able to grow on the New and Kerr selective media and utilize erythritol but not phenylalanine, tryptophan, and tartarate as a sole carbon source. Negative results were obtained from 3-keto-lactose production and oxidase test. The strain produced alkalifrom malonate and citrate and showed acid litmus milk reaction At least two large plasmids were detected in the cell lysate. According to all of these results, it could be concluded that subdivision of isolated strain was biovar 2.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.10
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• pp.1390-1394
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• 2004

We have achieved efficient transformation system for forage-type tall fescue plants by Agrobacterium tumefaciens. Mature seed-derived embryogenic calli were infected and co-cultivated with each of three A. tumefaciens strains, all of which harbored a standard binary vector pIG121Hm encoding the neomycin phosphotransferase II (NPTII), hygromycin phosphotransferase (HPT) and intron-containing $\beta$-glucuronidase (intron-GUS) genes in the T-DNA region. Transformation efficiency was influenced by the A. tumefaciens strain, addition of the phenolic compound acetosyringone and duration of vacuum treatment. Of the three A. tumefaciens strains tested, EHA101/pIG121Hm was found to be most effective followed by GV3101/pIG121Hm and LBA4404/pIG121Hm for transient GUS expression after 3 days co-cultivation. Inclusion of 100 $\mu$M acetosyringone in both the inoculation and co-cultivation media lead to an improvement in transient GUS expression observed in targeted calli. Vacuum treatment during infection of calli with A. tumefaciens strains increased transformation efficiency. The highest stable transformation efficiency of transgenic plants was obtained when mature seed-derived calli infected with A. tumefaciens EHA101/pIG121Hm in the presence of 100 $\mu$M acetosyringone and vacuum treatment for 30 min. Southern blot analysis indicated integration of the transgene into the genome of tall fescue. The transformation system developed in this study would be useful for Agrobacterium-mediated genetic transformation of tall fescue plants with genes of agronomic importance.

• Journal of Microbiology and Biotechnology
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• v.18 no.4
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• pp.754-758
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• 2008

Agrobacterium tumefaciens-mediated transformation (ATMT) was successfully applied to Monascus ruber. The optimum cocultivation time was 84 h with an efficiency of 900 to 1,000 transformants when $1{\times}10^6$ spores were used with the same volume of bacteria. The stability of transform ants was over 98% after five generations. When M. ruber was transformed with A. tumefaciens YL-63 containing the green fluorescent protein gene (egfp), the green fluorescent signal was observed throughout hyphae, confirming expression of the gene. This efficient transformation and expression system of M. ruber by ATMT will facilitate the study of this fungus at a molecular genetic level.

• Journal of Ginseng Research
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• v.10 no.1
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• pp.45-54
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• 1986

These studies were carried out to obtain the basic information about transformation of ginseng plant by potential vector system, utilization of opine compound by Agrobacterium sap. , and initiation of crown gall tumor callus. Crown gall tumors were induced from stem of Panax ginseng C.A. Meyer by infection of Agrobacterium tumefaciens. Therefore, it was clarified that transformation of ginseng by Ti plasmid was possible. The crown gall tumors induced by Agrobacterium tumefaciens isolated. from the soil were different in a shape, size, and growth rate. Especially, infection of ginseng by Agrobacterium tumefaciens Y104 led to the amorphic tumor, Tumor tissue derived from stem crown gall could not be continuously cultured on the medium which did not contain phytohormone, and did not form the callus even on the medium supplemented with 2,4-D. On the other hand, the root crown gall tumors formed the calli but the formation rate of callus was quite low. As for the utilization of octopine and nopaline, it was found that 3 strains of Agrobacterium app., Y104, Y110 and C58, utilized nopaline only, Y109 utilized octopine, and Y101 failed to utilize either compound.

• The Korean Journal of Pesticide Science
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• v.2 no.2
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• pp.97-101
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• 1998

Agrobacterium vitis causes a crown gall disease in grapevine and that is one of the major hindrances for the wide cultivation and production of grapevine. We studied the possibility of biological control using selected biological control agent. One isolate from the infected soil, named as strain 27, was able to inhibit the biovar 1; A. tumefaciens C58 and Ach5, biovar 2; A. rhizogenes 13264, and biovar 3; A. vitis, in vitro and in vivo test. The putative biological control agent, A. radiobacter strain 27 was carrying the plasmid and the size of isolated plasmid was very similar to that of pAgK84 of A. radiobacter K84.