• 미생물학회지
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• 제25권1호
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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.

• 식물조직배양학회지
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• 제22권4호
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• pp.195-200
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• 1995

동물유전자인 mouse adenosine deaminase (ADA)유전자가 안정적으로 연초 형질전환체에서 발현되었다. ADA cDNA 을 연초에서 강력히 발현시키기 위해서 35S/35S/AMV promoter을 부착시켰으며 식물세포에 도입을 위해서 binary vector인 pRD400을 이용하였다. 연초의 형질전환은 Tri- parental mating에 의해서 도입된 ADA 유전자 함유 binary vector와 disarmed Ti-plasmid을 함유하고 있는 Agrobacterium tmefacience MP9O을 사용하였다. 동시배양은 연초의 잎 disc을 이용해서 kanamycin이 첨가된 배지로부터 직접 shoots을 선발하여 형질전환체로 유도하였다. 형질전환 체에 ADA 유전자의 삽입여부는 PCR을 이용하였으며, 실험결과 ADA 유전자를 확인할 수 있었다. 또한 도입된 mouse ADA 유전자로부터 mRNA 및 단백질 합성여부를 각각 northern blot 및 immunoblot 분석한 결과 형질전환체 에서는 공히 확인되었다.

• The Plant Pathology Journal
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• 제39권2호
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• pp.220-227
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• 2023

Rose crown gall caused by Agrobacterium tumefaciens is a major disease that damages the production of cutroses in Korea. The effective prevention methods for this disease include the use of resistant varieties. This study was conducted to evaluate the resistance of 58 Korean cultivars and six foreign cultivars to crown gall disease with nodal explants in vitro. Among 180 A. tumefaciens strains, pathogenic strain RC12 was selected as an inoculant strain. The strain RC12 was identified based on characteristics of some selective media, pathogenicity test, and polymerase chain reaction analysis. Forty rose cultivars formed tumors on explants inoculated with A. tumefaciens RC12. However, 24 cultivars, including 22 Korean cultivars and 2 foreign cultivars, showed resistance to A. tumefaciens RC12 without forming any tumors. Six cultivars with tumor formation rates of over 30% formed initial tumors within 23 days after inoculation. Six cultivars with low tumor formation rates of around 5% formed initial tumors after 28 days of inoculation. It was found that gall formation rate was highly correlated with the initial gall formation period. Thus, the relationship between the period of gall formation and the rate of gall formation could be useful for assessing resistance to crown gall disease. In vitro inoculation methods could be used to evaluate resistance of cut-rose cultivars to crown gall diseases.

• Journal of Plant Biotechnology
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• 제2권3호
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• pp.143-149
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• 2000

An Efficient in vitro regeneration system and an optimized Agrobacterium mediated transformation protocol are described, based on the use of young seedling cotyledons of Capsicum annuum L. Optimal regeneration efficiency can be obtained by cultivating cotyledon explants on media containing 4 mg/L benzyladenine and 0.1 mg/L indolacetic acid. The effect of antibiotics used to eliminate Agrobacteria, as well as the toxic level of some generally used selection agents (kanamycin, geneticin, hygromycin, phosphinotricin and methotrexate) in regenerating pepper tissues were determined. To enable the comparison of different selection markers in identical vector background, a set of binary vectors containing the marker genes for NPTII, HPT, DHFR and BAR respectively, as well as the CaMV 35S promoter/enhancer-GUS chimaeric gene was constructed and introduced into four different Agrobacterium host strains.

• 한국환경과학회지
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• 제15권12호
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• pp.1185-1191
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• 2006

The high surface area polymer, AquaMats$^{circledR}$ was used for the leachate purification process originated from the Samsan Dong Landfill in Ulsan Metropolitan City. And then, three species of dominant microbes were isolated and identified from AquaMats$^{circledR}$. Gram staining revealed these microbes to be Gram-negative rod strains: They were identified as Agrobacterium radiobacter, Pseudomonas cepacia, Flavobacterium indologenes. All they showed no growth on media in which the leachate was added alone, but a rapid proliferation rate on media with yeast extract as nutrient requirements.

• Journal of Plant Biology
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• 제39권3호
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• pp.161-166
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• 1996

The hairy root cultures of Centella asiatica were established by infection leaf explants with Agrobacterium rhizogenes A4, 15834 in 1/2 Murashing and skoog liquid medium supplemented with 50 $\mu$M acetosyringone. The induced hairy roots were subjected to paper electrophoresis for the detection of opine and opine-positive clones which were considered to have been transformed. Five hairy root clones were selected according to the different bacterial strains used, growth rate and pattern. Among media tested, MS basal medium substituted phosphate concentration by 2.5mM K2HPO4 showed the highest growth rate in the dark condition.

• Mycobiology
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• 제43권1호
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• pp.1-8
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• 2015

Mushroom transformation requires a series of experimental steps, including generation of host strains with a desirable selective marker, design of vector DNA, removal of host cell wall, introduction of foreign DNA across the cell membrane, and integration into host genomic DNA or maintenance of an autonomous vector DNA inside the host cell. This review introduces limitations and obstacles related to transformation technologies along with possible solutions. Current methods for cell wall removal and cell membrane permeabilization are summarized together with details of two popular technologies, Agrobacterium tumefaciens-mediated transformation and restriction enzyme-mediated integration.

• Journal of Plant Biotechnology
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• 제38권4호
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• pp.308-314
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• 2011

Efficient transformation and regeneration methods are a priority for successful application of genetic engineering to vegetative propagated plants such as grape. In this study, methods for Agrobacterium tumefaciens-mediated transformation and plant regeneration of grapevine (Vitis vinifera) were evaluated. Tamnara, Heukgoosul, Heukbosek, Rizamat were co-cultivated with Agrobacterium strains, LBA4404 containing the vector pBI121 carrying with CaMV 35S promoter, GUS gene as reporter gene and resistance to kanamycin as selective agent. Seven percent of the maximum regeneration frequency was obtained from co-cultivated with explants from Rizamat with LBA4404 strain on selection medium with kanamycin. The addition of acetosyringone, 200 ${\mu}m$ in virulence induction step was a key factor for successful GUS reporter gene expression in grapevine transformation. Transgenic plants showed resistance to kanamycin and the GUS positive response in leaf ($T_0$) stem ($T_0$) and petiole ($T_0$).

• Journal of Microbiology and Biotechnology
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• 제9권5호
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• pp.637-645
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• 1999

Two putative regulator genes, mocR and mocS, of the moc (mannityl opine catabolism) operons in pTi15955 of the octopine-/mannityl opine-type Agrobacterium tumefaciens strain 15955, were tested for their possible roles as repressors in the moc operons. The regions upstream of macC and mocD, the first structural genes in the two divergently oriented moc operons, were transcriptionally fused into the promoterless lacZ reporter gene. Each of the lacZ-fusions was introduced into Agrobacterium strain UIA5, a Ti plasmid-cured derivative, harboring either a mocR or a mocS clone. The resulting strains were grown in media containing various sugar sources, and the $\beta$-galactosidase activities were quantitatively measured. The results suggested that MocR repressed the expression of macC and macD. The expression of the fused $\beta$-galactosidase was not induced by mannopine (MOP) or possible catabolic intermediates of the opine, e.g. santhopine (SOP), glucose, mannose, or glutamine. However, the repression was significantly relieved by the supplementation of MOP and the concomitant introduction of the agcA gene encoding MOP cyclase that catalyzes the lactonization of MOP to agropine (AGR). These results suggested that AGR, rather than MOP or the other catabolic intermediates, is the inducer for the expression of the operon. On the contrary to previous report showing that the induction levels of macC and macD were lowered by the supplementation of inorganic nitrogen in media, the expression of these genes was not affected by the level of nitrogen in our reporter system. MocS did not strongly repress the expressions of macC and mocD. It is possible that MocS may be involved in the regulation of the operons present downstream of the moc operon, which are responsible for the utilization of mannopinic acid and agropinic acid.

• 한국토양비료학회지
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• 제20권4호
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• pp.359-367
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• 1987

수종(數種)의 유기산(有機酸), 당류(糖類)(단당(單糖), 복합당(複合糖)) 및 수도근계분비물질(水稻根系分泌物質)에 대(對)한 협생질소고정균(協生窒素固定菌)의 화학주화성(化學走化性)을 검토(檢討)하기 위하여 우리나라 서해간척지(西海干拓地)의 잡초(雜草) 및 수도(水稻) 근조직(根組織)에서 분리동정(分離同定)한 균(菌)을 공시(供試)하여 시험(試驗)하였다. 공시(供試)한 협생질소고정균중(協生窒素固定菌中) Azospirillum lipoferum Ecc 3-1은 유인물질(誘引物質)이 있는 방향(方向)으로 이동(移動)하기 위한 천이환(遷移環)을 만들었으나 Agrobacterium radiobacter Ecc 1-1과 Pseudomonas sp Ecc 4-1은 천이환(遷移環)을 만들지 않했다. 당류(糖類) 및 유기산(有機酸)에 대(對)한 공시세균(供試細菌)의 화학주화성(化學走化性)은 D(+)-glucose나 D(+)-fructose 보다 malic acid나 citric acid에 대(對)한 주화성(走化性)이 높았다. 특(特)히 복합당(複合糖)인 D-galactouronic acid와 L-aspartate는 공시(供試)한 타종(他種)의 당류(糖類)나 유기산(有機酸)보다 가장 높은 주화력(走化力)을 보였다. 수도근분비물질(水稻根分泌物質)에 대(對)한 공시협생질소고정균(供試協生窒素固定菌)의 주화성(走化性)을 수도품종(水稻品種)과 세균간(細菌間)에 서로 상이(相異)한 교호관계(交互關係)를 보였다.