• Title/Summary/Keyword: Agrobacterium strains

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Formation of Crown Gall Tumor in Panax ginseng C.A. Meyer (인삼의 Crown Gall Tumor형성에 관한 연구)

  • 최광태;양덕춘
    • 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.

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Identification of Agrobacterium tumefaciens from Soil and Transformation of Maize (토양으로부터의 Agrobacterium tumefaciens의 분리, 동정 및 옥수수의 형질전환에 이용)

  • 노광수;강봉중
    • KSBB Journal
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    • v.7 no.3
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    • pp.191-200
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    • 1992
  • Several strains of Agrobacterium tumefaciens were isolated from soil in the Taegu area and characterized to develop some useful vector systems for higher plant genetic engineering. The selected colonies had a unique form, and strains from the colonies were capable of tumor formation on the sunflower leaf surface. They had a large plasmid. The restriction analysis showed that they were another kinds of Ti plasmic compared with C58 and Ach5. The isolated strains were identified as the nopaline type and also as biovar 1 A. tumefaciens, according to their tumor morphology, blophyslcal and biochemical characteristics. One of the isolated strains, AK204 was transformed with binary vector (pGA642), having selectable marker (Kmr, Tcr). Furthermore, maize tissue cells were transformed by cocultivation with AK204/pGA642, and the transformants were selected on the selective medium and identified using PAGE patterns of their soluble proteins.

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Transformation of Populus Species by an Agrobacterium Binary Vector System (Agrobacterium Binary Vector에 의한 포플러 형질전환(形質轉換)을 위한 기초연구(基礎研究))

  • Chun, Young Woo;Klopfenstein, Ned B.;McNabb, Harold S. Jr.;Hall, Richard B.
    • Journal of Korean Society of Forest Science
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    • v.77 no.2
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    • pp.199-207
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    • 1988
  • Three clones of Populus alba ${\times}$ P. grandidentata have been tested for susceptibility to Agrobacterium tumefaciens strains A281 and A348. We determined the optimum concentration of kanamycin sulfate for effective selection of leaf disc-derived, transgenic tissues transformed using Agrobacterium binary vector pGA472 containing a neomycin phosphotransferase gene (NPT-II) which confers kanamycin resistance. Of the wild type Ti plasmids contained by the two Agrobacterium strains, pTiBo542 of strain A281 appears to be best suited to serve as a helper plasmid for binary vector systems. A relatively low concentration (10mg/l) of kanamycin sulfate inhibited adventitious shoot initiation from leaf discs on regeneration medium. Transformed kanamycin-resistant calli were obtained by culturing Agrobacterium inoculated leaf discs on selective regeneration medium. The transformed kanamycin-resistant calli continued to grow on regeneration media supplemented with kanamycin sulfate to levels of 50 and 200mg/l. The growth of non-co-cultivated control calli was severely inhibited on regeneration medium containing 50mg/l kanamycin sulfate.

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Production of Transgenic Melon from the Cultures of Cotyledonary-Node Explant Using Agrobacterium-Mediated Transformation (Agrobacterium 공동 배양을 통한 자엽절 절편 배양으로부터 멜론 형질전환체 생산)

  • Cho Mi-Ae;Song Yun-Mi;Park Yun-Ok;Ko Suck-Min;Min Sung-Ran;Liu Jang-Ryol;Lee Jun-Haeng;Choi Pil-Son
    • Journal of Plant Biotechnology
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    • v.32 no.4
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    • pp.257-262
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    • 2005
  • Agrobacterium tumefaciens-mediated cotyledonary-node explants transformation was used to produce transgenic melon. Cotyledonary-node explants of melon (Cucumis melo L. cv. Super VIP) were co-cultivated with Agrobacterium strains (LBA4404, GV3101, EHA101) containing the binary vector (pPTN289) carrying with CaMV 35S promoter-gus gene as reporter gene and NOS promoter-bar gene conferring resistance to glufosinate (herbicide Basta) as selective agent, and the binary vector (pPTN290) carrying with Ubiquitin promoter-GUS gene and NOS promoter-nptll gene conferring resistance to paromomycin as selective agent, respectively. The maximum transformation efficiency (0.12%) was only obtained from the cotyledonary-node explants co-cultivated with EHA101 strain (pPTN289) on selection medium with 5 mg/L glufosinate and not produced a transgenic melon from the cotyledon or cotyledonary-node co-cultivated with other strains. Finally, five plants transformed showed the resistance in glufosinate antibiotic and the GUS positive response in leaf ($T_0$), flower ($T_0$), seeds ($T_1$) and plantlet ($T_1$). Southern blot analysis revealed that the gus gene integrated into each genome of transgenic melon.

Simple Detection of Opines by Paper Electrophoresis for Hairy Roots Transformed with Agrobacterium rhizogenes Strains

  • Cho, Hyeon-Je;Ha, Hyo-Cheol;Lee, Jae-Sung;Widholm, Jack M.
    • Journal of Applied Biological Chemistry
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    • v.44 no.2
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    • pp.92-94
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    • 2001
  • A simple protocol for the detection of opines, cucumopines and mikimopines using a general horizontal or vertical get electrophoresis system for protein or DNA separation in the laboratory are demonstrated. This electrophoresis method can also be applied to other opines as long as correct detection reagent and buffer system are used.

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Isolation and identification of tumor-inducing (Ti) Plasmids (식물종양유도 플라스미드의 분리 및 동정)

  • 음진성;김계현;심웅섭
    • Korean Journal of Microbiology
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    • v.24 no.1
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    • pp.1-6
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    • 1986
  • Tumor-inducing (Ti) plasmid which may be used in genetic engineering of higher plants, was isolated from Agrobacterium tumefaciens in Korea. Among the 7 strains of A. tumefaciens isolated from various regions in Korea, KU12, KU13, KU14, and KU49 strains were confirmed to contain plasmid. The isolated Ti plasmids were digested with 4 kinds of restriction enzymes, respectively. According to the result of the cleavage patterns, we confirmed that these plasmids in the KU12, KU13, KU14 and KU49 strains are different.

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A Comparison of the Phenotypic and Genetic Stability of Recombinant Trichoderma spp. Generated by Protoplast- and Agrobacterium-Mediated Transformation

  • Cardoza Rosa Elena;Vizcaino Juan Antonio;Hermosa Maria Rosa;Monte Enrique;Gutierrez Santiago
    • Journal of Microbiology
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    • v.44 no.4
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    • pp.383-395
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    • 2006
  • Four different Trichoderma strains, T. harzianum CECT 2413, T. asperellum T53, T. atroviride T11 and T. longibrachiatum T52, which represent three of the four sections contained in this genus, were transformed by two different techniques: a protocol based on the isolation of protoplasts and a protocol based on Agrobacterium-mediated transformation. Both methods were set up using hygromycin B or phleomycin resistance as the selection markers. Using these techniques, we obtained phenotypically stable transformants of these four different strains. The highest transformation efficiencies were obtained with the T. longibrachiatum T52 strain: 65-70 $transformants/{\mu}g$ DNA when transformed with the plasmid pAN7-1 (hygromycin B resistance) and 280 $transformants/l0^7$ spores when the Agrobacterium-mediated transformation was performed with the plasmid pUR5750 (hygromycin B resistance). Overall, the genetic analysis of the transform ants showed that some of the strains integrated and maintained the transforming DNA in their genome throughout the entire transformation and selection process. In other cases, the integrated DNA was lost.

Screening Resistance to Crown Gall Disease by Pathogen Inoculation in Muscadine and Florida Hybrid Grapes (병원균 접종에 의한 Muscadine 및 플로리다 교잡종 포도의 줄기혹병 저항성 검정)

  • Noh, Jung-Ho;Park, Kyo-Sun;Lu, Jiang;Yun, Hae-Keun
    • Korean Journal of Organic Agriculture
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    • v.23 no.3
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    • pp.497-507
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    • 2015
  • It is necessary to evaluate the resistance to disease among genetic resources for development of disease resistant grapes. This study was conducted to screen the resistance to crown gall in muscadine and Florida hybrid bunch grapes by pathogen inoculation. In order to compare the responses to infection with different pathogen strains, muscadine and Florida hybrid grapes were inoculated with 3 strains of Agrobacterium vitis. Although there were different levels crown gall formation among grape cultivars, there little variation in response to inoculated strains. Among 29 muscadine cultivars tested by inoculation of A. vitis 'C4612', most of them were shown to be susceptible, and 'Gold Isle' and 'Africa Queen' were highly susceptible, and two cultivars, 'Welder' and 'Jumbo' were found to be resistant to crown gall disease. Among Florida hybrid grapes, 'Daytona', 'Stover', and 'Swanee' were susceptible and 'Blanc du Bois' was moderately susceptible to crown gall. Because muscadine grapes have been actively utilized as useful genetic resources for development of new grape varieties by intersub-genus cross, this result from the screening of resistance among muscadine grapes can provide valuable information in breeding programs of grape resistant to crown gall.

Cytogenetic and Histological Characteristics of Ginseng Hairy Root Transformed by Agrobacterium rhizogenes (Agrobacterium rhizogenes에 의하여 형질전환된 인삼 모상근의 세포 유전학적 및 조직학적 특성)

  • 고경민
    • Journal of Plant Biology
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    • v.36 no.1
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    • pp.75-81
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    • 1993
  • Ginseng (Panax ginseng C.A. Meyer) hairy root transformed with Agrobacterium rhizogenes (strain A4) was examined cytogenetically and histologically to assess its characteristics. The optimum growth of hairy root obtained in hormone-free MS medium (sucrose 30 g/L, pH5-6) for long period cultures. All hairy root strains (HB1, HB2, HB3) had the 2n diploid number (2n=48) of chromosomes in root tip cells. There were no alteration in chromosome structure except in one cell of HB3 strain. Results of SDS-PAGE showed a few difference in pattern and number of bands between normal and hairy root of ginseng. The root anatomy of normal root and hairy root differed each other. The hairy root had a clearly defined vascular strand, and the morphology of cortical cells were disorganised with large intercellular spaces.

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