• 제목/요약/키워드: agrobacterium tumefaciens

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당근 절편을 이용한 Agrobacterium spp. 분리, 동정과 선발균주에 의한 연초의 형질전환 (Isolation and identification of Agrobacterium spp. using carrot disc and transformation of Nicotiana gluca by selected strains)

  • 양덕춘;양덕조;최광태
    • 한국연초학회지
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    • 제13권1호
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    • pp.43-52
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    • 1991
  • These studies were carred out to obtain the transformant from tobacco cells by Agrobacterium spp. from crown gall and soil at the natural field in Korea, and identified their virulence. Kodo's and Clark's selective media were used for isolation of Agrobacterium spp. In these media, total of 99 strains were characterized based on the morphological characteristics of colonies. Among them 34 strains were able to induce on carrot discs. And hypervirulent strains C23-1 and K29-1 were identified as Agrobacterium tumefaciens biotype 1 and biotype 2, respectively. These strains formed fast growing, larger gall as compared to those induced by other strains on the carrot discs. Transformed tobacco callus was initiated on the phytohormone free MS medium with 250$\mu\textrm{g}$/ml carbenicillin after co-cultivation of tobacco stem explants and Agrobacteria. On the phytohormone free media, shoot was rarely formed from transformed callus. However, these shoot were teratoma shoots which were not grown as normal shoot, and teratoma shoot from transformant by C23-1 was smaller than that of K29-1.

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Organogenesis and Production of Some Transgenic Legume Plants by Agrobacterium tumefaciens-mediated Herbicide Resistance Gene Transformation

  • Kantayos, Vipada;Lee, Hyo-Yeon;Bae, Chang-Hyu
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2018년도 추계학술대회
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    • pp.52-52
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    • 2018
  • Development of herbicide resistant transgenic legume plants through Agrobacterium-mediated transformation has been worked in many previous studied. Plant regeneration after infection is the important step to obtain successful transgenic plants. Many attempts try to find the optimum media condition for plant regeneration after infection. However, the transformation efficiency of legume plants is still low. In this study, regeneration of some Korean legume species including two soybean cultivars (Dawon and Pungsan) and pea have been done with organogenesis which is used various kind of explants such as cotyledonary-nodes in soybean and bud-containing tissue in pea. We developed the optimum media condition for plant regeneration regulators under Agrobacterium-mediated transformation using different kind and various concentration of plant growth. As the results, B5 medium containing 2 mg/L of 6-benzylaminopurine was selected in this study for the optimum plant regeneration media. The segments were inoculated with Agrobacterium suspension harbored an IG2 vector containing bar gene which confers resistance to phosphinotricin (PPT) in 3, 5 and 7 days. The transformation efficiency was achieved in Dawon 3.03 % and pea 1.46 % with co-cultivation period of 7 days which is showed a high number of GUS positive expression period.

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Agrobacterium을 이용한 고추의 Transient Expression 시스템 (Development of an Agrobacterium-mediated Transient Expression System for Intact Leaves of Chili Pepper)

  • 성은수;정영희;최도일
    • Journal of Plant Biotechnology
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    • 제31권3호
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    • pp.185-190
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    • 2004
  • Agrobacterium을 이용한 GUS 유전자를 효과적으로 발현시키기 위하여 수행되어진 실험 결과를 요약하면 다음과 같다. 박테이라 농도별 실험을 수행한 결과 균의 전처리 배양 농도 O $D_{600nm}$ 0.3일때 원심분리한 후 얻어진 균을 희석한 후의 최종 농도는 O $D_{600nm}$ 0.8로 맞춘 실험 처리구에서 GUS 유전자 발현율이55%로 가장 높게 나타났다. 병원성 유도 배지 내에 Acetosyringone (AS)이 첨가되지 않은 경우 GUS 유전자가 발현된 고추 잎을 얻을 수 없었으나, 200$\mu$M을 첨가했을 때 90%의 가장 높은 GUS 유전자 발현율을 나타내어 많은 수의 GUS spots을 관찰할 수 있었다. Agrobacterium에 의한 고추 잎의 감염 정도를 조사한 바 Agrobacterium으로 감염시킨지 3일째부터는 박테리아 에한 감염 정도가 심해져서 GUS 유전자 발현 정도가 약해지므로 Agrobacterium으로 감염시킨지 2일째 되었을 때 GUS 유전자 발현이 가장 강하게 나타난 것을 확인하였다. 이 같은 결과는 박테리아에 의한 감염이 일어난지 3일째 부터는 식물체의 감염부위 고사를 일으키는 것과 관련된 것으로 보인다.