• Title/Summary/Keyword: Agrobacterium tumefaciens.

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Studies on the crown gall disease of chestnut nurse seedling grafts (밤나무 유태 접목묘의 뿌리 흑병 ( 근두암종병 ) 에 관한 연구)

  • La, Yong-Joon;Hyong-Bin Im;Kwang-Yeun Lee;Du-Hyung Lee
    • The Korean Journal of Ecology
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    • v.2 no.1
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    • pp.15-20
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    • 1978
  • Pathological and anatomical studies on the cause of the crown gall like symptoms associated with the chestnut nurse grafts were undertaken. The crown gall bacterium, Agrobacterium tumefaciens, was isolated from the gall tissues of chestnut nurese grafts by using selective media developed by S초개소 et al. and kado and Heskett. Typical crown gall symptoms appeared on tomato, castor bean and geranium plants 10~21 days following inoculation with the bacterium isolated from the gall tissues of chestnut nurse grafts. Agrobacterium tumefaciens was reisolated from crown gall tissues of tomato, castor bean and geranium. Anatomical studies on the origin, growth and differentiation of the gall tissues of the chestnut nurse grafts confirmed that the gall tissues are of crown gall origin. Masses of Agrobacterium tumefaciens were observed from gall tissues of chestnut nurse grafts, so it could be confirmed that the crown gall symptoms prevalent on chestnut nurse grafts are caused by the crown gall bacterium, Agrobacterium tumefaciens.

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Characterization and biovar. cetermination of agrobacterium tumefaciens T7 isolated in Korea (한국에서 분리한 agrobacterium tumefaciens T7의 특성과 biovar.결정)

  • Rhee, Y.;Kim, C. J.;Kim, S. H.;Yoo, I. D.;Mheen, T. I.
    • 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.

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Oxalate Decarboxylase from Agrobacterium tumefaciens C58 is Translocated by a Twin Arginine Translocation System

  • Shen, Yu-Hu;Liu, Rui-Juan;Wang, Hai-Qing
    • 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.

Agrobacterium tumefaciens-Mediated Genetic Transformation: Mechanism and Factors

  • Kumar, Nitish;Vijayanand, K.G.;Reddy, Myppala P.;Singh, Amritpal S.;Naraynan, Subhash
    • Journal of Forest and Environmental Science
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    • v.25 no.3
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    • pp.195-204
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    • 2009
  • Agrobacterium-mediated genetic transformation has been widely used for the production of genetically modified transgenic plants to obtain specific desired traits. Most of the molecular mechanisms that underlie the transformation steps have been well elucidated over the years. However, a few steps, such as nuclear targeting, T-DNA integration, and Agrobacterium-plant proteins involved remain largely obscure and are still under extensive studies. This review describes the major steps involved in the molecular mechanism of Agrobacterium-mediated transformation and provides insight in the recent developments in studies on the Agrobacterium-mediated genetic transformation system. Some factors affecting the transformation efficiency are also briefly discussed.

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Development of Transgenic Tall Fescue Plants from Mature Seed-derived Callus via Agrobacterium-mediated Transformation

  • Lee, Sang-Hoon;Lee, Dong-Gi;Woo, Hyun-Sook;Lee, Byung-Hyun
    • 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.

Effective Combination of Agrobacterium tumefaciens Strains and Ti Plasmids for the Construction of Plant Vector System

  • Kim, Mi-Suk;Park, Jeong-Du;Eum, Jin-Seong;Sim, Woong-Seop
    • 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.

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Genetic Transformation of Lettuce (Lactuca sativa L.) with Agrobacterium tumefaciens (Agrobacterium tumefaciens에 의한 상추 (Lactuca sativa L.)의 형질전환)

  • 최언옥;양문식;김미선;은종선;김경식
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.1
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    • pp.55-58
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    • 1994
  • Agrobacterium tumefaciens LABA4404 harboring plant binary vector, pBI121, was used for genetic transformation of lettuce (Lactuca sativa t.). Cotyledon segments were infected with A. tumefaciens LBA4404 by cocultivation method and regenerated. Regenerated letture was subject to molecular analyses for integration into plant nuclear genome and expression of ${\beta}$-glucumnidase (GUS) gene. Southern and Northern blot analyses demonstrated that GUS gene was integrated into plant nuclear genome and expressed into its mRNA. The expression of GUS gene into its protein was confirmed by specetrophotometric assay of GUS activity.

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Transformation of A Plant by Ascorbate Peroxidase Gene using Agrobacterium tumefaciens (Ascorbate Peroxidase 유전자의 도입에 의한 식물의 형질전환)

  • 이인애;이효신;배은경;김기용;이병현;손대영;조진기
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.22 no.2
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    • pp.101-106
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    • 2002
  • This study was conducted to obtain the transformed tobacco (Nicotiana tubacum) plants with cytosolic ascorbate peroxidase gene(ApxSC7) using Agrobacterium tumefaciens LBA4404. A cDNA encoding the cytosolic ascorbate peroxidase of strawberry, ApxSC7, was introduced into tobacco plants via Agrobacterium-mediated gene transfer system. The expression vector, pIG-AP8, harboring ApxSC7 gene was used for production of transgenic tobacco plants. A large number of transgenic plants were regenerated on a medium containing hygromycin. Integration of ApxSC7 gene was confirmed by PCR and Southern blot analyses with genomic DNA. Northern blot analyses revealed that the pIGap8 gene was constitutively expressed.

Agrobacterium-Mediated Co-transformation of Multiple Genes in Metarhizium robertsii

  • Padilla-Guerrero, Israel Enrique;Bidochka, Michael J.
    • Mycobiology
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    • v.45 no.2
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    • pp.84-89
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    • 2017
  • Fungi of the Metarhizium genus are a very versatile model for understanding pathogenicity in insects and their symbiotic relationship with plants. To establish a co-transformation system for the transformation of multiple M. robertsii genes using Agrobacterium tumefaciens, we evaluated whether the antibiotic nourseothricin has the same marker selection efficiency as phosphinothricin using separate vectors. Subsequently, in the two vectors containing the nourseothricin and phosphinothricin resistance cassettes were inserted eGFP and mCherry expression cassettes, respectively. These new vectors were then introduced independently into A. tumefaciens and used to transform M. robertsii either in independent events or in one single co-transformation event using an equimolar mixture of A. tumefaciens cultures. The number of transformants obtained by co-transformation was similar to that obtained by the individual transformation events. This method provides an additional strategy for the simultaneous insertion of multiple genes into M. robertsii.

Studies of Plant Tumor Induction (Pat 1) Experiments on the Inoculation of Agrobacterium tumefaciens in out Field (식물의 암종유발에 관한 연구 1 (제 1 ) Agrobacterium tumefaciens 의 에 관하여)

  • 이민재;홍순우;최영길
    • Korean Journal of Microbiology
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    • v.4 no.2
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    • pp.1-4
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    • 1966
  • As a part of studies of plant tumor induction, this experiment was prepared for the purpose of studying the ability of tumor induction and the tendency of tumor initiation in some Korean plants using the various Agrobacterium tumefaciens strains. Results obtained from this experiment are as follows. The virulences of five strains used in this experiment were gradually decreased in order of strain A6Kl, B6, 11BV7, T37 and 11 BNV6. Especially strain T37 which is known to the host limited strain showed virulent effect to the most of plants given for the materials as well as strain A6Kl, B 6 and 11BV7. Concerning the grade of tumor development, in plants which has tough stem, for example, Glycine max Meer, tumor induction was not well developed after the inoculation of all strains. Particullary in Ricinus communes Linne all strains showed virulent effect but tumor tissues were declined in relation to the development of lignification. It was also confirmed that the induction of tumor tissues on plants is to delay according to the increase of the age of host plants.

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