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Characterization of Agrobacterium species by capillary isoelectric focusing

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Abstract

Tumorigenic and non-tumorigenic strains of Agrobacterium tumefaciens, A. rhizogenes, A.rubi, and A.vitis were examined using capillary isoelectric focusing, phenotypic determinative tests, PCR and fatty acid analysis. The isoelectric points (pI) of the 40 strains investigated clearly differentiated the strains according to their respective species. The different species were characterized with the following pI values: A. tumefaciens 2.2, A. rhizogenes 4.0, A. rubi 2.15, and A. vitis 2.6. This differentiation corresponded to the phenotypic, PCR and fatty acid characterizations. Strains with the similar chromosomal background but different plasmid content, e.g. A.vitis strain S4, and F2/5 gave the same pI values. Strains of Rhizobium species differed from Agrobacterium strains in their pI values. The advantage of capillary isoelectric focusing over the phenotypic determinative tests, PCR and fatty acid analysis is its speed (15 min), relative simplicity, and the very small amount of chemicals used. This rapid and simple method is a major improvement over the classical methods of separation of Agrobacterium species and should prove useful for rapid characterization of Agrobacterium-like colonies isolated from plant tumours for epidemiological and generic diversity studies.

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Acknowledgements

This work was supported by the Grant Agency of the Academy of Sciences of the Czech Republic No. IAAX00310701 and by the Institutional research plan AVO Z40310501. We are grateful to Dr. J.M. Young (New Zealand) for critical reading and comments on the manuscript. We thank Dr I. Dusha (Biological Research Centre, Hungarian Academy of Sciences, Szeged) providing Rhizobium strains.

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Authors and Affiliations

  1. Plant Protection Institute, Hungarian Academy of Sciences, 1525, Budapest, P.O.Box 102, Hungary

    Sándor Süle

  2. Institute of Analytical Chemistry of the ASCR, v. v. i., Veveří 97, 602 00, Brno, Czech Republic

    Marie Horká, Anna Kubesová & Jiří Salplachta

  3. State Phytosanitary Administration, Division of Diagnostics, 779 00, Olomouc, Czech Republic

    Hana Matoušková & Jaroslav Horký

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  1. Sándor Süle
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  2. Marie Horká
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  3. Hana Matoušková
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  4. Anna Kubesová
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  5. Jiří Salplachta
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  6. Jaroslav Horký
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Correspondence to Sándor Süle.

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Süle, S., Horká, M., Matoušková, H. et al. Characterization of Agrobacterium species by capillary isoelectric focusing. Eur J Plant Pathol 132, 81–89 (2012). https://doi.org/10.1007/s10658-011-9850-y

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  • DOI: https://doi.org/10.1007/s10658-011-9850-y

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