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Cloning and characterization of gene-resistant analogs (RGAs) involved in rust (Puccinia psidii) resistance in Eucalyptus grandis

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Abstract

Disease-resistant genes play an important role in defending against a variety of pathogens and insect pests in plants. Most of the disease-resistant genes encode proteins with conserved leucine rich repeat and nucleotide binding site domains. In this study, we cloned and characterized gene-resistant analogs (RGAs) from Eucalyptus grandis using degenerate PCR, with primers specifically targeting these two domains. The amplified fragments were cloned into the pGEM-T vector and transformed into Escherichia coli. Among the 90 clones obtained, 13 were sequenced and compared with each other and with previously identified gene-resistant diseases. A BLASTX search in GenBank revealed high similarities among the conserved domains of these cloned genes with RGA genes. Some clones, however, showed no significant similarity with DNA sequences in GenBank. Southern blotting analysis identified several polymorphic RFLP loci between distinct genotypes. However, none of them co-segregated with the Puccinia psidii Winter resistance gene 1 (Ppr1) in a population study.

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Acknowledgments

We thank the Suzano Celulose S/A for logistical support and the “Conselho Nacional de Desenvolvimento Científico e Tecnológico” (CNPq), the “Coordenação de Aperfeiçoamento de Pessoal de Nível Superior” (CAPES) and the “Fundação de Amparo à Pesquisa do Estado de Minas Gerais” (FAPEMIG) for financial support to this study.

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Authors and Affiliations

  1. Departamento de Engenharia Florestal, Universidade Federal dos Vales do Jequitinhonha e Mucuri, Diamantina, Minas Gerais, 39100-000, Brazil

    Marcelo Luiz Laia, Inaê Mariê de Araújo Silva, Janaína Fernandes Gonçalves & Ariadne Marques

  2. Departamento de Fitopatologia, Universidade Federal de Viçosa, Viçosa, Minas Gerais, 36570-900, Brazil

    Acelino Couto Alfenas & Sergio Hermínio Brommonschenkel

  3. Suzano Bahia Sul Celulose S/A, Suzano, São Paulo, Brazil

    Shinitiro Oda & Eduardo José de Melo

Authors
  1. Marcelo Luiz Laia
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  2. Acelino Couto Alfenas
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  3. Sergio Hermínio Brommonschenkel
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  4. Shinitiro Oda
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  5. Eduardo José de Melo
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  6. Inaê Mariê de Araújo Silva
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  7. Janaína Fernandes Gonçalves
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  8. Ariadne Marques
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Corresponding author

Correspondence to Acelino Couto Alfenas.

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The online version is available at http://www.springerlink.com

Corresponding editor: Chai Ruihai

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Laia, M.L., Alfenas, A.C., Brommonschenkel, S.H. et al. Cloning and characterization of gene-resistant analogs (RGAs) involved in rust (Puccinia psidii) resistance in Eucalyptus grandis . J. For. Res. 26, 711–718 (2015). https://doi.org/10.1007/s11676-015-0049-5

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  • DOI: https://doi.org/10.1007/s11676-015-0049-5

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