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Cultivar-specific kinetics of gene induction during downy mildew early infection in grapevine

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Abstract

The oomycete pathogen Plasmopara viticola (Berk. et Curt.) Berl. et de Toni is the causing agent of the destructive downy mildew disease in grapevine. Despite the advances towards elucidation of grapevine resistance mechanisms to downy mildew, increased knowledge of the biological and genetic components of the pathosystem is important to design suitable breeding strategies. Previously, a cDNA microarray approach was used to compare two Vitis vinifera genotypes Regent and Trincadeira (resistant and susceptible to downy mildew, respectively) in field conditions. The same cDNA microarray chip was used to confirm field-based results and to compare both genotypes under greenhouse conditions at 0, 6, and 12 h post-inoculation with P. viticola. Results show that when comparing both cultivars after pathogen inoculation, there is a preferential modulation of several defense, signaling, and metabolism associated transcripts in Regent. Early transcriptional changes are discussed in terms of genetic background and resistance mechanism. This study is the first to directly compare resistant and susceptible cultivars responses as early as 6 hpi with P. viticola, providing several candidate genes potentially related to the expression of resistance traits.

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Abbreviations

cDNA:

Complementary DNA

EST:

Expressed sequence tag

NCBI:

National Center for Biotechnology Information

MIPS:

Munich information center for protein sequences

hpi:

Hours post-inoculation

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Acknowledgements

This work was developed and supported within the frame of the project “Genomic Research-Assisted breeding for Sustainable Production of quality GRAPEs and WINE”, ERA-NET Plant Genomics granted research project (0313996A), ERA-PG/0004/2006, and by the Portuguese Foundation for Science and Technology with the fellowship SFRH/BPD/33281/2008 and SFRH/BPD/63641/2009.

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Authors and Affiliations

  1. Plant Systems Biology Lab, Center of Biodiversity, Functional & Integrative Genomics (BioFIG), Science Faculty of Lisbon University, 1749-016, Lisbon, Portugal

    Andreia Figueiredo, Filipa Monteiro, Ana Margarida Fortes & Maria Salomé Pais

  2. Julius Kuehn Institute, Federal Research Centre for Cultivated Plants, Institute for Grapevine Breeding Geilweilerhof, 76833, Siebeldingen, Germany

    Martina Bonow-Rex & Eva Zyprian

  3. Department of Statistics and Operations Research, FCUL, CEAUL, Lisbon, Portugal

    Lisete Sousa

Authors
  1. Andreia Figueiredo
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  2. Filipa Monteiro
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  3. Ana Margarida Fortes
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  4. Martina Bonow-Rex
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  5. Eva Zyprian
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  6. Lisete Sousa
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  7. Maria Salomé Pais
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Corresponding author

Correspondence to Andreia Figueiredo.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Online Resource 1

Technical details of Array Design and Spotting provided as MIAME (PDF 135 kb).

Online Resource 2

List of the unigenes differentially regulated in Regent and Trincadeira prior (0 h) and after inoculation with P. viticola (6 and 12 hpi). For each unigene is reported: (1) tentative consensus; (2) Genbank accession number obtained by sequence submission to dbEST; (3) expression profile at 0 h, 6 and 12 hpi; (4) description of putative function (if available) together with EST/TC and protein accession numbers obtained by nblast/nrblast analysis (March 2011) against the public databases: NCBI (Altschul et al. 1997), VitisEXPDB (Doddapaneni et al. 2008), DFCI Grape Gene Index (release 6.0) (Lee et al. 2005; Quackenbush et al. 2001), and UNIREF50 (Suzek et al. 2007) (PDF 161 kb).

OnlineResource 3

Transcript gene primer sequences and amplicon characteristics used for quantitative real-time PCR validation of the expression profiles of seven P. viticola-responsive genes following MIQE guidelines. Reaction efficiency calculated from the slope of the standard curve (Efficiency = −1 + 10(−1/slope)) is presented (PDF 110 kb).

Online Resource 4

Quantitative PCR efficiency plots. Median quantification cycle (Cq) values of each set of tenfold serial dilution plotted against the logarithm of cDNA concentration. Reaction efficiency is given by [10(1/−S) − 1] × 100%, where S represents the slope of the linear regression line (PDF 106 kb).

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Figueiredo, A., Monteiro, F., Fortes, A.M. et al. Cultivar-specific kinetics of gene induction during downy mildew early infection in grapevine. Funct Integr Genomics 12, 379–386 (2012). https://doi.org/10.1007/s10142-012-0261-8

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