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PacCl, a pH-responsive transcriptional regulator, is essential in the pathogenicity of Colletotrichum lindemuthianum, a causal agent of anthracnose in bean plants

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Abstract

In fungi, the expression of genes encoding proteins related to parasitism is regulated by several factors, including pH. This study reports the structural and functional characterization of the pacCl gene, which encodes the transcription factor PacC of C. lindemuthianum. The pacCl gene showed reduced expression in acidic pH, and its transcription was activated by elevated extracellular pH. The importance of this gene was demonstrated by the development of a pacC1 disruption mutant line of C. lindemuthianum. The mutant line was able to penetrate the host tissue through differentiation of primary hyphae. However, it was not able to cause maceration on the infected plant tissue. The results suggest that PacCl is a regulator of gene activation, and its expression is required for fungal growth in alkaline conditions, as well as for the transcription of genes necessary for the passage from the biotrophic to the necrotrophic phase.

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Acknowledgments

The authors would like to thank the Research Foundation of the State of Minas Gerais (FAPEMIG), the Brazilian Federal Agency for the Support and Evaluation of Graduate Education (CAPES), and the National Council for Scientific and Technological Development (CNPq) for their financial support.

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Authors and Affiliations

  1. Departamento de Botânica e Ecologia, Universidade Federal do Mato Grosso, CEP: 78060-900, Cuiabá, MT, Brazil

    Marcos Antônio Soares

  2. Departamento de Microbiologia, Instituto de Biotecnologia Aplicada a Agropecuária (BIOAGRO), Universidade Federal de Viçosa, CEP: 36570-000, Viçosa, MG, Brazil

    Guilherme Bicalho Nogueira, Denise Mara Soares Bazzolli, Elza Fernandes de Araújo & Marisa Vieira de Queiroz

  3. Institut de Biotechnologie des Plantes, UMR-CNRS 8618, Universite’ Paris-Sud, 1405 cedex, Orsay, France

    Thierry Langin

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  1. Marcos Antônio Soares
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Correspondence to Marisa Vieira de Queiroz.

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Electronic Supplementary Material A1

pacCl inactivation analysis. (a) Determining the copy number of pacCl gene by DNA hybridization. (b) The pBKSpac::hph2 plasmid used in the transformation. The clear rectangle represents the pacCl cDNA fragment. The trans-primer (gray triangle) was inserted into the region between the annealing sequences of the oligonucleotides Opac1 and Opac2; it amplified a 1,079-bp fragment of the pBKSpac::hph2 and 1221-bp of the genomic DNA. (c) PCR results using the group of oligonucleotide pair Opac1/Opac2 or Ohph3/Ohph4 and DNA from lines Mutpac2, Tr1, Tr2, Tr3, and Tr4 of C. lindemuthianum; M: 100-bp marker. (d) Molecular characterization of the pacCl − mutant by DNA hybridization using as a probe a 1.8-kb fragment of the pacCl gene and the empty pBluescriptSK+. (M) - Lambda bacteriophage DNA digested with Hind III; (Mut): mutant line Mutpac2; (WT): wild-type line. (JPEG 2059 kb)

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Soares, M.A., Nogueira, G.B., Bazzolli, D.M.S. et al. PacCl, a pH-responsive transcriptional regulator, is essential in the pathogenicity of Colletotrichum lindemuthianum, a causal agent of anthracnose in bean plants. Eur J Plant Pathol 140, 769–785 (2014). https://doi.org/10.1007/s10658-014-0508-4

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