Two novel Venturia inaequalis genes induced upon morphogenetic differentiation during infection and in vitro growth on cellophane
Venturia inaequalis is a hemibiotrophic ascomycete that causes apple scab. Germ tubes, from conidia or ascospores, penetrate the leaf or fruit surface directly via appressoria-like swellings; subsequently the hyphae divide laterally to form a stroma between the cuticle and the outer wall of the epid...
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Published in: | Fungal genetics and biology Vol. 45; no. 10; pp. 1329 - 1339 |
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Abstract | Venturia inaequalis is a hemibiotrophic ascomycete that causes apple scab. Germ tubes, from conidia or ascospores, penetrate the leaf or fruit surface directly via appressoria-like swellings; subsequently the hyphae divide laterally to form a stroma between the cuticle and the outer wall of the epidermal cells. This morphological switch can be mimicked by growing the fungus in vitro on cellophane discs. The aim of this work was to identify genes upregulated in planta using growth on cellophane as a model. Four cDNA clones were found to be induced by growth on cellophane, and qRT-PCR showed two of these genes were up-regulated over a thousand fold in infected apple leaves compared to liquid culture. The predicted proteins for both genes possess putative signal peptides for secretion but have no similarity to sequences in publicly available databases. Both genes encode proteins with novel, imperfect repeat domain structures, the number of which vary in an isolate-specific fashion. Cin1 has seven or eight repeats of about 60 amino acids with four conserved cysteine residues per repeat, while Cin3 has four or five repeats of 32 amino acids with no cysteines. Both proteins appear to have evolved through internal duplication. Cin3, in particular, shows considerable between-strain variation in domain structure, indicating a high degree of recombination at this locus and revealing that the repeat structure has most likely arisen by unequal crossing-over. Results of this study support the hypothesis that cellophane-grown V. inaequalis mimics aspects of biotrophic infection and provide the first insights into novel fungal genes expressed during apple scab infection and their mechanisms of evolution. |
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AbstractList | Venturia inaequalis is a hemibiotrophic ascomycete that causes apple scab. Germ tubes, from conidia or ascospores, penetrate the leaf or fruit surface directly via appressoria-like swellings; subsequently the hyphae divide laterally to form a stroma between the cuticle and the outer wall of the epidermal cells. This morphological switch can be mimicked by growing the fungus in vitro on cellophane discs. The aim of this work was to identify genes upregulated in planta using growth on cellophane as a model. Four cDNA clones were found to be induced by growth on cellophane, and qRT-PCR showed two of these genes were up-regulated over a thousand fold in infected apple leaves compared to liquid culture. The predicted proteins for both genes possess putative signal peptides for secretion but have no similarity to sequences in publicly available databases. Both genes encode proteins with novel, imperfect repeat domain structures, the number of which vary in an isolate-specific fashion. Cin1 has seven or eight repeats of about 60 amino acids with four conserved cysteine residues per repeat, while Cin3 has four or five repeats of 32 amino acids with no cysteines. Both proteins appear to have evolved through internal duplication. Cin3, in particular, shows considerable between-strain variation in domain structure, indicating a high degree of recombination at this locus and revealing that the repeat structure has most likely arisen by unequal crossing-over. Results of this study support the hypothesis that cellophane-grown V. inaequalis mimics aspects of biotrophic infection and provide the first insights into novel fungal genes expressed during apple scab infection and their mechanisms of evolution. |
Author | Conolly, Joshua J. Templeton, Matthew D. Kucheryava, Nataliya Sutherland, Paul W. Bowen, Joanna K. Plummer, Kim M. Rikkerink, Erik H.A. Hahn, Matthias Mesarich, Carl H. Kemen, Eric |
Author_xml | – sequence: 1 givenname: Nataliya surname: Kucheryava fullname: Kucheryava, Nataliya organization: University of Kaiserslautern, Department of Biology, Phytopathology, Postbox 3049, 67653 Kaiserslautern, Germany – sequence: 2 givenname: Joanna K. surname: Bowen fullname: Bowen, Joanna K. organization: Horticulture and Food Research Institute of New Zealand, Mt Albert Research Centre, PB 92-169, Auckland 1142, New Zealand – sequence: 3 givenname: Paul W. surname: Sutherland fullname: Sutherland, Paul W. organization: Horticulture and Food Research Institute of New Zealand, Mt Albert Research Centre, PB 92-169, Auckland 1142, New Zealand – sequence: 4 givenname: Joshua J. surname: Conolly fullname: Conolly, Joshua J. organization: Horticulture and Food Research Institute of New Zealand, Mt Albert Research Centre, PB 92-169, Auckland 1142, New Zealand – sequence: 5 givenname: Carl H. surname: Mesarich fullname: Mesarich, Carl H. organization: Horticulture and Food Research Institute of New Zealand, Mt Albert Research Centre, PB 92-169, Auckland 1142, New Zealand – sequence: 6 givenname: Erik H.A. surname: Rikkerink fullname: Rikkerink, Erik H.A. organization: Horticulture and Food Research Institute of New Zealand, Mt Albert Research Centre, PB 92-169, Auckland 1142, New Zealand – sequence: 7 givenname: Eric surname: Kemen fullname: Kemen, Eric organization: University of Konstanz, Department of Biology, Universitätstr. 10, Konstanz, 78457, Germany – sequence: 8 givenname: Kim M. surname: Plummer fullname: Plummer, Kim M. organization: La Trobe University, Department of Botany, Bundoora, Vic. 3086, Australia – sequence: 9 givenname: Matthias surname: Hahn fullname: Hahn, Matthias organization: University of Kaiserslautern, Department of Biology, Phytopathology, Postbox 3049, 67653 Kaiserslautern, Germany – sequence: 10 givenname: Matthew D. surname: Templeton fullname: Templeton, Matthew D. email: MTempleton@hortresearch.co.nz organization: Horticulture and Food Research Institute of New Zealand, Mt Albert Research Centre, PB 92-169, Auckland 1142, New Zealand |
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Keywords | Apple black spot Malus X domestica Unequal crossing-over Stroma Apple scab Malus pumila Repeat-domain proteins Haplotype |
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Snippet | Venturia inaequalis is a hemibiotrophic ascomycete that causes apple scab. Germ tubes, from conidia or ascospores, penetrate the leaf or fruit surface directly... |
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SubjectTerms | Amino Acid Sequence Apple black spot Apple scab Ascomycetes Ascomycota - chemistry Ascomycota - genetics Ascomycota - growth & development Ascomycota - metabolism Cellophane Fungal Proteins - chemistry Fungal Proteins - genetics Fungal Proteins - metabolism Gene Expression Regulation, Fungal Haplotype Malus Malus - microbiology Malus pumila Malus X domestica Models, Biological Molecular Sequence Data Morphogenesis Plant Diseases - microbiology Plant Leaves - chemistry Plant Leaves - genetics Plant Leaves - growth & development Plant Leaves - metabolism Protein Sorting Signals Protein Structure, Tertiary Repeat-domain proteins Sequence Alignment Stroma Unequal crossing-over Up-Regulation Venturia inaequalis |
Title | Two novel Venturia inaequalis genes induced upon morphogenetic differentiation during infection and in vitro growth on cellophane |
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