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      Use of a non-homologous end-joining-deficient strain (delta-ku70) of the biocontrol fungus Trichoderma virens to investigate the function of the laccase gene lcc1 in sclerotia degradation

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          Abstract

          The aim of this study was to apply a generated Δtku70 strain with increased homologous recombination efficiency from the mycoparasitic fungus Trichoderma virens for studying the involvement of laccases in the degradation of sclerotia of plant pathogenic fungi. Inactivation of the non-homologous end-joining pathway has become a successful tool in filamentous fungi to overcome poor targeting efficiencies for genetic engineering. Here, we applied this principle to the biocontrol fungus T. virens, strain I10, by deleting its tku70 gene. This strain was subsequently used to delete the laccase gene lcc1, which we found to be expressed after interaction of T. virens with sclerotia of the plant pathogenic fungi Botrytis cinerea and Sclerotinia sclerotiorum. Lcc1 was strongly upregulated at early colonization of B. cinerea sclerotia and steadily induced during colonization of S. sclerotiorum sclerotia. The Δtku70Δlcc1 mutant was altered in its ability to degrade the sclerotia of B. cinerea and S. sclerotiorum. Interestingly, while the decaying ability for B. cinerea sclerotia was significantly decreased, that to degrade S. sclerotiorum sclerotia was even enhanced, suggesting the operation of different mechanisms in the mycoparasitism of these two types of sclerotia by the laccase LCC1.

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          Most cited references 45

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          Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.

          A new method of total RNA isolation by a single extraction with an acid guanidinium thiocyanate-phenol-chloroform mixture is described. The method provides a pure preparation of undegraded RNA in high yield and can be completed within 4 h. It is particularly useful for processing large numbers of samples and for isolation of RNA from minute quantities of cells or tissue samples.
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            Single step method of RNA isolation by acid guanidium thiocyanate phenol chloroform extraction. Anal

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              Fungal laccases - occurrence and properties.

               Petr Baldrian (2006)
              Laccases of fungi attract considerable attention due to their possible involvement in the transformation of a wide variety of phenolic compounds including the polymeric lignin and humic substances. So far, more than a 100 enzymes have been purified from fungal cultures and characterized in terms of their biochemical and catalytic properties. Most ligninolytic fungal species produce constitutively at least one laccase isoenzyme and laccases are also dominant among ligninolytic enzymes in the soil environment. The fact that they only require molecular oxygen for catalysis makes them suitable for biotechnological applications for the transformation or immobilization of xenobiotic compounds.
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                Author and article information

                Affiliations
                [1]Department of Tree Science, Entomology and Plant Pathology G. Scaramuzzi, Plant Pathology Section, Faculty of Agriculture, University of Pisa, Via del Borghetto 80, 56124 Pisa, Italy
                [2]Research Area Gene Technology and Applied Biochemistry, Institute of Chemical Engineering, Vienna University of Technology, Gumpendorferstrasse 1a, 1060 Vienna, Austria
                [3]Scuola Normale Superiore di Pisa, Piazza dei Cavalieri 7, 56126 Pisa, Italy
                Author notes

                Communicated by U. Kueck.

                Contributors
                +43-1-58801166554 , vseidl@mail.tuwien.ac.at
                Journal
                Curr Genet
                Current Genetics
                Springer-Verlag (Berlin/Heidelberg)
                0172-8083
                1432-0983
                26 September 2010
                26 September 2010
                February 2011
                : 57
                : 1
                : 13-23
                3023040
                20872221
                322
                10.1007/s00294-010-0322-2
                © The Author(s) 2010
                Categories
                Research Article
                Custom metadata
                © Springer-Verlag 2011

                Genetics

                mycoparasitism, ku70, laccase, trichoderma virens, sclerotia, gene targeting

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