ArticleCurrent genetics2010
Agrobacterium-meditated gene disruption using split-marker in Grosmannia clavigera, a mountain pine beetle associated pathogen.
Article in Current genetics, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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13 citing papers in PubMed.
- Establishment of RNA Interference Genetic Transformation System and Functional Analysis of FlbA Gene inInternational journal of molecular sciences · 2023Article
- Phylogeny of Leptographium qinlingensis cytochrome P450 genes and transcription levels of six CYPs in response to different nutrition media or terpenoids.Archives of microbiology · 2021Article
- Targeted Disruption of Scytalone Dehydratase Gene UsingJournal of fungi (Basel, Switzerland) · 2020Article
- A Negative Regulator of Carotenogenesis inApplied and environmental microbiology · 2020Article
- An efficient gene disruption method using a positive-negative split-selection marker and Agrobacterium tumefaciens-mediated transformation for Nomuraea rileyi.World journal of microbiology & biotechnology · 2018Article
- MFS Transporters and GABA Metabolism Are Involved in the Self-Defense Against DON inFrontiers in plant science · 2018Article
- A silver bullet in a golden age of functional genomics: the impact ofFungal biology and biotechnology · 2017Review
- Targeted Disruption of Melanin Biosynthesis Genes in the Human Pathogenic Fungus Lomentospora prolificans and Its Consequences for Pathogen Survival.International journal of molecular sciences · 2016Article
- Gene discovery for enzymes involved in limonene modification or utilization by the mountain pine beetle-associated pathogen Grosmannia clavigera.Applied and environmental microbiology · 2014Article
- Flavan-3-ols in Norway spruce: biosynthesis, accumulation, and function in response to attack by the bark beetle-associated fungus Ceratocystis polonica.Plant physiology · 2014Article
- Construction of a Kluyveromyces lactis ku80⁻ host strain for recombinant protein production: extracellular secretion of pectin lyase and a streptavidin-pectin lyase chimera.Molecular biotechnology · 2014Article
- The genome and transcriptome of the pine saprophyte Ophiostoma piceae, and a comparison with the bark beetle-associated pine pathogen Grosmannia clavigera.BMC genomics · 2013Article
- Genome and transcriptome analyses of the mountain pine beetle-fungal symbiont Grosmannia clavigera, a lodgepole pine pathogen.Proceedings of the National Academy of Sciences of the United States of America · 2011Article
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5 authors.
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Abstract
Grosmannia clavigera is a fungal pathogen associated with the mountain pine beetle (Dendroctonus ponderosae) which is devastating large areas of western Canada's conifer forests. This fungus also produces a dark melanin pigment that discolors pine sapwood. We have generated the draft genome of G. clavigera. However, functional characterization of genes identified in the genome sequence requires an efficient gene disruption method. In this work, we report a gene replacement strategy for G. clavigera using the Agrobacterium-mediated transformation in conjunction with linear or split-marker deletion cassettes. In addition, we used long flanking regions up to 3 kb from both sides of the targeted genes in our deletion cassettes. We assessed this gene disruption method with two genes from the melanin biosynthesis pathway that produce easily detectable white and red/brown mutant phenotypes: polyketide synthase and scytalone dehydratase. The approach yielded G. clavigera gene replacements with homologous recombination rates between 65 and 82%. For filamentous fungi, this is the first report showing that split-markers can be used with Agrobacterium-mediated transformation to generate appropriate mutants. This method can now be applied to efficiently identify genes involved in G. clavigera fungal pathogenicity and will facilitate understanding how the fungus overcomes the host defence system.
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