ArticlePloS one2014
A high-throughput gene disruption methodology for the entomopathogenic fungus Metarhizium robertsii.
Article in PloS one, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 35 papers.
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Who cites it
35 citing papers in PubMed, 60 citations in OpenAlex.
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- Engineered Metarhizium fungi produce longifolene to attract and kill mosquitoes.Nature microbiology · 2025Article
- Mr-lac3 and Mr-lcc2 inJournal of fungi (Basel, Switzerland) · 2025Article
- Histone deacetylase HDAC3 regulates ergosterol production for oxidative stress tolerance in the entomopathogenic and endophytic fungusmSystems · 2024Article
- The secretory protein COA1 enables Metarhizium robertsii to evade insect immune recognition during cuticle penetration.Communications biology · 2024Article
- Characterization of regulatory genesFrontiers in microbiology · 2024Article
- Ethanol mediates the interaction betweenMicrobiology spectrum · 2023Article
- Hyphal Growth and Conidia Germination Are Induced by Phytohormones in the Root Colonizing and Plant Growth Promoting FungusJournal of fungi (Basel, Switzerland) · 2023Article
- The Growth and Conidiation ofJournal of fungi (Basel, Switzerland) · 2023Article
- The SWC4 subunit of the SWR1 chromatin remodeling complex is involved in varying virulence ofVirulence · 2022Article
- Bioremediation of mercury-polluted soil and water by the plant symbiotic fungusProceedings of the National Academy of Sciences of the United States of America · 2022Article
- New Downstream Signaling Branches of the Mitogen-Activated Protein Kinase Cascades Identified in the Insect Pathogenic and Plant Symbiotic FungusFrontiers in fungal biology · 2022Review
- Slt2-MAPK/RNS1 Controls Conidiation via Direct Regulation of the Central Regulatory Pathway in the FungusJournal of fungi (Basel, Switzerland) · 2021Article
- The Sugar Transporter MST1 Is Involved in Colonization of Rhizosphere and Rhizoplane by MetarhiziummSystems · 2021Article
- A novel cascade allows Metarhizium robertsii to distinguish cuticle and hemocoel microenvironments during infection of insects.PLoS biology · 2021Article
- A Novel Nitrogen and Carbon Metabolism Regulatory Cascade Is Implicated in Entomopathogenicity of the Fungus Metarhizium robertsii.mSystems · 2021Article
- Updates inJournal of fungi (Basel, Switzerland) · 2021Review
- An Efficient Strategy for Obtaining Mutants by Targeted Gene Deletion inFrontiers in microbiology · 2021Article
- Transcriptional Differences Guided Discovery and Genetic Identification of Coprogen and Dimerumic Acid Siderophores inFrontiers in microbiology · 2021Article
- An invasive beetle-fungus complex is maintained by fungal nutritional-compensation mediated by bacterial volatiles.The ISME journal · 2020Article
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Authors and funding
8 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Systematic gene disruption is a direct way to interrogate a fungal genome to functionally characterize the full suite of genes involved in various biological processes. Metarhizium robertsii is extraordinarily versatile, and it is a pathogen of arthropods, a saprophyte and a beneficial colonizer of rhizospheres. Thus, M. robertsii can be used as a representative to simultaneously study several major lifestyles that are not shared by the "model" fungi Saccharomyces cerevisiae and Neurospora crassa; a systematic genetic analysis of M. robertsii will benefit studies in other fungi. In order to systematically disrupt genes in M. robertsii, we developed a high-throughput gene disruption methodology, which includes two technologies. One is the modified OSCAR-based, high-throughput construction of gene disruption plasmids. This technology involves two donor plasmids (pA-Bar-OSCAR with the herbicide resistance genes Bar and pA-Sur-OSCAR with another herbicide resistance gene Sur) and a recipient binary plasmid pPK2-OSCAR-GFP that was produced by replacing the Bar cassette in pPK2-bar-GFP with a ccdB cassette and recombination recognition sites. Using this technology, a gene disruption plasmid can be constructed in one cloning step in two days. The other is a highly efficient gene disruption technology based on homologous recombination using a Ku70 deletion mutant (ΔMrKu70) as the recipient strain. The deletion of MrKu70, a gene encoding a key component involved in nonhomologous end-joining DNA repair in fungi, dramatically increases the gene disruption efficiency. The frequency of disrupting the conidiation-associated gene Cag8 in ΔMrKu70 was 93% compared to 7% in the wild-type strain. Since ΔMrKu70 is not different from the wild-type strain in development, pathogenicity and tolerance to various abiotic stresses, it can be used as a recipient strain for a systematic gene disruption project to characterize the whole suite of genes involved in the biological processes of M. robertsii.
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