PlourdeJacobiBernier2008b

Reference

Plourde, K., Jacobi, V., Bernier, L. (2008) Use of insertional mutagenesis to tag putative parasitic fitness genes in the Dutch elm disease fungus Ophiostoma novo-ulmi subsp. novo-ulmi. Canadian Journal of Microbiology, 54(9):797-802.

Abstract

We used insertional mutagenesis to produce genetically tagged mutants of the Dutch elm disease fungus Ophiostoma novo-ulmi subsp. novo-ulmi. We first optimized transformation of O. novo-ulmi protoplasts by the restriction enzyme mediated integration method. A concentration of 80 U of HindIII with 10^8 fungal protoplasts and 5 ug of plasmid DNA was the most efficient for generating a high number of O. novo-ulmi mutants carrying a single insertion in their genome. Mycelium- and yeast-like growth kinetics of 24 O. novo-ulmi mutants were evaluated in vitro. Flanking sequences were succesfully recovered in 8% of the transformants analyzed. Some mutant phenotypes appeared to result from gene disruption events, whereas others likely involved modifications of noncoding regions. Several nuclear loci that control vegetative growth and could potentially impact parasitic fitness were successfully tagged.

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@ARTICLE { PlourdeJacobiBernier2008b,
    AUTHOR = { Plourde, K. and Jacobi, V. and Bernier, L. },
    TITLE = { Use of insertional mutagenesis to tag putative parasitic fitness genes in the Dutch elm disease fungus Ophiostoma novo-ulmi subsp. novo-ulmi },
    JOURNAL = { Canadian Journal of Microbiology },
    YEAR = { 2008 },
    VOLUME = { 54 },
    PAGES = { 797-802 },
    NUMBER = { 9 },
    MONTH = { September },
    ABSTRACT = { We used insertional mutagenesis to produce genetically tagged mutants of the Dutch elm disease fungus Ophiostoma novo-ulmi subsp. novo-ulmi. We first optimized transformation of O. novo-ulmi protoplasts by the restriction enzyme mediated integration method. A concentration of 80 U of HindIII with 10^8 fungal protoplasts and 5 ug of plasmid DNA was the most efficient for generating a high number of O. novo-ulmi mutants carrying a single insertion in their genome. Mycelium- and yeast-like growth kinetics of 24 O. novo-ulmi mutants were evaluated in vitro. Flanking sequences were succesfully recovered in 8% of the transformants analyzed. Some mutant phenotypes appeared to result from gene disruption events, whereas others likely involved modifications of noncoding regions. Several nuclear loci that control vegetative growth and could potentially impact parasitic fitness were successfully tagged. },
    KEYWORDS = { Dutch elm disease, Ophiostoma novo-ulmi, REMI, gene tagging, parasitic fitness, vegetative growth, insertional mutagenesis },
    OWNER = { gagnea },
    TIMESTAMP = { 2008.10.29 },
}

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