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United States Department of Agriculture

Agricultural Research Service

Title: Transient Expression and Insertional Mutagenesis of Puccinia Triticina Using Biolistics

Authors
item Webb, Craig
item Szabo, Les
item Bakkeren, Guus - AGRI FOOD - CANADA
item Garry, Clark - DEPT BIOLOGY - UNIV WISCO
item Staples, Richard - BOYCE THOMPSON INSTITUTE
item Fellers, John

Submitted to: Functional and Integrative Genomics
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: October 4, 2005
Publication Date: May 2, 2006
Citation: Webb, C., Szabo, L.J., Bakkeren, G., Garry, C.E., Staples, R.C., Fellers, J.P. 2006. Transient expression and insertional mutagenesis of puccinia triticina using biolistics. Functional and Integrative Genomics.

Interpretive Summary: The rust fungi are very important pathogens in most crop species. However, little is known about how the rust fungi infect plants. In this manuscript we describe a technique that can be used to induce mutations as well as introduce new genes into the wheat leaf rust fungus. This method will be useful for future studies of the genes that control the infection process in rust fungi.

Technical Abstract: The genus Puccinia contains more than 4,000 species all of which are fungal plant pathogens. Little is known about the molecular biology of this group and tools for understanding gene function have not yet been established. The wheat leaf rust pathogen Puccinia triticina is an economically significant, biotrophic, basidiomycetous fungus. A set of parameters was established for the transformation of urediniospores of P. triticina and used to analyze the expression of three heterologous promoters. When transferred into P. triticina urediniospores, transient expression was observed across four helium pressures using one size of gold and three sizes of tungsten microprojectiles. Each of the three promoters displayed strong transient expression in germinated urediniospores, however higher numbers of GUS-positive urediniospores were observed when either the actin or Hss1 promoters were used. The information gleaned from testing transient expression variables such as particle size, type of projectile, helium pressure, and multiple heterologous promoters will be of benefit for further studies of Puccinia triticina.

Last Modified: 12/21/2014
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