Query (optional)   in Class  All Allele Author Colleague Collection Assembly Gene Gene Class Gene Product Germplasm Image Journal Keyword Library Locus Map Map Data Marker Pathology Polymorphism Probe Protein QTL Rearrangement Reference Sequence Species Trait Trait Study Two Point Data  

---

GrainGenes Reference Report: MPM-12-647

[Submit comment/correction]

Reference	MPM-12-647
Title	A transient assay system for the functional assessment of defense-related genes in wheat.
Journal	Molecular Plant-Microbe Interactions
Year	1999
Volume	12
Pages	647-654
Author	Schweizer P Pokorny J Abderhalden O Dudler R
Abstract	Summary: The generation and characterization of transgenic wheat plants is a tedious and time-consuming process that limits the number of putatively important transgenes that can be tested. We therefore established a transient assay system based on wheat leaves to study the effect of transiently expressed genes on the interaction with the wheat powdery mildew fungus Erysiphe (syn. Blumeria) graminis f. sp. tritici. Young wheat leaves were bombarded with tungsten particles coated with a mixture of plasmids carrying the beta-glucuronidase (GUS) reporter gene and a test gene. Leaves were subsequently challenge inoculated with E. graminis and the fungus was allowed to develop for 40 h. After being stained for GUS enzymatic activity as well as for epiphytic fungal structures, the phenotype of transformed epidermal cells was evaluated by bright-field microscopy. The fungus was routinely found to penetrate cells transiently expressing GUS with an efficiency of approximately 35%, which should suffice to detect putative transgene effects. Transgenes encoding a low-molecular weight cell-wall protein of wheat (WIR1), a thaumatin-like protein, and a glucanase had no effect on fungal penetration of transformed epidermal cells. On the other hand, transgenes encoding a pathogen-induced wheat protein of unknown function (WCI5), a chitinase, a glucose oxidase, and a putative peroxidase significantly reduced fungal penetration
Keyword	[ Hide all but 1 of 28 ] 40 assays beta glucuronidase biochemical techniques biolistics cell wall cell wall components chitinase defense mechanisms enzyme activity epidermis erysiphe erysiphe graminis f fungi gene gene expression histoenzymology molecular weight phenotypes plant proteins plasmid vectors reporter gene system transgenic plant tritici triticum aestivum wheat leaves wheat protein

GrainGenes is a product of the Agricultural Research Service of the US Department of Agriculture.