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United States Department of Agriculture

Agricultural Research Service

Title: A Selection and Tagging System for Monitoring the Release of Fungal Weed Biocontrol Agents

item Luster, Douglas
item Berthier, Yvette
item Bailey, Bryan
item Nelson, A

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: October 12, 1996
Publication Date: N/A

Technical Abstract: Release of microbial agents into the environment for control of weeds will require methods for unambiguous identification and detection of the released microbe. We have designed a system for transformation of fungal weed pathogens which incorporates a mechanism for selection of transformants avoiding the use of antibiotic resistance genes. Isolates of Fusarium oxysporum f. sp. erythroxyli from Peru and Hawaii, which are pathogens of Erythroxylum coca in South America, will be transformed with coding sequence for green fluorescent protein (GFP) from the bioluminescent jellyfish, Aequorea victoria. Introduction of the GFP sequence into filamentous fungi will ultimately provide a unique target sequence for sensitive PCR detection in terrestrial habitats. We have made plasmid constructs incorporating GFP into the filamentous fungal vector pHRC [Kistler, H.R.C. and U.K. Benny, Curr. Genet (1988) 13:145-149] under control of transcriptional elements of the Aspergillus nidulans trpC gene. Preliminary results indicate that transformants are expressing the GFP-hph fusion construct, as evidenced by hygromycin resistance to 500 ug/ml and immunoblot detection of GFP protein expression. The hygromycin resistance was stably passed to progeny. We are currently examining the pathogenicity of initial transformants, and analyzing the copy number and stability of the fusion sequence in progeny from the original transformants

Last Modified: 4/18/2015
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