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Title: INVESTIGATING THE EFFECTS OF BT CORN ON SOIL MICROBIAL COMMUNITIES USING PARAMAGNETIC BEAD-ENABLED T-RFLP

Author
item Blackwood, Christopher
item Buyer, Jeffrey

Submitted to: American Society for Microbiology
Publication Type: Abstract Only
Publication Acceptance Date: 2/21/2003
Publication Date: 5/20/2003
Citation: BLACKWOOD, C.B., BUYER, J.S. INVESTIGATING THE EFFECTS OF BT CORN ON SOIL MICROBIAL COMMUNITIES USING PARAMAGNETIC BEAD-ENABLED T-RFLP. AMERICAN SOCIETY FOR MICROBIOLOGY 103RD GENERAL MEETING ABSTRACTS. 2003. ABSTRACT N-196.

Interpretive Summary:

Technical Abstract: The effects of expression of Cry protein in transgenic Bt corn on soil microbial communities was investigated using terminal restriction fragment length polymorphism (T-RFLP), PLFA, and Biolog profiles. Two lines of Bt-corn and near-isogenic non-Bt corn were grown in three soils of differing textures. To facilitate extraction of phylogenetic information directly from T-RFLP profiles, a suite of PCR primers was designed to amplify major taxonomic groups of soil microorganisms separately. A SAS computer program was written to assist selection of restriction enzymes after performing theoretical restrictions on a sequence database using all known restriction enzymes. Primer specificity was tested by random sequencing in clone libraries generated from direct soil DNA extracts. Paramagnetic bead-enabled T-RFLP was developed to accommodate a larger number of samples and primers at lower cost than previous methods. Streptavidin-coated paramagnetic beads were used to physically capture terminal restriction fragments, which could then be separated by standard gel electrophoresis. Beta proteobacterial, PLFA, and Biolog profiles of soil microbial communities grown with Bt corn lines were not significantly different from those grown with near-isogenic non-Bt lines. Communities from different soil types were significantly different.