Genome-wide analysis of T-DNA integration into the chromosomes of Magnaporthe oryzae

Mol Microbiol. 2007 Oct;66(2):371-82. doi: 10.1111/j.1365-2958.2007.05918.x. Epub 2007 Sep 10.

Abstract

Agrobacterium tumefaciens-mediated transformation (ATMT) has become a prevalent tool for functional genomics of fungi, but our understanding of T-DNA integration into the fungal genome remains limited relative to that in plants. Using a model plant-pathogenic fungus, Magnaporthe oryzae, here we report the most comprehensive analysis of T-DNA integration events in fungi and the development of an informatics infrastructure, termed a T-DNA analysis platform (TAP). We identified a total of 1110 T-DNA-tagged locations (TTLs) and processed the resulting data via TAP. Analysis of the TTLs showed that T-DNA integration was biased among chromosomes and preferred the promoter region of genes. In addition, irregular patterns of T-DNA integration, such as chromosomal rearrangement and readthrough of plasmid vectors, were also observed, showing that T-DNA integration patterns into the fungal genome are as diverse as those of their plant counterparts. However, overall the observed junction structures between T-DNA borders and flanking genomic DNA sequences revealed that T-DNA integration into the fungal genome was more canonical than those observed in plants. Our results support the potential of ATMT as a tool for functional genomics of fungi and show that the TAP is an effective informatics platform for handling data from large-scale insertional mutagenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agrobacterium tumefaciens / genetics
  • Chromosomes, Fungal / genetics*
  • DNA, Bacterial / genetics*
  • Genetic Vectors / genetics
  • Genome, Fungal*
  • Magnaporthe / genetics*
  • Polymerase Chain Reaction
  • Sequence Analysis, DNA
  • Transformation, Genetic

Substances

  • DNA, Bacterial
  • T-DNA