A genomic approach to understanding the cause and effect of annual ryegrass toxicity

Annual Ryegrass Toxicity (ARGT) is a potentially lethal disease affecting livestock grazing on pastures or consuming fodder that include annual ryegrass (Lolium rigidum) contaminated with corynetoxins. The corynetoxins (CTs), among the most lethal toxins produced in nature, are produced by the bacterium Rathayibacter toxicus that uses a nematode vector to attach to and infect the seedheads of L.rigidum. There is little known of the factors that control toxin production. Several studies have speculated that a bacteriophage specific to R.toxicus may be implicated in CT production. We have developed a PCR-based assay to test for both bacterium and phage in ryegrass material and results indicate that there is a correlation between phage and bacterial presence in all toxic ryegrass samples tested so far. This PCR-based technique may ultimately allow for a rapid, high-throughput screening assay to identify potentially toxic pastures and feed in the field. Currently, ~80% of the 45 Kb genome has been sequenced an investigation to further elucidate its potential role in toxin production.Furthermore, specific alterations in gene expression as a result of exposure to CTs or the closely related tunicamycins (TMs), which are commercially available and considered biologically indistinguishable from CTs, will be evaluated for use as biomarkers of exposure. The effects of both toxins will be analysed in vitro using a rat hepatocyte cell line and screened on a low-density DNA micro array “CT-Chip” that contains <100 selected rat hepatic genes. The results are expected to further define the bioequivalence of CTs and TMs and to identify levels of exposure that are related to specific toxic effects or have no adverse effect on livestock.<br />