The symbiotic relationship between perennial ryegrass and its endophytic fungus, Neotyphodium lolii, is of considerable agronomic significance in New Zealand. Livestock ingesting N. lolii-infected perennial rye grass may succumb to a neuromuscular condition, ryegrass staggers, the probable cause of which has been identified as the endophyte-produced indole-diterpenoid mycotoxin lolitrem B. The effects of plant component, plant reproductive development, and plant management (clipping) on the concentration and accumulation of lolitrem B were investigated. Eight different naturally occurring perennial ryegrass-N. lolii associations were cloned. They were either trimmed regularly to maintain their vegetative state (V clones) or not trimmed at all to enable full plant reproductive development (R clones). At times that corresponded approximately with the start, middle, and end of the plant reproductive phase, V and R clones of each grass-endophyte association were destructively harvested. After dissection into a number of plant components, harvested material was analyzed for lolitrem B content by HPLC. Analyses were also performed on all clippings taken from the V clones. Highest lolitrem B concentrations in vegetative parts were found in older leaf sheaths and dead leaves. Towards the end of the plant reprodutive phase, dead leaves also contained a large proportion of the total lolitrem B content of the V clones. The highest concentration of lolitrem B was found in the seed, which accounted for nearly 60% of the total quantity in R clones. Leaf blades, young leaf sheaths, roots, and crowns generally contained low to moderate concentrations. In late spring, mean concentrations in R and V clones were similar. By the time mature seed was present (mid-summer), mean concentrations in R clones were three times higher than in V clones. This difference was attributed largely to the contribution by the seed in the R clones. The implications of these findings in relation to ryegrass staggers in livestock are discussed.