Quantitative PCR Method To Measure the Fungal Endophyte in Locoweeds

Abstract

A fungal endophyte ( Undifilum oxytropis ) has been implicated in the synthesis of swainsonine in Oxytropis and Astragalus species, commonly known as locoweeds. A quantitative PCR method has been developed to measure the amount of endophyte in Oxytropis and Astragalus species. The limit of quantitation was estimated to be 0.2 pg of endophyte/ng of total DNA. This method of analysis was used to quantify the amount of endophyte in 10 plants each of Oxytropis sericea (white point locoweed), Astragalus mollissimus (wooly locoweed), and Astragalus lentiginosus (spotted locoweed). A significant amount of individual plant variability was observed in endophyte content among individuals in all three species. In one O. sericea and one A. lentiginosus plant swainsonine concentrations were near or below the limit of detection. These plants also had the lowest amounts of endophyte when compared to the other specimens. This method will be a useful tool in further investigating the role the endophyte plays in swainsonine production in various locoweed species.