Abstract
BackgroundChoke, caused by the endophytic fungus Epichloë typhina, is an important disease affecting orchardgrass (Dactylis glomerata L.) seed production in the Willamette Valley. Little is known concerning the conditions necessary for successful infection of orchardgrass by E. typhina. Detection of E. typhina in plants early in the disease cycle can be difficult due to the sparse distribution of hyphae in the plant. Therefore, a sensitive method to detect fungal infection in plants would provide an invaluable tool for elucidating the conditions for establishment of infection in orchardgrass. Utilization of a marker gene, such as the green fluorescent protein (GFP), transformed into Epichloë will facilitate characterization of the initial stages of infection and establishment of the fungus in plants.FindingsWe have developed a rapid, efficient, and reproducible transformation method using electroporation of germinating Epichloë conidia isolated from infected plants.ConclusionsThe GFP labelled E. typhina provides a valuable molecular tool to researchers studying conditions and mechanisms involved in the establishment of choke disease in orchardgrass.
Highlights
Orchardgrass (Dactylis glomerata L.) is an important forage grass species
The green fluorescent protein (GFP) labelled E. typhina provides a valuable molecular tool to researchers studying conditions and mechanisms involved in the establishment of choke disease in orchardgrass
The digested DNA was precipitated with 1/10 volume of of hygromycin to be used for selection, conidia (~5000 conidia/plate) were placed on increasing concentrations of hygromycin B, from 0-400 mg/L, on Corn Meal Malt Agar (CMMA - g corn meal agar, 2 g yeast extract, g malt extract and 5 g agar per liter)
Summary
About 97% of orchardgrass seed used for pastures and hay in North America is produced in the Willamette Valley in western Oregon. Choke disease, caused by the endophytic fungus Epichloë typhina, was first reported in the Willamette Valley during the mid 1990s [1]. Fertilization requires the transfer of conidia of one mating type to stroma of the opposite mating type. This is typically accomplished by flies in the genus Botanophila [2]. Choke, caused by the endophytic fungus Epichloë typhina, is an important disease affecting orchardgrass (Dactylis glomerata L.) seed production in the Willamette Valley. Utilization of a marker gene, such as the green fluorescent protein (GFP), transformed into Epichloë will facilitate characterization of the initial stages of infection and establishment of the fungus in plants
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
