2010 Annual Report 1a.Objectives (from AD-416) The objective of this cooperative research project is to develop technologies associated with grass endophytes and evaluate plant germplasm for use in irrigated and non-irrigated pastures. 1b.Approach (from AD-416) Collaborative experiments will be designed and implemented either in the laboratory or the open-field depending upon the objective. Laboratory experiments will be conducted jointly in Logan, Utah, and Christchurch, New Zealand, where endophytes are involved. Where experiments involve the construction of molecular markers for endophyte identification, work will be performed in Logan, Utah. Germplasm evaluation of Cropmark grass accessions (cultivars and lines) will be conducted in the Great Basin region of Utah in appropriate growing areas. This research will attempt to: . 1)Develop methodologies associated with the identification and mutagenesis of endophytes and their inoculation into grass plants;. 2)Construct markers which will identify specific endophytes; and. 3)Evaluate agronomic performance of endophyte and non-endophyte containing Cropmark grass germplasm along with appropriate + or - endophyte containing control germplasm. 3.Progress Report Plant endophyte infection is found in a wide array of grasses. While some endophyte infections produce toxic effects to livestock and wildlife, some have demonstrated agricultural benefits. For example, some endophyte infections confer resistance to insect, nematodes, and fungi to the host plant through the production of alkaloids compounds. One of the purposes of this project is to develop strain-specific DNA marker that will be useful in identifying specific strains of beneficial Neotyphodium endophytes. Multiple PCR primer pairs were designed from the the Neotyphodium uncinatum loline alkaloid synthesis gene clusters, LOL-1 and LOL-2. There primers were used to amplify gene sequences from multiple Neotyphodium strains, as well as other endophytes found in FRRL grasses. Poor quality of endophyte genomic DNA has made it difficult produce high quality, reproducible DNA amplifications. Analysis of coding and non-coding DNA sequences from some successful PCR amplifications has yielded minimal gene sequence data that have not yet led to the development of strain-specific markers. Efforts to improve the quality of extracted genomic DNA from the Neotyphodium endophytes are ongoing. ADODR monitoring is done via e-mail and phone calls.