2011 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 The main objective of this project will be 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. During FY-2011: Plant endophytes are found in a wide array of grasses. Some endophytes produce toxic effects to livestock and wildlife, while some have demonstrated agricultural benefits such as increased 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. Recent advances in DNA sequencing technologies (i.e. pyrosequencing) makes it feasible to sequence multiple fungi genomes in timely fashion. Obtaining high quality Neotyphodium DNA at suitable quantities has proven difficult. Efforts to improve the methodology of DNA extraction from the endophytes have been successful. Preparation of cultures and DNA samples for genomic sequencing using Roche 454 technology is ongoing. A series of experiments were conducted to develop a protocol for endophyte inoculation to include varying light intensities and duration, and growing media. Data were taken from water stressed endophyte + and – cloned Festuca rubra and Poa pratensis plants under a rainout shelter (first year data) to include biomass and sampling for chemical composition of plants. Monitoring Methods: • Email correspondence for the planning and execution of jointly planned experiments was successfully completed. • Visit by Nick Cameron, the Research Director of Cropmark Seeds to the Forage and Range Research Laboratory was planned and executed.