Details for: W6 36676, Phleum alpinum L., Phleum commutatum(-)2009

ERP planted 2004 increase seed on September 20, 2007. Three endophyte positive plants were harvested to produce the 2008 increase endo positive seed. Four endophyte negative plants were harvested to produce 2008 increase endo negative seed.

ERP planted the two types of 2008 increase seed on March 5, 2009. Eighteen endo positive plants were harvested to produce the 2009 increase endo positive seed. Fifteen plants were harvested to produce the 2009 increase endo negative seed.

All testing for the presence of viable endophyte was done in the ERP lab. Five to 6 weeks after PI test plants from E+ and E- source plants? were rooted in pots, the endophyte status of each plant was determined by isolating Neotyphodium fungi on potato dextrose agar (PDA) supplemented with streptomycin sulfate and tetracycline hydrochloride (50?g each per ml PDA). Following procedures in Clement et al. (2001), basal stem sections (~1 cm) from at least 2 tillers per plant were surface disinfected and placed on PDA in sealed petri dishes and incubated in a laboratory (complete darkness, room temperature). Petri dishes were examined for mycelial growth from plant tissue at 2-3 day intervals for 45 days. A PI test plant was scored E- if Neotyphodium mycelia did not appear during this period of time. This time consuming isolation method generates reliable data on the presence of viable Neotyphodium in plant tissue.

see: Clement, S. L., Elberson, L. R., Youssef, N. N., Davitt, C. M., and Doss, R. P. 2001. Incidence and diversity of Neotyphodium fungal endophytes in tall fescue from Morocco, Tunisia, and Sardinia. Crop Science 41:570-576.