Description

Mutualistic relationships between endophytic fungi and grasses have shown to improve the hardiness of the host. This relationship is common in grasses, including native cool-season grasses that are important in both forage and grassland ecosystems. Elymus genus members, such as Canada wildrye (CAWR), commonly host the endophytic fungi, Epichloё, while southeastern wildrye (SEWR) may not. In this study, seed of ten Elymus accessions and seed and leaves from local SEWR were assessed for endophyte infection. Infection status was confirmed via seed squash and leaf peel techniques and assessed using microscopy. Seed of one SEWR and nine CAWR accessions were assessed for endophyte infection by softening seeds in NaOH. Softened seeds were stained and squashed between a microscope slide and coverslip. Infection status was determined by scanning for mycelia within each seed. Leaf peels were excised from SEWR, stained and observed. Infection status was determined by scanning for mycelia in epidermal cells. Following assessment, all SEWR germplasm was E-, while six CAWR accessions were E+. Endophyte-infected seeds were germinated and pseudostems were used to isolate the endophyte by sterilizing the pseudostems and placing them on PDA in a dark germination chamber until endophyte growth. Isolated endophyte was used to infect E- SEWR. Infection status of SEWR was determined using leaf peels and PCR. This research will help determine if forced endophyte infection can be performed in SEWR.

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Isolation and Culture of Epichloё sp. for Re-Infection of Endophyte-Free Southeastern Wildrye (Elymus glabriflorus)

Mutualistic relationships between endophytic fungi and grasses have shown to improve the hardiness of the host. This relationship is common in grasses, including native cool-season grasses that are important in both forage and grassland ecosystems. Elymus genus members, such as Canada wildrye (CAWR), commonly host the endophytic fungi, Epichloё, while southeastern wildrye (SEWR) may not. In this study, seed of ten Elymus accessions and seed and leaves from local SEWR were assessed for endophyte infection. Infection status was confirmed via seed squash and leaf peel techniques and assessed using microscopy. Seed of one SEWR and nine CAWR accessions were assessed for endophyte infection by softening seeds in NaOH. Softened seeds were stained and squashed between a microscope slide and coverslip. Infection status was determined by scanning for mycelia within each seed. Leaf peels were excised from SEWR, stained and observed. Infection status was determined by scanning for mycelia in epidermal cells. Following assessment, all SEWR germplasm was E-, while six CAWR accessions were E+. Endophyte-infected seeds were germinated and pseudostems were used to isolate the endophyte by sterilizing the pseudostems and placing them on PDA in a dark germination chamber until endophyte growth. Isolated endophyte was used to infect E- SEWR. Infection status of SEWR was determined using leaf peels and PCR. This research will help determine if forced endophyte infection can be performed in SEWR.