Poster

         Symbioses: Plant, Animal, and Microbe Interactions

Antagonistic activity is observed from seed endophytes: A potential tool to aid seeds in establishment during restoration

Presenting Author
Alexandrite Greenhouse
Description
Ecological restoration of damaged habitats is of major concern for many ecosystems today. In the arid Inland Northwest, harsh conditions and microbial diseases can reduce the rate of establishment of seedlings used in restoration. Seed endophytes are potential tools that have yet to be explored in restoration. The goal of our research was to isolate and identify seed endophytes from plant species often used in restoration in the Inland Northwest and to test the antagonistic interactions between endophytic bacteria and fungi. To isolate seed endophytes, we surface-sterilized seeds from two species commonly used in field restoration in the Inland Northwest, the common burnet (Sanguisorba minor, abbreviated as Sami) and a species of needle and thread grass (Hesperostipa comata, abbreviated as Heco). We plated these sterilized seeds on four different agar media (PDA, TSA, R2A, and LB) in order to isolate a more diverse set of endophytes, both bacteria and fungi. Once we isolated the endophytes, we then chose four bacteria and four fungi from each seed species and set them up on agar plates to test the degree of their antagonism. We set up our antagonism plates by taking 4mm plugs of the fungi from a pure culture as well as a swab of bacteria and placing them on PDA plates approximately 2 cm apart. We then compared the degree of inhibition of the bacteria and the fungi to our controls. From our census of endophytes on our seed plates, we discovered that there was a higher frequency and greater richness of fungal morphotypes than bacterial morphotypes (72% more fungal morphotypes than bacterial for S. minor and 28% more fungal morphotypes than bacterial for H. comata). We also found that there was a greater total richness of endophytes for H. comata seeds than for S. minor seeds (58% more morphotypes for H. comata than for S. minor). From our antagonism data, we found that three out of four of the bacteria isolated from H. comata seeds and one out of four from the S. minor seeds were at least somewhat antagonistic to the fungi. Meanwhile, one of four H. comata bacteria and two of four bacteria for S. minor were completely overtaken by all fungi. These results imply that the antagonistic interactions were driven by the antifungal properties of the different bacterial morphotypes rather than by the fungal morphotypes. This is relevant to our research goal in that it could mean endophytic bacteria could be more effective at promoting early seedling establishment in the face of fungal pathogens.