A suite of rare microbes interacts with a dominant, heritable, fungal endophyte to influence plant trait expression

Endophytes are microbes that live, for at least a portion of their life history, within plant tissues. Endophyte assemblages are often composed of a few abundant taxa and many infrequently-observed, low-biomass taxa that are, in a word, rare. The ways in which most endophytes affect host phenotype are unknown; however, certain dominant endophytes can influence plants in ecologically meaningful ways–including by affecting growth and immune system functioning. In contrast, the effects of rare endophytes on their hosts have been unexplored, including how rare endophytes might interact with abundant endophytes to shape plant phenotype. Here, we manipulate both the suite of rare foliar endophytes (including both fungi and bacteria) and Alternaria fulva–a vertically-transmitted and usually abundant fungus–within the fabaceous forb Astragalus lentiginosus. We report that rare, low-biomass endophytes affected host size and foliar %N, but only when the heritable fungal endophyte (A. fulva) was not present. A. fulva also reduced plant size and %N, but these deleterious effects on the host could be offset by a negative association we observed between this heritable fungus and a foliar pathogen. These results demonstrate how interactions among endophytic taxa determine the net effects on host plants and suggest that the myriad rare endophytes within plant leaves may be more than a collection of uninfluential, commensal organisms, but instead have meaningful ecological roles.

Localization of endophytic Undifilum fungi in locoweed seed and influence of environmental parameters on a locoweed in vitro culture system

Endophytic Undifilum oxytropis (Q. Wang, Nagao & Kakish) Pryor, Creamer, Shoemaker, McLain-Romero, & Hambleton found within toxic locoweeds ( Astragalus and Oxytropis spp.) produces the alkaloid swainsonine, which is responsible for locoism in grazing animals. We sought to determine the location of U. oxytropis within locoweed seed, develop endophyte free plants, and assess the influence of environmental stresses on locoweed and endophyte cultures. Undifilum was identified within the parenchyma layers in seeds of Astragalus lentiginosus M.E. Jones using light microscopy and polymerase chain reaction. Astragalus lentiginosus and Oxytropis sericea Nutt. seedlings produced in embryo culture without seed coats did not contain swainsonine or fungus. Plants produced from whole seed contained U. oxytropis in both foliage and root tissues. When the in-vitro cultured plants of O. sericea and U. oxytropis cultures were subjected to environmental stresses including high temperature, low and high pH media, nutrient deficient media, and polyethylene glycol (PEG) amended media to simulate water deficit, both dry mass and swainsonine levels were affected. Swainsonine levels were greatest for O. sericea and Undifilum cultures in PEG or hydrochloric acid amended media. Plants grown in PEG-amended media had significantly greater dry mass, while Undifilum grown in PEG-amended media had lower dry mass than other treatments.