Potential of endophytic fungi collected from Cucurbita pepo roots grown under three different agricultural mulches as antagonistic endophytes to Verticillium dahliae in western Washington.

Verticillium dahliae is a significant pathogen in cucurbit cropping systems for which there are limited control options outside of soil fumigation. Endophytes, fungi and bacteria that live within plant hosts without impacting the host negatively, have exhibited antagonism to V. dahliae. The objectives of this study were to survey potential V. dahliae-antagonistic endophytes from roots of 'Cinnamon Girl' pumpkin (Cucurbita pepo) grown under either polyethylene (PE), an experimental polylactic acid/ poly(hydroxalkanoate) (PLA/PHA) mulch, Weed Guard Plus, or no mulch, as well as from 'Sugar Baby' watermelon (Citrullus lanatus), and 'Tetsukabuto' squash (C. maxima x C. moschata). Four selected endophytes were screened for antagonism against V. dahliae in the laboratory, greenhouse, and field. A total of 777 isolates of potential fungal endophytes were recovered from pumpkin, watermelon, and squash roots between 2015 and 2016 of which 198 isolates were identified down to the genus level. Of those isolates, frequency of isolation was greatest for Dichotomopilus/Chaetomium spp. (5%), Cladosporium spp. (15.2 %), Clonostachys spp. (5.6 %), Epicoccum spp. (22.2 %), and Fusarium spp. (24.7 %). All five genera only weakly associated with roots grown under a particular mulch treatment (Cramer's V = 0.22) or cucurbit host (Cramer's V = 0.1925). In a laboratory culture plate assay, V. dahliae isolate JAW-113 was plated against one of four prospective endophytes (Dichotomopilus sp., Epicoccum sp., Microdochium sp., or Schizothecium sp.). The area under the Verticillium culture growth curve (AUVGC) was significantly highest (P < 0.0001) when V. dahliae was by Schizothecium sp. or Dichotomopilus sp. In a greenhouse study using a Mason jar assay with V. dahliae amended potting mix, pumpkin plant vigor, plant fresh weight, root fresh weight, and root dry weight were significantly higher (P < 0.05) for plants inoculated with Dichotomopilus sp., Epicoccum sp., Microdochium sp., and Schizothecium sp. compared to plants without endophyte inoculation. Subsequent field trials in 2017 and 2018 showed no significant differences in foliar disease severity or fruit yield, regardless of whether plants were inoculated with an endophyte or not. However, recovery of V. dahliae colony forming units from pumpkin stem sap was significantly lower (P < 0.0001) for plants inoculated with either Dichotomopilus sp. or Schizothecium sp. in 2017 or Dichotomopilus sp. in 2018.