LAUSR

Simple Summary Observations in the animal room have shown that the seeds stored by harvester ants, although in a damp environment, are less likely to mold. It was hypothesized that harvester ants may use actinomycetes to protect their seed stores, given that leafcutter ants use actinomycetes as producers of defensive substances. Two actinomycetes were isolated from the harvester ant Messor orientalis. The fermentation broth of the actinomycetes showed significant inhibitory effects on the three indicator fungi. Coculture experiments supported the observed inhibitory effects. The antifungal activities of actinomycetes in harvester ants were revealed. This research provides a significant theoretical reference for the abovementioned hypothesis and for the potential agricultural applications of these actinomycetes for multiple crops. Abstract Observations have shown that seeds collected by harvester ants are less likely to mold. Based on evolutionary analysis and other research, it was hypothesized that harvester ants could apply actinomycetes to protect seeds, similar to the protection of mutualistic fungi by leafcutter ants. Two actinomycetes were successfully isolated from the harvester ant Messor orientalis. The taxonomic status of the actinomycetes was determined by 16S rRNA sequence analysis and biochemical experimental observations. Their inhibitory effects on plant pathogens were measured. One of the bacteria was identified as Brachybacterium phenoliresistens and denoted as B. phenoliresistens MO. The other belonged to the genus Microbacterium. It was named Microbacterium sp. Growth rate determination and coculture experiments were performed to explore the inhibitory effect of actinomycetes on indicator plant pathogens. The inhibition rates of the actinomycetes toward Peronophythora litchii and Rhizoctonia solani were 100% in media containing 30% or more fermentation broth, and they also showed an inhibitory effect on Colletotrichum siamense. The coculture experiment supported this result by showing that the growth of P. litchii and R. solani was inhibited in the presence of actinomycetes. Therefore, the results of this study show the agricultural application potential of these bacteria and may provide a reference for research on the symbiosis of harvester ants with actinomycetes.