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Summary  Systemic fungicides and antifungals are used as frontline treatments for fungal diseases in plants and humans. It is generally accepted that fungicides will bring significant negative side‐effects to the environment and result in fungicide resistance in the pathogenic fungi. Although previous research has focused on fungicide application rates and fungal resistance for a long time, little attention has been paid to fungicide residues after treatment, especially their potential role in fungal growth and sporulation. Here we investigated the effect of fungicides at sublethal concentrations on fungal sporulation. The results showed that two kinds of 14α‐demethylase inhibitors (DMIs) fungicides increased the number of isolates of  Colletotrichum  spp. to sporulate on PDA. Both on PDA medium and plant tissue, low concentration of DMI fungicides could promote spore production of  Colletotrichum  spp., whereas pyraclostrobin, a quinone outside inhibitor (QoIs), had no significant effects on sporulation of  Colletotrichum  spp. Transcriptomic analysis suggested that the DMIs fungicide stress signal may be transmitted to the central regulatory pathway through the FluG‐mediated signalling pathway, and further confirmed the morphological effect of DMI fungicide on promoting sporulation of  Colletotrichum . To our knowledge, this is the first study to provide insights into the reproductive response of fungi in response to fungicide stress. Our findings indicate that fungicides have two‐way effects on the growth and reproduction of pathogenic fungi and provide a new basis for the scientific and rational use of fungicides.

Reproduction response of Colletotrichum fungi under the fungicide stress reveals new aspects of chemical control of fungal diseases