Novel cellular functions of Cys ₂ -His ₂ zinc finger proteins in anthracnose development and dissemination on pepper fruits by <i>Colletotrichum scovillei</i>

<i>Colletotrichum</i> species are notorious for causing anthracnose on many fruits, leading to significant economic losses worldwide. As a model, we functionally characterized cys2-his2 (C₂H₂) zinc finger proteins (CsCZFs) in <i>Colletotrichum scovillei</i>, a major causal agent of pepper fruit anthracnose in many countries. In all, 62 CsCZFs were identified by <i>in silico</i> genomic analysis. Twelve were selected based on their expression profiles to generate targeted deletion mutants for functional investigation. <i>ΔCsczf1</i> markedly reduced conidiation and constitutive expression of <i>CsCZF1</i> partially recovered conidiation in an asexual reproduction-defective mutant, <i>ΔCshox2</i>. Deletion of <i>CsCZF12</i>, orthologous to the calcineurin-responsive transcription factor <i>Crz1</i>, impaired autophagy in <i>C. scovillei. ΔCsczf9</i> was defective in surface recognition, appressorium formation, and suppression of host defenses. <i>CsCZF9</i> was identified as an essential and novel regulator under the control of the mitogen-activated protein kinase (CsPMK1) in an early step of appressorium development in <i>C. scovillei</i>. This study provides novel insights into <i>CsCZF</i>-mediated regulation of differentiation and pathogenicity in <i>C. scovillei</i>, contributing to understanding the regulatory mechanisms governing fruit anthracnose epidemics.IMPORTANCEThe phytopathogenic fungus <i>Colletotrichum scovillei</i> is known to cause serious anthracnose on chili pepper. However, the molecular mechanism underlying anthracnose caused by this fungus remains largely unknown. Here, we systematically analyzed the functional roles of cys2-his2 zinc finger proteins (CsCZFs) in the dissemination and pathogenic development of this fungus. Our results showed that CsCZF1 plays an important role in conidiation and constitutive expression of CsCZF1 restored conidiation in an asexual reproduction-defective mutant, <i>ΔCshox2</i>. The CsCZF9, a novel target of the mitogen-activated protein kinase (CsPMK1), is essential for surface recognition to allow appressorium formation and suppression of host defenses in <i>C. scovillei</i>. The CsCZF12, orthologous to the calcineurin-responsive transcription factor Crz1, is involved in the autophagy of <i>C. scovillei</i>. Our findings reveal a comprehensive mechanism underlying CsCZF-mediated regulation of differentiation and pathogenicity of <i>C. scovillei</i>, which contributes to the understanding of fruit anthracnose epidemics and the development of novel strategies for disease management.