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Chitin is the main component of fungal cell walls, which can be recognized by pattern recognition receptors (PRRs) as pathogen-associated molecular patterns (PAMP). Chitinase in filamentous fungi has been reported to degrade immunogenic chitin oligomers, thereby preventing chitin-induced immune activation. In this study, we identified the chitinase families in 10 fungal genomes. A total of 131 chitinase genes were identified. Among the chitinase families, 16 chitinase genes from <i>Puccinia striiformis</i> f. sp. <i>tritici</i> (<i>Pst</i>) were identified, and the expression of <i>PstChia1</i> was the highest during <i>Pst</i> infection. Further studies indicated that <i>PstChia1</i> is highly induced during the early stages of the interaction of wheat and <i>Pst</i> and has chitinase enzyme activity. The silencing of <i>PstChia1</i> revealed that <i>PstChia1</i> limited the growth and reduced the virulence of <i>Pst</i>. The expression level of TaPR1 and TaPR2 was induced in <i>PstChia1</i> knockdown plants, suggesting that <i>PstChia1</i> is involved in regulating wheat resistance to <i>Pst</i>. Our data suggest that <i>PstChia1</i> contributes to pathogenicity by interfering with plant immunity and regulating the growth of <i>Pst</i>.