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During an ongoing research survey of saprobic fungi in Thailand, four coelomycetous strains were isolated from decaying leaves in Chiang Mai and Phitsanulok Provinces. Morphological characteristics demonstrated that these taxa are typical of <i>Cytospora</i> in forming multi-loculate, entostromatic conidiomata, branched or unbranched conidiophores, with enteroblastic, phialidic conidiogenous cells and hyaline, allantoid, aseptate conidia. Multiloci phylogeny of ITS, LSU, ACT, RPB2, TEF1-α and TUB2 confirmed these taxa are distinct new species in <i>Cytospora</i> in Cytosporaceae (Diaporthales, Sordariomycetes), viz., <i>Cytospora</i> <i>chiangmaiensis</i>, <i>C. phitsanulokensis</i> and <i>C</i>. <i>shoreae</i>. <i>Cytospora</i> <i>chiangmaiensis</i> has a close phylogenetic relationship with <i>C</i>. <i>shoreae</i>, while <i>C. phitsanulokensis</i> is sister to <i>C. acaciae</i>. These three novel species were also preliminary screened for their antagonistic activity against five plant pathogenic fungi: <i>Colletotrichum</i><i>fructicola</i>, <i>Co. siamense</i>, <i>Co. artocarpicola</i>, <i>Co. viniferum</i> and <i>Fusarium sambucinum</i>. <i>Cytospora</i> <i>shoreae</i> and <i>C. phitsanulokensis</i> showed >60% inhibition against <i>Co. viniferum</i> and <i>F. sambucinum,</i> while <i>C. chiangmaiensis</i> had moderate inhibition activity against all pathogens.