This study aims to explore the diversity and to identify some ascomycetes macrofungal species found in a para rubber plantation, analysis of the internal transcribed spacer (ITS) region and nuclear large subunit rDNA sequences (LSU) morphology by Eastern Thailand. The species identification of the collection numbers RP2, RP3, RP4, and RP5 by both ITS and LSU sequences was consistent. The RP2, RP3, RP4, and RP5 were identified as Daldinia eschscholtzii, Cookeina sulcipes, Cookeina garethjonesii, and Cookeina tricholoma, respectively. Two unidentified species, Trichoderma sp. (RP1) and Xylaria sp.(RP6) maybe require additional molecular markers other than ITS and LSU. This study suggested that macrofungal identification required a combination of morphological and molecular biology approaches for specificity and accuracy. In some fungal taxa, sequence analysis of ITS and LSU could not discriminate fungal species. Although most of the ascomycetes in this study have previously been described in Thailand, this is the first report of ascomycetes macrofungi from para rubber plantations.