Two novel species of Aspergillus section Nigri from Thai coffee beans are described as Aspergillus aculeatinus sp. nov. and Aspergillus sclerotiicarbonarius sp. nov. Their taxonomic status was determined using a polyphasic taxonomic approach with phenotypic (morphology and extrolite profiles) and molecular (β-tubulin, internal transcribed spacer and calmodulin gene sequences) characteristics. A. aculeatinus sp. nov. is a uniseriate species with a similar morphology to Aspergillus aculeatus and Aspergillus japonicus, but producing smaller conidia (2–5 μm). A. aculeatinus sp. nov. produced neoxaline, secalonic acid D and F, and aculeacins. A. sclerotiicarbonarius sp. nov. is a biseriate species similar to Aspergillus carbonarius and Aspergillus ibericus, but produces abundant sclerotia and some unique indol-alkaloids. The type strain of Aspergillus sclerotiicarbonarius sp. nov. is CBS 121057T (=IBT 28362T) and the type strain of Aspergillus aculeatinus sp. nov. is CBS 121060T (=IBT 29077T).