Abstract Microbial trace fossils on terrestrial basalts can be used as an analogue in the search for traces of life on other terrestrial planets. This study reports on microbial trace fossils within Permian greenstones in the Maizuru Terrane, southwest Japan, which is recognized as back-arc basin oceanic crust that consists mainly of metabasalt and metagabbro. The trace fossils have been studied by means of morphology, mineralogy, elemental mapping, and carbon isotope analysis. Although minute original textures of trace fossils are recrystallized in these rocks, Granulohyalichnus vulgaris isp., Tubulohyalichnus spiralis isp., and Tubulohyalichnus annularis isp. were identified. Significant concentration of C within the trace fossils implies these are organic remnants from microbes. The δ13CPDB values <−7‰ of calcite within the greenstones indicates that the bacterial activity took place prior to the formation of calcite veins. The results support that microbial trace fossils within low-grade metamorphic basalt can be reliably identified based on their morphology and chemical composition, as reveled by elemental mapping. In this context, glassy Martian basalt may be the best rock type to investigate in terms of searching for signs of microbial activity on Earth and other planets.