Epiphytic diatoms on seagrass and seaweed were collected from tropical (e.g., Siladen Island, Celebes Sea, Indonesia and Phú Bài, China Sea, Vietnam), subtropical (e.g., Sharm el-Sheikh, Red Sea, Egypt), and temperate regions (e.g., Patmos Island, Greece) in 2000, 2005, and 2006. Eight species of Mastogloia, belonging to the section Sulcatae, are described mainly through scanning electron microscopy, including two new species to science, M. oculoides and M. sergiana. These species show a differently shaped median depression on the external valve face between the raphe-sternum and the valve margin. Moreover, they lack a developed conopeum or pseudoconopeum, which covers the median depression in other species of the section Sulcatae. This study gives new insights on the ultrastructure of the Mastogloia's valves and provides an update of their current geographical distribution.