Volatile oil, as one of the important functional components in Perilla frutescens leaves, plays a significant role in anti-bacteria, anti-inflammatory and anti-oxidation. In this study, by taking use of 18 perilla germplasm resources in Guizhou province, we extracted the volatile oil with the steam distillation followed by GC-MS analysis. The significant differences in essential oil content and composition among different P. frutescens materials were observed. For instance, the content of essential oil was ranged from 0.46% and 1.44%. Seven-five chemical constituents were identified in the essential oils, mainly including: perillaketone, piperidone, elemicin, perillaldehyde, myristicin, caryophyllene, perillane, apioland acetophenone . Based on synthesis pathways, the tested Perilla materials can be classified into six chemotypes including F-type, PA-type, PK-type, PL-type, PT-type, PP-type. In addition, we also identified some special substances, such as citral, citronellene, eucalyptus oil essence, orange flower tertiary alcohol, the presence of which may provide a hypothesis on explaining a unique aroma of some Perilla plants in Guizhou. Therefore, this work showed abundant diversity on content and chemotype composition of P. frutescens resources in Guizhou, and also might provide theoretical reference for quality control and clinical application in the future.