Abstract: 【Objective】To study the phenotype and genetic diversity of Machilus gamblei King ex Hook. f. germplasm in Guangdong and provide the basic data for conservation, effective utilization of M. gamblei gemplasm and the breeding of new varieties.【Method】Five typical populations of M. gamblei in Guangdongwere selected, such as Guangzhou Wangzishan Forest Park, Conghua Shimen National Forest Park, Huizhou Nankunshan Resort and so on. Phenotypic traits were determined and RAPD markerswere used in this study to analyze DNA fragment diversity, to explore the distribution and characteristics of phenotypicand genetic diversity between the populations and inside the populations.【Result】Among the 15 phenotypic traits of M. gamblei, the coefficient variation(CV) of 12 phenotypic traits were lower than its average;and the genetic diversity index(H') of 11 traits were higher than the average. The result showed that M. gamblei variation degree of phenotypic traits was low, but the variation types were abundant. The high resolving power(Rp) primers, OPAA-16, OPBH-10 and OPB-17, could effectively evaluate the genetic diversity among different samples of M. gambleiin Guangdong. Based on phenotypic traits, 30 samples of M. gamblei were clustered when genetic distance was 10, and the characterization relationship of samples which from Guangzhou Wangzishan Forest Park, Conghua Shimen Forest Park and Shantou Nanao Island Resorts was close. At the genetic distance of 15, based on the RAPD markers, 30 samples could be divided into 5 groups, samples of Huizhou Nankunshan Resorts, Shimen Forest Park and Wangzishan Forest Park had close genetic distance, samples of Nanao Island Resorts and Shenzhen Maluanshan Country Park showed similar genetic background. By comparing the cluster of phenotypic traits and RAPD markers, poor consistency of samples in the two groups was found.【Conclusion】The phenotypic traits and genetic diversity of M. gamblei in Guangdong has rich variation between the populations and inside the populations. Samples from Huizhou Nankunshan Resort show the richest variation in both phenotypic traits and genetic diversity, which would be helpful for germplasm resources species improvement, effective utilization and other related researches.