Wild Brassica juncea is a widespread weed in China with increasingly great impact on the yield of many crops. This study aimed to develop microsatellite markers for assessing the genetic diversity and population genetic structure of B. juncea, and to provide basic information for biological and chemical control of the weed. The compound microsatellite marker technique was used to develop markers for investigating population genetics of wild B. juncea. Twelve loci were obtained, each of which showed high polymorphisms when tested in two populations in Sichuan and Jiangsu Provinces. The number of alleles per locus ranged from 4 to 27, with an average of 15.2 alleles per locus. The newly developed microsatellite loci will be informative for further investigations of the population genetics and evolutionary patterns of wild B. juncea.