Saccharum spontaneum L. is the closest relative wild species to the modern sugarcane cultivar. Considering the capacity on resistance to a variety of adverse environments, it was popularly used as donors in sugarcane breeding for resistant cultivars. To date, only a few of S. spontaneum clones and their resistance genes have been incorporated effectively into sugarcane commercial varieties, and the most of present cultivars have a single or weak resistance capacity and a narrow resource of resistance genes. It is of great significance to enhance the researches on screening of superior breeding parents with high stress resistance and mining of stress resistance genes from the wild germplasm such as S. spontaneum. This review summarized the past and current research progresses on stress tolerance identification, cloning of resistant genes as well as functional validation from different S. spontaneum genotypes, for instance in fields of drought/chill tolerance and insects/pathogen diseases infections. The current problems and future prospects of the highly resistant genotypes screening and gene mining are also discussed. We expect to provide better understanding on the resistant variance from genotypes, and insights in efficient use of the rich resistant gene pool for future sugarcane breeding of multiple resistance polymerization.