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Sugarcane (Saccharum officinarum L.) is among the important commodities of the commercial sector since it accounts for 65% of the worlds sugar production. Indonesia is the worlds eighth largest country in terms of productivity, planting area and yield of sugarcane. Environmental stresses due to climate change are among factors that may cause negative impacts on the production of sugarcane. The purpose of the present study was to determine sugarcane cultivars tolerant to environmental stress based on a multiplex analysis of SCDR 1 and P5CS, as well as sugarcane defense mechanisms against environmental stress conditions. The methods used in this study were quantitative test with spectrophotometry, qualitative test with electrophoresis, DNA amplification of the target SCDR 1 and P5CS by the PCR method. SCDR 1 is a gene involved in the responses to drought, salinity and oxidative stresses in sugarcane, while P5CS is involved in the responses to drought stress and shows a negative correlation to sucrose productivity under drought stress conditions. The presence of these two genes in the tested sugarcane cultivars indicates a tolerant trait of environmental stress. The study found that 24 tested sugarcane cultivars are tolerant to environmental stress due to the presence of those two genes.