【Objective】The present experiment was conducted to develop the optimized cDNA-AFLP protocol and an efficient silver staining system to study the differential expressions of genes regulated by ethephon， and to understand the molecular mechanism of ethephon regulated growth and development in sugarcane. 【Method】Forty-day old plants of sugarcane variety ROC16 were foliarly sprayed with ethephon (200 mg/L)， and the leaves were sampled at 0，3，6，12，24 and 48 hours to extract the total RNA and establish the cDNA-AFLP protocol and silver staining system. Using combinations of 193 primer pairs， some transcript derived fragments (TDFs) were isolated， cloned， detected with reverse Northern blotting and sequenced.【Result】The cDNA-AFLP analysis results showed high level of polymorphism amongst the treatment and control. 35 to 80 bands were amplified in each sample. Eleven samples with 24 TDFs did not display any differential expression，and the ratio of hypocrite positive TDFs was recorded as 46%. The chitinase I(CHI），glutathione S-transferase(GST），auxin-responsive protein(ARP），light harvesting chlorophyll a/b-binding protein（LHC），nuclear binding protein gene and a set of unknown genes were differentially expressed in the ethephon treatments. CHI and GST have an important role in resistance to stress and various diseases，while LHC and nuclear binding protein are related to light harvesting and CO2 fixation in photosynthesis，respectively. 【Conclusion】The cDNA-AFLP technique was found to be very effective and feasible to study the differential expression of genes in sugarcane. Ethophen was found to regulate the expression of genes related to the primary metabolism of sugarcane and resistance to disease and stress by regulating the expression of related genes.