Among the agricultural crop residues, wheat straw is the second largest biomass feedstock in the world after rice straw. The recent study was done to evaluate fermentable sugars production potential of wheat varieties, mostly grown in northern and central a part of India, which has not been studied before with respect to hydrogen production. Characterization of wheat varieties on the basis of its chemical constituents i.e. fibre (cellulose, hemicellulose and lignin), sugar content and total solids, showed it as a potential feedstock for hydrogen production. Pretreatment is considered the most important step in determining the cost and success of the process of enzymatic saccharification of lignocellulosic biomass. The effectiveness of pretreatment using microwave assisted dilute acid to produce maximum fermentable sugars in terms of total soluble sugar from the straw was investigated. The results showed that microwave assisted acid pretreatment is a promising approach to enhance total soluble sugar yields for hydrogen production. The temperature 160?C and time period 10 min was best for total soluble sugar recovery from wheat straw. The wheat variety HD 2967 was found to be the best followed by WR 544, C 306 and K 68 with respect to the release of soluble sugars from the straw. HD 2967 had significantly higher value in terms of sugar than other varieties.