Cocklebur (Xanthium strumarium L.) seed oil is available for biodiesel production as a wild plant species. In the present study, the cocklebur seeds collected from Muslubelen, Yozgat, Turkey, at 1440 m in 2014 were sown in the Bozok University Agriculture Faculty Topcu Research Area during 2015 and 2016. The maximum crude oil ratio was obtained as 35%. Oleic and linoleic acid contents of cocklebur seed oil were determined as 11.37% and 76.97%, respectively. Some important fuel properties such as density, flash point, water content, copper strip corrosion, iodine value, linolenic acid, methyl ester, and sulfated ash content values were measured and compared to the EN 14214 and ASTM D6751 standards. These values were found to be within the standard ranges. Cost analysis of the cocklebur plant was also performed and compared to other field crops. According to our results, it seems that agronomic production of cocklebur will be possible in marginal areas because its genetic properties enable cocklebur to grow in dry areas. Therefore, production of cocklebur as an industrial plant may contribute to energy shortages without competing with food production. In areas devoid of sufficient irrigation possibilities, agriculture based on grain and cereal decreases the biodiversity. Production of drought-tolerant plants such as cocklebur in these areas will have positive effects on conservation and development of biodiversity in the ecosystem.