Now days, production of fuels and petrochemicals from renewable lignocellulosic biomass is an indispensable issue to meet the growing energy demand. Meanwhile, the changes in the climate and soil topography influence the growth and development as well as canopy level of the lignocellulosic biomass. In this study, Zilla spinosa Turr (Zilla) plants with similar age and size were collected from three main sectors (upstream, midstream, and downstream) of Wadi Hagul during spring (April) and summer (July) seasons. Environmental stresses evoked reduction in the energy trapping pigments concomitant with increments in chlorophyll fluorescence in summer harvested plants particularly at downstream. Furthermore, the biofuels generating compounds including carbohydrate, lignin, and lipid making the plant biomasses are greatly affected by environmental conditions. Greater amount of lignin was estimated in summer harvested Z . spinosa shoots particularly at downstream. Moreover, the total oil content which is a promising source of biodiesel was considerably decreased during summer season particularly at downstream. The physical properties of the lipids major constituent fatty acid methyl esters determine the biofuel properties and contribute in the adaptation of plants against environmental stresses. Hence, the analysis of fatty acid profile showed significant modifications under combined drought and heat stress displayed in the summer season. The maximum increase in saturated fatty acid levels including tridecanoic acid (C13:0), pentadeanoic acid (C15:0), palmitic acid (C16:0), and stearic acid (C18:0) were estimated in spring harvested Z. spinosa aerial portions particularly at midstream. In spite of the reduction in the total oil content, a marked increase in the value of unsaturated to saturated fatty acids ratio and thereby the unsaturation index were achieved during the dry summer period. Henceforth, these seasonal and spatial variations in fatty acids profiles may contribute in the acclimatization of Z. spinosa plants to soil water scarcity associated with heat stress experienced during summer. In addition, the alterations in the fatty acid profiles may match biofuel requirements. In conclusion, the most adequate growing season (spring) will be decisive for achieving high lipid productivity associated with improved biofuel quality in terms of high saturated fatty acids percentage that improves its cetane number. However, the dry summer season enhanced the accumulation of greater amount of lignin that may enhance the biodiesel quantity.