Agricultural and agro-industrial lignocellulosic residues represent an important renewable resource for the production of fuels and chemicals towards a bio-based economy. Olive pruning, vineyard pruning and almond shells are important residues from agricultural activities in Mediterranean-type ecosystems. In the current work, bioethanol production from the above three types of agro-residues was studied, focusing on the effect of different pretreatment methods on enzymatic saccharrification efficiency of cellulose and production of second-generation bioethanol. Dilute acid, hydrothermal and steam explosion pretreatments were compared in order to remove hemicellulose and facilitate the subsequent enzymatic hydrolysis of the hemicellulose-deficient biomass to glucose. Enzymatic hydrolysis was performed in a free-fall mixing reactor enabling high solids loading of 23% w/w. This allowed hydrolysis of up to 67% of available cellulose in almond shells and close to 50% in olive pruning samples, and facilitated high ethanol production in the subsequent fermentation step; the highest ethanol concentrations achieved were 47.8 g/L for almond shells after steam explosion and 42 g/L for hydrothermally pretreated olive pruning residue.