In this paper, we explore the role of biofuel production on deforestation in low- and middle-income countries. Since the 2000s, biofuel production has been rapidly developing to address issues of economic development, energy poverty and reduction of greenhouse gas (GHG) emissions. However, the sustainability of biofuels is being challenged, particularly at the environmental level, due to their impact on deforestation and the GHG emissions they can generate as a result of land-use changes. In order to isolate the impact of bioethanol and biodiesel production among classic determinants of deforestation, we used a fixed effects panel model of 112 low- and middle-income countries between 2001 and 2012. Firstly, we found a positive relationship between bioethanol production and deforestation in these countries, among which we highlighted the specificity of Upper-Middle-Income Countries (UMICs). An acceleration of incentives for the production of biofuels from 2006 onwards enabled us to highlight higher marginal impacts for the production of bioethanol in the case of low- and middle-income countries and UMICs. Secondly, for low- and middle-income countries, these results are not significant before 2006. Thirdly, biodiesel production appears to have an impact on deforestation before 2006 on both subsamples. These last two results seem surprising and could be related to the role of biofuel production technologies and the crop yields used in their production.