ABSTRACT Climate and land-use change profoundly affect plant species distribution (SD) and composition, and the impact of these processes is expected to increase in the coming years. As a proxy of global changes, knowledge of SD and diversity along climatic gradients is essential to determine the efforts needed for species conservation. Plant spectral diversity is an emerging approach used as a proxy for species diversity based on remote sensing. Thus, the research aim was to develop a comprehensive methodology based on spectral diversity for SD and richness mapping and to study their relations with environmental and human-derived factors, demonstrated along Mediterranean to semi-arid climatic gradient. The study addresses two main knowledge gaps regarding spectral diversity: (1) improving the accuracy of woody species classification by features extraction and selection, and by using texture analysis in an ecosystem characterized by high spatial variability and relatively small-sized and sparse woody vegetation; and (2) developing a better estimate of the local species richness and their response to environmental and human-derived factors (i.e. climate, topography, substrate, and land cover factors) across a transition zone between Mediterranean woodlands and semi-arid dwarf shrublands. A hyperspectral image was acquired for a 43-km strip along the study area using an airborne flight of AISA-FENIX (380–2500 nm, 420 bands) at the end of the 2017 rainy season. The dominant species were surveyed, with a total number of 247 trees and shrubs, to train a machine learning support vector machine (SVM) classification for species distribution mapping, which yielded an overall accuracy of 86.1%. A feature extraction and selection methodology was developed, combining principal component analysis and neighborhood component analysis techniques, facilitating the identification of 33 spectral diagnostic bands out of 330 spectral bands. The classification accuracy was decreased by about 2% to 84.2% using only 33 spectral bands. The classification accuracy improved by about 7.1% for the seven large crown species (93.3%) by adding texture information. Later, the local species richness was calculated by utilizing the alpha diversity index (i.e. the Shannon Index) for 30-m grid cells and was tested in response to environmental (i.e. climate, substrate, and topography) and human-derived factors (i.e. land cover). The highest sensitivity to alpha diversity factors was mean annual precipitation, slope, and land surface temperature. The alpha diversity showed higher richness in the natural Mediterranean shrubland and the guarrigue located in the northern part of the climate gradient. We suggest that the approach presented here significantly improves the estimation of woody species distribution and diversity in areas characterized by high spatial heterogeneity along steep climatic gradients.