The potential of near infrared (NIR) reflectance spectroscopy to predict various physical, chemical and biochemical properties in Mediterranean soils from SE Spain was evaluated. Soil samples (n=393) were obtained by sampling thirteen locations during three years (2003-2005 period). These samples had a wide range of soil characteristics due to variations in land use, vegetation cover and specific climatic conditions. Biochemical properties also included microbial biomarkers based on phospholipid fatty acids (PLFA). Partial least squares (PLS) regression with cross validation was used to establish relationships between the NIR spectra and the reference data from physical, chemical and biochemical analyses. Based on the values of coefficient of determination (r(2)) and the ratio of standard deviation of validation set to root mean square error of cross validation (RPD), predicted results were evaluated as excellent (r(2)>0.90 and RPD>3) for soil organic carbon, Kjeldahl nitrogen, soil moisture, cation exchange capacity, microbial biomass carbon, basal soil respiration, acid phosphatase activity, β-glucosidase activity and PLFA biomarkers for total bacteria, Gram positive bacteria, actinomycetes, vesicular-arbuscular mycorrhizal fungi and total PLFA biomass. Good predictions (0.81<r(2)<0.90 and 2.5<RPD<3) were obtained for exchangeable calcium and magnesium, water soluble carbon, water holding capacity and urease activity. Resultant models for protozoa and fungi were not accurate enough to satisfactorily estimate these variables, only permitting approximate predictions (0.66<r(2)<0.80 and 2.0<RPD<2.5). Electrical conductivity, pH, exchangeable phosphorus and sodium, metabolic quotient and Gram negative bacteria were poorly predicted (r(2)<0.66 and RPD<2). Thus, the results obtained in this study reflect that NIR reflectance spectroscopy could be used as a rapid, inexpensive and non-destructive technique to predict some physical, chemical and biochemical soil properties for Mediterranean soils, including variables related to the composition of the soil microbial community composition.