Biodiversity loss in tropical forests is rapidly increasing, which directly influence the biomass and productivity of an ecosystem. In situ methods for species diversity assessment and biomass in synergy with hyperspectral data can adeptly serve this purpose and hence adopted in this study. Quadrat sampling was carried out in Shoolpaneshwar Wildlife Sanctuary (SWS), Gujarat, which was used to compute Shannon–Weiner Diversity Index (H′). Above ground biomass (AGB) was calculated measuring the Height and Diameter at Breast Height (DBH) of different trees in the sampling plots. Four spectral indices, namely Normalized Difference Vegetation Index (NDVI), Enhanced Vegetation Index (EVI), Photochemical Reflectance Index (PRI), and Structure Insensitive Pigment Index (SIPI) were derived from the EO-1 Hyperion Data. Spearman and Pearson’s correlation analysis was performed to examine the relationship between H′, AGB and spectral indices. The best fit model was developed by establishing a relationship between H′ and AGB. Fifteen models were developed by performing multiple linear regression analysis using all possible combinations of spectral indices and H′ and their validation was performed by relating observed H′ with model predicted H′. Pearson’s correlation relation showed that SIPI has the best relationship with the H′. Model 15 with a combination of NDVI, PRI and SIPI was determined as the best model for retrieving H′ based on its statistics performance and hence was used for generating species diversity map of the study area. Power model showed the best relationship between AGB and H′, which was used for the development of AGB map. © 2020, International Society for Tropical Ecology.