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Aim: Morpho-molecular analyses for taxonomic characterization of nine predominant Penicillium species present in the soil of different parts of India. Methodology: Fifteen Penicillium isolates were isolated from the soil samples collected from the experimental field of ICAR-Indian Agricultural Research Institute (IARI), New Delhi. Another twenty-six isolates were procured from Indian Type Culture Collection (ITCC), Division of Plant Pathology, ICAR-IARI, New Delhi which were isolated from the soil of different parts of India. Total 41 isolates were characterized following distinct macroscopic (colony texture, colony colour exudate production; soluble pigmentation; reverse coloration and mycelial growth) and microscopic observations (type of penicillus; shape of phialides; conidial shape, size and pigmentation). Molecular characterization was done using partial β-tubulin gene sequence which is considered an excellent marker in differentiating Penicillium species. Results: The morphological data in species description perfectly matched with molecular data generated using β-tubulin gene marker and nine different species viz., P. aethiopicum, P. chrysogenum, P. crustosum, P. janthinellum, P. mononematosum, P. oxalicum, P. polonicum, P. singorense and Talaromyces pinophilus (Syn. P. pinophilum) were confirmed. Interpretation: Above nine Penicillium species are found predominantly in the soil collected from different parts of India. The β-tubulin gene can be considered as an excellent marker to differentiate Penicillium species as confirmed from this study. The combined morpho- molecular analyses can be further utilized to delineate morphologically similar Penicillium species and also helpful to establish new species of Penicillium.