Isolation of a potential rock phosphate solubilizing Aspergillus sp. towards development of biofertilizer

Abstract

Phosphorus (P) is one of the key bio elements that limits agricultural production. Although Sri Lanka is endowed with rock phosphate deposits, practical means to utilize this resource are limited. Phosphate-solubilizing fungi (PSF) play an important role in enhancing the bioavailability of soil phosphorus to plants. In this study, fungi from eight soil samples in Kaikawala, Sri Lanka, were isolated using spread plate method and analyzed for its phosphate solubility capacity. One fungal isolate with significant halo zones on Pikovskaya’s agar (PVK) plates, containing 0.5% tricalcium phosphate, was identified. After purification, the isolate was transferred to three media setups: PVK with 0.5% apatite (ERP) as the phosphate source, PVK with 0.5% tricalcium phosphate as a positive control, and PVK without P source as a negative control and analyzed for the solubilization index (SI). DNA sequences of the internal transcribed spacers with 5.8S region of ribosomal DNA (ITS1-5.8S-ITS2) was analyzed with universal primer pair ITS4-F/ITS5-R to determine the identity of the species. Fungal isolate PSF01 showed a phosphate solubilizing activity on both PVK media, ERP (1.07 ± 0.03 cm SI) and tricalcium phosphate (1.07 ± 0.01 cm), with similar effects (p > 0.05). However, the SI in the negative control was 1.00 ± 0.00 cm without halo zone (p < 0.05). The fungal strain is fast growing with initially white but quickly changing to black colonies after producing conidial spores on potato dextrose agar with distinctive conidial heads and pale-yellow lower surface characterized as Aspergillus. Sanger DNA sequencing identified the fungus as Aspergillus niger (99%) and a phylogenetic tree was constructed using MEGA11 software using reference data obtained from NCBI GenBank data to identify the isolated PSF01. Consequently, our preliminary studies demonstrate the importance of examining more soil samples to identify PSF for sustainable agricultural applications in the future.