A Novel Granular Formulation of Filamentous Fungi (Aspergillus tubingensis and Trichoderma virens): Development, Characterization, and Evaluation for Enhanced Phosphorus Availability in Agricultural Soils

Phosphorus (P) is an essential nutrient in plant development, but its availability in the soil is often limited due to chemical fixation and poor solubility. This study presents the development, characterization and evaluation of a novel granular bioinoculant formulated with Aspergillus tubingensis (P-solubilizing) and Trichoderma virens (P-mineralizing) using clinoptilolite (CZ) as a carrier to improve P bioavailability. The formulation process included the evaluation of the proposed components, the standardization of conidia production in different media cultures and conditions, the elaboration and characterization of the bioinoculant and its evaluation in plants. In this study, in vitro analysis demonstrated the synergistic effect of the components, showing that in all treatments with dual inoculation and CZ, the amount of soluble phosphorus (SP) was higher than in their counterparts (from 27.8 to 36.8 mg·L−1). A concentration greater than 1 × 109 CFU·mL−1 was obtained by standardizing the production of conidia in different media (PDA, V8-Agar and Molasses Agar), which were then used to produce granular batches containing at least 2 × 107 CFU·g−1. Furthermore, the size (88% of the granules measured <4.5 mm), purity (<2 CFU·g−1 in 10−4 dilution), and moisture content of the prototype granules (3.3–3.8%) were confirmed to be within established international quality parameters. Plant evaluations in chili and tomato demonstrated the formulation efficacy, showing an increase in both soluble and foliar P content (with at least 30% more than controls), alongside improvements in all parameters evaluated that are related to plant growth promotion (with at least 15% more growth than controls). The development of this formulation prototype represents a focused effort toward process standardization and optimization required to validate developed formulations, thus promoting the advancement of applied biotechnology.