Exogenous Nutrient Bag Formulations Affect Soil Fertility and Microbial Communities in Morchella sextelata Cultivation.

The application of exogenous nutrient bags (ENBs) is critical for achieving high yields in morel mushroom cultivation; however, the high cost and lack of locally adapted formulations have become major constraints for the sustainable expansion of this industry. This study aimed to evaluate the effects of ten treatments, including a conventional formulation control, incorporating locally available agricultural by-products (rapeseed straw, organic fertilizer, alfalfa, and oats), on soil fertility, enzyme activity, microbial community structure, and ultimately the yield and quality of cultivated Morchella sextelata in Jianzha County, Qinghai. A randomized complete block design was established, and soil physicochemical properties, microbial biomass carbon/nitrogen/phosphorus (MBC/MBN/MBP), and enzyme activities (urease, phosphatase, and catalase) were measured at 38, 62, and 137 days post-application, while bacterial and fungal community compositions were characterized via high-throughput sequencing of 16S rRNA and ITS2 genes. Our results demonstrated that the wa-60 treatment (30% wheat + 60% alfalfa) outperformed all the other formulations, delivering the highest yield (1.32 ± 0.08 kg/m2) and a 28.6% increase over the control (1.03 ± 0.06 kg/m2; p < 0.01), while maintaining a high total amino acid content (19.8 g/100 g). Soil analysis revealed that wa-60 notably enhanced urease activity (peaking at 85 µg/g/h at day 137) and alkaline phosphatase activity (322.55 µg/g/h), alongside significant increases in MBC and MBN. Microbiome profiling further demonstrated that wa-60 selectively increased the relative abundance of various taxa, particularly the bacterial phylum Bacteroidota and the fungal phylum Mortierellomycota. Correlation analyses indicated strong positive associations among these enriched taxa (primarily at the genus level), enhanced enzyme activities, and improved soil nutrient availability. Collectively, these findings establish that the wa-60 formulation, leveraging locally sourced alfalfa, represents a cost-effective and high-performance strategy for morel cultivation in the Qinghai Plateau, providing a microbial-ecological basis for optimizing ENB design and offering a practical pathway for recycling agricultural waste in edible mushroom production.