# Characteristics of Plant Community, Soil Physicochemical Properties, and Soil Fungal Community in a 22-Year Established Poaceae Mixed-Sown Grassland

## TL;DR

This study shows that mixing three to four grass species in artificial grasslands improves plant growth, soil quality, and fungal diversity over 22 years on the Qinghai–Tibet Plateau.

## Contribution

The study identifies optimal grass mixtures for grassland restoration by linking plant diversity to soil and fungal community improvements.

## Key findings

- Mixing three grass species (HC) increased plant biomass and soil nutrients significantly compared to monoculture.
- Mixing four grass species (HD) improved fungal diversity and community structure.
- Soil electrical conductivity (SEC) was a key driver of fungal community assembly and correlated with other soil properties.

## Abstract

This study aims to evaluate the restoration effect of artificially mixed-sown grasslands by investigating the characteristics of plant communities and soil fungal communities in long-term (22-year-established) artificial grasslands under six Poaceae mixture combinations. The experiment took mixed-sown grasslands of grass species established in 2002 on the Qinghai–Tibet Plateau as the research object. It employed ITS gene high-throughput sequencing technology to construct a fungal community distribution map and combined it with FUNGuild (Functional Guilds of Fungi) functional predictions to analyze fungal species abundance, structural diversity, molecular co-occurrence networks, and functional characteristics. By integrating Mantel test and RDA (redundancy analysis), we identified key environmental factors driving soil microbial community structure in mixed-sown grasslands and revealed the plant–soil–microbe interaction mechanisms in a Poaceae mixture grassland. The results showed that the HC treatment (a mixture of three grass species) significantly enhanced plant biomass and soil nutrient accumulation. In 2023 and 2024, its aboveground biomass increased by 66.14% and 60.91%, respectively, compared to the HA treatment (monoculture). Soil organic matter increased by 52.32% and 48.35%, while electrical conductivity decreased by 48.99% and 51.72%, respectively. The fungal community structure improved under the HD treatment (a mixture of four grass species), with an increased abundance of the dominant phylum Ascomycota and a 14.44% rise in the Shannon index compared to the HA treatment. The network complexity under the HF treatment (a mixture of six grass species) increased (with edge numbers reaching 494), while the functional abundance of plant pathogen was significantly lower than that under the HA treatment. Mantel test and RDA revealed that SEC (soil electrical conductivity) was significantly positively correlated with pH, while both exhibited negative correlations with other plant and soil physicochemical indicators. Moreover, SEC emerged as the core factor driving fungal community assembly. Mixed sowing of three to four grass species effectively regulated soil electrical conductivity, simultaneously enhancing plant biomass, soil nutrients, and fungal community diversity, representing an optimal strategy for artificial restoration of degraded grasslands.