To understand how light intensity influences plant morphology and photosynthesis in the forage crop alfalfa (Medicago sativa L. cv. Zhongmu 1), we investigated changes in leaf angle orientation, chlorophyll fluorescence, parameters of photosynthesis and expression of genes related to enzymes involved in photosynthesis, the Calvin cycle and carbon metabolism in alfalfa seedlings exposed to five light intensities (100, 200, 300, 400 and 500 μmol m⁻² s⁻¹) under hydroponic conditions. Seedlings grown under low light intensities had significantly increased plant height, leaf hyponasty, specific leaf area, photosynthetic pigments, leaf nitrogen content and maximal PSII quantum yield, but the increased light-capturing capacity generated a carbon resource cost (e.g., decreased carbohydrates and biomass accumulation). Increased light intensity significantly improved leaf orientation toward the sun and upregulated the genes for Calvin cycle enzymes, thereby increasing photosynthetic capacity. Furthermore, high light (400 and 500 μmol m⁻² s⁻¹) significantly enhanced carbohydrate accumulation, accompanied by gene upregulation and increased activity of sucrose and starch-synthesis-related enzymes and those involved in carbon metabolism. Together, these results advance our understanding of morphological and physiological regulation in shade avoidance in alfalfa, which would guide the identification of suitable spatial planting patterns in the agricultural system.

WRKY transcription factors, so named because of the WRKYGQK heptapeptide at the N-terminal end, are widely distributed in plants and play an important role in physiological changes and response to biotic and abiotic stressors. Many previous studies have focused on the evolution of WRKY transcription factors in a given plant; however, little is known about WRKY evolution in legumes. The gene expression pattern of duplicated WRKY transcription factors remains unclear. We first identified the WRKY proteins in 12 legumes. We found that the WRKYGQK heptapeptide tended to mutate into WRKYGKK. The Q site in WRKYGQK preferentially mutated, while W, K, and Y were conserved. The phylogenetic tree shows that the WRKY proteins in legumes have multiple origins, especially group IIc. For example, WRKY64 from Lupinus angustifolius (LaWRKY64) contains three WRKY domains, of which the first two clustered together in the N-terminal WRKY domain of the group I WRKY protein, and the third WRKY domain grouped in the C-terminal WRKY domain of the group I WRKY protein. Orthologous WRKY genes have a faster evolutionary rate and are subject to constrained selective pressure, unlike paralogous WRKY genes. Different gene features were observed between duplicated WRKY genes and singleton WRKY genes. Duplicated Glycine max WRKY genes with similar gene features have gene expression divergence. We analyzed the WRKY number and type in 12 legumes, concluding that the WRKY proteins have multiple origins. A novel WRKY protein, LaWRKY64, was found in L. angustifolius. The first two WRKY domains of LaWRKY64 have the same origin. The orthologous and paralogous WRKY proteins have different evolutionary rates. Duplicated WRKY genes have gene expression divergence under normal growth conditions in G. max. These results provide insight into understanding WRKY evolution and expression.

High soil salinity is the main factor that limits soil microbial activity in the Yellow River Delta (YRD); however, its effects on fungal community and ecological function are unknown. Here, we comparatively investigated the diversity and structures of soil fungal communities targeting the internally transcribed fungal spacer gene using Illumina MiSeq sequencing methods under a salt gradient with five levels, namely, Low: low-salinity soil, Medium: medium-salinity soil, High: high-salinity soil, Extreme: extreme-salinity soil, and a non-salted site as the control (Non-saline). The results show that bulk density (BD) values significantly increased (p < 0.05), while significantly lower values of soil total carbon (TC), total nitrogen (TN), and fungal Shannon and Chao indexes were observed as the salinization gradient increased (p < 0.05). The relatively high levels of the families Nectriaceae and Cladosporiaceae distinguished two of the clusters, indicating two enterotypes of low (Non-saline and Low) and high (Medium, High, and Extreme) salinity soils, respectively. The family Nectriaceae was most abundant in the networks, and the positive correlations were more pronounced than negative correlations; however, Cladosporiaceae was the family most negatively correlated with others based on the network analysis. At the ecological function level, plant saprotrophs and litter saprotroph were significantly less abundant in extremely saline soil than non-saline soil. The change in soil properties (TC, TN, and BD) caused by soil salinization [salt and electrical conductivity (EC)] regulated the diversity of soil fungal communities, and ecological function, as indicated by Pearson correlation analyses. We suggest further investigation into the ecological functions of soil microorganisms in the extremely saline-alkaline soils of the YRD.