Alfalfa (Medicago sativa L.) is a nutritious forage plant in dryland regions with strong drought tolerance and broad adaptability. To understand the architectural characters and physiological functions of the root systems in response to different drought stress and rehydration, two experiments were conducted in a glasshouse. Experiment 1 used three alfalfa cultivars and four soil moisture treatments in a split-plot design with four replicates to investigate the response of root systems of different alfalfa cultivars to drought stress and subsequent rehydration. Experiment 2 used one alfalfa cultivar and four soil moisture treatments in a randomized complete block (RCB) design with three replicates to measure reactive oxidant species and antioxidant defense systems of alfalfa root. The soil moisture treatments included (1) 75–80% of the water-holding capacity (CK); (2) 60–65% of the water-holding capacity and subsequent rehydration to 75–80% of the water-holding capacity; (3) 45–50% of the water-holding capacity and subsequent rehydration to 60–65% and 75–80% of the water-holding capacity; and (4) 30–35% of the water-holding capacity and subsequent rehydration to 45–50%, 60–65% and 75–80% of the water-holding capacity. Our results demonstrated that plants had higher root biomasses (mean = 0.77 g), fractal dimensions (1.47), fractal abundances (3.84), and branching ratios (0.39) under severe water deficit and rehydration than under less stressful soil moisture conditions. Plant root systems tended to exhibit herringbone branching (Topological Index = 0.63) when soil moisture was at 60–65% water-holding capacity with the soil being then rehydrated, whereas they tended to have a smaller lateral root length, root surface area, root volume, and branching ratio and larger root diameter under more favorable soil moisture conditions for less stressful soil moisture treatments. Severe water stress and subsequent considerably improved superoxide (O₂⁻) generation rate, the content of hydrogen peroxide (H₂O₂) and Malondialdehyde (MDA), and non-enzyme antioxidant glutathione (GSH), and reduced the content of ascorbate (AsA) and activity of catalase (CAT), ascorbate peroxidase (APX), and glutathione peroxidase (GPX). It is concluded that Medicago sativa L. cv Zhaodong (ZD) and cv Aohan (AH) exhibited higher drought resistances than cv Golden empress (GE) using topological and fractal analyses. This could be a key contributor to altering optimum soil moisture conditions and screening alfalfa cultivars based on root system architecture and antioxidant defense systems. The molecular functions warrant further investigation which could help explain the observed root phenotypes after drought conditions and rehydration.

Belowground root systems under pasture intercropping exhibit complex interactions, and the root interactions of different intercropping combinations are still poorly understood. Therefore, in this work, two perennial and annual herbages were intercropped in pairs and evaluated at a ratio of 1:1. The root morphology and topological structure differed significantly with intercropping combinations. (1) Compared with other cropping patterns, the mean root diameter (RD) of intercropped alfalfa (Medicago sativa L.) and common vetch (Vicia sativa L.) increased notably. The root surface area (RSA), root volume (RV), and mean RD increased significantly when oat (Avena sativa L.) was intercropped with alfalfa. Similarly, the RSA and RV increased in intercropped oat, intercropping relative to monocropping. (2) The forage topological index of the intercropping system was close to one, which was close to that of the herringbone branching. Additionally, the intercropping system had a lower intensity of underground root competition. The root system of the different forage intercropping combinations tended to transition to dichotomous branching. (3) The correlations between root parameters differed according to forage species. Therefore, different intercropping combinations had different belowground root levels of competitiveness and interactions, thereby changing the resource competition environment.

Alfalfa (Medicago sativa L.) is a perennial leguminous plant, with a strong tap root system that plays an important role in alfalfa’s adaptation to drought stress. However, a better understanding of root functional traits and how these root traits are related to whole plant responses in order to improve pasture productivity under water deficit. Two greenhouse experiments were conducted: Experiment 1 used three alfalfa cultivars and four levels of soil water content treatments to investigate herbage productivity, growth point density, residual shoot weight, and root weight. Experiment 2 assessed relationships among root-sourced abscisic acid (ABA), root system architecture and plant biomass in response to water deficit. The results demonstrated that root system was used as a useful tool to improve tolerant and adaptation when alfalfa copied with lower levels of soil water content. On average, maintaining 60–65% soil water-holding capacity alfalfa had the highest herbage accumulation (6.7 g DM pot-1), growing point density (46.5 pot-1), and residual shoot biomass (1.8 g DM pot-1). At the level of water stress, Medicago sativa L. cv Zhaodong (ZD) and cv Aohan (AH) tended to exhibit a herringbone branching pattern with less root tips, root forks, altitude, and magnitude than cv Golden empress (GE). Principal component analysis and structural equation modeling revealed that root-sourced ABA positively regulated the altitude and magnitude of root system architecture, root tips and root forks, and was closely associated with plant root biomass and herbage biomass. It was concluded that these findings can contribute to developing optimum irrigation strategies and help alfalfa breeders in the development of new cultivars with improved drought tolerance based on root system architecture, plant hormone, and plant growth.

Leymus chinensis is a grass species in the family Triticeae that is found in the Eurasian grassland region and is known for its outstanding ecological advantages and economic value. However, the increasing adoption of photovoltaic agriculture has modified the light environment for the grass, markedly inhibiting its photosynthesis, growth, and yield. This study used physiological and transcriptomic analyses to investigate the complex response mechanisms of two L. chinensis genotypes (Zhongke No. 3 [Lc3] and Zhongke No. 5 [Lc5]) under shading stress. Growth phenotype analysis revealed the superior growth performance of Lc3 under shading stress, evidenced by enhanced plant height and photosynthetic parameters. Additionally, differentially expressed genes (DEGs) were predominantly enriched in starch and sucrose metabolism and glycolysis/gluconeogenesis pathways, which were the most consistently enriched in both L. chinensis genotypes. However, the flavonoid biosynthesis and galactose metabolism pathways were more enriched in Lc3. Weighted gene co-expression network analysis identified the LcGolS2 gene, which encodes galactinol synthase, as a potential hub gene for resistance to shade stress in comparisons across different cultivars and shading treatments. The use of qRT-PCR analysis further validated the genes involved in these pathways, suggesting that they may play critical roles in regulating the growth and development of L. chinensis under shading conditions. These findings provide new insights into the molecular mechanisms underlying the growth and development of L. chinensis under different shading stress conditions.