Plant breeding for resistance to agricultural pests is an essential element in the development of integrated crop management systems; however, the molecular and genetic mechanisms underlying resistance are poorly understood. In this pilot study, a transcriptomic analysis of a resistant (R) vs. a susceptible (S) variety of alfalfa, with (+T) or without (−T) thrips (= 4 treatments) was conducted, ‘GN-1’ (China) was defined as the resistant cultivar, and ‘WL323’ (America) was defined as the susceptible cultivar. A total of 970 mRNAs were differentially expressed, of which 129 up- and 191 down-regulated genes were identified in the R + T/R-T plants, while 413 up- and 237 down-regulated genes were identified in the S + T/S-T plants. KEGG analysis mapped 33 and 80 differentially expressed genes to 11 and 14 substantially enriched pathways for GN-1 and WL323, respectively. Five shared pathways were linked to plant resistance traits, including beta-Alanine metabolism, fatty acid degradation, chloroalkane and chloroalkene degradation, flavonoid biosynthesis, and phenylalanine metabolism. Results indicated both thrips resistant and susceptible alfalfa cultivars can regulate gene expression in the salicylic acid (SA) and flavonoid biosynthesis pathways to induce defensive genes and protein expression (e.g. polyphenol oxidase, protease inhibitor), which enhances plant defence capacity.

The grasslands in North China are rich in fungal resources. However, the knowledge of the structure and function of fungal communities and the role of microbial communities in vegetation restoration and succession are limited. Thus, we used an Illumina HiSeq PE250 high-throughput sequencing platform to study the changing characteristics of soil fungal communities in degraded grasslands, which were categorized as non-degraded (ND), lightly degraded, moderately degraded, and severely degraded (SD). Moreover, a correlation analysis between soil physical and chemical properties and fungal communities was completed. The results showed that the number of plant species, vegetation coverage, aboveground biomass, and diversity index decreased significantly with increasing degradation, and there were significant differences in the physical and chemical properties of the soil among the different degraded grasslands. The dominant fungal phyla in the degraded grassland were as follows: Ascomycota, 44.88%–65.03%; Basidiomycota, 12.68%–29.91%; and unclassified, 5.51%–16.91%. The dominant fungi were as follows: Mortierella, 6.50%–11.41%; Chaetomium, 6.71%–11.58%; others, 25.95%–36.14%; and unclassified, 25.56%–53.0%. There were significant differences in the microbial Shannon–Wiener and Chao1 indices between the ND and degraded meadows, and the composition and diversity of the soil fungal community differed significantly as the meadows continued to deteriorate. The results showed that pH was the most critical factor affecting soil microbial and fungal communities in SD grasslands, whereas soil microbial and fungal communities in ND grasslands were mainly affected by water content and other environmental factors.

The double selection of environment adaptation and host specificity forced the diversification of rhizobia in nature. In the tropical region of China, Medicago polymorpha and Medicago lupulina are widely distributed, particularly in purple soil. However, the local distribution and diversity of rhizobia associated with these legumes has not been systematically investigated. To this end, root nodules of M. polymorpha and M. lupulina grown in purple soil at seven locations in Yunnan Province of China were collected for rhizobial isolation. The obtained rhizobia were characterized by RFLP of 16S–23S rRNA intergenic spacer, BOXAIR fingerprinting, and phylogeny of housekeeping and symbiosis genes. As result, a total of 91 rhizobial strains were classified into species Sinorhizobium medicae and S. meliloti, while three nodC gene types were identified among them. S. medicae containing nodC of type I was dominant in farmlands associated with M. polymorpha; while S. meliloti harboring nodC of type III was dominant in wild land nodulated by M. lupulina. For both rhizobial species, greater genetic diversity was detected in the populations isolated from their preferred host plant. A high level of genetic differentiation was observed between the two Sinorhizobium species, and gene flow was evident within the populations of the same species derived from different soil types, indicating that rhizobial evolution is likely associated with the soil features. To examine the effects of environmental features on rhizobial distribution, soil physicochemical traits and rhizobial genotypes were applied for constrained analysis of principle coordinates, which demonstrated that soil features like pH, nitrogen and sodium were the principle factors governing the rhizobial geographical distribution. Altogether, both S. medicae and S. meliloti strains could naturally nodulate with M. polymorpha and M. lupulina, but the rhizobium-legume symbiosis compatibility determined by both the host species and soil factors was also highlighted.

Drought is a major abiotic stress that limits the growth and yield of alfalfa, a vital forage legume. The plant metalloproteinase Filamentation temperature-sensitive H (FtsH) is an ATP- and Zn²⁺-dependent enzyme that plays a significant character in the plant’s response to environmental stress. However, its functional role in drought resistance remains largely unexplored. This study investigates the drought tolerance role of alfalfa MsFtsH8 by analyzing the growth, physiology, and gene expression of overexpressing plants under drought conditions. The results demonstrated that both MsFtsH8-overexpressing Arabidopsis and alfalfa plants exhibited superior growth condition and enhanced membrane stability. The overexpressing alfalfa plants also showed reduced MDA levels, higher proline content, lower H₂O₂ accumulation, an increased activity of antioxidant-related enzymes (SOD, POD, and CAT) activity, and an elevated expression of antioxidant-related genes. These results indicated that the overexpression of MsFtsH8 enhanced growth, improved osmotic regulation, reduced ROS levels, and increased antioxidative capacity, ultimately leading to greater drought tolerance in alfalfa. Our findings suggest that MsFtsH8 mitigates oxidative damage caused by drought by modulating the plant’s antioxidant system, thus improving drought tolerance in alfalfa. This study provides a molecular basis and candidate genes for enhancing drought resistance in alfalfa through genetic engineering.

To elucidate the photosynthetic physiological mechanisms influencing alfalfa (Medicago sativa L.) yield and quality under varying planting densities, the cultivar ‘Zhongmu No.1’ was used as experimental material. The effects of different row spacing (R1, R2, R3) and seeding rate (S1, S2, S3, S4, S5) combinations on chlorophyll content (ChlM), nitrogen flavonol index (NFI), chlorophyll fluorescence parameters, forage quality, and hay yield were systematically analyzed. Results showed that alfalfa under R1S3 treatment achieved peak values for ChIM, NFI, EE, and hay yield, whereas R1S4 treatment yielded the highest Fv/Fm and CP content. Redundancy analysis further indicated that yield was most strongly associated with ChlM, NFI, Y (II), and qP. Y (II), and qP significantly influenced alfalfa forage quality, exerting negative effects on ADF and NDF, while demonstrating positive effects on CP and EE. In conclusion, narrow row spacing (15 cm) with moderate seeding rates (22.5–30 kg·hm⁻²) optimizes photosynthetic performance while concurrently enhancing both productivity and forage quality in alfalfa cultivated, establishing a theoretical foundation for photosynthetic regulation in high-quality and high-yield alfalfa cultivation.