Fusarium root rot, caused by Fusarium spp. in alfalfa (Medicago sativa L.), adversely impacts alfalfa by diminishing plant quality and yield, resulting in substantial losses within the industry. The most effective strategy for controlling alfalfa Fusarium root rot is planting disease-resistant varieties. Therefore, gaining a comprehensive understanding of the mechanisms underlying alfalfa’s resistance to Fusarium root rot is imperative. In this study, we observed the infection process on alfalfa seedling roots infected by Fusarium acuminatum strain HM29-05, which is labeled with green fluorescent protein (GFP). Two alfalfa varieties, namely, the resistant ‘Kangsai’ and the susceptible ‘Zhongmu No. 1’, were examined to assess various physiological and biochemical activities at 0, 2, and 3 days post inoculation (dpi). Transcriptome sequencing of the inoculated resistant and susceptible alfalfa varieties were conducted, and the potential functions and signaling pathways of differentially expressed genes (DEGs) were analyzed through gene ontology (GO) classification and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Meanwhile, a DEG co-expression network was constructed though the weighted gene correlation network analysis (WGCNA) algorithm. Our results revealed significant alterations in soluble sugar, soluble protein, and malondialdehyde (MDA) contents in both the ‘Kangsai’ and ‘Zhongmu No. 1’ varieties following the inoculation of F. acuminatum. WGCNA analysis showed the involvement of various enzyme and transcription factor families related to plant growth and disease resistance, including cytochrome P450, MYB, ERF, NAC, and bZIP. These findings not only provided valuable data for further verification of gene functions but also served as a reference for the deeper explorations between plants and pathogens.

Oat (Avena sativa) is a significant cereal crop that is extensively cultivated in temperate regions and is suitable for growth at higher elevations. The damage degree and epidemic area of oat spikelet rot caused by Dactylobotrys graminicola are generally increasing and spreading. Mycelium growth rate method was used to identify biological characteristics and in vitro fungicide screenings of D. graminicola. The results show that sorbitol and casein tryptone were the best carbon and nitrogen source for the D. graminicola, respectively. The optimal temperature for mycelium growth and conidia production of the D. graminicola was 20 °C; meanwhile, an acidic environment was shown to be conducive to mycelia growth, and alkaline facilitated conidia production. Among the ten tested fungicides, 20% Pydiflumetofen showed the best inhibition rate, with an EC₅₀ (concentration for 50% of maximal effect) value of 0.005 mg/L; 30% Difenoconazole Propiconazole and 35% Metalaxyl-M Fludioxonil also showed sufficient inhibitory effects against D. graminicola, with EC₅₀ value of 0.05 mg/L and 0.04 mg/L. Furthermore, we used artificial inoculation to determine the effectiveness of fungicide control in field, Trifloxystrobin 10%-Tebuconazole 20% with more than 90% control effectiveness, followed by 20% Pydiflumetofen. The results of this study not only revealed the biological characteristics of D. graminicola, but also provided effective candidate fungicides for the prevention and control of oat spikelet rot disease.

Syrian rue (Peganum harmala L) belongs to the family Zygophyllaceae and is mainly distributed in arid and semi-arid areas of Xinjiang, Gansu, Ningxia, Qinghai, and Inner Mongolia in China. It serves as a pioneer species in soil and water conservation, as well as in reclamation of wastelands, playing a crucial role in soil preservation and stabilization against sand encroachment. In 2023, a disease resembling root rot and wilt affected the quality and yield of Syrian rue in Xilingol (122.67°N, 42.78°E), Inner Mongolia, China. Symptoms ranged from yellowing to wilting, accompanied by darkening of the xylem in the roots. This disease was observed in approximately 50% of Syrian rue plants in desertification-prone grassland. A total number of 45 symptomatic root samples were collected from Syrian rue plants to isolate the pahogen. Small pieces of diseased tissue were surface sterilized with 75% alcohol for five seconds, rinsed twice with sterilized water, dried, and then placed on water agar and incubated for 72 h at 25℃ in darkness. A total of 45 isolates were obtained, showing identical morphology and sequences of the internal transcribed spacer region (ITS), translation elongation factor 1-alpha (EF1-α) and RNA polymerase II second largest subunit (RPB2) genes. Therefore, the isolate LTP-5 was used as the representative for further study. On potato dextrose agar (PDA), fungal colonies of LTP-5 exhibited yellow to light brown colore with white aerial mycelia and irregular growth after 7 days of incubation at 25℃. Conidia and chlamydospre development were observed in carboxymethylcellulose sodium medium. Chlamydospores were abundant, terminal or intergrown between hyphae, rough brown walls of 5.9 to 19.82 μm in diameter (n = 75) were observed in the culture. Macroconidia had 5 to 7 septa, thick in the middle and thin at both ends, measured 28.17 to 62.81 μm in length and 1.50 to 4.19 μm in width (n = 128). Microconidia were absent. The morphological characteristics were consistent with the descriptions of Fusarium equiseti (Nelson et al., 1983). The pathogen was further confirmed to be F. equiseti by sequence analysis of the ITS, EF1-α and RPB2 genes amplified using polymerase chain reaction (PCR) with the primers ITS4/ITS5 (White et al., 1990), EF-1/EF-2 (O'Donnell et al., 1998), and RPB2-5F2 /fRPB2-7cR (Zhen et al., 2017). The sequences showed 100% similarity to the F. equiseti strain in the NCBI GenBank with accession numbers MH054914.1, KJ396323.1, and KT213286.1 for ITS, EF1-α, and RPB2, respectively. The sequences of PCR products were submitted to GenBank with accession numbers PP814863.1 (ITS), PP831628.1 (EF1-α) and PP826937.1 (RPB2). Pathogenicity tests were conducted through a pot assay. Five Syrian rue seedlings were inoculated by immersing plant roots in a suspension of 1×107 spores/mL for 30 min and transplanting to pots containing autoclaved soil. Control plants were immersed in sterile water. The inoculated plants showed yellow and root rot but no symptoms were observed on control plants. The same fungus was successfully re-isolated and identified based on the morphological characterization and sequence analyses from the infected root tissue but not from non-inoculated plants. In addition, F. equiseti has been reported to cause disease on Sugar beet (Khan et al., 2021), Watermelon (Rahman et al., 2021), potato (Cui et al., 2021), and other plants. This is the first report confirming F. equiseti as causal agent of Syrian rue wilt and root rot in China.

Alfalfa Verticillium wilt, caused by Verticillium alfalfae, is a globally significant disease with increasing incidence and expanding epidemic areas. This study surveyed six major alfalfa-producing regions in Inner Mongolia, China—Chifeng, Tongliao, Ulanqab, Ordos, Bayannur, and Hohhot—and successfully isolated V. alfalfae exclusively from samples collected in Hohhot and Bayannur. Based on morphological characterization, multi-locus phylogenetic analysis (act, tef1-α, gapdh, and ts genes), and pathogenicity tests fulfilling Koch’s postulates, all 33 isolates were consistently identified as V. alfalfae, with disease severity levels ranging from 3.04 to 4.79 on the susceptible cultivar Zhongmu No. 1. As a preliminary assessment, the in vitro sensitivity of a representative strain, Va8, to eight commercial fungicides was evaluated using the mycelial growth inhibition method. Among the tested fungicides, 30% difenoconazole–propiconazole exhibited the strongest inhibitory effect (EC₅₀ = 0.14 μg/mL), followed by 10% trifloxystrobin & 20% tebuconazole (EC₅₀ = 0.20 μg/mL). However, given the substantial virulence variation observed among isolates, these sensitivity data should be interpreted with caution, as population-level differences may exist. These findings represent the first confirmed report of V. alfalfae in Inner Mongolia and provide a preliminary yet critical reference for prioritizing candidate fungicides for future multi-isolate and field evaluations.