Rheum palmatum L. is an important traditional Chinese medicinal herb now in demand worldwide. Recently, the theoretical framework suggested that sucrose triggers colonization of PGPM (plant growth-promoting microbes) in the rhizosphere, but their interactions on the plant remain largely unknown. Here, we applied three concentrations of both Bacillus amyloliquefaciens EZ99 inoculant (1.0 × 10⁵, 1.0 × 10⁶, and 1.0 × 10⁷ colony-forming units (CFU)/mL, denoted as LB, MB, and HB, respectively) and sucrose (0.15, 1.5, and 15 g/L, denoted as LS, MS, and HS, respectively) to investigate their co-effects on R. palmatum in a field experiment. The results showed that LB + MS (1.0 × 10⁵ CFU/mL Bacillus + 1.5 g/L sucrose) and LB + LS (1.0 × 10⁵ CFU/mL Bacillus + 0.15 g/L sucrose) treatments significantly increased root fresh weight (p ≤ 0.05). Metabolite analysis revealed that the treatment LB + LS significantly increased the relative content of major active components in rhubarb, namely anthraquinones and phenolic compounds, by 1.5% and 2.3%. Although high sucrose addition increased the activities of certain soil enzymes, the LB + LS treatment significantly increased total potassium (TK), whereas it decreased available potassium (AK), which facilitated the potassium utilization in rhizosphere soil. Furthermore, rhizosphere microbiomes revealed that fungal diversity was augmented in LB + LS treatment, in which the common causative fungal pathogen Fusarium spp. showed an effective suppression. Additionally, the redundancy analysis and Spearman correlations revealed a positive relationship of Sphingomonas associated with change in potassium bioavailability. Altogether, our findings suggest that the combined application of a bacterial inoculant and sucrose can improve the growth and quality of R. palmatum, and stimulate uptake of plant nutrients that contribute to alter the microbial community for biocontrol potential. Hence, this work not only has broad application prospects across economical plants, but also emphasizes agroecological practices for sustainable agriculture.

Codonopsis pilosula Franch., also known as Dangshen, is an important medicinal plant in China. It is widely cultivated for a major income of local farmers in Dingxi, Gansu Province. Its dried roots have the effects of supplementing vital energy, nourishing spleen and lung, enhancing organic immunity, helping depressurization, and improving microcirculation, etc., for humans. In June to October, 2018-2020, root rot disease was observed on C. pilosula with incidences up to 20% in the Dingxi region. We collected ten diseased and healthy plants from Dingxi (35°06'N, 104°29'E, 2206 m a.s.l.) in October 2019. The rotting root tissues were sterilized with 70% ethanol for 30 s and 3% NaOCl for 5 min and placed on potato dextrose agar (PDA) plates incubated at 25℃to isolate the pathogen (Shang et al. 2014). From the similar fungal cultures isolated after 7 days on PGA, isolate B17 was purified for morphological and molecular characterization. Its colony appeared light purple and produced long aerial hyphae. Slightly curved macroconidia (12.3 to 31.7 × 3.1 to 5.1 μm, n=40) and oval-ellipsoid and cylindrical microconidia (6.1 to 9.9 × 2.8 to 4.5 μm, n=30) were observed. The internal transcribed spacer region (ITS) and the translation elongation factor-1 alpha (TEF-1α) gene were amplified using primers ITS1/ITS4 and EF-1/EF-2 (Uwaremwe et al. 2020), respectively. The 489 bp (ITS) and 631 bp (TEF-1α) sequences were deposited in GenBank (Accession No. MN744360 and MN786974, respectively). The ITS sequence had 100% homology to isolate JJF2 (No. MN626452, ITS) (Ma et al. 2020), and the TEF-1α sequence had 100% homology to isolate Fo353 (No. KM065860) (Koyyappurath et al. 2016) of Fusarium oxysporum Schlecht. emend. Snyder & Hansen, which caused root rot of Panax ginseng and Vanilla planifolia, respectively. A phylogenetic tree was generated using the unweighted pair-group method with arithmetic average in the MycoBank database (O'Donnell et al. 2015), which clustered isolate B17 in the F. oxysporum species complex. Twenty 1-year-old plants of C. pilosula were inoculated with were inoculated by dipping the washed roots in a conidial suspension (2 ×106 conidia/ml added with 0.2% Tween 20) for 20 min before transplanted into pots (16 × 16 × 23 cm) with four plants per pot filled with sterilized peat and soil mixture (2:1 v/v) and grown in a greenhouse at 26oC with >70% humidity and 16 h light. Sterilized water added with 0.2% Tween 20 was used as a control. One week after inoculation, the leaves of pathogen-inoculated plants became yellow, and wilting occurred at the leaf tips 18 days later. Some of the inoculated plants died 45 days after inoculation, and the low part of roots had dark brown to black lesions and became rotting. The control plants did not show symptoms. The pathogenicity test was repeated three times with the same fungus isolated from the infected root tissue. To the best of our knowledge, this is the first report that F. oxysporum causes root rot on C. pilosula in China. F. oxysporum is a serious threat to C. pilosula cultivation, and the finding of this pathogen provides a clear target for root rot control.

Potato black scurf, caused by Rhizoctonia solani, is a severe soil-borne disease that affects the quality and production of potatoes worldwide. In our study, we used Paenibacillus polymyxa YF and Bacillus amyloliquefaciens HT to determine the antagonistic ability of R. solani, with a particular focus on the antagonistic action of P. polymyxa YF to R. solani and its biocontrol effect on potato black scurf. In fermentation, filtrate assay, 50% filtrate of P. polymyxa YF and B. amyloliquefaciens HT inhibited the growth of R. solani by 85.55% and 82.86%, respectively. Microscopic observations showed notable morphological changes with mycelial collapse, atrophy, and deformation following treatment with the antagonistic filtrates. Moreover, cell membrane permeability results showed increased conductivity in bacteria-treated samples compared to the control. P. polymyxa YF exhibited stable colonization on potato plants and secreted various extracellular enzymes (protease, amylase, and cellulase), along with the synthetic substances with growth-promoting effects, such as siderophores and Indole-3-acetic acid (IAA). Whether it is in the excised tissue inoculation or potted experiment, the negative control showed the highest rank of disease symptoms. In the pot experiment, after YF treatment, physiological parameters showed remarkable changes in plant height, root length, stem thickness, and dry and fresh weight. Compared to blank control, the activities of the four resistant enzymes increased significantly in the P. polymyxa YF treatment group. The upregulation in the P. polymyxa YF group was 4.04, 0.54, 0.46, and 3.10 times, respectively. PCR analysis identified genes in both bacterial strains coding for antimicrobial lipopeptides, including fenB, ituC, and srfAA, which are associated with fengycin, iturin, and surfactin synthesis. These findings demonstrated that P. polymyxa YF has a prominent antagonistic effect on R. solani, suggesting its potential as an effective biological control agent for controlling potato black scurf.

Root rot is a very destructive soil-borne disease, which severely affects the quality and yield of Angelica sinensis in major planting areas of Gansu Province, China. Twelve Fusarium strains were identified from root rot tissue and infected soil in the field by comparing each isolate strain internal transcribed spacer, translation elongation factor 1-α sequence and RNA polymerase second largest subunit gene with the sequences of known fungal species in the NCBI database. Of these isolates, four were F. acuminatum, followed by three F. solani, two F. oxysporum, and one each of F. equiseti, F. redolens, and F. avenaceum. Under greenhouse conditions, pathogenicity testing experiment was carried out using five strains: two F. acuminatum, one F. solani, one F. oxysporum, and one F. equiseti. Among them, the incidence of F. acuminatum–induced root rot on A. sinensis was 100%; hence, it was the most aggressive. Liquid chromatography was used to show that F. acuminatum could produce neosolaniol (NEO), deoxynivalenol, and T-2 toxins. Of these, the level of NEO produced by F. acuminatum was high compared with the other two toxins. By isolating Fusarium spp. and characterizing their toxin-producing capacity, this work provides new information for effectively preventing and controlling A. sinensis root rot in the field as well as improving the quality of its medicinal materials.

Fusarium avenaceum is the predominant fungal pathogen responsible for root rot in Angelica crops and poses a serious threat to their commercial quality and yield in China. This fungus produces enniatin B (ENN B), a toxin that could be a pathogenicity and virulence factor in plant–pathogen interactions. Yet whether ENN B exacerbates host infection and the onset of root rot in Angelica spp. caused by F. avenaceum is surprisingly understudied. Pathogenicity assays revealed that ENN B co-inoculation with F. avenaceum significantly increased the root rot disease index in Angelica sinensis from 83.33% (pathogen alone) to 92.86% (p < 0.05). Toxin degradation experiments showed that the bacteria Paenibacillus polymyxa and Bacillus tequilensis were capable of degrading 60.69% and 70.02% of ENN B, respectively. Response surface optimization (24.5 °C, 22.01 mg/L ENN B, 0.99% inoculum) enhanced degradation by B. tequilensis to 81.94%, a 11.74% improvement. Three ester compounds were identified by LC-HRMS as potential degradation products of ENN B. In planta trials demonstrated that the disease index was 50.01% for the group co-inoculated with ENN B degradation products and F. avenaceum, a 42.85% reduction compared to the group co-inoculated with ENN B and F. avenaceum. This study provides a new microbial strategy for controlling root rot in Angelica crops from a mycotoxin degradation perspective, which can be applied to promote sustainable agricultural production.

Soil salinization impacts over one billion hectares, threatening global food security. Here, a salt-tolerant bacterial strain, Pseudomonas fluorescens G3, was isolated from the rhizosphere of maize (Jinongyu-719) growing in saline–alkali soils in Gansu Province, China. This strain demonstrated the ability to secrete indole-3-acetic acid (IAA), 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase, and extracellular polysaccharides. It also exhibited notable phosphate-solubilizing activity and robust siderophore production capabilities. Under salt stress conditions (200 mM NaCl), the P. fluorescens G3 strain significantly improved maize’s growth parameters, namely its plant height, root length, and dry weight. Further, it enhanced antioxidant enzyme activity while reducing the accumulation of malondialdehyde (MDA), mitigating stress-induced oxidative damage. In P. fluorescens G3-inoculated plants, leaf and root Na⁺ contents decreased by 34.90% and 33.91%, while their K⁺ contents increased by 40.20% and 33.47%, respectively. Inoculation with P. fluorescens G3 enhanced taxonomic richness (ACE, Chao1) and evenness (Shannon, Simpson) in the rhizosphere bacterial community, leading to a significantly greater relative abundance of several bacterial genera: Pseudomonas, Methylophaga, Enhygromyxa, Desulfuromonas, and Devosia. These shifts in the microbial community composition suggest a potential restructuring of functional profiles, possibly enhancing processes beneficial to plant salt tolerance, such as ion homeostasis and stress mitigation: the biosynthesis of cofactors and secondary metabolites; bacterial secretion and two-component systems; porphyrin metabolism; flagellar assembly; biofilm formation; and bacterial chemotaxis. Redundancy analysis revealed positive correlations between microbial composition at both the phylum and genus levels and the activity of stress resistance enzymes after treatment with Pseudomonas fluorescens. This study provides important theoretical foundations and microbial resources for utilizing microbial community regulation in saline–alkali soil bioremediation.