Yongqiang Ma

1. As significant constraints in agricultural production, farmland weeds cause substantial annual yield losses. In rice production alone, weeds lead to losses of approximately 10 million tons annua...

To clarify the genomic information of strain Botrytis euroamericana HZ-011 and to screen the relevant functional genes for weed control. The whole genome of the DNA of strain HZ-011 was sequenced in this study using the third-generation single-molecule sequencing technology. The splicing and assembly of the three generations of sequencing data were performed using Hifiasm software, the gene assembly results were predicted using GeneMarkS software, and the predicted gene protein sequences were compared with the four major databases for gene annotation using the BLASTTx program. The results showed that the total gene length of strain HZ-011 was 4,402,230 bp, with 41.72% GC content. In total, 12,654 coding genes and 385 non-coding genes were predicted for this strain by whole genome sequencing. The basic annotation results showed that 12,250 genes were annotated, accounting for 96.88% of the total number of coding genes. The advanced genome annotation results showed that 1,629 genes were annotated in the CAZy database, and 1,864 genes were annotated in the PHI database, of which 54 genes were associated with the pathogenicity of pathogenic bacteria. In addition, 59 clusters of secondary metabolite synthesis genes and 12,654 signaling proteins were predicted, of which 1,088 secreted proteins were predicted, accounting for 8.6% of the total number of predicted signal peptide proteins. The whole-genome data of strain HZ-011 screened a large number of genes related to cell wall degradation enzymes and secondary metabolite synthesis regulation. The screened naphthalene gene cluster may represent a new pathogenic substance and is worthy of further in-depth research. The genome-wide data of strain HZ-011 provide a good foundation for in-depth studies of the pathogenicity-regulating genes of this strain and for investigating the pathogenicity mechanism of this strain.

In order to establish a rapid and sensitive LAMP visual detection method for Cherry Virus A on-site, this study used the conserved fragment of the CVA coat protein (CP) sequence as a template for primer design. The rapid visual LAMP detection method for Cherry Virus A was successfully established by optimizing the reaction system components (concentration ratio of internal and external primers, and concentrations of loop primers, Bst DNA, Mg²⁺, dNTPs and betaine) and reaction conditions (temperature and time). This method enables specific detection of Cherry Virus A and facilitates visual inspection of crude nucleic acid extracts within 40 min, significantly reducing the diagnostic turnaround time. The limit of detection is 67.54 pg μL⁻¹ (cDNA), which is 100 times more sensitive than PCR. Analysis of 70 field sweet cherry samples revealed an RT-LAMP positivity rate of 91.42%, significantly surpassing the 71.42% achieved by RT-PCR. This method is suitable for the rapid on-site detection of Cherry Virus, and can also provide a theoretical reference for the early diagnosis of cherry viral diseases.

The herbicidal efficacy and crop safety of Fusarium acuminatum strain HL-29, an endophytic fungus isolated from infected Amaranthus retroflexus in Qinghai Province, were evaluated. In vitro leaf assays demonstrated its pathogenicity against four broadleaf weeds, with efficacy ranked as follows: Elsholtzia densa = Senecio vulgaris = Chenopodium album > Malva verticillata. Pot trials further confirmed that the HL-29 fermentate caused 100% disease incidence in S. vulgaris, C. album, and E. densa. Notably, the strain showed no pathogenicity toward seven major local crops, indicating excellent selectivity. Scanning electron microscopy (SEM) revealed key morphological changes during the infection process on C. album leaves. These results establish F. acuminatum HL-29 as a promising biocontrol candidate against broadleaf weeds in the Qinghai–Tibet Plateau, providing a theoretical foundation for developing alpine-adapted mycoherbicides.

The herbicidally active Botrytis strain HZ-011 was isolated from naturally infected leaves of Rumex patientia L., and previous studies found that strain HZ-011 exhibits highly effective herbicidal effects against the broadleaf weeds Amaranthus...

Microbial herbicides, recognized for their target specificity, environmental compatibility, and simple production processes, hold promising potential for sustainable agriculture. This study isolated a strain of Alternaria gaisen (designated GD-011) from infected Medicago sativa L. in Qinghai Province, China, and evaluated its herbicidal potential through systematic development and efficacy assessment. Using single-factor and orthogonal experimental designs, the optimal sporulation substrate was identified as wheat bran, and the fermentation medium was optimized to consist of 14.5 g wheat bran, 19.4 g wheat middlings, 1.5 g rapeseed cake, and 14.6 g corn flour. Based on colony diameter and OD₆₀₀ measurements, diatomite was selected as the most suitable carrier, while bentonite, humic acid, and polyvinyl alcohol were chosen as the stabilizer, protectant, and dispersant, respectively. Pot trials under controlled conditions demonstrated strong herbicidal activity of GD-011 against three common weed species: Chenopodium album L., Elsholtzia densa Benth., and Galium aparine L. The highest efficacy was observed against C. album, with disease incidence and fresh weight inhibition reaching 80.83% and 79.87%, respectively. Inhibition rates for both E. densa and G. asparine exceeded 60%. A wettable powder formulation developed from GD-011 showed particularly effective control of C. album and E. densa, providing a practical foundation for the application of GD-011 as a novel bioherbicide.

In order to develop a new environmentally friendly microbial herbicide for the field of weed control, this study used the metabolite butyl hydroxybenzoate (BP) of the HY-02 strain of Alternaria as the research object. The BP emulsion formula was determined to be a mixture of BP, methanol, and Tween-20 in a ratio of 1:1:2 g/mL. The seed germination inhibition effect, the phytotoxicity of living plants, crop safety, and the field effect of the emulsion were studied. Research has found that adding 0.75% BP emulsion to the seed culture medium inhibits the germination of weed seeds such as Amaranthus retroflexus L., Malva verticillata L. var., and Chenopodium album L. While Brassica campestris L. seeds were unaffected, Triticum aestivum L and Hordeum vulgare L. stem and leaf growth were inhibited. Cucumis sativus L., Lactuca sativa L. var. asparagina, Spinacia oleracea L., and Capsicum annuum L. seeds are significantly inhibited, with germination rates below 20%. We sprayed 0.75% BP emulsion onto live potted plants; among the weeds, the incidence of Amaranthus retroflexus L., Lepyrodiclis holosteoides, Thlaspi arvense L, Galium spurium L., Malva verticillata L. var. Crispa, Chenopodium album L., and Avena fatua L reached 100%. The Pisum sativum L. and Triticum aestivum L. crops were not affected (NS), and they had slight plant height inhibition and slight susceptibility (LS) to highland Hordeum vulgare L. and peppers. They were highly phytotoxicity to Cucumis sativus L. and Spinacia oleracea L. Some plant leaves became infected and died, with incidences of 85% and 82%, respectively. The field experiment showed that after diluting the BP emulsifiable concentrate, the seedling stage spray was inoculated into the Triticum aestivum L. field, and it was found that the BP emulsifiable concentrate at the concentration of 1.00%~0.75% had a herbicidal effect on weeds such as Chenopodium album L., Elsholtzia densa Benth, and Amaranthus retroflexus L. in the Triticum aestivum L. field, and it was safe for Triticum aestivum L. crops in the field. These results indicate that BP emulsion could be developed into a new environmentally friendly microbial herbicide for field application in grass (Triticum aestivum L. and Hordeum vulgare L.) crops. At the same time, BP’s excellent antibacterial, low-toxicity, hydrolysis, and other effects can promote diversification in herbicide development.

This study conducted a transcriptome sequencing analysis of the interaction between Chenopodium album and Botrytis strain HZ-011 to identify genes involved in the response to fungal infections and elucidate the molecular mechanisms underlying the interaction. High-throughput RNA-seq technology was employed to analyze the transcriptomes of C. album leaves at 1, 4, and 5 days post-inoculation (dpi) with Botrytis strain HZ-011. The results revealed 11,645 differentially expressed genes (DEGs) at 1 dpi, including 7399 upregulated and 4246 downregulated genes; 11,285 DEGs at 4 dpi (7801 upregulated and 3484 downregulated); and 9976 DEGs at 5 dpi (7723 upregulated and 2253 downregulated). GO functional analysis indicated that downregulated DEGs were significantly enriched in chloroplast and plastid functional expression at 1, 4, and 5 dpi. Following infection by Botrytis strain HZ-011, downregulated genes were significantly enriched in pathways related to photosynthesis, including photosynthetic pathways, light-harvesting antenna proteins, and carotenoid biosynthesis. This suggests that the photosynthetic process in C. album was markedly inhibited, disrupting nutrient supply and leading to herbicidal effects. Notably, genes such as PSB28, PSBP, CAP10A, and CRTL-E-1 were significantly enriched in these pathways, indicating their potential roles in the herbicidal mechanism. These findings provide a foundation for understanding the herbicidal activity of strain HZ-011 and identifying potential targets for developing novel microbial herbicides.

Weeds compete with crops for resources, posing multiple negative impacts for agricultural production systems and triggering degradation of ecosystem services (e.g., alterations in the soil microbial community structure). Under the guidance of green plant protection, the development of efficient biocontrol strains with environmentally friendly characteristics has become a crucial research direction for sustainable agriculture. This study aimed to develop a fungal bioherbicide by isolating and purifying a pathogenic fungal strain (JZ-5) from infected redroot pigweed (Amaranthus retroflexus L.). The strain exhibited pathogenicity rates ranging from 23.46% to 86.25% against six weed species, with the most pronounced control efficacy observed against henbit deadnettle (Lamium amplexicaule L.), achieving a pathogenicity rate of 86.25%. Through comprehensive characterization of cultural features, morphological observations, and molecular biological identification, the strain was taxonomically classified as Fusarium oxysporum. Scanning electron microscopy revealed that seven days post-inoculation, F. oxysporum JZ-5 formed dense mycelial networks on the leaf surfaces of cluster mallow (Malva verticillata L.), causing severe tissue damage. Safety assessments demonstrated that the spore suspension (10⁴ spores/mL) had no adverse effects on three crops: hulless barley (Hordeum vulgare var. coeleste L.), wheat (Triticum aestivum L.), and potato (Solanum tuberosum L.). These findings suggest that F. oxysporum strain JZ-5 warrants further investigation as a potential bioherbicide for controlling three problematic weed species—Chenopodium album L. (common lambsquarters), Elsholtzia densa Benth. (dense-flowered elsholtzia), and Lamium amplexicaule L. (henbit deadnettle)—in cultivated fields of hulless barley (Hordeum vulgare var. coeleste L.), wheat (Triticum aestivum L.), and potato (Solanum tuberosum L.). This discovery provides valuable fungal resources for ecologically sustainable weed management strategies, contributing significantly to the advancement of sustainable agricultural practices.

Potato blackleg disease is caused by Pectobacterium atrosepticum, can seriously destroy potatoes’ growth and development. To accurately evaluate the expression levels of genes involved in potato (Solanum tuberosum) responses to P. atrosepticum infection, seven candidate reference genes (EF1α, eIF5A3, Tubulin, Ubiquitin, GAPDH, Actin, and CYP3) were systematically assessed for their expression stability at 1, 3, and 5 days after inoculation (dai) using the geNorm, NormFinder, BestKeeper, ∆Ct, and RefFinder algorithms. The results demonstrated that EF1α exhibited the highest stability among all experimental conditions, followed by eIF5A3 and Tubulin, whereas Ubiquitin displayed the least stability. To validate the screening outcomes, the expression patterns of four disease resistance-related genes (RBOHC, WRKY24, MPK3, and CPK32) were analyzed in both resistant and susceptible potato cultivars using the EF1α as the most stable and Ubiquitin as the least stable. Validation experiments revealed that the expression levels of disease resistance-related genes were stable and consistent with the RNA-Seq data when EF1α was used as a reference gene. In contrast, using Ubiquitin as a reference gene led to significant variability. Therefore, EF1α can be employed as the reference gene when studying the interaction between the potato and P. atrosepticum, providing a standardized reference for the subsequent studies on screening of disease resistance genes and exploring of disease resistance mechanism in potato.