Ya Zhang

Soybean (Glycine max) is a crucial source of plant protein worldwide, fungal diseases are one of the significant factors affecting productivity during the soybean cultivation process (Boufleur et a...

BACKGROUND: Phenazine‐1‐carboxamide (PCN) is a new fungicide; however, the number of its inhibitory targets is unknown. The target genes were identified by high‐throughput screening, reverse molecular docking, drug‐target binding, molecular dynamics, RNA interference and nanotechnology. RESULTS: The results identified three novel target genes among 400 differentially expressed genes. These genes were then subjected to a series of analyses to determine their potential as drug targets. This included evaluation of their hydrophobicity, hydrogen bonding, π–π interactions, and binding free energy. Additionally, the stability and flexibility of the target proteins were assessed using the GROMACS software, and the results indicated that they were stable. An RNA interference system was constructed to investigate the potential of these proteins as drug targets. An RNA interference system was constructed for the three novel target genes of PCN. The double‐stranded RNAs (dsRNAs) were combined with nano and mesoporous silica nanoparticles (MSNs) that significantly enhanced the control effect of dsRNA‐PCN on Rhizoctonia solani and increased the adsorption of dsRNAs on the leaf surface. The growth of R. solani was slowed by dual treatment with dsRNA@MSNs. The dsP450‐FAH treatment demonstrated the strongest effect, with the dsRNA@MSNs‐treated group exhibiting a reduction of 50.43% compared to the control group and 44.44% compared to the PCN treatment. CONCLUSION: In this study, three novel target genes were identified, and their involvement in the inhibition of R. solani by PCN was confirmed through RNA interference and nanotechnology. The results provide a foundation for the development of novel biopesticides. © 2025 Society of Chemical Industry.

The antifungal targets of the new fungicide N-(naphthalen-1-yl)-phenazine-1-carboxamide (NNPCN) are still incomplete, limiting its application. To identify potential new targets of NNPCN and facilitate target hunting, a suite of techniques was employed to conduct experiments on Rhizoctonia solani. Nine potential targets were identified, exhibiting strong binding affinity to NNPCN, as indicated by binding free energies below -100.000 kJ/mol. Notably, pectin lyase, glycosyl hydrolase, fumarate transporter, and cytochrome monooxygenase showed exceptionally strong binding. The mRNA expression analysis revealed significant downregulation in certain target genes: E3 ubiquitin ligase (AG1IA_02506), aldehyde dehydrogenase (AG1IA_03762), fumarate transporter (AG1IA_03944), and pectin lyase (AG1IA_03046) decreased by 42%, 66%, 83%, and 69%, respectively, while other key genes were upregulated. Pectin lyase protein was obtained through prokaryotic expression at 0.4 mg/mL concentration. A novel thiobarbituric acid test system verified pectin lyase as a potential NNPCN target, with the enzyme activity multiple being only 0.169 after NNPCN treatment. These findings enhance our understanding of NNPCN's mode of action and could guide its improved application.

Lodging has a negative effect on rice production and leads to a great loss in yield and quality. It is necessary to clarify the effects of straw return measures coupled with rice-duck co-culture on lodging and to explore a measure that can improve lodging resistance. A randomized block experiment with six treatments (rice monoculture (RNN), rice-duck co-culture (RND), direct straw return and rice monoculture (RSN), direct straw return coupled with rice-duck co-culture (RSD), straw carbon and rice monoculture (RBN), and straw carbon coupled with rice-duck co-culture (RBD)) was conducted to investigate the mechanism of the change in lodging resistance. RNN’s rice yield was 6258.02 kg ha⁻¹. The yield of RND, RSN, RSD, and RBN increased by 15.51, 3.06, 10.23, and 1.59%, respectively, while RBD decreased by 5.01% relative to RNN. Direct straw return and straw biochar return coupled with rice-duck co-culture has both negative and positive effects on lodging resistance because of its properties. The stem’s mechanical properties were mainly decided by weight, length, plumpness, and culm anatomy. The increased bending moment at breaking, lodging strength, and bending strength with the RND, RSN, RSD, RBN, and RBD treatments increased, indicated an increase in lodging resistance. Our results clearly demonstrate that direct straw return and straw biochar return coupled with rice-duck co-culture could increase the lodging resistance. In total, rice-duck co-culture could increase the lodging resistance with a higher yield. Compared to straw biochar application, straw return can stabilize the yield and improve the lodging resistance of rice. Thus, direct straw return coupled with rice-duck co-culture should be explored for improving lodging resistance under the condition of ensuring yield.

Replant disease caused by continuous cropping obstacles commonly occurs in a Lilium brownii consecutive monoculture. To reveal the mechanisms contributing to the continuous cropping obstacles of L. brownii, four cropping patterns (fallow, L. brownii-rice rotation, newly planted L. brownii, and 2-year L. brownii consecutive monoculture) were designed, and Illumina MiSeq (16S rDNA and ITS) was utilized to detect shifts in the microbial community in the rhizosphere. Our result showed that planting of L. brownii significantly reduced soil pH. Consecutive monoculture of L. brownii can significantly decrease the diversity and abundance of soil bacteria, but markedly increase the diversity and abundance of soil fungi. Under the four planting pattern treatments, the changes in soil pH were consistent with the changes in the Shannon diversity index of soil bacterial communities, whereas we observed a negative correlation between soil pH and Shannon diversity index for fungi. The relative abundance of Lactobacillales significantly increased in soils of L. brownii consecutive monoculture, while Acidobacteriales, Solibacterales, and Xanthomonadales increased in soils of L. brownii-rice rotation and newly planted L. brownii. Collectively, this work aimed to elucidate the relationship between the L. brownii planting patterns and soil microbiome, thereby providing a theoretical basis for screening new biological agents that may contribute to resolving continuous cropping obstacles of L. brownii.

The aim of the present investigation was to determine the active ingredients in Amaranthus tricolor L. leaves and develop a biological pesticide. Organic solvent extraction, column chromatography, liquid chromatography, ODS-C18 reverse elution, Sephadex LH-20 gel filtration, H spectrum, and C spectrum were used to isolate the pure product for an assessment of the agricultural activity and bacteriostatic mechanisms. The results showed that the activity of the crude extract following carbon powder filtration was 1.63-fold that of the non-filtered extract. Further isolation was performed to obtain two pure products, namely, hydroxybenzoic acid (HBA) and benzo[b]furan-2-carboxaldehyde (BFC), and their molecular formulas and molecular weights were C₇H₆O₃ and 138.12, and C₉H₆O₂ and 146.12, respectively. Our study is the first to determine that HBA has bacteriostatic activity (MIC 125 μg/mL) and is also the first to isolate BFC from A. tricolor. The ultrastructure observation results showed that HBA caused the bacteria to become shriveled, distorted, and deformed, as well as exhibit uneven surfaces. After HBA treatment, 70 differentially expressed metabolites were detected in the bacteria, of which 9 were downregulated and 61 were upregulated. The differentially expressed metabolites were mainly strigolactones, organic acids and derivatives, fatty acids, benzene and substituted benzene derivatives, amino acids and associated metabolites, and alcohols and amines. Among all of the downregulated differentially expressed metabolites, MEDP1280 was the most critical, as it participates in many physiological and biochemical processes. The enrichment analysis showed that the differentially expressed metabolites mainly participate in tyrosine metabolism, biosynthesis of amino acids, cysteine and methionine metabolism, and arginine and proline metabolism. Additionally, HBA was found to disrupt cell membrane permeability and integrity, causing the leakage of substances and apoptosis. The physiological and biochemical test results showed that HBA could increase the pyruvate levels in bacteria but could decrease the activities of respiratory enzymes (malate dehydrogenase (MDH) and NADH oxidase) and antioxidant enzymes (superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX)). Inverse molecular docking was used to study the binding between HBA and respiratory and antioxidant enzymes. The results showed that HBA could bind to MDH, NADH oxidase, SOD, and GSH-PX, suggesting that these enzymes may be the effector targets of HBA. Conclusion: The optimal active ingredient in A. tricolor that can inhibit Acidovorax avenae subsp. citrulli was identified as HBA. HBA mainly disrupts the cell membrane, damages the metabolic system, and inhibits respiration and antioxidant enzyme activity to control bacterial growth. These results provide a reference for the further development of biological pesticides.

To explore the molecular mechanism through which the novel fungicide N-(naphthalen-1-yl) phenazine-1-carboxamide (NNPCN) inhibits Rhizoctonia solani, we clarified the target and mode of action, explored lead compounds, and developed novel fungicides. Methods: Growth observation, scanning electron microscopy, transmission electron microscopy, transcriptome sequencing technology, quantitative real-time PCR (qRT-PCR), physiological and biochemical determination, and reverse molecular docking technology were used to study the effects of this compound on the microscopic morphology of R. solani. The differentially expressed genes (DEGs), functions, and metabolic pathways were analyzed. The genes displaying significant differences were randomly selected for qRT-PCR verification and confirmed by physiological and biochemical determination to construct their binding mode with key targets. The results showed that the mycelium treated with NNPCN produced a red secretion and exhibited progressive creeping growth. Under a scanning electron microscope, hyphal swelling, uneven thickness, fractures, deformities, and hyphal surface warts increased. Under a transmission electron microscope, the cell wall was separated, the subcellular organelles were disintegrated, and the septum disappeared. Furthermore, there were 6838 DEGs under NNPCN treatment, including 291 significant DEGs, of which 143 were upregulated and 148 downregulated. Ten DEGs were randomly selected for qRT-PCR verification, and the gene expression trend was consistent with the transcriptome sequencing results. Gene Ontology enrichment analysis showed that the DEGs were significantly enriched in cell wall glucan decomposition and metabolism, cell membrane synthesis, metabolism, composition, organic hydroxyl compounds, oxidoreductase activity, and transition metal ion binding. Metabolic pathway enrichment analysis showed that there were 16 significant metabolic pathways, such as steroid biosynthesis and ABC transporters. Further study found that genes, such as the glycosyl hydrolase family 10 domain-containing protein, which is related to glucan catabolic process function as tied to the cell wall, were downregulated. Lipid oxidation, modification, and other genes related to the cell membrane were also downregulated. Secondly, genes related to lipid modification, lipid metabolism processes, integral components of the membrane, and other ABC transporters were downregulated. Fatty-acid oxidation and carbohydrate metabolic processes, which are related to antioxidant and metabolic functions, displayed significant differences in their target genes. Nitrite reductase [NADH] activity and mitochondrial organization gene expression were downregulated. These results revealed that target genes may involved in the cell wall, cell membrane, antioxidant and metabolism, nitrogen metabolism, and mitochondria. The results of the physiological and biochemical tests showed that NNPCN decreased the β-1,3-glucanase, malondialdehyde, and ATPase activities and nucleic acid leakage but increased the activity of nitrate reductase. The results of the reverse molecular docking showed that NNPCN could freely bind to target proteins such as β-1,3-glucanase, ABC transporter, and NADPH nitrate reductase, whereby NNPCN could bind to glucanase via van der Waals and electrostatic forces and to ABC transporter and NADPH nitrate reductase via hydrogen bonding. Conclusion: The mechanism via which NNPCN inhibits R. solani may be related to the cell wall structure, cell membrane damage, antioxidant activity, and metabolism.