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Article

Biocontrol Activity of Volatile Organic Compounds Emitted from Bacillus paralicheniformis 2-12 Against Fusarium oxysporum Associated with Astragalus membranaceus Root Rot

by
Yan Wang
1,
Jiaqi Yuan
1,
Rui Zhao
1,
Shengnan Yuan
1,
Yaxin Su
1,
Wenhui Jiao
1,
Xinyu Huo
1,
Meiqin Wang
1,
Weixin Fan
2 and
Chunwei Wang
1,*
1
Shanxi Key Laboratory of Bioagent Utilization and Eco-Pesticide Innovation, College of Plant Protection, Shanxi Agricultural University, Jinzhong 030801, China
2
Experiment Teaching Center, Shanxi Agricultural University, Jinzhong 030801, China
*
Author to whom correspondence should be addressed.
Microorganisms 2025, 13(8), 1782; https://doi.org/10.3390/microorganisms13081782
Submission received: 10 June 2025 / Revised: 18 July 2025 / Accepted: 25 July 2025 / Published: 31 July 2025
(This article belongs to the Section Microbial Biotechnology)
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Abstract

Root rot, mainly caused by Fusarium oxysporum, is one of the most destructive diseases and leads to significant economic loss of Astragalus membranaceus. To develop an effective strategy for the management of this serious disease, a bacterial strain 2-12 was screened from A. membranaceus rhizosphere soil and identified as Bacillus paralicheniformis based on the phylogenetic analyses of gyrase subunit B gene (gyrB) and RNA polymerase gene (rpoB) sequences. Interestingly, the volatile organic compounds (VOCs) produced by B. paralicheniformis 2-12 exhibited potent antifungal activities against F. oxysporum, as well as fifteen other plant pathogens. Under scanning electron microscopy observation, hyphae treated with the VOCs exhibited abnormal variation such as distortion, twist, and vesiculation, leading to distinctive protoplasm shrinkage. After treatment with B. paralicheniformis 2-12 VOCs, the lesion diameter and disease incidence both reduced significantly compared to control (p < 0.05), thus demonstrating prominent biological efficiency. Moreover, B. paralicheniformis 2-12 VOCs were composed of 17 VOCs, including 9 alkanes, 3 alcohols, 3 acids and esters, 1 aromatic compound, and 1 alkyne compound. A total of 1945 DEGs, including 1001 up-regulated and 944 down-regulated genes, were screened via transcriptome analysis. These DEGs were mainly associated with membranes and membrane parts, amino acid metabolism, and lipid metabolism. The findings in this work strongly suggested that B. paralicheniformis 2-12 VOCs could be applied as a new candidate for the control of A. membranaceus root rot.

1. Introduction

Astragalus membranaceus (Fisch.) Bge var. mongholicus (Bge.) Hsiao (Leguminosae), also named Huangqi, belonging to the Astragalus genus in the Leguminosae family, is one of the most important Chinese medicinal herbs native to Northern China [1,2]. It has been well known for its various pharmacological activities such as antioxidant, anti-tumor, diuretic, vital-energy-tonifying, and tissue-generative effects for more than 2000 years [3,4]. Owing to an increasing demand, wild A. membranaceus sharply decreased and could not meet the needs in the market [5]. Recently, A. membranaceus has been widely artificially planted in Shanxi, Gansu, and Inner Mongolia in China [2,5]. However, the occurrence of various diseases has attracted more attention due to serious losses. Particularly, root rot, one of the most economically severe diseases, was considered as a key constraint to commercial production of A. membranaceus. Based on the survey, root rot can cause a severe yield loss of more than 30% [6]. Several Fusarium spp., such as F. oxysporum, F. acuminatum, F. solani, and F. redolens, have been reported to cause A. membranaceus root rot [2]. Nevertheless, F. oxysporum are found to be the dominant pathogen associated with root rot on A. membranaceus [7].
Due to their low cost, convenient operation, and high control effects, several chemical fungicides such as hymexazol, carbendazim, and azoxystrobin have always been used to prevent and control Fusarium root rot on Chinese medical herbs in the field [5,8]. Previous studies also indicated that carbendazim, prochloraz, and fludioxonil had strong inhibition effects on Fusarium spp. and could be considered as candidates for the management of A. membranaceus root rot [9]. However, the long-term and extensive use of chemicals may cause adverse problems such as antibiotic resistance, environmental pollution, and even harm to human health [10,11]. Thus, new sustainable measures are required to control A. membranaceus root rot in the field.
Biological control is usually regarded as one of the most promising methods that could exhibit broad antifungal spectrum, minimal residual toxicity, and environmental friendliness [10,12]. Biological control agents (BCAs) and their products were widely used to control harmful pathogens on crops [11]. The mechanisms of BCAs mainly include nutrient and space competition, induced resistance, and the production of antimicrobial substances [10]. Among the mechanisms, volatile organic compounds (VOCs) produced by BCAs, the important natural microbial antagonists, have attracted more great attention because of their renewability, biodegradability, and environmental safety [13]. Particularly, VOCs with a low molecular weight (<300 Da) have been proven to disperse easily in air and could play an important role in fungal inhibition and disease suppression without physical contact with pathogens or crops [2,14]. Previous studies determined that some BCAs, belonging to the genera of Bacillus, Streptomyces, and Pseudomonas, could produce antifungal VOCs and have been widely applied to control root rot in the field [15,16,17,18]. However, to our best knowledge, few reports were available on the biological effects of microbial VOCs on A. membranaceus root rot.
In this work, the aims are to screen and identify new BCAs, evaluate the inhibition activities of the VOCs to plant pathogens, investigate the biological efficiency of VOCs on A. membranaceus root rot, determine the compositions of the VOCs, and elucidate the response of VOCs to F. oxysporum at the molecular level. Our findings would be expected to provide a theoretical basis for the development of novel biofumigants in the future.

2. Materials and Methods

2.1. Plant Pathogens and Culture Media

F. oxysporum H1 was isolated from a diseased sample of A. membranaceus in Hunyuan county, Shanxi Province of China. Fifteen other plant pathogens (listed in Table 1) were provided by the plant pathology laboratory of Shanxi Agricultural University.

2.2. Isolation and Screening of Antagonistic Bacterial Strain

A total of 10 g of rhizosphere soil sample collected from a healthy plant was added to 100 mL of sterile distilled water and shaken at 28 °C and 220 r/min for 10 min. Antagonistic bacteria exhibiting good inhibition effects on F. oxysporum were isolated from the soil bacterial solution by the serial dilution method and further purified from single colonies on nutrient agar (NA) medium at 28 °C for 48 h. All the bacterial isolates were stored at 4 °C before use.
The antagonistic effect of the test isolate was determined using the dual culture method previously described by Alijani et al. [19] and Wang et al. [20]. A mycelial disk (6 mm diameter) from a freshly prepared 7-day-old culture was transferred to the center of a 20 mL potato dextrose agar (PDA) medium plate. A total of 100 μL of the bacterial inoculum with 1 × 106 colony-forming units (CFUs)/mL was inoculated on another plate with 15 mL of NA medium. Then, the two plates were rapidly sealed with polyethylene (PE) film. A PDA medium plate containing pathogen and another plate containing NA medium alone were used as control. The experiment was repeated three times. After the VOC treatment, mycelial growth was evaluated in two cross directions each day for 7 days. The percentage of mycelial inhibition of the antagonistic bacterial isolate was calculated based on the formula: Percentage of mycelial inhibition (%) = (dc − dt)/(dc − d0) × 100, where dc represents the colony diameter of the control, dt represents the colony diameter of the treatment, and d0 is the diameter of the mycelial plug [21]. To further determine the inhibition effects of the VOCs on mycelial growth, the wet and dry weights were also measured at 7 days after treatment.

2.3. Identification of Antagonistic Bacterial Isolate

The colony characteristics of the tested isolate were described according to Bergey’s Manual of Determinative Bacteriology. The physiological and biochemical characteristics of the tested isolate were determined according to Bergey’s Manual of Determinative Bacteriology. For accurate identification, molecular features, including gyrase subunit B gene (gyrB) and RNA polymerase gene (rpoB) gene sequences, were amplified using the polymerase chain reaction (PCR). PCR amplification for the target gene sequences was performed using the described program with minor modifications [22,23,24,25]. The PCR amplification products were resolved on 1% agarose gels and further sequenced by Sangon Biotech Co., Ltd. (Shanghai, China).
The target gene sequences were aligned with the reference sequences using BLAST-N tool in the GenBank database (https://novopro.cn/blast/tblastn.html, accessed on 9 June 2025). The known sequences with high similarity were downloaded and used to analyze the evolutionary trajectory of the tested isolate. Multiple sequence alignments were performed using the CLUSTAL X Ver. 2.0 program. Phylogenetic trees were constructed to reveal the phylogenetic relationship using MEGA 7.0.14 in accordance with the neighbor-joining statistical method.

2.4. Scanning Electron Microscopy (SEM) Analysis

After exposure to the VOCs for 5 days, fungal hyphae taken from the inhibition areas were used to reveal the external morphological changes of F. solani and F. oxysporum. Samples without VOC treatments were used as control. The fungal hyphae were fixed with 1.5% glutaraldehyde and 1% osmium tetroxide [20]. Then, the tested samples were sputter-coated with gold palladium and observed using a JSM-6490LV scanning electron microscope (SEM, JEOL Company, Tokyo, Japan).

2.5. VOC Activities of Antagonistic Bacterial Isolate Against Other Plant Pathogenic Fungi

To confirm the effects of VOCs from antagonistic bacterial isolates on plant pathogenic fungi, the inhibition activities of antagonistic bacterial isolates against fifteen plant pathogenic fungi were measured using the dual culture method as described in Section 2.2. Percentage of mycelial inhibition was also calculated using the formula described above. Each experiment was conducted in five replicates.

2.6. Biological Efficiency of the VOCs on A. membranaceus Root Rot

To confirm biological potential, the effects of VOCs emitted from antagonistic bacterial isolates on A. membranaceus root rot were assessed by a previous method with slight modifications [20,26]. The tested isolate was placed on an NA plate and incubated at 23 °C for 36 h. A single colony was transferred into 20 mL of sterile LB broth and shaken at 28 °C and 180 r min−1 for 15 h. Next, 100 μL of the cultural broth was placed on the surface of the LA plate. Five A. membranaceus roots were wounded with a sterilized needle (2 mm in diameter and 3 mm in depth) and dripped with 20 μL of conidia suspension (106 conidia/mL). Subsequently, the LA plate and inoculated A. membranaceus roots were placed into a sealed container, with high relative humidity maintained (approximately 90%). A total of 100 μL of sterile water placed on the LA plate was used as control. After incubation at 25 °C for 7 and 14 d, the biological efficiency was evaluated based on the disease incidence and lesion diameter [20].

2.7. Extraction and Analysis of VOCs from Antagonistic Bacterial Strain

The tested strain was transferred into Luria–Bertani (LB) medium for 14 h at 25 °C. After addition of 5 mL of Luria–Bertani agar (LA) medium containing 5 mg/L of heptyl acetate, a solid-phase micro-extraction (SPME) tube was placed at a 45-degree angle to increase the inoculation area [27,28]. Next, 100 μL of bacterial suspension (106 colony-forming units (CFUs)/mL) was added onto the surface of the LA medium in the SPME tube and then incubated at 25 °C for 1, 3, 5, and 7 days. The tube containing only LA medium was used as control. For VOC extraction, SPME fiber (75 μm CAR/PDMS) was inserted into the tube and exposed to the headspace at 40 °C for 30 min.
The extracted analytes were further separated and detected using gas chromatography–mass spectrometry (GC-MS). The GC-MS system consisted of a Trace-1300 gas chromatograph, equipped with a Trace-ISQ quadruple mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA). A TR-5MS capillary column (30 m × 0.25 mm) with 0.25 μm film thickness (Thermo Fisher Scientific, Waltham, MA, USA) was used for separation of the analytes with a split–splitless injector. Helium (99.999% purity) was used as the carrier gas at a flow rate of 1.0 mL/min. The starting temperature of the oven was 40 °C for 3 min, at a heating rate of 10 °C/min to 180 °C, and then ramped at 40 °C/min to 270 °C, held for 4 min. Mass spectrometry was performed in electron ionization mode at 70 eV. Mass spectra scan ranged from 40 to 400 m/z. Mass spectral data of the VOCs from bacterial samples were compared with those of the standard compounds in the National Institutes of Standards and Technology (NIST) databases. The compounds were further verified based on their direct match (SI), reverse match (RSI), and similarity in NIST database [26,29,30,31].

2.8. Transcriptome Sequencing

F. oxysporum mycelial plug (6 mm diameter) was placed onto PDA plate and treated with 100 μL of B. paralicheniformis 2-12 bacterial inoculum (1 × 106 CFU/mL) using the dual culture method as described in Section 2.2. After incubation at 25 °C for 5 days, 1000 mg hyphae of fungal colony were collected from the control groups and the treatment groups, placed into a 1.5 mL tube, and immediately frozen in liquid nitrogen. The collected hyphae were stored at −80 °C before transcriptomic analysis.
Then, the total RNA of hyphae was extracted using TRIzol Reagent Invitrogen (Invitrogen Life Technologies, Carlsbad, CA, USA). The RNA integrity was measured using a 2100 Bioanalyzer (Agilent Technologies, Inc., Santa Clara, CA, USA) and quantified using a NanoDrop 2000 Spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA). Total RNA was combined with Oligo (dT) magnetic beads to purify mRNA, and fragmentation buffer was used to fragment mRNA. A set of 6-mer primers was added, and reverse transcriptase was used to synthesize double-stranded cDNA using mRNA as the template. The ends of the double-stranded cDNA were flattened at the 3′ end, and an A tail was added. Next, the sequencing libraries were constructed using the TruSeq™ RNA sample preparation kit (San Diego, CA, USA). Subsequently, transcriptome sequencing was performed using the Illumina HiSeq×TEN platform [32]. Differential expression genes (DEGs) were screened by DESeq2 software (Version 1.42.0) based on the threshold of |log2 (fold change)| ≥ 1 and false discovery rate (FDR) < 0.05 [32,33]. Then, all the DEGs were mapped to Gene Ontology (GO) terms in the GO database [10]. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was further conducted using KEGG Orthology Based Annotation System (KOBAS) software (Version 3.0) [34].

2.9. Data Analysis

Each experiment was repeated three times, and the data were expressed as mean ± standard deviation (SD). For normality assessment, the experimental data were evaluated using the Shapiro–Wilk test. Statistically significant difference was determined by one-way analysis of variance (ANOVA) via Duncan’s multiple range test at p < 0.05 using SPSS software version 19.0 (IBM Corp., New York, NY, USA).

3. Results

3.1. Isolation and Screening of Antagonistic Bacterial Isolate with Antifungal VOCs

To screen new biocontrol agents, a total of 559 strains were isolated from 111 soil samples. Moreover, 58 strains with strong antagonistic activities in vitro were screened using the dual culture method. As a result, a new bacterial isolate was designated as 2-12 and exhibited a potent percentage of mycelial inhibition of 60.41% at 7 days after VOC treatment (Figure 1a). The wet weight and dry weight of mycelia were further assessed in VOC treatment. Within 6 and 7 days, the wet weights were 0.0345 and 0.0203 g, while the dry weights were 0.0040 and 0.0124 g, which all exhibited significant differences compared to the control (p < 0.05, Figure 1b,c).

3.2. Identification of Strain 2-12

The colony of strain 2-12 is circular, slightly convex, opaque, and milky white on the NA plate (Figure 2a). Strain 2-12 is a Gram-positive, aerobic cell (Figure 2b). Under the SEM observation, strain 2-12 is rod-shaped and straight, measuring 0.5 to 0.8 μm × 1.5 to 3.0 μm (n = 50) (Figure 2c). In the phylogenetic trees based on gyrB and rpoB gene sequences, isolate 2-12 was grouped into the same clade with B. paralicheniformis strains and distantly separated from other Bacillus species (Figure 3). Isolate 2-12 was identified as Bacillus paralicheniformis based on morphological characteristics and phylogenetic analyses. Subsequently, B. paralicheniformis 2-12 was deposited in China Center for Type Culture Collection (CCTCC) under accession number CCTCC M 20232598.

3.3. SEM Analysis of F. oxysporum Hyphae

F. oxysporum hyphae were observed under SEM observation. Without the VOCs from B. paralicheniformis 2-12, the control hyphae exhibited normal cylindrical shapes and smooth surfaces (Figure 4a–c). However, F. oxysporum hyphae treated with the VOCs have many abnormal variations such as distortion, twist, and vesiculation (Figure 4d–f). In particular, the hyphae surfaces were significantly shriveled, thus leading to distinctive protoplasm shrinkage compared to the control groups (Figure 4).

3.4. VOC Activities of B. paralicheniformis 2-12 Against Fifteen Plant Pathogenic Fungi

Table 1 illustrates the VOC activities from B. paralicheniformis 2-12 against fifteen plant pathogenic fungi. The VOCs exhibited potent inhibitory effects on the fifteen fungi with percentages of mycelial inhibition of 31.88~94.96%. Most of the variables (except F. graminearum and Botrytis cinerea) presented high Shapiro–Wilk p-values (greater than 0.05), indicating the normal distribution of our variables.

3.5. Biocontrol Effects of B. paralicheniformis 2-12 VOCs on A. membranaceus Root Rot

Biocontrol effects of the VOCs were measured at 7 and 14 d after treatment (Figure 5). At 7 d after treatment, the disease incidence significantly decreased to 13.33%, while that of the control groups maintained at 100%. The lesion diameter on treatment groups reduced to 0.20 cm, while those of the control groups were 2.20 cm. At 14 d after treatment, the disease incidence still maintained low levels (23.33%), and the lesion diameter reduced to 0.78 cm. Moreover, the experimental data exhibited significant difference (p < 0.05) compared to the control groups (Figure 5).

3.6. Determination of B. paralicheniformis 2-12 VOCs

The VOCs emitted from B. paralicheniformis 2-12 were determined using the SPME-GC-MS method based on the retention time (RT), direct match (SI), and reverse match (RSI). As a result, 17 VOCs, including 9 alkanes, 3 alcohols, 3 acids and esters, 1 aromatic compound, and 1 alkyne, were found in B. paralicheniformis 2-12 (Table 2). Of these VOCs, alkanes were found to be the most diverse class and might be the most important component of antifungal VOCs.

3.7. Transcriptomic Analysis

3.7.1. Quality Control of Data

The control groups and treatment groups were set with three biological replicates for each treatment. A total of 361 million pieces for raw reads and 360 million pieces for clean reads were obtained through RNA-seq analysis. The Q20 data of the control and treatment groups ranged from 99.54% to 99.58%, while the Q30 data ranged from 97.23% to 97.42%, thus indicating good sequencing quality for bioinformatic analysis.

3.7.2. Analysis of Differentially Expressed Genes (DEGs)

Under B. paralicheniformis 2-12 VOC stress, the distribution of DEGs in F. oxysporum was intuitively visualized in a volcano plot (Figure 6a). A total of 1945 DEGs, including 1001 up-regulated and 944 down-regulated genes, were identified compared to the control groups based on the criteria of p-value < 0.05 and fold change ≥ 2. Next, a heatmap was used to reveal the expression levels for the tested samples. Consequently, three treatment samples were clustered in one clade and separated from the control samples (Figure 6b).

3.7.3. GO and KEGG Annotation Analyses

GO annotation analysis showed that these DEGs were mainly classified into three main functional categories: biological processes, cellular components, and molecular functions. Of these categories, the most significantly enriched GO terms were related to catalytic activity, membranes, metabolic processes, and membrane parts (Figure 5b). The results of KEGG analysis showed that these DEGs were involved in metabolism, genetic information processing, environmental information processing, and cellular processes. Among the 20 KEGG pathways, carbohydrate metabolism, amino acid metabolism, and lipid metabolism were found to be the most important pathways, accounting for large percentages of 14.65%, 20.41%, and 20.41%, respectively (Figure 5b).

3.7.4. GO and KEGG Enrichment Analyses

To illustrate the functions of the DEGs, GO and KEGG enrichment analyses were conducted in this work. The DEGs were classified into 170 GO terms, mainly including transmembrane transport, transmembrane transporter activity, transporter activity, and oxidoreductase activity (Figure 7a). The KEGG enrichment analysis showed that the DEGs were mostly enriched in tryptophan metabolism, glycine, serine and threonine metabolism, steroid biosynthesis, phenylalanine metabolism, and pentose and glucuronate interconversions (Figure 7b).

4. Discussion

In this study, a new bacterial isolate 2-12 exhibited a potent mycelial inhibition effect on F. oxysporum, and its VOCs were also found to present noticeable inhibition activities against F. oxysporum, as well as fifteen other plant pathogens. Isolate 2-12 was further identified as Bacillus paralicheniformis. Although B. paralicheniformis has been used to control various diseases such as root rot, anthracnose, leaf spot on Quinoa, pearl millet blast, and tomato wilt [35,36,37], this is the first study demonstrating the antifungal activity of VOCs produced by B. paralicheniformis. Therefore, this study will provide a promising candidate for the development of new biocontrol agents.
Numerous reports have demonstrated that producing VOCs from microorganisms is a promising biological strategy owing to the prominent advantages of VOCs, including easy volatilization, non-physical contact, low residual, and high inhibitory activity [2,10,11,38]. Bacillus spp. have been reported to present strong antifungal activities against plant pathogens and are increasingly applied as the important biological control agents [34]. It is proved that the VOCs produced from B. subtilis, B. siamensis, B. velezensis, and B. aryabhattai could exhibit excellent biocontrol potential to various fungal pathogens in the field [39,40,41,42]. In this work, a new biological control agent, B. paralicheniformis 2-12, was found to produce antifungal VOCs. The percentage of mycelial inhibition was up to 60.41%. Moreover, B. paralicheniformis 2-12 VOCs also exhibited potent inhibitory effects on the other fifteen plant pathogenic fungi, indicating significantly antifungal activity and broad-spectrum inhibition characteristics. To reveal the morphological variations, F. oxysporum hyphae was further observed under SEM after treatment with B. paralicheniformis 2-12 VOCs. SEM analysis demonstrated that, compared to the control groups, the hyphae were distinctly twisty, vesicular, and shriveled on the surface, which is in agreement with previous reports [2,33]. In this study, new evidence suggested that VOCs produced by B. paralicheniformis could destroy the hyphae structure, thus leading to the inhibition of F. oxysporum hyphal growth.
Several reports have determined that the composition of bacterial VOCs mainly included alcohols, alkenes, aldehydes, acids, esters, ketones, aromatic compounds, and sulfides [2,42,43]. For instance, Calvo et al. [41] highlighted that three VOCs (diacetyl, benzaldehyde, and isoamyl alcohol) released from B. velezensis strains were proved to have high inhibitory activities against various kinds of post-harvest fungal pathogens. Wang et al. [2] revealed that B. siamensis YJ15 could produce 8 VOCs, including 4 alcohols, 1 ketone, 1 ester, 1 acid, and 1 aromatic compound, with high antagonistic activity against B. cinerea. Song et al. [42] reported that B. aryabhattai AYG1023 could produce 6 VOCs, including 4 ketones, 1 alcohol, and 1 aldehyde. Similarly, in this work, the VOCs produced by B. paralicheniformis were categorized as 9 alkanes, 3 alcohols, 3 acids and esters, 1 aromatic compound, and 1 alkyne compound. Although ethyl 2-methylbutanoate, one of the VOCs produced by B. siamensis YJ15, has been proved to exhibit antifungal activity against B. cinerea [2], the antifungal activities of other VOCs still remain largely unknown. So, further studies are strongly recommended to evaluate the activities of each volatile compound and to help reveal the antifungal mechanism.
To reveal the influence of VOCs on F. oxysporum at the molecular level, transcriptomic analysis was performed to investigate differential gene expression. Under B. paralicheniformis 2-12 VOC stress, a total of 1945 DEGs were found compared to the control groups. GO annotation analysis indicated that many DEGs were involved in catalytic activity, membranes, metabolic processes, and membrane parts. Cell membranes are of importance for cell integrity and fungal survival [33]. This finding showed that membrane and membrane components might be the main targets of the VOCs, which is in good agreement with previous studies as described by Yue et al. [10] and Duan et al. [33]. Additionally, the main KEGG pathways, such as amino acid metabolism and lipid metabolism, were also found through KEGG annotation analysis. Nishida et al. [44] indicated that amino acid metabolism could serve as a major nutrient source for fungi and may represent a new target for antifungal mechanisms. Mccarthy and Walsh [45] also reported that amino acid transport and metabolism were identified as potential drug targets to treat fungal infections. Lipids could increase cell membrane stability and regulate its fluidity and permeability [39]. Chen et al. [46] noted that citral causes the disorder of lipid metabolism and further disrupts the biological membrane functions of Alternaria alternata. Thus, amino acid metabolism and lipid metabolism might play important roles in the growth inhibition of F. oxysporum.

5. Conclusions

This study identified a new BCA, B. paralicheniformis 2-12, that could produce antifungal VOCs. B. paralicheniformis 2-12 VOCs exhibited prominent mycelial inhibition activity against F. oxysporum, as well as fifteen other plant pathogens. SEM analysis demonstrated that the hyphae treated with the VOCs were distinctly twisty, vesicular, and shriveled on the surface. Additionally, B. paralicheniformis 2-12 VOCs also demonstrated potent biological efficiency. B. paralicheniformis 2-12 VOCs were composed of 17 VOCs, including 9 alkanes, 3 alcohols, 3 acids and esters, 1 aromatic compound, and 1 alkyne compound. Through transcriptomic analysis, a total of 1945 DEGs were obtained and might be closely related to membranes and membrane parts, amino acid metabolism, and lipid metabolism. Although more studies are required to explain how the VOCs affect the growth of F. oxysporum, the findings will give new insights into the suppression of A. membranaceus root rot in the field.

Author Contributions

Conceptualization, writing—original draft, Y.W.; investigation, J.Y.; methodology, R.Z., S.Y., Y.S. and W.J.; data curation, X.H.; validation, M.W. and W.F.; supervision, writing—review and editing, C.W. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by Modern Agro-industry Technology Research System of Shanxi Province (2024CYJSTX16), Fundamental Research Program of Shanxi Province (20210302123404), and Graduate Education Innovation Project of Shanxi Province (2023SJ115).

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The original contributions presented in this study are included in the article. Further inquiries can be directed to the corresponding author.

Conflicts of Interest

The authors declare no conflicts of interest.

References

  1. Song, Y.; Li, P.; Wang, D.; Cheng, Y. Micellar electrokinetic chromatography for the quantitative analysis of flavonoids in the Radix of Astragalus membranaceus var. Mongholicus. Planta Med. 2008, 74, 84–89. [Google Scholar] [CrossRef]
  2. Wang, Y.; Wang, C.; Ma, Y.; Zhang, X.; Yang, H.; Li, G.; Li, X.; Wang, M.; Zhao, X.; Wang, J.; et al. Rapid and specific detection of Fusarium acuminatum and Fusarium solani associated with root rot on Astragalus membranaceus using loop-mediated isothermal amplification (LAMP). Eur. J. Plant Pathol. 2022, 163, 305–320. [Google Scholar] [CrossRef]
  3. Zhou, R.; Chen, H.; Chen, J.; Chen, X.; Wen, Y.; Xu, L. Extract from Astragalus membranaceus inhibit breast cancer cells proliferation via PI3K/AKT/mTOR signaling pathway. BMC Complement. Altern. Med. 2018, 18, 83. [Google Scholar] [CrossRef] [PubMed]
  4. Ny, V.; Houka, M.; Pavela, R.; Tříska, J. Potential benefits of incorporating Astragalus membranaceus into the diet of people undergoing disease treatment: An overview. J. Funct. Foods 2021, 77, 104339. [Google Scholar] [CrossRef]
  5. Wang, J.; Wang, Y.; Wang, Y.; Li, Y.; Yang, L.; Lu, B.; Chen, C.; He, R.; Wang, X.; Gao, J. Species and distribution of Fusarium causing ginseng root rot and antifungal efficacy of captan and hymexazol against F. spp. in China. Plant Dis. 2025. [Google Scholar] [CrossRef] [PubMed]
  6. Wang, Y.; Luo, J.; Shan, S.; Jiang, J.; Quan, X.; Wu, S.; Huang, S. Isolation and identification of root rot fungus of Astragalus membranaceus. Agric. Biotechnol. 2018, 7, 250–253. [Google Scholar] [CrossRef]
  7. Gao, F.; Xu, Y.; Zhao, L.; Chao, J.; Zhao, Z. Analysis of key differential metabolites in Astragalus membranaceus var. mongholicus against Fusarium oxysporum and assessment of sucrose as putative resistance-related metabolite. Physiol. Mol. Plant Pathol. 2025, 138, 102671. [Google Scholar] [CrossRef]
  8. Wang, C.; Hao, D.; Jiao, W.; Li, J.; Yuan, J.; Ma, Y.; Wang, X.; Xu, A.; Wang, Q.; Wang, Y. Identification and fungicide sensitivity of Fusarium spp. associated with root rot of Scutellaria baicalensis in Shanxi Province, China. Phytopathology 2024, 114, 1533–1541. [Google Scholar] [CrossRef]
  9. Ma, G.; Duan, X.; Xu, W.; Zhou, Y.; Ma, H.; Ma, W.; Qi, H. Identification and laboratory screening of chemical agents of root rot pathogens of Astragalus membranaceus var. mongholicus. Acta Agrestia Sin. 2022, 30, 1122–1130. [Google Scholar]
  10. Yue, Y.; Wang, Z.; Zhong, T.; Guo, M.; Huang, L.; Yang, L.; Kan, J.; Zalán, Z.; Hegyi, F.; Takács, K.; et al. Antifungal mechanisms of volatile organic compounds produced by Pseudomonas fluorescens ZX as biological fumigants against Botrytis cinerea. Microbiol. Res. 2023, 267, 127253. [Google Scholar] [CrossRef]
  11. Ni, H.; Kong, W.L.; Zhang, Q.Q.; Wu, X.Q. Volatiles emitted by Pseudomonas aurantiaca ST-TJ4 trigger systemic plant resistance to Verticillium dahliae. Microbiol. Res. 2024, 287, 127834. [Google Scholar] [CrossRef]
  12. Shi, X.; Zhu, D.; Chen, J.; Qin, Y.; Li, X.; Qin, S. Growth promotion and biological control of fungal diseases in tomato by a versatile rhizobacterium, Pseudomonas chlororaphis subsp. aureofaciens SPS-41. Physiol. Mol. Plant Pathol. 2024, 131, 102274. [Google Scholar] [CrossRef]
  13. Mari, M.; Bautista-Banos, S.; Dharini Sivakumar, D. Decay control in the postharvest system: Role of microbial and plant volatile organic compounds. Postharvest Biol. Technol. 2016, 122, 70–81. [Google Scholar] [CrossRef]
  14. Netzker, T.; Shepherdson, E.M.F.; Zambri, M.P.; Elliot, M.A. Bacterial volatile compounds: Functions in communication, cooperation, and competition. Annu. Rev. Microbiol. 2020, 74, 409–430. [Google Scholar] [CrossRef] [PubMed]
  15. Dinango, V.N.; Eke, P.; Youmbi, D.Y.; Kouokap, L.R.K.; Kouipou, R.M.T.; Tamghe, G.G.; Mabou, L.C.N.; Wakam, L.N.; Boyom, F.F. Endophytic bacteria derived from the desert-spurge (Euphorbia antiquorum L.) suppress Fusarium verticillioides, the causative agent of maize ear and root rot. Rhizosphere 2022, 23, 100562. [Google Scholar] [CrossRef]
  16. Manikandan, A.; Jaivel, N.; Johnson, I.; Krishnamoorthy, R.; Senthilkumar, M.; Raghu, R.; Gopal, N.O.; Mukherjee, P.K.; Anandham, R. Suppression of Macrophomina root rot, Fusarium wilt and growth promotion of some pulses by antagonistic rhizobacteria. Physiol. Mol. Plant Pathol. 2022, 121, 101876. [Google Scholar] [CrossRef]
  17. Tu, M.; Zhu, Z.; Zhao, X.; Cai, H.; Zhang, Y.; Yan, Y.; Yin, K.; Sha, Z.; Zhou, Y.; Chen, G.; et al. The versatile plant probiotic bacterium Bacillus velezensis SF305 reduces red root rot disease severity in the rubber tree by degrading the mycelia of Ganoderma pseudoferreum. J. Integr. Agric. 2024. [Google Scholar] [CrossRef]
  18. Xie, S.; Si, H.; Wang, S.; Duan, Y.; Wang, Z.; Niu, J. Biocontrol activities and mechanisms of endophytic Bacillus licheniformis SY41 against Atractylodes chinensis root rot. Sci. Hortic. 2025, 341, 113995. [Google Scholar] [CrossRef]
  19. Alijani, Z.; Amini, J.; Ashengroph, M.; Bahramnejad, B. Antifungal activity of volatile compounds produced by Staphylococcus sciuri strain MarR44 and its potential for the biocontrol of Colletotrichum nymphaeae, causal agent strawberry anthracnose. Int. J. Food Microbiol. 2019, 307, 108276. [Google Scholar] [CrossRef]
  20. Wang, C.; Duan, T.; Shi, L.; Zhang, X.; Wang, M.; Wang, J.; Ren, L.; Zhao, X.; Wang, Y. Characterization of volatile organic compounds produced by Bacillus siamensis YJ15 and their antifungal activity against Botrytis cinerea. Plant Dis. 2022, 106, 2321–2329. [Google Scholar] [CrossRef]
  21. Tian, J.; Huang, B.; Luo, X.; Zeng, H.; Ban, X.; He, J.; Wang, Y. The control of Aspergillus flavus with Cinnamomum jensenianum Hand.-Mazz essential oil and its potential use as a food preservative. Food Chem. 2012, 130, 520–527. [Google Scholar] [CrossRef]
  22. Yamamoto, S.; Harayama, S. PCR amplification and direct sequencing of gyrB genes with universal primers and their application to the detection and taxonomic analysis of Pseudomonas putida strains. Appl. Environ. Microb. 1995, 61, 1104–1109. [Google Scholar] [CrossRef] [PubMed]
  23. Ghazouani, A.; El-Gayar, K.; Abada, E. Molecular characterization of gene encoding halo tolerant amylase of Bacillus paralicheniformis isolated from Jazan region. Mod. J. Microb. Biol. 2020, 2, 1–8. [Google Scholar] [CrossRef]
  24. Clerck, D.E.; Vanhoutte, T.; Hebb, T.; Geerinck, J.; Devos, J.; De Vos, P. Isolation, characterization, and identification of bacterial contaminants in semifinal gelatin extracts. Appl. Environ. Microb. 2004, 70, 3664–3672. [Google Scholar] [CrossRef] [PubMed]
  25. Cao, F.; Yang, X.; Ma, M.; Chen, H.; Shen, D.; Li, J. Advances in the identification of Bacillus subtilis and closely related species. Microbiol. China 2014, 41, 968–974. [Google Scholar] [CrossRef]
  26. Zhang, X.; Gao, Z.; Zhang, X.; Bai, W.; Zhang, L.; Pei, H.; Zhang, Y. Control effects of Bacillus siamensis G-3 volatile compounds on raspberry postharvest diseases caused by Botrytis cinerea and Rhizopus stolonifer. Biol. Control 2020, 141, 104135. [Google Scholar] [CrossRef]
  27. Robles, C.A.; Ceriani-Nakamurakarea, E.; Slodowicz, M.; González-Audino, P.; Carmarán, C.C. Granulobasidium vellereum (Ellis & Cragin) Jülich, a promising biological control agent. Biol. Control 2018, 117, 99–108. [Google Scholar] [CrossRef]
  28. Wang, C.; Wang, Y.; Wang, L.; Fan, W.; Zhang, X.; Chen, X.; Wang, M.; Wang, J. Biocontrol potential of volatile organic compounds from Pseudomonas chlororaphis ZL3 against postharvest gray mold caused by Botrytis cinerea on Chinese cherry. Biol. Control 2021, 159, 104613. [Google Scholar] [CrossRef]
  29. Guo, Y.; Chen, D.; Dong, Y.; Ju, H.; Wu, C.; Lin, S. Characteristic volatiles fingerprints and changes of volatile compounds in fresh and dried Tricholoma matsutake Singer by HS-GC-IMS and HS-SPME-GC-MS. J. Chromatogr. B 2018, 1099, 46–55. [Google Scholar] [CrossRef]
  30. Xu, Y.X.; Zhang, M.; Fang, Z.X.; Sun, J.C.; Wang, Y.Q. How to improve bayberry (Myrica rubra Sieb. et Zucc.) juice flavour quality: Effect of juice processing and storage on volatile compounds. Food Chem. 2014, 151, 40–46. [Google Scholar] [CrossRef]
  31. Wang, S.S.; Zhou, F.Y.; Wang, B.; Xu, D.D.; Cao, Q.J.; Lu, M.; Sun, J.H. Volatiles produced by bacteria alleviate antagonistic effects of one associated fungus on Dendroctonus valens larvae. Sci. China Life Sci. 2017, 60, 924–926. [Google Scholar] [CrossRef]
  32. Huang, Y.; Shan, X.; Zhang, C.; Duan, Y. Pseudomonas protegens volatile organic compounds inhibited brown rot of postharvest peach fruit by repressing the pathogenesis-related genes in Monilinia fructicola. Food Microbiol. 2024, 122, 104551. [Google Scholar] [CrossRef] [PubMed]
  33. Duan, W.Y.; Zhu, X.M.; Zhang, S.B.; Lv, Y.Y.; Zhai, H.C.; Wei, S.; Ma, P.A.; Hu, Y.S. Antifungal effects of carvacrol, the main volatile compound in Origanum vulgare L. essential oil, against Aspergillus flavus in postharvest wheat. Int. J. Food Microbiol. 2024, 410, 110514. [Google Scholar] [CrossRef] [PubMed]
  34. Wang, K.; Qin, Z.; Wu, S.; Zhao, P.; Zhen, C.; Gao, H. Antifungal mechanism of volatile organic compounds produced by Bacillus subtilis CF-3 on Colletotrichum gloeosporioides assessed using omics technology. J. Agric. Food Chem. 2021, 69, 5267–5278. [Google Scholar] [CrossRef] [PubMed]
  35. Jinal, H.N.; Sakthivel, K.; Amaresan, N. Characterisation of antagonistic Bacillus paralicheniformis (strain EAL) by LC–MS, antimicrobial peptide genes, and ISR determinants. Antonie Leeuwenhoek 2020, 113, 1167–1177. [Google Scholar] [CrossRef]
  36. Xie, T.; Shen, S.; Hu, R.; Li, W.; Wang, J. Screening, identification, and growth promotion of antagonistic endophytes associated with Chenopodium quinoa against quinoa pathogens. Phytopathology 2023, 113, 1839–1852. [Google Scholar] [CrossRef]
  37. Kushmitha, B.; Ashraf, S.; Nakkeeran, S.; Johnson, I.; Saranya, N.; Mahendra, K.; Raish, M. Whole genome sequencing of rice endophyte Bacillus paralicheniformis NB stem 4: A potential biocontrol agent for the suppression of pearl millet blast disease. Physiol. Mol. Plant Pathol. 2025, 138, 102663. [Google Scholar] [CrossRef]
  38. Chaouachi, M.; Marzouk, T.; Jallouli, S.; Elkahoui, S.; Gentzbittel, L.; Ben, C.; Djébali, N. Activity assessment of tomato endophytic bacteria bioactive compounds for the postharvest biocontrol of Botrytis cinerea. Postharvest Biol. Technol. 2021, 172, 111389. [Google Scholar] [CrossRef]
  39. Gao, Z.; Zhang, B.; Liu, H.; Han, J.; Zhang, Y. Identification of endophytic Bacillus velezensis ZSY-1 strain and antifungal activity of its volatile compounds against Alternaria solani and Botrytis cinerea. Biol. Control 2017, 105, 27–39. [Google Scholar] [CrossRef]
  40. Jiang, C.H.; Liao, M.J.; Wang, H.K.; Zheng, M.Z.; Xu, J.J.; Guo, J.H. Bacillus velezensis, a potential and efficient biocontrol agent in control of pepper gray mold caused by Botrytis cinerea. Biol. Control 2018, 126, 147–157. [Google Scholar] [CrossRef]
  41. Calvo, H.; Mendiara, I.; Arias, E.; Gracia, A.P.; Blanco, D.; Venturini, M.E. Antifungal activity of the volatile organic compounds produced by Bacillus velezensis strains against postharvest fungal pathogens. Postharvest Biol. Technol. 2020, 166, 111208. [Google Scholar] [CrossRef]
  42. Song, C.; Zhang, Y.; Zhao, Q.; Chen, M.; Zhang, Y.; Gao, C.; Jia, Z.; Song, S.; Guan, J.; Shang, Z. Volatile organic compounds produced by Bacillus aryabhattai AYG1023 against Penicillium expansum causing blue mold on the Huangguan pear. Microbiol. Res. 2024, 278, 127531. [Google Scholar] [CrossRef]
  43. Martins, S.J.; Faria, A.F.; Pedroso, M.P.; Cunha, M.G.; Rocha, M.R.; Medeiros, F.H.V. Microbial volatiles organic compounds control anthracnose (Colletotrichum lindemuthianum) in common bean (Phaseolus vulgaris L.). Biol. Control 2019, 131, 36–42. [Google Scholar] [CrossRef]
  44. Nishida, I.; Watanabe, D.; Tsolmonbaatar, A.; Kaino, T.; Ohtsu, I.; Takagi, H. Vacuolar amino acid transporters upregulated by exogenous proline and involved in cellular localization of proline in Saccharomyces cerevisiae. J. Gen. Appl. Microbiol. 2016, 62, 132–139. [Google Scholar] [CrossRef] [PubMed]
  45. Mccarthy, M.W.; Walsh, T.J. Amino acid metabolism and transport mechanisms as potential antifungal targets. Int. J. Mol. Sci. 2018, 19, 909. [Google Scholar] [CrossRef] [PubMed]
  46. Chen, Y.; Zeng, Y.; Li, Y.; Ye, Z.; Li, L.; Zhou, Z. Antifungal effects of citral against Alternaria alternata in postharvest pear fruit and its potential mechanism. Postharvest Biol. Technol. 2025, 223, 113424. [Google Scholar] [CrossRef]
Microorganisms 13 01782 g001
Figure 1. The antifungal activity of VOCs produced by isolate 2-12 against Fusarium oxysporum. (a) Mycelial growth; (b) mycelial wet weight; (c) mycelial dry weight. The same letter indicates no significant difference based on Duncan’s multiple range test.
Figure 1. The antifungal activity of VOCs produced by isolate 2-12 against Fusarium oxysporum. (a) Mycelial growth; (b) mycelial wet weight; (c) mycelial dry weight. The same letter indicates no significant difference based on Duncan’s multiple range test.
Microorganisms 13 01782 g001
Microorganisms 13 01782 g002
Figure 2. The morphological characteristics of isolate 2-12. (a) Colony on NA plate at 28 °C for 2 d; (b) Gram stain; (c) bacterial cell under transmission electron microscope.
Figure 2. The morphological characteristics of isolate 2-12. (a) Colony on NA plate at 28 °C for 2 d; (b) Gram stain; (c) bacterial cell under transmission electron microscope.
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Microorganisms 13 01782 g003
Figure 3. Phylogenetic tree based on gyrB (a) and rpoB (b) gene sequences.
Figure 3. Phylogenetic tree based on gyrB (a) and rpoB (b) gene sequences.
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Microorganisms 13 01782 g004
Figure 4. Scanning electron microscopy (SEM) analysis of Fusarium oxysporum hyphae. (a–c) hyphae treated without VOCs as controls; (d–f) hyphae treated with the VOCs. Scale bar = 5 μm.
Figure 4. Scanning electron microscopy (SEM) analysis of Fusarium oxysporum hyphae. (a–c) hyphae treated without VOCs as controls; (d–f) hyphae treated with the VOCs. Scale bar = 5 μm.
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Microorganisms 13 01782 g005
Figure 5. Biocontrol effects of Bacillus paralicheniformis 2-12 VOCs on Astragalus membranaceus root rot. (a) Disease incidence; (b) lesion diameter. The different letters mean significant difference based on Duncan’s multiple range test.
Figure 5. Biocontrol effects of Bacillus paralicheniformis 2-12 VOCs on Astragalus membranaceus root rot. (a) Disease incidence; (b) lesion diameter. The different letters mean significant difference based on Duncan’s multiple range test.
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Microorganisms 13 01782 g006
Figure 6. Transcriptomic analysis in Fusarium oxysporum under Bacillus paralicheniformis 2-12 VOC stress. (a) The scatter plot; (b) heatmap; (c) GO functional annotation analysis; (d) KEGG pathway annotation analysis.
Figure 6. Transcriptomic analysis in Fusarium oxysporum under Bacillus paralicheniformis 2-12 VOC stress. (a) The scatter plot; (b) heatmap; (c) GO functional annotation analysis; (d) KEGG pathway annotation analysis.
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Microorganisms 13 01782 g007
Figure 7. Enrichment analyses of DEGs. (a) GO enrichment analysis; (b) KEGG enrichment analysis.
Figure 7. Enrichment analyses of DEGs. (a) GO enrichment analysis; (b) KEGG enrichment analysis.
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Table 1. The antifungal activity of VOCs emitted from Bacillus paralicheniformis 2-12 against plant pathogenic fungi.
Table 1. The antifungal activity of VOCs emitted from Bacillus paralicheniformis 2-12 against plant pathogenic fungi.
FungiHostPercentage Mycelial
Inhibition (%)		Shapiro–Wilk p
Fusarium oxysporumPhaseolus radiatus69.04 ± 0.63 de0.244
Fusarium solaniAstragalus membranaceus75.89 ± 1.93 bc0.258
Exserohilum turcicumZea mays77.22 ± 2.57 bc0.550
F. semitectumZea mays54.66 ± 2.00 h0.843
F. graminearumZea mays62.12 ± 2.89 fg0.040
Fusarium oxysporumMusa paradisiaca63.06 ± 2.87 fg0.420
Botrytis cinereaCerasus pseudocerasus94.96 ± 2.28 a0.042
F. acuminatumAstragalus membranaceus72.97 ± 6.15 cd0.323
F. avenaceumVitis vinifera31.88 ± 4.65 j0.796
F. equisetiAstragalus membranaceus47.89 ± 4.45 i0.453
Fusarium graminearumTriticum aestivum76.07 ± 2.03 bc0.245
Alternaria alternataChrysanthemum morifolium64.57 ± 2.88 ef0.534
Colletotrichum gloeosporioidesCapsicum annuum58.39 ± 3.47 gh0.063
F. oxysporumSolanum melongena81.10 ± 0.32 b0.828
F. oxysporumCucumis sativus59.39 ± 5.00 fgh0.328
The same letter indicates no significant difference based on Duncan’s multiple range test.
Table 2. The analysis of VOCs from Bacillus paralicheniformis 2-12 by SPME-GC-MS.
Table 2. The analysis of VOCs from Bacillus paralicheniformis 2-12 by SPME-GC-MS.
CompoundsChemical
Formula		MW 1RT 2 (min)SI 3RSI 4
Ethyl 2-methylbutanoateC7H14O2130.186.09693748
Benzeneethanaminea-methylC9H13N135.236.28705720
Propanoic acid, 3-amino-3-oxo-C3H5NO3103.086.31603952
N-Acetyl-L-alanineC5H9NO3131.136.32592981
2,4-OctadiyneC8H10106.176.92744848
TridecaneC13H28184.369.9825865
Undecane,5-methylC12H26170.339.91801876
Decane,2,6-dimethylC12H26170.3310.66620901
3,5-dimethylheptan-3-olC9H20O144.2510.67603734
1-Butanol,2,2-dimethylC6H14O102.1710.67670775
2-MethylundecaneC12H26170.3313.32835864
3,3-DimethylhexaneC8H18114.2314.00742786
2,3,4-TrimethylpentaneC8H18114.2314.14555764
Heptane,1-propoxyC10H22O158.2814.36580671
2,5-Dimethyl-3,4-hexanediolC8H18O2146.2314.36603691
Decane,2,9-dimethylC12H26170.3316.19675818
2,4,6-TrimethyloctaneC11H24156.3116.2688808
1 Retention time. 2 Molecular weight. 3 Direct match. 4 Reverse match.
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MDPI and ACS Style

Wang, Y.; Yuan, J.; Zhao, R.; Yuan, S.; Su, Y.; Jiao, W.; Huo, X.; Wang, M.; Fan, W.; Wang, C. Biocontrol Activity of Volatile Organic Compounds Emitted from Bacillus paralicheniformis 2-12 Against Fusarium oxysporum Associated with Astragalus membranaceus Root Rot. Microorganisms 2025, 13, 1782. https://doi.org/10.3390/microorganisms13081782

AMA Style

Wang Y, Yuan J, Zhao R, Yuan S, Su Y, Jiao W, Huo X, Wang M, Fan W, Wang C. Biocontrol Activity of Volatile Organic Compounds Emitted from Bacillus paralicheniformis 2-12 Against Fusarium oxysporum Associated with Astragalus membranaceus Root Rot. Microorganisms. 2025; 13(8):1782. https://doi.org/10.3390/microorganisms13081782

Chicago/Turabian Style

Wang, Yan, Jiaqi Yuan, Rui Zhao, Shengnan Yuan, Yaxin Su, Wenhui Jiao, Xinyu Huo, Meiqin Wang, Weixin Fan, and Chunwei Wang. 2025. "Biocontrol Activity of Volatile Organic Compounds Emitted from Bacillus paralicheniformis 2-12 Against Fusarium oxysporum Associated with Astragalus membranaceus Root Rot" Microorganisms 13, no. 8: 1782. https://doi.org/10.3390/microorganisms13081782

APA Style

Wang, Y., Yuan, J., Zhao, R., Yuan, S., Su, Y., Jiao, W., Huo, X., Wang, M., Fan, W., & Wang, C. (2025). Biocontrol Activity of Volatile Organic Compounds Emitted from Bacillus paralicheniformis 2-12 Against Fusarium oxysporum Associated with Astragalus membranaceus Root Rot. Microorganisms, 13(8), 1782. https://doi.org/10.3390/microorganisms13081782

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