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The antagonistic activity of Streptomyces spiroverticillatus (No. HS1) against of poplar canker pathogen Botryosphaeria dothidea

Abstract

Background

Poplar canker caused by Botryosphaeria dothidea is one of the most severe plant disease of poplars worldwide. In our study, we aimed to investigate the modes of antagonism by fermentation broth supernatant (FBS) of Streptomyces spiroverticillatus HS1 against B. dothidea.

Results

In vitro, the strain and FBS of S. spiroverticillatus HS1 significantly inhibited mycelial growth and biomass accumulation, and also disrupted the mycelium morphology of B. dothidea. On the 3rd day after treatment, the inhibition rates of colony growth and dry weight were 80.72% and 52.53%, respectively. In addition, FBS treatment damaged the plasma membrane of B. dothidea based on increased electrical conductivity in the culture medium, and malondialdehyde content of B. dothidea mycelia. Notably, the analysis of key enzymes in glycolysis pathway showed that the activity of hexokinase (HK), phosphofructokinase (PFK), and pyruvate kinase (PK), Ca2+Mg2+-ATPase were significantly increased after FBS treatment. But the glucose contents were significantly reduced, and pyruvate contents were significantly increased in B. dothidea after treatment with FBS.

Conclusions

The inhibitory mechanism of S. spiroverticillatus HS1 against B. dothidea was a complex process, which was associated with multiple levels of mycelial growth, cell membrane structure, material and energy metabolism. The FBS of S. spiroverticillatus HS1 could provide an alternative approach to biological control strategies against B. dothidea.

Peer Review reports

Background

Poplars (Populus L.) are the main plants in the plains and sandy areas of northern China [1]. They contribute to protect and improve environment, such as protecting farmland from soil erosion caused by wind and water, providing forest products, and park land for recreation [2, 3]. Poplar canker is a major disease affecting poplars worldwide. It mainly involves infections in the trunk and branches, which can lead to tree death in severe cases. At present, there are more than ten pathogens that can cause poplar canker worldwide [4]. Among them, B. dothidea is widely distributed worldwide and has been identified as a pathogen of many woody plants. It also shows high genetic diversity in the same host, which is one of the major reasons for the outbreak of poplar canker [5, 6]. Using chemical fungicides, such as benomyl, tebuconazole, carbendazim, thiophanate-methyl, is effective against pathogens [7, 8]. However, chemical control is not a sustainable solution owing to the potential human health hazards, environmental pollution, and enhanced pathogen resistance [9]; therefore, the use of biological control has attracted more attention in recent years.

At present, many studies mainly focus on the development of biocontrol agents (BCAs) and antagonistic mechanisms. Some studies have shown that the extracts of plants such as Cymbopogon, Curcuma longa, and Notopterygium incisum, as well as bioactive components such as cuminaldehyde, geraniol, β-citronellol, and lemon essential oil can inhibit B. dothidea mycelial growth, and the germination and formation of spores [10,11,12]. The screening of antagonistic microorganisms generally focuses on Bacillus spp. and Streptomyces spp. from soil or healthy hosts. Fengycin is produced by Bacillus subtilis isolated from healthy apples, and it has a good control effect on apple ring rot caused by B. dothidea [13]. B. amyloliquefaciens subsp. plantarum (CGMCC 11640) isolated from soil can inhibit B. dothidea growth and provides a bio-resource for the biocontrol of Carya cathayensis canker [14]. In addition, a few studies provide a new strategy of using varieties of effective antagonistic substances in combination for inhibiting pathogen growth instead of using a single biocontrol strain metabolite. For example, ε-polylysine, chitosan oligomers, their conjugates, S. rochei, S. lavendofoliae culture filtrates, and their secondary metabolites with chitosan oligomers have been used to inhibit the growth of B. dothidea, Neofusicoccum parvum and Diplodia seriata that cause grapevine trunk diseases [15].

Streptomyces are the most widely studied and notable species among the Actinomycetes and they produce many naturally occurring antibiotics [16], which are widely used as BCAs owing to their ability to produce various secondary metabolites. S. violaceusniger has antagonistic activity against Fusarium oxysporum that causes banana wilt; antagonistic activity involves disrupting cell membranes and inhibiting mycelial growth and spore germination [17]. A low-concentration culture filtrate (diluted to 10−3) of Streptomyces spp. can inhibit the spore germination of Botrytis cinerea [18]. It was reported that antibiotic compounds consisting of heterologous small-molecular weight molecules, could affect pathogen growth and metabolism at relatively low concentrations and play a significant role in plant disease control [19, 20]. Also, some studies have reported that the antibiotic production of Streptomyces sp. had a negative impact on normal pathogen metabolism by altering the key enzyme activity [21]. In addition to the antagonistic effect of Streptomyces applied to inhibit plant pathogen, Streptomyces could promote plant growth [22,23,24] and induce systemic resistance against pathogen invasion [25]. For example, S. albus CAI-21 can promote the growth of rice, sorghum, and certain legumes, and it can also inhibit the growth of Macrophomina phaseolina in sorghum anthracnose infections [26]. In summary, Streptomyces and their secondary metabolites are important sources of biocontrol agents, because it could affect pathogen growth and cell metabolism, and also may induce systemic resistance.

In our previous study, S. spiroverticillatus isolated from forest soil in Jilin Province, China, has been found to display broad-spectrum antifungal activity against a range of plant pathogens. The preventive and therapeutic effects of S. spiroverticillatus HS1 fermentation broth supernatant (FBS) on poplar canker have been reported to be 60.87% and 71.74%, respectively. It efficiently prevented and control the occurrence and spread of canker [27]. However, the antifungal mechanism of S. spiroverticillatus HS1 against B. dothidea are unclear. In our study, we aimed to systematically test the antagonistic mechanisms of S. spiroverticillatus HS1, it will provide a new method to control the poplar canker disease.

Results

Antagonistic activity of S. spiroverticillatus HS1 against B. dothidea in vitro

Live S. spiroverticillatus HS1 exerted a strong inhibitory effect on B. dothidea with an inhibition band of 20.66 mm (Fig. 1a), and the FBS displayed an inhibition zone diameter of 40.26 mm (Fig. 1b). The mycelia of B. dothidea in the control group showed fast growth and covered the entire petri dish (Fig. 1a-C). B. dothidea mycelia in the treatment group showed slow growth away from the S. spiroverticillatus HS1, and the growth area was less than 1/3rd of the petri dish area (Fig. 1a-T). Compared with the control (Fig. 1b-C), the inhibition zone created by FBS of S. spiroverticillatus HS1 was large and clear (Fig. 1b-T). The anti-fungal effect of live strain and FBS of S. spiroverticillatus HS1 were long-lasting and stable, and the experiment was reproducible.

Fig. 1
figure 1

Antagonistic activity of S. spiroverticillatus HS1 vs. B. dothidea in vitro (T-treatment, C-control). Dual cultures of S. spiroverticillatus HS1 and B. dothidea; b Dual cultures of S. spiroverticillatus HS1-FBS and B. dothidea

Antagonistic effect of S. spiroverticillatus HS1-FBS against B. dothidea mycelia

Effect of FBS on the mycelial morphology of B. dothidea

Microscopic observation revealed that the mycelia of B. dothidea in the control group showed dense and vigorous growth with smooth colonies (Fig. 2-CK). In contrast, the mycelia were deformed, sparse, and shrunk, and the internodes were shortened, twisted, enlarged and broken after treatment with FBS (Fig. 2-Treatment).

Fig. 2
figure 2

Monitoring of mycelial morphology of B. dothidea using an optical microscope. The mycelia of B. dothidea exposed to FBS (Treatment) or to equivalent sterile fermentation medium (CK). The magnification was 400 times

Effect on the dry weight of B. dothidea mycelia

At 1–3 d after FBS treatment, the dry weight of mycelia in all treatment groups was significantly lower than that in the control group. Undiluted FBS showed the best antagonistic effect on B. dothidea mycelium biomass, and the rates of inhibition of B. dothidea growth based on dry weight of mycelia were 58.42% (1 d) and 52.53% (3 d). The dry weight of mycelia in the control group was 0.42 g and 0.46 g for 1d and 3d respectively, and 0.31 g and 0.32 g after treatment with 400 times diluted fermentation solution separately. The FBS diluted by 400 time retained the antagonistic effect, and treated B. dothidea was significantly different from that in the control group (Fig. 3).

Fig. 3
figure 3

The influence of the S. spiroverticillatus HS1-FBS on the dry weight of B. dothidea. Data represent the mean ± standard deviation (n = 3). The numbers on X axis represent the different dilution ratio of S. spiroverticillatus HS1-FBS treatments. CK, equivalent volume of sterile fermentation medium. The different letters indicate significant differences among treatments using Duncan’s test at p < 0.05. The lower-case letters and the upper-case letters represent the significant difference on Day1and Day 3, respectively

Effect of FBS on growth of B. dothidea

The effects of different concentrations of S. spiroverticillatus HS1-FBS on B. dothidea colony growth are shown in Table 1. The growth rate of B. dothidea in medium containing FBS was slow. The growth rate gradually decreased with an increase in inhibition rate as the FBS concentration increased. The inhibition rate after treatment with undiluted FBS was the highest at 80.72% on day 3, followed by 67.99% and 72.11% on days 5 and 7, respectively, when compared to that of diluted FBS treatments (P < 0.05). The inhibition rate on the 7th day was higher than those on the 3rd and 5th days when the dilution was less than 100-fold, indicating that the antagonistic effect of FBS was efficient, durable, and stable (Table 1).

Table 1 The effect of S. spiroverticillatus HS1 fermentation broth on the growth of mycelia of B.dothidea

Antagonistic effect of FBS on B. dothidea cell membrane

Effect of FBS on membrane permeability of B. dothidea

After treatment of B. dothidea with FBS, the conductivity was significantly higher than that of the control (P < 0.001), indicating that the permeability of B. dothidea mycelial cell membrane was increased and electrolyte leakage occurred. In the FBS treatment group, electrolyte leakage continued as the experiment was prolonged, and there was no decrease. The highest value was 3.693 ms/cm at 12 h, which was 5.77 times that of the control group, indicating that electrolyte leakage was severe (Fig. 4a). It can be presumed that FBS had a considerably negative impact on the cell membrane permeability of mycelia.

Effect of FBS on membrane lipid peroxidation of B. dothidea

Malondialdehyde (MDA) is the product of cell membrane lipid peroxidation, and its level reflects the degree of damage caused to the cell membrane. The MDA content in the mycelia of B. dothidea treated with FBS was significantly higher than that in the control group (P < 0.05), and lipid peroxidation of the cell membrane was increased, indicating that the cell membrane was severely damaged. The degree of lipid peroxidation gradually increased with an increase in treatment time. At 48 h, the MDA content in the FBS treatment group was 2.88 times that of the control group (Fig. 4b), further indicating that the FBS had a considerably adverse effect on the mycelial cell membrane.

Fig. 4
figure 4

Influence of S. spiroverticillatus HS1-FBS on the membrane of B. dothidea. a Cell membrane conductivity of B. dothidea; b The level of MDA in B. dothidea. Data are expressed as mean ± SD of three biological replicates. Differences between samples were determined by a two-tailed Student’s t-test at P < 0.05. *, 0.01 < P < 0.05; **, 0.01 < P < 0.001; ***, P < 0.001

Antagonistic effect of S. spiroverticillatus HS1-FBS on glycolysis pathway in B. dothidea

Effect of FBS on HK, PFK, PK, and Ca2+Mg2+-ATP activity

The activities of three key enzymes (HK, PFK, and PK) in the glycolysis pathway were determined to evaluate the effect of FBS on the metabolism of B. dothidea. The HK activity in all groups of B. dothidea showed a downward trend in general (Fig. 5a). However, the activity of HK in the FBS treatment group was slightly higher than that in the control, especially within 2–4 h, and the difference was considerable (P < 0.001). After treatment with FBS for 4 h, the activity of PFK in the cells was higher than that in the control group, and the difference was significant at 6, 12, and 48 h (P < 0.001). Notably, PFK activity in the FBS treatment group was 5.2 times that of the control group at 6 h (Fig. 5b). Similarly, PK activity in the FBS treatment group was slightly higher than that in the control group, and the difference was evident at 4, 8, 10, and 24 h (P < 0.05) (Fig. 5c). The activity of Ca2+Mg2+-ATPase reflected the energy metabolism of B. dothidea. As time increased, the enzyme activity of all groups showed a downward trend. Moreover, the Ca2+Mg2+-ATPase activity in the FBS treatment group was higher than that of the control group, especially within 2–10 h (Fig. 5d), and the difference was highly significant (P < 0.001). FBS treatment enhanced Ca2+ Mg2+-ATPase activity, accelerated the utilization of ATP, and affected the regular energy supply of cells.

Fig. 5
figure 5

Influence of S. spiroverticillatus HS1-FBS on HK (a), PFK (b), PK (c), Ca2+ Mg2+-ATPase (d) activity in B. dothidea. Differences between samples were determined by two-tailed Student’s t-test at P < 0.05. *, 0.01 < P < 0.05; **, 0.01 < P < 0.001; ***, P < 0.001

Effect of FBS on total glucose and total pyruvate levels

The total glucose level in the control group was stable at first and then decreased sharply at 12 h. The FBS treatment group showed an upward trend as the total glucose level increased sharply at 10 h, and the two groups showed opposite trends (Fig. 6a). The total glucose level of the control group was markedly higher than that of the FBS treatment group for 2–8 h (P < 0.05). Especially at 4 h, the total glucose level in the control group was 3-fold higher than that in the FBS treatment group. Therefore, glucose metabolism in the cells of B. dothidea was accelerated via FBS treatment at this time, and the glucose content could not accumulate, which was mutually verified based on the enhanced activity of the three carbohydrate-metabolizing enzymes. In contrast, the total pyruvate level in the FBS treatment group was significantly higher than that in the control group (P < 0.001). Notably, the pyruvate level in the FBS treatment group was 3.9 times that of the control group at 24 h (Fig. 6b). This indicated that pyruvate gradually accumulated after FBS treatment, which further indicated abnormal glucose metabolism and confirmed the severe adverse effects on the growth of the B. dothidea.

Fig. 6
figure 6

Influence of S. spiroverticillatus HS1-FBS on total glucose and total pyruvate level in B. dothidea. a Total glucose level; b Total pyruvate level. Differences between samples were determined by two-tailed Student’s t-test at P < 0.05. *, 0.01 < P < 0.05; **, 0.01 < P < 0.001; ***, P < 0.001

Materials and methods

The identification of S. spiroverticillatus HS1 in vitro

S. spiroverticillatus HS1 was previously isolated from the soil at Hengshan Protection Station, Changbai Mountain National Nature Reserve, Jilin Province, China. It was identified based on the morphology, physiological and biochemical characteristics, and 16S rDNA sequence (GenBank accession number: MN636764). B. dothidea, the causal agent of poplar canker, was isolated, identified, maintained, and stored by the Jilin Provincial Academy of Forestry Sciences. The S. spiroverticillatus HS1 fermentation medium and FBS has been optimized and widely used [28].

Antagonistic activities of S. spiroverticillatus HS1-FBS against mycelial growth of B. dothidea in vitro

Antifungal activity

In vitro antifungal activity of S. spiroverticillatus HS1 against B. dothidea was tested by the dual culture technique. The mycelia plug (5 mm in diameter) of B. dothidea taken from 5-day-old target fungus on Potato Dextrose Agar (PDA) was placed 5 cm away from the bacterial streak line of S. spiroverticillatus HS1. The dual culture plates were incubated at 28 °C, and the inhibition of fungal growth antifungal activity was measured every day until 7 days after inoculation.

Antifungal activity of FBS

In vitro antifungal activity of S. spiroverticillatus HS1-FBS against B. dothidea was also tested by the dual culture technique. The mycelia of B. dothidea was mixed with PDA which was under 45℃, and the oxford cups containing 200ul of S. spiroverticillatus HS1-FBS were placed in the center of the cooled PDA. The dual culture plates were incubated at 28 °C, and the inhibition of fungal growth antifungal activity was measured every day until 7 days after inoculation.

Mycelial morphology

FBS was added to potato dextrose agar (PDA) medium at 50 °C at a volume ratio of 1:4, and the mycelial plug of B. dothidea (5 mm in diameter) was placed in the center of the plate following culturing at 28 °C for 5 d. The medium supernatant without bacteria was used as a control. The mycelial morphology was observed and compared with that of the control using an optical microscope (BX53; Olympus, Tokyo, Japan).

Mycelial dry weight

Briefly, three mycelial plugs (5 mm) of B. dothidea were inoculated in 100 ml potato dextrose broth (PD) medium at 28 °C with shaking at 150 rpm for 48 h. Subsequently, 20 ml of the undiluted or different fold diluted FBS (10-, 25-, 50-, 100-, 200-, and 400-fold) were added into erlenmeyer flasks as different treatment. Cultures of pathogen conducted in PD and supplemented with equivalent fermentation medium were used as control. After 1 and 3 d of culture, the mycelia were collected and rinsed with sterile water and then dried in an oven at 80 °C for 3 h. The dry weight of mycelia was determined, and each treatment was performed in triplicate.

Mycelial growth

The PDA medium was adjusted with undiluted FBS or the FBS diluted by 10-, 25-, 50-, 100-, 200-, or 400-fold at 50 °C in a volume ratio of 1:4, and the mycelial plug of B. dothidea (5 mm in diameter) was placed in the center of the plate following culturing at 28 °C for 5 d. Cultures of pathogen grown in PDA mixing with equivalent fermentation medium were used as control. The diameter of the mycelial colony was measured using the crosshairs at 3, 5, and 7 d. Each treatment was performed in triplicate. Rate of inhibition of growth was calculated as follows:

$$\mathrm{Inhibition}\;\mathrm{rate}\;(\%)\;=\;(\mathrm{control}\;\mathrm{net}\;\mathrm{growth}\;-\;\mathrm{treatment}\;\mathrm{net}\;\mathrm{growth})\;/\;\mathrm{control}\;\mathrm{net}\;\mathrm{growth}\;\times\;100\%.$$

Effect of S. spiroverticillatus HS1-FBS on B. dothidea membrane

Electrolyte leakage

Briefly, 15 B. dothidea mycelial plugs (5 mm) were inoculated into PD medium (100 ml) and incubated at 28 °C with shaking at 150 rpm for 48 h. FBS solution (20 mL) was added separately. The conductivity was measured using a conductivity meter (FC20; OHAUS, Shanghai, China) at 0, 2, 4, 6, 8, 10, 12, 24, and 48 h [29]. Each treatment was performed in triplicate. Cultures of B. dothidea supplemented with equivalent fermentation medium were used as a control.

Malondialdehyde (MDA) content

B. dothidea mycelial plugs were cultured at 28 °C with shaking at 150 rpm. Briefly, 0.5 g of wet mycelia was collected at 0, 2, 4, 6, 8, 10, 12, 24, and 48 h and then rinsed with sterile water several times, after which it was ground in liquid nitrogen for homogenization [29]. Subsequently, 4 mL of 0.5% barbituric acid and 20% trichloroacetic acid was added to the mixture and mixed well. The suspension was centrifuged at 12,000 g for 20 min, followed by treatment in boiling water bath for 25 min and cooling in an ice bath. The absorbance at 450 nm, 532 nm, and 600 nm was measured using an ultraviolet spectrophotometer (UV-2100; UNICO, Shanghai, China), each treatment and control was replicated for three times. The MDA level was measured as follows:

$$\mathrm{MDA}\;(\mathrm{nmol}/\mathrm g)\:=\:6.45\ast(\mathrm{OD}532-\mathrm{OD}600)-0.56\ast\mathrm{OD}450$$

Determination of enzyme activity, total glucose, and pyruvate content in glycolysis metabolism pathway

B. dothidea mycelial plugs were cultured at 28 °C with shaking at 150 rpm. Briefly, 0.1 g of wet mycelia was collected at 0, 2, 4, 6, 8, 10, 12, 24, and 48 h [29], rinsed with sterile water and ground in liquid nitrogen. Then, the activities of hexokinase (HK), 6-phosphofructokinase (PFK), Ca2+ Mg2+-ATPase, and pyruvate kinase (PK) in the glycolysis pathway as well as the total glucose and total pyruvate levels were measured using commercial assay kits (Comin log, Suzhou, China, http://www.cominbio.com/index.html).

Statistical analysis

All statistical analyses were performed using the IMB SPSS statistics software (Version 20.0; IBM, Armonk, NY, United States); values of P < 0.05 were considered significant. One-way analysis of variance followed by Duncan’s post-hoc test was used to compare the means among treatments. A two-tailed Student’s t-test was used to determine significant differences between samples.

Discussion

It is essential to study the function of BCAs along with their limitations and requirements to exploit their potential in plant disease management. The mechanisms of action of BCAs vary extensively. As one of the most effective and popular BCAs, Streptomyces fermentation broth represents the main source of antibiotics or antagonistic substances that inhibit the growth of pathogenic bacteria and fungi [30,31,32,33]. By studying the appearance, cell membrane permeability, other physiological indicators of mycelia, and the impact on related enzymes in metabolic pathways, we can understand the specific mode of action of Streptomyces-induced inhibition of pathogen growth and clarify the disease resistance mechanism of the strain.

The FBS of S. spiroverticillatus HS1 exerted a significant inhibitory effect on the growth of B. dothidea mycelia. Morphologically, the mycelia treated with FBS were sparsely shrunk and broken, and the internodes were shortened, twisted, and enlarged, which is consistent with the findings of Jiang et al. [34], which showed that the mycelia of B. dothidea exhibit premature aging and wrinkling. Except for the shrinkage and deformation of mycelial morphology, the growth of mycelia was also inhibited with significant decrease in growth rate and dry weight of mycelia. The undiluted FBS of S. spiroverticillatus HS1 had the best antagonistic effect. The inhibition rates of B. dothidea growth volume were 58.42% (1 d) and 52.53% (3 d), and inhibition of growth rate were 80.72% (3 d), 67.99% (5 d), and 72.11% (7 d). The changes in mycelia morphology, mass and volume mutually verify that the growth of B. dothidea was seriously damaged, thus it can be inferred that the cell structure and metabolism of B. dothidea may also be affected. We suggested that the decrease of growth may indicate the mycelial cells may be disrupted.

By measuring the electrical conductivity of the culture medium of pathogens, we could understand the membrane permeability [35]. The FBS-treated mycelium of B. dothidea resulted in a significant increase in electrical conductivity. We suggested that the leakage of cell contents from the B. dothidea after FBS-treated, and the permeability of the B. dothidea cell membrane may be increased. Some studies proved that the antagonistic mycin N2 derived from Streptomyces sp. N2 exerted its antagonistic activity by inducing cell wall degradation and oxidative stress in Rhizoctonia solani, thus leading to fungal morphogenesis and autolysis [36]. MDA was accumulated in large amounts in the cells indicating severe lipid peroxidation of the cell membrane. The fluidity and permeability of cell membranes were altered, which ultimately led to changes in cell structure and function. The decreased ergosterol formation and increased MDA levels after treatment of F. oxysporum f. sp. niveum with S. corchorusii strain AUH-1, which is attributed to adverse effects on the structure and function of cell membranes [37]. Similarly, Woo et al. showed that an anti-pythium protein from Streptomyces sp. strain AP77 can significantly inhibit the hyphal growth of Pythium porphyrae due to alteration of the membrane permeability in P. porphyrae [38]. Therefore, we hypothesized that the cell membrane represents one of the sites of action of FBS in pathogenic mycelium characterized by disruption of the cell membrane structure and inability to maintain cell proliferation and growth, resulting in abnormal atrophy and fracture of the mycelium.

Glycolysis is the main process via which eukaryotes obtain energy. PFK and PK are often described as “key regulatory enzymes” in glycolysis since under physiological conditions, they catalyze irreversible reactions and their activities are controlled allosterically by ATP and ADP [39]. Saccharomyces cerevisiae Y-912 can significantly inhibit the growth, spore germination rate, and germ tube length of Fusarium graminearum [40]. Proteomics-based analyses have shown that few essential proteins and enzymes related to basal metabolism in the glycolytic pathway and tricarboxylic acid cycle, such as glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate mutase, enolase, fructose diphosphate aldolase, and other enzymes are downregulated. Similarly, the related proteins and genes involved in amino acid metabolism have been reported to be downregulated [40]. The glycolysis pathway involves a series of reactions that convert glucose into pyruvate and is accompanied by ATP generation. Therefore, to verify the inhibitory effect of S. spiroverticillatus HS1 on B. dothidea, it is necessary to evaluate the effect of FBS on the activity of enzymes involved in the glycolysis pathway. After treatment with the FBS, the three key enzymes of the glycolysis pathways in B. dothidea, including HK, PFK, and PK, showed significantly increased activities; the enzyme activity of the FBS group was several times that of the control group, such as that of PFK at 6 h. We speculated that the increased activity of key enzymes will accelerate the conversion of glucose. The Ca2+Mg2+-ATPase activity was also enhanced, accelerating energy consumption. The increase in activity of these key enzymes in the glycolysis pathway may seriously affect the accumulation of nutrients and energy in the mycelia. The reduction in the total glucose level and the accumulation of total pyruvate content in the cell will disrupt basal metabolism and reduce cell function, accelerating the senescence and death of pathogen cells.

Conclusions

B. dothidea treated with live S. spiroverticillatus HS1-FBS showed a clear zone of inhibition, retarded mycelial growth, and other antagonistic effects. B. dothidea showed abnormal and broken mycelia after treatment with FBS. The cell mass and growth rate of B. dothidea reduced after FBS treatment. Further, the mycelial cell membrane was damaged, the permeability increased, and lipid peroxidation was severe. In mycelial cells, the activities of key enzymes of the glycolysis pathway, including HK, PFK, PK, and Ca2+Mg2+-ATP, increased, the total glucose content decreased, and the total pyruvate content increased. Glycolysis was abnormal and accompanied by cell dysfunction.

Collectively, the inhibitory effect of S. spiroverticillatus HS1-FBS on B. dothidea was multifaceted and the different mechanisms complement each other. However, the effective antagonistic substances in FBS need to be further identified. The FBS of S. spiroverticillatus HS1 can effectively prevent the occurrence and damage of poplar canker, and has a good development and application prospects.

Availability of data and materials

No applicable.

Data availability

The authors will supply the relevant data in response to reasonable requests.

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Funding

This work was supported financially by the department of Science and Technology of Jilin Province [20230508105RC] and the department of Science and Technology of Jilin Province[20190301043NY].

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Q.L. and X.L. Writing - review & editing, Validation, Writing-original draft. H.M. and T.Z. Investigation, Writing - review, Y.Z. and T.G., Editing. J.C., Writing-review, Software and Formal analysis. L.L., Conceptualization, Writing-review & editing, Supervision, Funding acquisition.

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Liu, Q., Li, X., Mao, H. et al. The antagonistic activity of Streptomyces spiroverticillatus (No. HS1) against of poplar canker pathogen Botryosphaeria dothidea. BMC Microbiol 24, 343 (2024). https://doi.org/10.1186/s12866-024-03494-z

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