Acremonium persicae strain with hyperparasitism effect on caffeia spp. And application of acremonium persicae strain

Abstract

The invention discloses an acremonium persicae strain with a hyperparasitism effect on caffeia spp. And application of the acremonium persicae strain. The name of the hyperparasitic bacterium is Acremonium persicae HY85, the Acremonium persicae is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number of the Acremonium persicae HY85 is CGMCC No.41896. The invention further discloses a preparation method of the hyperparasitic bacterium. The acremonium persicae HY85 has an antibacterial effect on the puccinia caffei. The invention also discloses application of acremonium persicae HY85 in prevention and treatment of coffee rust.

Description

Technical Field

[0001] The invention belongs to the field of microbial applications, and particularly relates to a Acremonium persicae strain having a heavy parasitic effect on coffee rust fungus and an application thereof. Background Art

[0002] Coffee leaf rust (CLR), caused by the fungus Hemileia vastatrix, is one of the most devastating diseases in coffee plantations worldwide, significantly impacting both yield and quality (Salazar-Navarro et al., 2024). The pathogen invades through the stomata on the underside of coffee leaves. In the early stages, yellow, circular lesions appear on the upper surface of the leaf. As the disease progresses, these lesions age, forming brown patches in the center. In the later stages, the entire leaf turns yellow and eventually falls off. Infection by coffee leaf rust severely impacts coffee growth, leading to a significant drop in yield the following year (Fu Xingfei et al., 2024). Catimor 7963, the primary coffee variety cultivated in my country, has gradually lost its rust resistance due to years of cultivation.

[0003] Currently, there are multiple methods for controlling coffee rust, including physical methods, chemical methods, and planting rust-resistant varieties (Aristizábal and Johnson, 2022; Consonni et al., 2018; Sera et al., 2022). Physical methods are inefficient and labor-intensive, while chemical methods can cause coffee rust fungi to develop resistance and pollute the environment (Araaf et al., 2024; Martinho, 2020; Sera et al., 2022). Although planting disease-resistant varieties is more economical and environmentally friendly, after years of planting, new species often overcome their resistance. Currently, nine coffee rust resistance genes have been derived, using S H 1. S H 2. S H 3. S H 4. S H 5. S H 6. S H 7. S H 8. S H 9 said that due to the continuous emergence of new rust fungus species, the rust resistance gene S originally used to resist coffee rust H 1-S H 9 has been gradually conquered by newly emerged rust species (Bai Xuehui et al., 2018). As coffee rust fungi continue to evolve into new physiological species, the potential risk of coffee rust damage is increasing.

[0004] As a key biological control mechanism for plant diseases, hyperparasitism has attracted widespread attention from plant pathology researchers worldwide. Hyperparasitism is extremely common in plant rust control, playing an indispensable role in natural ecological disease control and in biocontrol strategies for plant rust (Sui Guoqiang et al., 2020). The widespread presence of hyperparasitic fungi in coffee rust provides new insights and strategies for its biological control. Summary of the Invention

[0005] The invention provides an Acremonium persicinum HY85 strain that heavily parasitizes coffee rust fungus and an application thereof.

[0006] The Acremonium persicinum of the present invention was isolated from a typical coffee leaf rust sample with heavy parasites collected from a Pu'er coffee plantation in Yunnan Province and identified as Acremonium persicinum HY85. It was deposited in the China General Microbiological Culture Collection Center (address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing) on ​​April 21, 2025, with a deposit number of CGMCC No. 41896. After culturing Acremonium persicinum HY85 on PDA medium for 14 days, the colony size was 38 mm, the colony was nearly round, and white carpet-like; the hyphae were dense and velvety ( Figure 1 A), the back is brown, gradually turning into light brown from the center to the edge ( Figure 1 B).

[0007] The Acremonium persicinum HY85 strain of the present invention has a significant inhibitory effect on Hemileia vastatrix. Therefore, the present invention provides the strain and its metabolites for biological control of coffee rust.

[0008] The present invention has the following good effects: after 96 hours, the germination rate of coffee rust uredia spores treated with the HY85 spore suspension was 5%, while the germination rate of untreated coffee rust uredia spores was 56.67%. This shows that the HY85 spore suspension has an inhibition rate of 91.18% on the germination of coffee rust uredia spores (Table 1).

[0009] After 16 days of cross-inoculation of leaf cakes of susceptible rust varieties, the test results showed that ( Figure 5), only coffee leaf cakes inoculated with coffee rust uredia developed visible chlorotic lesions. However, coffee leaves inoculated with strain HY85 alone, simultaneously with the hyperparasitic strain HY85 and the rust, inoculated with the rust 72 hours before the hyperparasitic strain HY85, or inoculated with the hyperparasitic strain HY85 72 hours before the coffee rust developed no visible chlorotic lesions following the invasion of coffee uredia. This indicates that strain HY85 is not only non-pathogenic to coffee but also inhibits the invasion of coffee leaf rust spores. Furthermore, the order of mycelial abundance of the hyperparasitic strain HY85, from strongest to weakest, is Hv0 + Hy0, Hv0 + Hy72, Hy0 + Hv72, and Hy. This indicates that mycelial growth of strain HY85 is positively correlated with the presence of coffee rust.

[0010] The qPCR detection system was used to detect the content of coffee rust in cross-inoculated leaf cakes. The test results showed that the coffee rust DNA was detected from the three leaf cakes inoculated with rust only, HV-1, HV-2, and HV-3, with average Ct values ​​of 17.733, 15.459, and 17.506, respectively, and the copy numbers were 1.41×10 8 , 7.59×10 8 , 1.66×10 8 However, no coffee rust fungus was detected in the DNA of the three leaf cakes inoculated with the heavy parasite HY85 72 hours after inoculation with the fungus. Figure 6 F). This indicates that the HY85 strain inhibits the invasion of C. coffeae.

[0011] In vitro lesion control tests showed that the yellow powdery uredia on the lesions of the control coffee rust fungus grew normally ( Figure 7 A). However, 96 hours after the coffee rust lesions were inoculated with the strain HY85, the original yellow powdery coffee rust uredia on the lesions completely disappeared, and were replaced by clearly visible white mycelium of the hyperparasite ( Figure 7 B). This indicates that strain HY85 can parasitize the uredia of the rust fungus and interfere with its growth and reproduction. Scanning electron microscopy (SEM) observations showed that the uredia on the rust lesions that were not inoculated with the heavy fungus grew and reproduced normally, and uredia with a hunchback shape and short spines on the back were observed ( Figure 7 C). However, the summer spores on the surface of coffee lesions treated with the HY85 bacterial solution were sunken and collapsed, and the spore structure was severely damaged ( Figure 7 D) It was confirmed that Acremonium persicinum has a significant inhibitory effect on coffee rust fungus and has the potential to prevent and control coffee rust. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1The colony morphology of strain HY85 on PDA medium in Example 1 (cultured for 14 days); Figure 1 Middle (A) is the positive morphology of strain HY85 PDA culture, Figure 1 Middle (B) is the morphological image of the back of PDA culture.

[0013] Figure 2 This is the conidia morphology of the strain HY85 under an optical microscope in Example 1;

[0014] Figure 3 This is a scanning electron micrograph of strain HY85 in Example 1;

[0015] Figure 4 Phylogenetic identification of strain HY85 and related strains in Example 2; Figure 4 A in the middle is the phylogenetic tree of ITS gene sequences, Figure 4 B in the middle is the phylogenetic tree of LSU gene sequences, Figure 4 Middle C is the phylogenetic tree of ITS and LSU double gene sequences.

[0016] Figure 5 Symptoms of strain HY85 in Example 4 16 days after cross-inoculation with coffee rust fungus; Figure 5 In the middle, Hv, inoculated with coffee rust spores; Hy, inoculated with HY85; Hv0+Hy0; inoculated with coffee rust and HY85 at the same time; Hv0+Hy 72 , first inoculate rust fungus, then inoculate HY85 72h later; Hy0+Hv 72 HY85 was inoculated first, and then rust fungus was inoculated 72 hours later.

[0017] Figure 6 The coffee rust qPCR detection technology system and the effect of strain HY85 on the biomass change of coffee rust in Examples 5 and 6; Figure 6 Middle, A, temperature gradient amplification curve; B, specific detection of different DNA amplification characteristics; C, concentration gradient melting curve; D, concentration gradient amplification curve; E, standard curve of H. vasitarrix qPCR quantitative detection; F, qPCR detection amplification curve of cross-inoculation H. vasitarrix.

[0018] Figure 7 The indoor control effect of the strain HY85 in Example 7 is shown; A, CK indicates no treatment; B, HY85 indicates inoculation of the strain HY85 on a pile of coffee rust spores; C, scanning electron microscopic morphology of the untreated rust fungus; D, scanning electron microscopic morphology 96 hours after inoculation with the strain HY85 bacterial solution.

[0019] Biomaterial Deposit

[0020] Name: Acremonium persicinum HY85

[0021] Classification name: Acremonium persicinum

[0022] Deposit date: April 21, 2025

[0023] Deposit unit: China General Microbiological Culture Collection Center (Address: No. 1, Beichen West Road, No. 3, Chaoyang District, Beijing)

[0024] Deposit number: CGMCC No. 41896. DETAILED DESCRIPTION

[0025] Example 1, Obtaining of Acremonium persicinum HY85

[0026] A typical coffee rust sample with heavy parasitic fungi was collected from a coffee plantation in Pu'er, Yunnan. In the laboratory, sterile forceps were used to pick up white mycelium from the rust spore pile of the diseased leaf lesion under a sterile operation table, and inoculate it on Potato Dextrose Agar (PDA) medium. The inoculated culture dish was placed in a constant temperature incubator at 28°C for culture. After 3 days of culture, the mycelium was cut from the edge of the white colony and transferred to a new PDA medium for purification culture at 28°C in a constant temperature incubator, numbered and preserved.

[0027] After 14 days of culture of strain HY85 on PDA medium, the colony size was 38 mm, the colony was nearly round and white carpet-like; the mycelium was dense and woolly, the back was brown, gradually changing to light brown from the center to the edge. Figure 1 Under an optical microscope (10x40), the spores were oval-shaped, about (1.8-2.6 μm) x (2.6-3.8 μm) in size. Figure 2 Under a scanning electron microscope (x3000), the spore surface was smooth and long oval-shaped. Figure 3

[0028] Example 2, Molecular identification of Acremonium persicinum HY85

[0029] ​DNA from the purified strain HY85 obtained in Example 1 was extracted from the hyperparasitic mycelium using a fungal DNA extraction kit (OMEGA, Beijing, China) according to the manufacturer's instructions and stored at -20°C. PCR amplification was performed using universal fungal primers ITS1 / ITS4 (White et al., 1990) and LROR (Rehner and Samuels, 1994) / LR5 (Moriya et al., 2005). The PCR reaction system (25 μL) consisted of 2.5 μL of 10x Ex Taq Buffer, 2 μL of dNTP Mix, 1 μL each of forward and reverse primers, 0.2 μL of Ex Taq enzyme (2 U), 17.3 μL of ddH₂O, and 1 μL of DNA template. The PCR amplification protocol was as follows: 94°C pre-denaturation for 4 min, denaturation at 94°C for 30 s, annealing at 54°C for 30 s, extension at 72°C for 1 min, 35 cycles of this, and 10 min of extension at 72°C, followed by storage at 4°C. PCR amplification products were ligated using the pMDTM18-T Vector Cloning Kit and transformed into competent Escherichia coli Trans 5a cells for cloning. Positive clones were identified and sent to Beijing Liuhe BGI Genomics Co., Ltd. for sequencing. Sequence fragments obtained were compared for homology with the NCBI database (https: / / www.ncbi.nlm.nih.gov / ) to preliminarily identify their species. Sequences of homologous strains and foreign strains were downloaded, and a phylogenetic tree was constructed using the neighbor-joining method (1000 bootstraps) using MEGA11.0 software.

[0030] Strain HY85 DNA was amplified using universal primers for the ITS and LSU genes, and sequencing yielded 583 and 919 bp nucleotide fragments, respectively. The ITS sequence is shown as Sequence 1 in the sequence listing, and the LSU sequence is shown as Sequence 2 in the sequence listing. A homology search of the NCBI database revealed that the ITS and LSU sequence fragments shared 97.75% and 99.33% similarity with Acremonium persicinum CBS:378.70A, respectively. Cluster phylogenetic trees revealed that strain HY85 and Acremonium persicinum CBS:378.70A resided on the same evolutionary branch, whether in the ITS and LSU single-gene phylogenetic tree or the ITS and LSU double-gene phylogenetic tree. Figure 4 ). Combining morphological and molecular characteristics, strain HY85 was identified as Acremonium persicinum.

[0031] The strain, named Acremonium persicinum HY85, was deposited in the China General Microbiological Culture Collection Center (address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing), with a deposit date of April 21, 2025, and a deposit number of CGMCC No. 41896.

[0032] Example 3: Effect of inhibiting uredospore germination of Hemileia vastatrix

[0033] Fresh uredia spores of C. coffeei were collected from coffee rust-susceptible varieties at the Ruili coffee base in Yunnan and placed in waterproof capsules. Fresh uredia spores of C. coffeei and spores of strain HY85 were prepared in the laboratory with sterile water to a concentration of 1 mg / mL and 1×10 6 Spore suspension (2 × 2 × 0.3 cm) was placed on a glass slide to inhibit uredia germination. The following steps were performed: ① 20 μL of the uredia spore suspension and 20 μL of sterile water were vortexed and spread onto the water agar block. ② 20 μL of the HY85 spore suspension and 20 μL of the uredia spore suspension were vortexed and spread onto the water agar block. Each treatment was repeated three times. The glass slide was placed in a Petri dish and incubated at 26-28°C for 96 h. The number of germinating spores was recorded under an optical microscope (NiKon, NI / E, Japan). The inhibition rate was calculated according to the method (Mudyiwa et al., 2017). The criterion for judging uredia germination was that the germ tube length was greater than half the spore diameter. The experiment on the inhibition of uredia spore germination of coffee rust showed that the germination rate of coffee rust uredia spores treated with HY85 spore suspension was 5% after 96 hours, while the germination rate of untreated coffee rust uredia spores was 56.67%. This shows that the inhibition rate of HY85 spore suspension on coffee rust uredia spore germination was 91.18% (Table 1).

[0034] Table 1. Inhibitory effect of HY85 spore suspension on spore germination of coffee rust

[0035]

[0036]

[0037] Example 4: Inhibitory effect of strain HY85 on the infection process of coffee rust (Hemileia vastatrix)

[0038] Three PDA cakes (5 mm) of the purified strain HY85 obtained in Example 1 were inoculated into 150 mL of liquid culture medium (10 g glucose, 3 g yeast powder, 1000 mL purified water, sterilized at 121°C for 20 min) and cultured at 28°C with shaking until the spore concentration was about 1 × 10 6 / mL. Fresh summer spores of coffee rust fungus were collected and prepared into a 1mg / mL spore suspension with sterile water. Healthy susceptible coffee leaves of Matari coffee variety were collected, leaf cakes were made with a 2cm diameter puncher, and then washed with sterile water. The cross-inoculation experiment was divided into 5 groups: Hv, inoculated with 25μL of coffee rust fungus spore suspension; Hy, inoculated with 25μL of strain HY85 spore suspension; Hv0+Hy0, inoculated with coffee rust fungus spore suspension and HY85 spore suspension at the same time, 25μL each; Hv0+Hy 72 , first inoculate 25 μL of coffee rust spore suspension, and then inoculate 25 μL of HY85 spore suspension 72 h later; Hy0+Hv 72 Plants were inoculated with 25 μL of a spore suspension of HY85 and then, 72 hours later, with 25 μL of a spore suspension of C. coffeae. Each experiment was repeated five times. After inoculation, the plants were incubated in a 24°C artificial climate chamber (MMM Climacell 707, Germany) at 100% humidity and darkness for 24 hours. After 24 hours, the cycle was adjusted to 12 hours of light / 12 hours of darkness (Silva et al., 2012). After 16 days of incubation, the disease progression of each treatment was observed.

[0039] The cross-inoculation leaf cake test showed that only the coffee leaf cake inoculated with coffee rust uredus spores had visible chlorotic spots ( Figure 5 A), while the susceptible coffee leaves inoculated with strain HY85 alone, inoculated with both the hyperparasitic strain HY85 and rust of C. coffeae, inoculated with rust of C. coffeae 72 hours after inoculation with the hyperparasitic strain HY85, and inoculated with the hyperparasitic strain HY85 72 hours after inoculation with rust of C. coffeae did not develop chlorotic spots after invasion by coffee uredia spores ( Figure 5 B~E). This shows that strain HY85 is not only non-pathogenic to host coffee, but also has an inhibitory effect on the invasion of spores of coffee leaf rust pathogen C. coffeae. On the other hand, from the perspective of the aerial mycelium of the heavy parasite attached to the surface of coffee leaves, the order of mycelial luxuriance from strong to weak is Hv0+Hy0, Hv0+Hy 72 、Hy0+Hv 72 , Hy. This shows that the mycelial growth of strain HY85 is positively correlated with the presence or absence of coffee rust ( Figure 5 ).

[0040] Example 5: Establishment of a qPCR detection system for C. coffeae

[0041] A set of primers and probes specific for the ITS2 sequence of C. coffeae were designed online (https: / / www.genscript.com / tools / real-time-pcr-taqman-primer-design-tool). These primers and probes were synthesized by Liuhe BGI, with a FAM group added to the 5' end and a BHQE group added to the 3' end of the probe.

[0042] DNA from fresh spores of the coffee rust fungus HVB1 was extracted using a fungal DNA extraction kit (OMEGA, Beijing, China) according to the manufacturer's instructions. The DNA was amplified using primers ITS1 (5'-TCCGTAGGTGAACCTGCGG-3') (White et al., 1990) / HV-ITS2-R (5'-CAAAATATGTCATACCTCTCATTCT-3') (Han et al., 2017) and transformed into Escherichia coli Trans 5a clones. Positive clones harboring the ITS2 sequence of the HVB1 strain were verified by sequencing and extracted using a plasmid extraction kit (OMEGA, Beijing, China). Primers and probes were screened for optimal temperature, tested for specificity and sensitivity, and a standard curve was established and optimized using the recombinant plasmid of the coffee rust fungus HVB1 strain as a template.

[0043] Screening of optimal temperature for primers and probes: The reaction system was 2xRealStar Fast Probe Mix 10 μL; upstream and downstream primers (10 μmoL·L -1 )I 0.5 μL each; probe (100 μmol·L -1 ) 0.5μL; High / Low Rox Reference Dye 0.4μL; DNA template 0.5μL; ddH2O to 20.0μL. The reaction procedure was: 95°C pre-denaturation for 30 seconds; 95°C denaturation for 15 seconds; 40 cycles of 8 temperature gradients between 48.3 and 68.7°C; annealing for 30 seconds; and extension at 72°C for 30 seconds. A positive result was determined when the amplification curve Ct value (the number of cycles required for the fluorescence signal of the amplified product to reach the set threshold) was <30 and the melting curve showed a single peak. The optimal temperature was screened and used in the subsequent qPCR quantitative detection system.

[0044] Specificity detection: qPCR detection was performed using DNA of C. coffeae strains HvL72, HvL86, HvL95, HvL119, C. coffeae hyperparasites HY72, HHV, HY48, HY22, HY83, and other strains CPE5, Zao 1, 21BS01-4, BEC419, and BEC366 as templates, recombinant plasmid DNA HVB1 of C. coffeae as a positive control, and ddH2O as a negative control using the optimal reaction system and conditions to verify the specificity of the probe.

[0045] Sensitivity detection and establishment of standard curve: The initial concentration of coffee rust recombinant plasmid DNA HVB1 was determined by spectrophotometer to be 74 ng / μL, and it was diluted in 10-fold concentration gradient to obtain 10 -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 - 7 qPCR amplification was performed with a series of template concentrations, such as ng / μL, and a standard curve was established based on the relationship between Ct value and template concentration.

[0046] A set of primers and probes (primers HVL F1 / R1: probe HVLT1) were designed from the ITS2 region. Through eight gradient annealing temperature comparison tests, the optimal annealing temperature of the HVL F1 / R1 primers was screened to be 54°C ( Figure 6 A). At this temperature, the primer combination and probe can specifically detect C. coffeae rust, while other non-rust fungal DNA templates cannot detect ( Figure 6 B) Serial dilution 10 -1 ~10 -7 Melting curve ( Figure 6 C) are all single peaks, the amplification curves are smooth and orderly, and the amplification efficiency is good, which shows that qPCR detection is highly sensitive to the rust fungus HVB1 plasmid. -1 ~10 -7 Amplification curve ( Figure 6 D), the obtained Ct value was used as the vertical axis and the logarithmic value of each gradient template DNA copy number was used as the horizontal axis to draw the standard curve for the quantitative detection of coffee rust ( Figure 6 E), the linear equation is y = -3.1061x + 43.041; R 2 =0.992, the amplification efficiency (E) was 109.9%, meeting the amplification requirements.

[0047] Example 6: Effect of strain HY85 on biomass changes of coffee rust fungus

[0048] Example 5 The coffee rust qPCR detection system was obtained. 16 days after cross-inoculation, the Hv group, Hy0+Hv 72 Three leaf cakes were randomly selected from the group to extract DNA, and the content of coffee rust in the leaf cakes was quantitatively detected using the qPCR quantitative detection method. The test results showed that the coffee rust DNA was detected in the three leaf cakes HV-1, HV-2, and HV-3 that were only inoculated with rust, with average Ct values ​​of 17.733, 15.459, and 17.506, respectively, and the copy number was 1.41×10 8 , 7.59×10 8 , 1.66×10 8 However, the coffee rust fungus was not detected in the DNA of the three leaf cakes inoculated with the heavy parasite 72 hours before the rust fungus. Figure 6 F).

[0049] Example 7: Growth inhibition of naturally occurring coffee rust by strain HY85

[0050] During the coffee rust season, leaves with coffee rust lesions of severity 5-6 were selected from coffee plantations in Dehong and Ruili, Yunnan Province [Ministry of Agriculture and Rural Affairs. (2016). Standards for the Description and Evaluation of Tropical Crop Germplasm Resources. Institute of Spice and Beverages, Chinese Academy of Tropical Agricultural Sciences, Dehong Institute of Tropical Agricultural Sciences, Yunnan Province]. Leaf cakes with lesions of relatively uniform size were punched using a 2 cm hole punch and subjected to an antibacterial test against hyperparasitic fungi. Each lesion on the leaf cake was inoculated with 25 μL of strain HY85. An equal number of lesions were not inoculated with strain HY85 as a control. Each experiment was repeated 10 times. After inoculation with HY85, the leaves were incubated in an artificial climate chamber (MMM Climacell 707, Germany) at 100% humidity and 24°C under 12 h light / 12 h dark conditions for 96 h. The experimental results were analyzed, and the inhibition of strain HY85 against rust fungi was observed using a scanning electron microscope.

[0051] In vitro lesion control tests showed that the yellow powdery uredia on the control coffee rust lesions grew normally ( Figure 7 A). However, 96 hours after the coffee rust lesions were inoculated with the strain HY85, the original yellow powdery uredus of the coffee rust fungus on the lesions completely disappeared, and were replaced by clearly visible white mycelium of the heavy parasite ( Figure 7 B). This indicates that HY85 can parasitize the uredia of the rust fungus and interfere with its growth and reproduction. Scanning electron microscopy (SEM) observations showed that the uredia on the lesions that were not inoculated with the heavy fungus grew and reproduced normally, and uredia with a hunchback shape and short spines on the back were observed ( Figure 7C). After the strain HY85 was parasitized, the summer spores at the coffee lesions became sunken and collapsed, and the spore structure was severely damaged ( Figure 7 D) Therefore, strain HY85 has a good effect in controlling coffee leaf rust caused by Puccinia camelus.

[0052] The above-described embodiments merely illustrate several implementations of the present invention. While the descriptions are relatively specific, they are intended to be illustrative rather than restrictive of the present invention. Those skilled in the art will readily appreciate that numerous modifications, variations, and improvements can be made without departing from the spirit and scope of the appended claims, all of which fall within the scope of protection of the present invention.

Claims

1. A strain of Acremonium persicae, characterized in that The classification name of the strain is Acremonium persicinum, and the name is HY85. The strain has been deposited in the General Microbiology Center of the China Culture Collection Administration of Microorganisms, and the deposit number is CGMCC No.41896.

2. Use of the Acremonium persicinum HY85 according to claim 1 in the preparation of a biocontrol agent, microbial fertilizer or biopesticide for inhibiting coffee rust.

3. Use of Acremonium persicinum HY85 according to claim 1 in inhibiting coffee rust.

4. A biocontrol agent for inhibiting coffee rust, wherein the active ingredient is the Acremonium persicinum HY85 described in claim 1.

5. A microbial fertilizer for inhibiting coffee rust, wherein the active ingredient comprises the Acremonium persicinum HY85 according to claim 1.

6. A biopesticide for inhibiting coffee rust, wherein the active ingredient comprises the Acremonium persicinum HY85 according to claim 1.

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