A kind of pine-loving tularensis CA5 and its application

Abstract

The present invention discloses Talaromyces pinophilus CA5 and its application, which belongs to the technical field of screening antagonistic strains of tea root rot pathogens. Talaromyces pinophilus CA5 was deposited in the China Center for Type Culture Collection on September 26, 2023, with a deposit number of CCTCC NO: M 20231797, and was classified and named Talaromyces pinophilus CA5. The fermentation stock solution, bacterial suspension and non-volatile metabolites of Talaromyces pinophilus CA5 can effectively inhibit the growth of tea root rot pathogens, and provide microbial resources and technical support for the prevention and treatment of tea root rot. In addition, the strain has a broad-spectrum antibacterial effect on 8 plant pathogens such as eggplant brown streak disease, potato early blight, wild rice root rot, corn leaf spot disease, citrus sand bark disease, pepper wilt, Polygonatum anthracnose, wild rice root rot, and strawberry blight, and has a wide range of application prospects.

Description

Technical Field

[0001] The invention relates to the technical field of screening antagonistic strains of tea tree root rot pathogens, and more specifically to a pine-loving Talaromyces CA5 and an application thereof. Background Art

[0002] Tea root rot is a soil-borne disease. The pathogen infects the roots of tea trees, causing the leaves to wither and fall off, and the roots to rot. The disease is very hidden and difficult to detect in the early stage of the disease. Later, the disease gradually develops. The aboveground part of the tea tree grows sparsely, the leaves turn yellow, and the buds are few and thin. In severe cases, the whole plant dies. The damage is irreversible, causing huge economic losses to tea production. In previous studies, a pathogen Fusarium cugenangense that causes tea root rot was reported, which belongs to a specific strain of the Fusarium oxysporum species complex (FOSC). It is well known that the Fusarium oxysporum species complex is a soil-borne pathogen with strong pathogenicity. It has a wide host distribution range and can cause hundreds of plants to be infected with root diseases, such as passion fruit stem base rot, wolfberry root rot, northern adenophora root rot, pepper root rot, etc. It can infect a wide range of important agricultural and horticultural plant hosts, causing crown or root rot, vascular wilt, etc., which has a huge negative impact on cash crop production. The pathogen mainly parasitizes in the rhizosphere soil of the plants, accumulates in the soil for a long time, and is highly infectious.

[0003] At present, the main methods for the prevention and control of tea tree root rot are agricultural, chemical and biological control, with chemical control being the main method. With the proposal of the zero growth plan for the use of chemical fertilizers and pesticides, finding safe and pollution-free disease prevention and control methods has become the main problem faced in agricultural production. Among them, biological control has become one of the current research hotspots with its green, pollution-free and environmentally friendly advantages. Biological control has the advantages of being environmentally friendly, protecting soil structure and not causing pathogens to develop drug resistance, which is in line with the national green and sustainable development strategy. Strains screened from the environment, including Bacillus, Pseudomonas, and Trichoderma, have been widely used in biological control. As a component of the plant microbial system, biocontrol fungi can promote plant growth through a variety of mechanisms, can minimize the ecological threats caused by chemical inputs in soil and crops, and have good prospects. There have been many reports on the use of biocontrol fungi in the prevention and control of crop root rot. For example, Zhang Fumei et al. isolated a strain of A. sphaerospora from blueberry tissue, which has a good control effect on blueberry root rot; Li Mengwei et al. isolated an endophytic fungus, Penicilium oxalicum, from the lateral roots of healthy Zanthoxylum bungeanum, which has a good control effect on Zanthoxylum bungeanum root rot; Han Zhongming et al. screened out a strain of Acremonium persicinum from the rhizosphere soil of Saposhnikovia divaricata, which has a good control effect on Saposhnikovia divaricata root rot; Yao Chenhui et al. screened out 11 fungi from the rhizosphere soil of tobacco root rot that have a good control effect on tobacco root rot; Shen Guanghui et al. screened out a Penicillium fungus and an Aspergillus terreus from the rhizosphere soil of strawberry, which have a high potential application value in the prevention and control of strawberry root rot.

[0004] Isolating and obtaining biocontrol bacteria with high inhibitory effects on pathogens is a necessary prerequisite for carrying out biocontrol work on tea root rot. Therefore, screening more biocontrol microorganisms that are highly effective in preventing and controlling tea root rot and providing biocontrol fungal resources with good prevention effects for tea root rot are technical problems that technicians in this field need to solve urgently. Summary of the invention

[0005] The present invention uses Fusarium cugenangense, the pathogen of tea tree root rot, as the target fungus, isolates and screens antagonistic fungi from the rhizosphere soil of tea tree root rot, and conducts morphological, physiological and biochemical characteristics and molecular biological identification on strains with better antagonistic effects, and conducts biological property determination, disease prevention and growth promotion index determination, antibacterial spectrum determination and antibacterial activity determination of different components of fermentation products, in order to provide new strain resources and technical support for the biological control of tea tree root rot.

[0006] One of the objectives of the present invention is to provide a Talaromyces pinophilus CA5, which was deposited at the China Center for Type Culture Collection on September 26, 2023, with the deposit number CCTCC NO: M 20231797, and is taxonomically named Talaromyces pinophilus CA5.

[0007] Another objective of the present invention is to provide the applications of the fermentation stock solution, bacterial suspension, and non-volatile metabolites of the Talaromyces pinophilus CA5 in inhibiting tea tree root rot pathogens.

[0008] Another objective of the present invention is to provide the applications of the Talaromyces pinophilus CA5 in inhibiting eight other plant pathogens to prove that the Talaromyces pinophilus CA5 has a broad-spectrum antibacterial effect. The eight other plant pathogens include the pathogens of eggplant brown spot, potato early blight, water bamboo root rot, maize northern leaf blight, citrus melanose, pepper fusarium wilt, polygonatum anthracnose, and strawberry phytophthora blight.

[0009] Another objective of the present invention is to provide the applications of the Talaromyces pinophilus CA5 in promoting plant growth.

[0010] In the above applications, the Talaromyces pinophilus CA5 has the abilities of decomposing organic phosphorus and inorganic phosphorus, potassium solubilization, siderophore production, nitrogen fixation, ammonia production, and IAA production.

[0011] Through the above technical solutions, the present invention uses the tea tree root rot pathogen as the target bacterium, isolates and screens antagonistic fungi from the rhizosphere soil of tea tree root rot, identifies the strains with better antagonistic effects by morphological, physiological and biochemical characteristics and molecular biology, and measures their biological characteristics, disease prevention and growth promotion indexes, antibacterial spectrum, and antibacterial activities of different components of the fermentation products, in order to provide new strain resources and technical support for the biological control of tea tree root rot. Description of the Drawings

[0012] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art. Obviously, the drawings in the following description are only the embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on the provided drawings.

[0013] Figure 1 The drawing shows the plate inhibition effect of strain CA5 on tea tree root rot pathogens; A. treatment group B. control;

[0014] Figure 2The attached figure shows the effect of strain CA5 on the mycelial growth of tea root rot pathogens; A. control B: treatment;

[0015] Figure 3 The attached figure shows the colony and microscopic characteristics of strain CA5; A, B colony front and back morphology C. hyphae and spore morphology;

[0016] Figure 4 The attached figure is a phylogenetic tree of strain CA5 constructed based on ITS sequences;

[0017] Figure 5 The attached figure shows the study of biological characteristics of strain CA5; A. different carbon sources; B. different nitrogen sources; C. different carbon-nitrogen ratios; D. different metal ions; E. different NaCl concentrations; F. different pH values;

[0018] Figure 6 The attached figure is a growth-promoting ability test diagram of strain CA5; A. Cellulase; B. β-1,3 glucanase; C. Ability to decompose inorganic phosphorus; D. Ability to decompose organic phosphorus; E. Ability to decompose potassium; F. Ability to produce siderophores; G. Ability to fix nitrogen; H. Ability to produce ammonia; I. Ability to produce IAA;

[0019] Figure 7 The attached figures show the antibacterial activity of volatile and less volatile metabolites of strain CA5; A. volatile metabolites (left: treatment, right: control); B. less volatile metabolites (left: treatment, right: control). DETAILED DESCRIPTION

[0020] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0021] Test materials in the embodiments:

[0022] The pathogen of tea tree root rot, Fusarium cugenangense, is a specific strain belonging to the Fusarium oxysporum species complex (FOSC) and was isolated and preserved in our laboratory.

[0023] The soil was collected from the rhizosphere soil of tea trees infected with root rot bacteria at the Chang'an Teaching Base of Tea Science of Hunan Agricultural University.

[0024] Example 1 Isolation and screening of antagonistic strains

[0025] According to the method of Wei Qi et al., 1 g of soil was added to 9 mL of sterile water. After oscillating for several minutes, the sterile water was diluted to 10 -1 ~10 -7 . Take 100 μL of each gradient and spread it on the PDA medium plate. Repeat each dilution gradient 3 times and invert it in a 25℃ constant temperature box for 7 days. After the colony grows on the plate, pick a single colony for purification and preservation. The isolated antagonistic bacteria were screened by the plate confrontation method. The tea tree root rot pathogen was used as the target bacteria, and the isolated fungi were screened by the plate confrontation method. Use a puncher to punch holes on the edges of the purified rhizosphere fungi and pathogens to obtain a 5mm diameter cake. The pathogen cake was inoculated into the center of a PDA culture dish with a diameter of 75mm, and the rhizosphere fungus cake was inoculated at two symmetrical points 20mm away from the pathogen cake. The pathogen was inoculated as a control, and repeated 3 times. It was placed in an inverted culture at 25℃ in the dark for 7 days. The fungi with antagonistic effects were selected for measurement and photography, and the inhibition rate of each strain was calculated.

[0026] Inhibition rate (%) = [(control colony diameter - treatment colony diameter) / control colony diameter] × 100% The results showed that a total of 38 fungal strains were isolated by the dilution coating method. The isolated fungi were subjected to a plate confrontation test with the tea root rot pathogens, and 5 antagonistic fungi with antibacterial effects on the tea root rot pathogens were screened out. Finally, one antagonistic fungus CA5 with obvious and stable inhibitory effect on the tea root rot pathogens was screened out (Table 1, Figure 1 ), the diameter of the pathogen colony after strain CA5 antagonized tea root rot pathogens was 15.81 mm, and the inhibition rate reached 77.39%, indicating that the antagonistic bacteria CA5 had a good antagonistic effect on tea root rot pathogens. Therefore, strain CA5 was selected for subsequent research.

[0027] Example 2 Antagonistic effect of strain CA5 on pathogenic bacteria

[0028] Place a glass slide in a culture dish and pour a thin layer of PDA plate. Inoculate pathogenic bacteria cake on one side and antagonistic bacteria cake on the other side. Use no antagonistic bacteria as the control. Set up 3 replicates for each treatment. After 7 days of culture, observe the morphological characteristics of pathogenic hyphae under an optical microscope.

[0029] The results showed that the mycelium morphology of the pathogen was observed under an optical microscope. The mycelium of the control group was smooth, uniform in thickness, slender, without swelling or breakage ( Figure 2 A); however, the mycelium of the tea tree root rot pathogen inhibited by strain CA5 grew abnormally, with fractures, twisting, and swelling ( Figure 2 B); This indicates that strain CA5 can achieve an antibacterial effect by affecting the hyphae growth of tea tree root rot pathogens.

[0030] Table 1 Inhibitory effect of strain CA5 on mycelium of tea root rot pathogens

[0031]

[0032] Note: Data are expressed as mean ± standard deviation. Different lowercase letters after the data in the same column indicate significant differences (P < 0.05).

[0033] Example 3 Identification of strain CA5

[0034] Morphological identification The strain was cultured in PDA at 25℃ in the dark for 7 days. The colony diameter was 46.80mm. The colony was a regular circle with neat edges and underdeveloped aerial hyphae. The front of the colony was pinkish white and the back of the colony produced pink pigment. Under the microscope, the spores were observed to be small oval particles connected in chains or clusters. According to the above colony culture growth morphological characteristics, it was preliminarily identified as Talaromyces pinophilus ( Figure 3 ).

[0035] Physiological and biochemical characteristics The results of physiological and biochemical tests showed that strain CA5 was positive for methyl red test MR, gelatin liquefaction test, hydrogen peroxidase test, and arginine decarboxylase test, and was positive for acetyl methyl test V.Ptest and hydrogen sulfide production H. 2 S production was negative; the strain was able to degrade glucose, lactose, maltose, fructose, sucrose, and mannitol (Table 2).

[0036] The ITS sequence length of strain CA5 was 530 bp. The sequence was uploaded to the GenBank database for homology comparison, and it was found that the similarity between strain CA5 and Talaromyces pinophilus was 98%. The ITS sequences of strains with higher similarity were selected to construct a phylogenetic tree ( Figure 4 ). Combining the morphological characteristics and molecular biological identification results, the strain CA5 was finally determined to be Talaromyces pinophilus. The Talaromyces pinophilus CA5 was deposited in the China Center for Type Culture Collection on September 26, 2023, with a deposit number of CCTCC NO: M 20231797 and a classification name of Talaromyces pinophilus CA5.

[0037] Study on the biological characteristics of strain CA5

[0038] Effects of different nutrients on the mycelial growth of strain CA5: The mycelial growth rates of strain CA5 in the medium containing fructose, sucrose, maltose, lactose and glucose were 5.24mm / d, 5.05mm / d, 5.64mm / d, 3.59mm / d and 4.75mm / d, respectively. Compared with the five carbon sources, its growth rate on the medium containing maltose was significantly higher than that on the medium containing fructose, sucrose, lactose and glucose; the mycelial growth rates of strain CA5 in the medium containing potassium nitrate, yeast extract, ammonium sulfate, peptone and sodium nitrate were significantly lower than those in the medium containing potassium nitrate, yeast extract, ammonium sulfate, peptone and sodium nitrate. The growth rates of the strain in the medium containing peptone were 4.18mm / d, 4.32mm / d, 3.79mm / d, 4.75mm / d and 4.38mm / d, respectively. Compared with the five nitrogen sources, the growth rate of the strain in the medium containing peptone was significantly higher than that in the medium containing other nitrogen sources. The growth rates of the strain in the basic medium with carbon-nitrogen ratios of 20:1, 40:1, 60:1 and 80:1 were 5.50mm / d, 5.07mm / d, 5.28mm / d and 4.74mm / d, respectively. The first three had significant differences compared with the carbon-nitrogen ratio of 80 / 1. + , Mn 2+ , K + , Fe 3+ and Mg 2+ The growth rates of the strains in the medium containing Mn were 4.36 mm / d, 5.33 mm / d, 4.38 mm / d, 4.17 mm / d and 4.75 mm / d, respectively. 2+ The growth rate of the culture medium was significantly higher than that of the other four metal ion mediums ( Figure 5 AD).

[0039] Effects of different NaCl concentrations on mycelial growth of strain CA5: NaCl had a significant effect on mycelial growth of strain CA5. When the NaCl concentration was 1%, mycelial growth was fastest, with a growth rate of 11.62 mm / d. As the salinity increased, mycelial growth was inhibited. When the concentration was 9%, the growth rate was 2.74 mm / d. When the concentration was 10%, the colony did not grow at all ( Figure 5 E).

[0040] Effect of different pH on mycelial growth of strain CA5: pH had a significant effect on mycelial growth of strain CA5. Its growth rate was 12.28-13.5 mm / d at pH 5.0-12, and the optimum pH was 6.0, with a growth rate of 13.5 mm / d ( Figure 5 F).

[0041] Table 2 Physiological and biochemical characteristics of strain CA5

[0042] Measurement items result Measurement items result Methyl red test + glucose + Acetyl methyl test - lactose + Gelatin liquefaction + maltose + Catalase + fructose + Arginine decarboxylase + sucrose + Hydrogen sulfide production - Mannitol +

[0043] Note: +: positive; -: negative.

[0044] Example 4 Determination of growth-promoting indexes of strain CA5

[0045] Strain CA5 produced transparent circles on the cellulase detection plate and β-1,3-glucanase detection plate, indicating that the strain has the ability to secrete cellulase and β-1,3-glucanase, while there were no transparent circles on the chitinase, amylase and protease detection plates, indicating that the chitinase, amylase and protease produced were insufficient in activity ( Figure 6 A, B); the strain produced transparent circles when grown on the organic phosphorus and inorganic phosphorus detection plates and the potassium solubilization detection plates, indicating that the strain has the ability to decompose organic phosphorus and inorganic phosphorus and the ability to solubilize potassium ( Figure 6 CE); the strain produced a red-purple halo when grown on the CAS double-layer medium, indicating that the strain has the ability to produce siderophores ( Figure 6 F); strain CA5 can still grow after 5 generations of subculture on nitrogen-free medium, indicating that the strain has the ability to fix nitrogen ( Figure 6 G); Taking the uninoculated culture medium as the control, Nessler's reagent was added to the control and the bacterial supernatant, and the supernatant turned orange-yellow, indicating that strain CA5 has the ability to produce ammonia ( Figure 6 H); using IAA standard solution as positive control, sterile water as blank control, and no tryptophan as negative control, after Salkowski colorimetric test, the color of the treated liquid turned pink, indicating that the strain has the ability to produce IAA ( Figure 6 I, Table 3).

[0046] Table 3 Growth-promoting ability test of strain CA5

[0047] Growth-promoting ability test result Growth-promoting ability test result Cellulase + Ability to decompose organic phosphorus + β-1,3-Glucanase + Potassium-dissolving capacity + Amylase - Siderophore production capacity + Protease - Nitrogen fixation capacity + Chitinase - Ammonia production capacity + Ability to decompose inorganic phosphorus + IAA production capacity +

[0048] Note: +: positive; -: negative.

[0049] Example 5 Determination of the antibacterial spectrum of strain CA5

[0050] The antibacterial spectrum of the antagonistic strain CA5 was determined by the plate confrontation method. A 5-mm diameter cake of each test pathogen was taken with a hole puncher and placed in the center of the culture dish. Antagonistic fungal cakes with a diameter of 5 mm were inoculated at two symmetrical points 20 mm away from the pathogen cake. Only the pathogens were inoculated as the control. Each treatment was repeated 3 times. After culturing in a constant temperature box at 25°C for 7 days, the diameter of the inhibition zone was measured and the inhibition rate was calculated.

[0051] The results of the antimicrobial spectrum determination test of antagonistic strains showed that strain CA5 had obvious inhibitory effects on eight plant pathogens, including eggplant brown streak disease pathogen P.vex ans, potato early blight pathogen A.alternate, wild rice root rot pathogen F.oxysporum, corn large blight pathogen E.turcicum, citrus sand bark disease pathogen D.citri, pepper wilt pathogen F.oxysporum, polygonatum anthracnose pathogen C.circinans, wild rice root rot pathogen F.oxysporum, and strawberry blight pathogen P.fragariae, with inhibition rates of 80.13%, 76.99%, 68.80%, 74.09%, 74.09%, 77.56%, 73.42%, and 86.85%, respectively, indicating that the antagonistic strain has good antagonistic effect and broad-spectrum antibacterial activity (Table 4).

[0052] Table 4 Inhibitory effect of strain CA5 on 8 plant pathogens

[0053] Pathogens Colony diameter Antibacterial rate Eggplant brown streak pathogen 1.14±015c 80.13±0.21b Potato early blight pathogen 1.79±0.09ab 76.99±1.40b Zizania root rot pathogen 1.61±0.09bc 68.80±1.29d Corn leaf blight pathogen 2.00±0.11a 74.09±1.69c Citrus scurf disease 1.98±0.07a 74.09±1.04c Fusarium wilt 1.68±0.03b 77.56±0.54b Polygonatum anthracnose pathogen 1.62±0.08b 73.42±1.15c Strawberry blight pathogen 1.36±0.04c 86.85±0.65a

[0054] Note: Data are expressed as mean ± standard deviation. Different lowercase letters after the data in the same column indicate significant differences (P < 0.05).

[0055] Example 6 Antibacterial activity of volatile and non-volatile metabolites of strain CA5

[0056] The antibacterial activity of volatile metabolites was determined by the double-button culture method: a 5-mm-diameter cake of antagonistic bacteria was taken with a cork puncher and inoculated in the center of a PDA plate, and a cake of the same size of the pathogen of tea root rot was taken and inoculated in the center of another PDA plate. The two plates with the inoculated bacteria were placed in a double-button culture at 25°C in the dark for 7 days; a pure PDA plate without pathogen inoculation was placed on the PDA plate inoculated with pathogens as a control, with three replicates for each treatment, the growth of pathogen hyphae was observed, the colony diameter of the pathogen was measured by the cross method, and the inhibition rate was calculated.

[0057] The antibacterial activity of the non-volatile metabolites was determined by the disc filter method: a sterilized double-layer glass paper was laid flat in the center of the poured PDA, and a 5 mm diameter antagonistic bacterial cake was taken with a hole puncher and inserted into the center of the filter membrane. After culturing at 25°C in the dark for 5 days, the filter membrane and the antagonistic bacterial hyphae were gently removed with tweezers, and a 5 mm diameter pathogenic bacterial block was inoculated in the original position, and cultured at 25°C in the dark for 5 days. Only the pathogenic bacterial block was inoculated on a new PDA plate as a control. Three replicates were set for each treatment to observe the colony growth, and the colony diameter of the pathogen was measured by the cross method to calculate the antibacterial rate.

[0058] The volatile metabolites and non-volatile metabolites of strain CA5 showed different degrees of inhibitory effects on tea root rot pathogens ( Figure 7), among which the antibacterial activity of volatile metabolites against tea root rot pathogens was 14.90%; the antibacterial activity of non-volatile metabolites against tea root rot pathogens was 44.33%.

[0059] Example 7 Antibacterial ability of different components of the fermentation product of strain CA5

[0060] Referring to the method of Zhang Yongzhi et al. and making some improvements, ten bacterial cakes were put into 200 ml of PDB medium and cultured at 180 r / min and 28°C for 7 days to obtain the fermentation liquid. The fermentation liquid was centrifuged at 12000 r / min and filtered with a 0.22 um sterile microporous filter membrane to obtain the sterile fermentation filtrate. The precipitate after centrifugation was washed three times with sterile water, and then the washed precipitate was mixed with an equal amount of sterile water to obtain a bacterial suspension. The fermentation liquid and bacterial suspension were prepared by the filter paper method, and the sterile fermentation filtrate was prepared by the mixed plate method. The inhibitory effect of the fermentation liquid, bacterial suspension and sterile fermentation filtrate of strain CA5 on pathogens was evaluated.

[0061] After 7 days of confrontation culture, the average colony diameter of the fermentation liquid group of strain CA5 was 1.86 cm, and the inhibition rate was 73.42%; the average colony diameter of the bacterial suspension group was 2.28 cm, and the inhibition rate was 67.32%; the average colony diameter of the sterile fermentation filtrate group was 5.19 cm, and the inhibition rate was 25.75%. The inhibition rate of the fermentation liquid group on the growth of pathogens was significantly higher than that of the other two groups. It can be seen that the fermentation products of strain CA5 have an inhibitory effect on tea root rot pathogens, and the fermentation liquid group has the strongest antibacterial ability. The bacteria and fermentation filtrate jointly play the main antibacterial effect, and the sterile fermentation filtrate has a weaker inhibitory effect on pathogens (Table 5).

[0062] Table 5 Antibacterial ability of different components of the fermentation product of strain CA5

[0063] Fermentation product type Colony diameter Antibacterial rate Fermentation liquid 1.86±0.06c 73.42±0.86a bacterial suspension 2.28±0.08b 67.32±1.20b Sterile fermentation filtrate 5.19±0.10a 25.75±1.54c

[0064] Note: Data are expressed as mean ± standard deviation. Different lowercase letters after the data in the same column indicate significant differences (P < 0.05).

[0065] Example 8 Potted Plant Prevention Test Evaluation

[0066] A two-year-old susceptible tea variety Quercus chinensis was selected as the test material, and a surfactant was added to the antagonistic bacteria fermentation liquid and mixed for use. Healthy tea potted plants with the same growth were selected and divided into 4 groups, namely the pathogen fermentation liquid treatment group, the strain CA5 fermentation liquid treatment group, the 10-fold strain CA5 fermentation liquid treatment group, and the 50-fold strain CA5 fermentation liquid treatment group. The specific operation was as follows: 20 mL of pathogen fermentation liquid was used to irrigate the roots of all 4 groups of tea seedlings; after 1 day of pathogen fermentation liquid treatment, 20 mL of strain CA5 fermentation liquid, 10-fold strain CA5 fermentation liquid, and 50-fold strain CA5 fermentation liquid were used to irrigate the roots of tea seedlings for 3 groups. The above operation was repeated once on the 14th day, and each treatment was repeated for 4 groups, with 4 potted plants in each group, a total of 64 potted plants, cultured under natural conditions, and their disease status was observed. After 28 days, the incidence rate survey and prevention and control effect evaluation were conducted. The incidence rate and control effect were calculated according to the following formulas: See Table 6.

[0067] Incidence rate (%) = (number of diseased plants / total number of plants) × 100%

[0068] Control effect (%) = (control incidence - treatment incidence) / control incidence × 100%

[0069] Table 6 Evaluation of potted plant protection effect test

[0070] deal with Incidence (%) Control effect (%) Pathogen fermentation broth 87.50％ — Strain CA5 fermentation stock solution 31.25％ 64.29％ 10 times strain CA5 fermentation broth 37.50％ 57.14％ 50 times strain CA5 fermentation broth 50.00％ 42.86％

[0071] The results of the potted plant prevention test showed (Table 6) that 28 days after the inoculation of the pathogen, the incidence rate of the pathogen fermentation liquid group was 87.5%, the incidence rate of tea seedling root rot in the groups treated with strain CA5 fermentation liquid, 10 times strain CA5 fermentation liquid and 50 times strain CA5 fermentation liquid were 31.25%, 37.50% and 50.00%, respectively, and the prevention and control effects of tea seedling root rot were 64.29%, 57.14% and 42.86%, respectively. The fermentation liquid had the best prevention and control effect. Strain CA5 has great application potential in the prevention and control of tea tree root rot.

[0072] Evaluation of growth promotion effect

[0073] A two-year-old susceptible tea variety Quercus chinensis was selected as the test material, and a surfactant was added to the antagonistic bacteria fermentation liquid and mixed for use. Healthy tea potted plants with the same growth were selected and divided into 4 groups, namely, the clean water treatment control group, the strain CA5 fermentation liquid treatment group, the 10-fold strain CA5 fermentation liquid treatment group, and the 50-fold strain CA5 fermentation liquid treatment group. The specific operation was as follows: 20 mL of clean water, strain CA5 fermentation liquid, 10-fold strain CA5 fermentation liquid, and 50-fold strain CA5 fermentation liquid were used to irrigate the tea seedlings with roots, and the treatment was repeated once on the 14th day. Each treatment was repeated in 4 groups, with 4 potted plants in each group, for a total of 64 potted plants, which were cultured under natural conditions. The plant height was measured at 0d and 28d, and the changes in plant height were analyzed, as shown in Table 7.

[0074] Table 7 Evaluation of potted plant growth promotion experiment

[0075] deal with Average plant height at 0d (cm) Average plant height at 28 days (cm) Average plant height growth (cm) Clear water control 26.02±0.41 26.39±0.43 0.37 Strain CA5 fermentation stock solution 26.82±0.48 28.28±0.51 1.46 10 times strain CA5 fermentation broth 26.41±0.35 27.56±0.39 1.15 50 times strain CA5 fermentation broth 26.23±0.39 27.07±0.42 0.84

[0076] The results showed (Table 7) that, compared with the control group, the average plant height increases of tea seedlings treated with strain CA5 fermentation solution, 10 times strain CA5 fermentation solution, and 50 times strain CA5 fermentation solution were 0.37 cm, 1.46 cm, 1.15 cm, and 0.84 cm, respectively. The fermentation solution had the best growth-promoting effect, indicating that strain CA5 can promote the growth of tea trees.

[0077] The various embodiments in this specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the various embodiments can be referenced to each other.

[0078] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A pine-loving Talaromyces CA5, characterized in that The pinophilic Talaromyces Talaromycespi nophilus )CA5 was deposited in the China Center for Type Culture Collection on September 26, 2023, with the deposit number CCTCC NO:M 20231797 and the classification name Talaromycespinophilus CA5.

2. Use of the pine-loving Talaromyces CA5 described in claim 1 in inhibiting tea tree root rot pathogens.

3. Use of the pine-loving Bacillaceae CA5 according to claim 1 in inhibiting 8 other plant pathogens, the other 8 plant pathogens include eggplant brown streak disease pathogen, potato early blight pathogen, wild rice root rot pathogen, corn leaf blight pathogen, citrus bark disease pathogen, pepper wilt pathogen, polygonatum anthracnose pathogen and strawberry blight pathogen.

4. Use of the pine-loving Talaromyces CA5 according to claim 1 in inhibiting tea tree root rot pathogens and thereby promoting the growth of tea trees.

5. The use according to claim 4, characterized in that: Talaromyces pineophilus CA5 has the ability to decompose organic and inorganic phosphorus, dissolve potassium, produce siderophores, fix nitrogen, produce ammonia and produce IAA.

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