A kind of Trichoderma aspergillus and bio-source synergistic fermentation liquid containing the metabolites of Trichoderma aspergillus and application thereof

Abstract

The present invention relates to a Trichoderma aspergillus and a biosource synergistic fermentation liquid containing the metabolites of Trichoderma aspergillus. The biosource fermentation liquid containing the metabolites of Trichoderma aspergillus CTCCSJ-WXH-CF002 can effectively prevent and control corn leaf blight, corn leaf blight and corn curvature leaf spot when used alone or in combination with low-toxic chemical pesticides through plot tests, and the combined use improves the control effect of chemical pesticides and plays a synergistic role. It is further verified through field test demonstration that the biosource fermentation liquid containing the metabolites of Trichoderma aspergillus CTCCSJ-WXH-CF002 has significant preventive effects on corn leaf blight, corn leaf blight, corn gray spot and corn ear rot when used alone. The biosource fermentation liquid containing the metabolites of Trichoderma aspergillus CTCCSJ-WXH-CF002 can be used alone or in combination with low-toxic chemical pesticides. It has been verified by plot and field tests that it can effectively prevent corn premature aging and has a significant yield-increasing effect.

Description

Technical Field

[0001] The invention belongs to the field of agricultural microorganisms, and relates to biological fermentation liquid technology, in particular to Trichoderma acanthosporum and a biological source synergistic fermentation liquid containing the metabolites of Trichoderma acanthosporum and its application. Background Art

[0002] Trichoderma belongs to the genus Trichoderma, family Trichoderma, class Hypomycetes, order Hypomycetes, and is a type of fungus widely distributed in soil and plants. According to reports, more than 250 species of Trichoderma have been discovered, which have antagonistic effects on 29 plant pathogenic fungi of 18 genera. Because of its good adaptability, strong colonization ability and outstanding biocontrol effect, it has great potential for biocontrol application in plant disease control. Common biocontrol Trichoderma include T.harzianum, T.viride, T.seudokoningii, T.Asperellum and T.longibrachiatum.

[0003] In recent years, due to the excessive use of chemical fertilizers and pesticides, ecological and environmental problems such as water and soil pollution, pesticide residues and frequent plant diseases have been caused. Biocontrol microorganisms are a good choice for improving soil ecological environment and preventing and controlling plant diseases. Trichoderma aculeatus has the advantages of high activity, strong reproduction, low price and convenient production. Solid-state fermentation is often used to produce agricultural microbial agents in industrial production. Many scholars have reported on the solid-state fermentation process level of common Trichoderma such as green Trichoderma, Trichoderma pseudokoningii, Trichoderma harzianum and Trichoderma longibrachiatum, and the technology is relatively mature. However, there are relatively few reports on the liquid fermentation process conditions of Trichoderma aculeatus, especially on the synergistic activity of the combination of fermentation liquid and chemical pesticides. The overall fermentation level is low. In industrial production, there are often prominent problems such as confusion of strains, weak functions, high rate of foreign bacteria and low fermentation level, which seriously hinder its industrial production.

[0004] Therefore, obtaining excellent biocontrol strains of Trichoderma aspergillus, refining operating procedures and high-level fermentation processes, and developing functions such as using fermentation liquid to enhance the control of crop diseases by combining with chemical pesticides will be of great help to my country in improving the industrial scale production of Trichoderma aspergillus and promoting the development of the agricultural economy. Summary of the invention

[0005] The purpose of the present invention is to overcome the shortcomings of the prior art, provide a kind of Trichoderma aspergillus and a bio-source synergistic fermentation liquid containing the metabolites of Trichoderma aspergillus, utilize the biological characteristics of Trichoderma aspergillus, and combine the liquid fermentation technology of Trichoderma aspergillus and the characteristics of the active ingredients of its metabolites to prepare a kind of Trichoderma aspergillus that can be mixed with green control chemical agents

[0006] The new spraying technology can promote crop growth and induce crop resistance to disease and stress. Whether the fermented liquid is used alone or in combination with chemical pesticides, it can reduce the occurrence of leaf spot disease and prevent premature aging. When used in combination with chemical pesticides, it can also improve the control effect of chemical pesticides.

[0007] The present invention solves the technical problem by adopting the following technical solutions:

[0008] In a first aspect, the present invention provides a Trichoderma asperellum CTCCSJ-WXH-CF002, whose deposit number is CCTCC NO: M2024435.

[0009] In a second aspect, the present invention provides a biogenic synergistic fermentation liquid, wherein the biogenic synergistic fermentation liquid comprises a liquid fermentation liquid of Trichoderma aspergillus CTCCSJ-WXH-CF002.

[0010] As an embodiment of the present invention, the bio-source enhanced fermentation liquid further comprises auxiliary agents; the auxiliary agents include preservatives and surface dispersants.

[0011] As an embodiment of the present invention, the preservative is xanthan gum, and the surface dispersant is tea saponin.

[0012] As an embodiment of the present invention, the biosource enhanced fermentation liquid contains 1-3 g / L xanthan gum and 3-7 g / L tea saponin.

[0013] As an embodiment of the present invention, the bio-source enhanced fermentation liquid is prepared by a method comprising the following steps:

[0014] S1. Strain culture: inoculating the activated Trichoderma aspergillus CTCCSJ-WXH-CF002 into a liquid culture medium for culture to obtain a seed solution of Trichoderma aspergillus CTCCSJ-WXH-CF002;

[0015] S2. Preparation of fermentation raw materials: Take the following components in parts by weight: 50-60 parts of corn flour, 2-4 parts of potassium dihydrogen phosphate, 1-3 parts of sodium nitrate, 0.5-2 parts of ammonium sulfate, 0.5-2 parts of sodium chloride, 0.1-1 parts of magnesium sulfate, add water to adjust the water content to 10%-40%; add trace element composition and chebulic acid; sterilize and cool;

[0016] S3, fermentation liquid preparation: taking the fermentation raw material obtained in step S2, adding the Trichoderma aspergillus CTCCSJ-WXH-CF 002 seed liquid, stirring evenly for fermentation; obtaining fermentation liquid.

[0017] As an embodiment of the present invention, the method further comprises step S4, preparation processing: sterilizing and centrifuging the fermentation broth, taking the supernatant; and adding an auxiliary agent.

[0018] As an embodiment of the present invention, in step S1, the characteristic value of the number of live bacteria in the seed liquid of Trichoderma aspergillus CTCCSJ-WXH-CF002 is 1.0×10 8 -6.0×10 8 cfu / mL.

[0019] As an embodiment of the present invention, step S1 specifically involves taking a bacterial disc of the activated Trichoderma aspergillus CTCCSJ-WXH-CF002 and placing it in a PD culture medium, culturing it at 25-30°C and 180-200r / min for 2-3 days to obtain a seed solution of Trichoderma aspergillus CTCCSJ-WXH-CF002.

[0020] As an embodiment of the present invention, in step S2, the trace element composition is composed of trace elements Mo, Mn, B, and Cu in a molar ratio of 1-3:1-3:0.5-2:0.5-2.

[0021] As an embodiment of the present invention, in step S2, based on the fermentation raw material adjusted to a water content of 10%-40% by adding water, the trace element composition is added in an amount of 0.05-0.2%.

[0022] As an embodiment of the present invention, in step S2, based on the fermentation raw material adjusted to a moisture content of 10%-40% by adding water, the added amount of chebulic acid is 0.05-0.2%.

[0023] As an embodiment of the present invention, in step S2, the sterilization is performed at 115-121° C. for 15-30 min, and the mixture is cooled and then used.

[0024] As an embodiment of the present invention, in step S3, the fermentation process of the fermentation is:

[0025] When the fermentation time is 0-24h, the fermentation temperature is 28°C, the rotation speed is 200rpm, the ventilation volume is 0.6vvm, and the pH is natural;

[0026] When the fermentation time is 24-96h, the fermentation temperature is 28°C, the rotation speed is 150rpm, the ventilation volume is 0.8vvm, and the pH is natural;

[0027] When the fermentation time is more than 96h, the fermentation temperature is 28℃, the rotation speed is 120rpm, the ventilation volume is 0.6vvm, and the pH is ≤5.5;

[0028] When the fermentation exceeds 96 hours and the pH is greater than 5 and less than 5.5, the fermentation is stopped, heated to 65-70℃ and maintained for 15 minutes to kill the Trichoderma, and then put into the tank.

[0029] As an embodiment of the present invention, in step S3, the inoculation amount is 0.3-1 v / v%.

[0030] In a third aspect, the present invention relates to a method for using a bio-source synergistic fermentation liquid, which can be applied alone or in combination with chemical pesticides.

[0031] As one embodiment of the present invention,

[0032] For single use: 300 mL / 667 m 2 Bio-source enhanced fermentation liquid;

[0033] Mixed use: Apply 300mL / 667m 2 Bio-source enhanced fermentation liquid +20mL / 667m 2 Pyraclostrobin.

[0034] In a fourth aspect, the present invention provides a use of Trichoderma aspergillus, or a bio-source enhanced fermentation liquid in at least one of the following aspects:

[0035] (1) Promote crop growth,

[0036] (2) Inducing crop disease resistance and stress resistance,

[0037] (3) Control corn plant diseases;

[0038] (4) Prevent premature senescence of corn leaves;

[0039] (5) Increase corn yield.

[0040] As an embodiment of the present invention, the corn plant disease includes at least one of corn leaf blight, corn leaf blight, corn curvularia leaf spot, corn gray leaf spot, and corn ear rot.

[0041] In the present invention, Trichoderma asperellum CTCCSJ-WXH-CF002, the preservation number is CCTCC NO: M2024435, the preservation date is: March 15, 2024, the preservation unit is: China Center for Type Culture Collection, and the preservation address is: Wuhan University, China.

[0042] Trichoderma asperellum GDFS1009, the deposit number is CGMCC NO.9512, the deposit date is August 13, 2014, the depository is China General Microbiological Culture Collection Center, and the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing.

[0043] The present invention has the following beneficial effects:

[0044] 1. The Trichoderma aspergillus CTCCSJ-WXH-CF002 of the present invention is isolated from a corn field in a protective cultivated land of Harqin Banner, Chifeng City, Inner Mongolia Autonomous Region.

[0045] 2. The liquid fermentation process of the present invention improves the fermentation raw material formula, adds a trace element composition and terminalia acid, enhances the efficiency of liquid fermentation and production of metabolites of Trichoderma spinulosa CTCCSJ-WXH-CF002, and ensures the optimal active ingredients of the fermentation liquid.

[0046] 3. The bio-source synergistic fermentation liquid of the present invention has been verified through small plot tests and large-scale demonstrations. It can effectively prevent and treat corn leaf blight, corn leaf blight, and corn leaf curvature leaf spot when used alone, and has the effect of preventing premature senescence of corn leaves and increasing yield;

[0047] 4. The bio-source synergistic fermentation liquid of the present invention, when used in combination with green chemical pesticides, can improve the control effect of chemical pesticides on corn leaf blight, corn leaf blight, and corn Curvularia leaf spot, and can also improve the effect of preventing premature aging of corn leaves and increasing yield, thereby playing a synergistic role. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Other features, objects and advantages of the present invention will become more apparent from the detailed description of non-limiting embodiments made with reference to the following drawings:

[0049] Figure 1 The colony morphology diagram of Trichoderma aspergillus CTCCSJ-WXH-CF002 of the present invention; wherein (a) is cultured at 28°C for 24h, (b) is cultured at 28°C for 48h, (c) is cultured at 28°C for 72h, and (d) is cultured at 28°C for 96h;

[0050] Figure 2 This is a schematic diagram of plant premature aging levels; in the figure, 1 means that the dead leaves account for 1%-10% of the total number of leaves, which is level 1; 3 means that the dead leaves account for 10%-30% of the total number of leaves, which is level 3; 5 means that the dead leaves account for 30%-50% of the total number of leaves, which is level 5; 7 means that the dead leaves account for 50%-70% of the total number of leaves, which is level 7; 9 means that the dead leaves account for more than 70% of the total number of leaves, which is level 9;

[0051] Figure 3 The control effect of Trichoderma fermentation liquid and pyraclostrobin on corn leaf blight;

[0052] Figure 4 The control effect of Trichoderma fermentation liquid and pyraclostrobin on corn leaf blight;

[0053] Figure 5The control effect of Trichoderma fermentation liquid and pyraclostrobin on corn Curvularia leaf spot;

[0054] Figure 6 The effect of Trichoderma fermentation liquid on preventing premature aging of leaves;

[0055] Figure 7 It is a phylogenetic tree diagram of Trichoderma aspergillus CTCCSJ-WXH-CF002 of the present invention;

[0056] in, Figure 3-6 a, b, and c indicate significant differences. DETAILED DESCRIPTION

[0057] The present invention is described in detail below in conjunction with embodiments. The following embodiments will help those skilled in the art to further understand the present invention, but are not intended to limit the present invention in any form. It should be noted that, for those of ordinary skill in the art, some adjustments and improvements can also be made without departing from the concept of the present invention. These all belong to the protection scope of the present invention.

[0058] Example 1: Isolation, cultivation and identification of strains

[0059] The present invention first isolates a strain of Trichoderma asperellum from the soil of a corn field in a protective cultivated land of Harqin Banner, Chifeng City, Inner Mongolia Autonomous Region. The strain is named: CTCCSJ-WXH-CF002, classified and named: Trichoderma asperellum, with a preservation number of CCTCC NO: M2024435, a preservation date of: March 15, 2024, a preservation unit: China Center for Type Culture Collection, and a preservation address: Wuhan University, China.

[0060] The strain of Trichoderma acanthosporum was identified, and the ITS full sequence information of the strain CTCCSJ-WXH-CF002 is as follows (SEQ ID NO.1):

[0061] .

[0062] Phylogenetic tree of Trichoderma aspergillus CTCCSJ-WXH-CF002 Figure 7 .

[0063] The culture characteristics of this strain are as follows:

[0064] like Figure 1 As shown in the figure, the strain of Trichoderma spinulosa CTCCSJ-WXH-CF002 grew well when cultured at 28℃ on PDA medium. The colonies of 24h grew in concentric circles and were transparent and fluffy. The colonies of 48h began to produce spores, and yellow-white color appeared in the middle spore-producing cluster area, yellow in the center, and white in the extension area. The colonies of 72h could cover a 9cm culture dish. The edge of the culture dish was observed to have produced spores around 96h, showing black particles, dark yellow, yellow, yellow-white, and white from the center to the edge, and yellow or yellow-green on the back. The main axis of the spore stalk is straight and slender, with opposite branches. The bottle-shaped stalk is slightly thin at the bottom, swollen in the middle, and thin at the top. The conidia are spherical or nearly spherical, and can produce ovoid thick-walled spores.

[0065] Example 2: Preparation of biogenic enhanced fermentation liquid and analysis of metabolites

[0066] 2.1. The process of preparing the bio-source enhanced fermentation liquid after strain isolation and cultivation includes the following steps:

[0067] (1) Strain culture: A 5 mm diameter bacterial disc of Trichoderma aspergillus CTCCSJ-WXH-CF002 activated on a PDA plate was cut and placed in a PD culture medium. The culture was cultured at 28°C and 180 rpm for 2 days to obtain a seed solution of Trichoderma aspergillus CTCCSJ-WXH-CF002. The characteristics of the live bacteria in the seed solution were that the top of the hyphae expanded and spore production began to occur.

[0068] (2) Preparation of fermentation raw materials: Take the following components in respective mass proportions: 55 parts of corn flour, 3.82 parts of potassium dihydrogen phosphate, 1.42 parts of sodium nitrate, 1.1 parts of ammonium sulfate, 1 part of sodium chloride, and 0.5 parts of magnesium sulfate, add water to adjust the water content to 40%, add 0.1% of a trace element composition and 0.1% of chebulic acid respectively, the composition and molar ratio of the trace element composition are: Mo, Mn, B, Cu = 2:2:1:1, then sterilize at 121°C for 30 min, cool and set aside.

[0069] (3) Preparation of fermentation liquid: Take the fermentation raw material prepared in step (2), inoculate the Trichoderma spinulosa CTCCSJ-WXH-CF002 seed liquid prepared in step (1) at an inoculum rate of 0.3% by volume, stir evenly and ferment; the fermentation process is as follows: when the fermentation time is 0-24h, the fermentation temperature is 28°C, the rotation speed is 200rpm, the ventilation volume is 0.6vvm, and the pH is natural; when the fermentation time is 24-96h, the fermentation temperature is 28°C, the rotation speed is 150rpm, the ventilation volume is 0.8vvm, and the pH is natural; when the fermentation time is more than 96h, the fermentation temperature is 28°C, the rotation speed is 120rpm, the ventilation volume is 0.6vvm, and the pH is ≤5.5; when the fermentation exceeds 96h, the fermentation is stopped when the pH is greater than 5 and less than 5.5, and the mixture is heated to 65-70°C for 15 minutes to kill the Trichoderma, and then put into the tank.

[0070] (4) Preparation processing: The sterilized fermentation broth was centrifuged at 8000 rpm, and the supernatant was taken and 2 g / L of xanthan gum as a preservative and 5 g / L of tea saponin as a surface dispersant were added.

[0071] 2.2. Detection and analysis of metabolites in bio-source enhanced fermentation liquid

[0072] The control Trichoderma aspergillus GDFS1009 (deposited in China General Microbiological Culture Collection Center, deposit date is August 13, 2014, deposit number is CGMCC NO.9512) was selected, and the activation method, fermentation process and auxiliary agent addition were the same as 2.1.

[0073] The metabolites in the bio-source enhanced fermentation liquid of the product of the embodiment of the present invention and the control Trichoderma aspergillus GDFS1009 fermentation liquid with a preservation number of CGMCC NO.9512 were detected and analyzed.

[0074] As shown in Tables 1 and 2, through the analysis of the full components of the fermentation broth metabolites, it can be seen that the metabolites in the fermentation broth of Trichoderma aspergillus CTCCSJ-WXH-CF002 are rich and diverse, and most of them are related to cytotoxic activity, antibacterial activity, etc., including heterocyclic compounds acetamiprid, allantoin, indole, indole-3-butyric acid, etc.; benzene and its derivatives propamide, myclobutanil, etc.; alcohols and amines curcumin, actinomycin, etc.; nucleotides and their metabolites adenine, etc.; organic acids and their derivatives salicylic acid, gallic acid, etc.; the content is higher than that of the control Trichoderma aspergillus GDFS1009 with the preservation number CGMCC NO.9512.

[0075] Table 1 Metabolite analysis

[0076]

[0077] Table 2

[0078]

[0079] Example 3: Preparation of comparative examples and analysis of antibacterial activity

[0080] 3.1 Preparation of different comparative examples

[0081] (1) Comparative Example D1 was prepared without adding the trace element composition Mo, Mn, B, and Cu. The rest of the preparation method was the same as Example 2.1.

[0082] (2) Comparative Example D2 was prepared without adding chebulic acid. The rest of the preparation method was the same as Example 2.1.

[0083] (3) Comparative Example D3 was prepared without adding either the trace element composition or terminalia chebula acid. The rest of the preparation method was the same as that of Example 2.1.

[0084] (4) Comparative Examples D4-D7 were prepared. The compositions of different trace elements were shown in Table 3. The rest of the preparation method was the same as that of Example 2.1.

[0085] Table 3

[0086]

[0087] 3.2 Antibacterial activity analysis

[0088] This example tests the inhibitory effects of the bio-source enhanced fermentation liquid prepared in Implementation 2.1 (hereinafter referred to as "Trichoderma fermentation liquid") and the fermentation liquid prepared in Comparative Examples D1-D7 on Helicosporium umbilical cordifolium, Helicosporium longifolium, Cercosporium zeae, and Fusarium graminearum.

[0089] On a clean bench, the Trichoderma fermentation liquid, D1-D7 and PDA medium were mixed in a ratio of 1:10, shaken evenly, poured into a plate (9 cm in diameter), and 15-20 mL of the mixed medium was poured into each sterilized culture dish. After the medium solidified, the target pathogens (Helminthosporium of Large Spot Disease, Helminthosporium, Cercosporium of Corn, Fusarium graminearum) were inoculated with a bacterial disc (0.5 cm) in the center of the medium; the pathogens were inoculated in the center of the PDA plate mixed with sterile water at a ratio of 1:10 as a control, and placed in a constant temperature incubator at 28°C for inversion culture for 5 days; 3 replicates were set for each treatment, the size of the colony was measured, and the inhibition rate was calculated.

[0090] Table 4

[0091]

[0092]

[0093] The main pathogens causing corn leaf blight, corn leaf blight, corn gray leaf blight and corn ear rot are shown in Table 4. The antibacterial test shows that the Trichoderma fermentation liquid prepared in Example 2.1 of the present invention can effectively inhibit the above four pathogens, and the effect is better than that of the control example, which provides a basis for further field application.

[0094] Example 4: Efficacy verification - cell trial

[0095] The bio-source synergistic fermentation liquid of Example 2.1 was used alone or in combination with green chemical pesticides to test the effects on corn diseases, premature aging, yield increase, etc. in a plot experiment.

[0096] (1) Experimental materials and methods

[0097] Tested corn varieties: Zhengdan 958, Xianyu 335;

[0098] Location: Research Base of Shenyang Agricultural University;

[0099] Test agents: the biogenic synergistic fermentation liquid of Example 2.1 (hereinafter referred to as "Trichoderma fermentation liquid") and 30% pyraclostrobin (Henan Yongguan Qiaodi Agricultural Technology Co., Ltd.).

[0100] Plot design: row width 0.6m, row length 10m, 40 plants per row, two rows for one treatment.

[0101] Field management: Fertilization on April 30, 40kg / 667m 2 ; Sow on May 7 and spray herbicide after sowing.

[0102] Experimental treatment: There are 6 treatments in the experiment, namely blank control, 100mL / 667m 2Trichoderma fermentation liquid, 300mL / 667m 2 Trichoderma fermentation liquid, 100mL / 667m 2 Trichoderma fermentation liquid + 20mL / 667m 2 Pyraclostrobin, 300mL / 667m 2 Trichoderma fermentation liquid + 20mL / 667m 2 Pyraclostrobin, 20mL / 667m 2 Pyraclostrobin.

[0103] Survey time: Leaf diseases were surveyed on September 9, 2023.

[0104] Survey standards: refer to the relevant standards of the Ministry of Agriculture.

[0105]

[0106] Control effect (%) = (disease index of blank control area - disease index of drug-treated area) / disease index of blank control area × 100%

[0107] Premature aging index:

[0108] If the dead leaves account for 1%-10% of the total leaves, it is level 1; 10%-30% is level 3; 30%-50% is level 5; 50%-70% is level 7; and more than 70% is level 9 (such as Figure 2 ).

[0109] Premature senescence index = [(number of plants at the dead leaf level × representative value of that level) / total number of plants × representative value of the heaviest dead leaf level)] × 100.

[0110] Anti-premature aging effect (%) = (premature aging index of blank control area - premature aging index of drug-treated area) / premature aging index of blank control area × 100%

[0111] (2) Test results and analysis

[0112] ① The control effect of Trichoderma fermentation liquid and pyraclostrobin on corn leaf blight

[0113] As shown in Table 5 and Figure 3 As shown in the figure, the control effect of Zhengdan 958 and Xianyu 335 when Trichoderma fermentation liquid was used alone was 5.4-19.97%, the control effect of pyraclostrobin was 7.18-19.16%, and the control effect of the combination of Trichoderma fermentation liquid and pyraclostrobin was 15.44-51.77%. The combination of Trichoderma fermentation liquid and pyraclostrobin had a synergistic effect on corn leaf blight. And the control effect of Trichoderma fermentation liquid and pyraclostrobin on the susceptible variety Xianyu 335 increased with the increase of the application amount of Trichoderma fermentation liquid, and the synergistic effect on green chemical pesticides was more obvious.

[0114] Table 5 Control effect of Trichoderma fermentation liquid and pyraclostrobin on corn leaf blight

[0115]

[0116] ②The control effect of Trichoderma fermentation liquid and pyraclostrobin on corn leaf blight

[0117] As shown in Table 6 and Figure 4 As shown, 300mL / 667m 2 Trichoderma fermentation liquid + 20mL / 667m 2 The control effect of Zhengdan 958 treated with pyraclostrobin was 55.91%. 300mL / 667m 2 Trichoderma fermentation liquid and 300mL / 667m 2 Trichoderma fermentation liquid + 20mL / 667m 2 The protective effects of Xianyu 335 treated with pyraclostrobin were 36.94% and 44.44% respectively.

[0118] Table 6 Control effect of Trichoderma fermentation liquid and pyraclostrobin on corn leaf spot

[0119]

[0120] ③The control effect of Trichoderma fermentation liquid and pyraclostrobin on corn Curvularia leaf spot

[0121] As shown in Table 7 and Figure 5 As shown, Zhengdan 958 and Xianyu 335 are at 300mL / 667m 2 The control effect of Trichoderma fermentation liquid was 27.53-34.54%, 100mL / 667m 2 The control effect of Trichoderma fermentation liquid treatment was 16.21-30.34%.

[0122] Table 7 Control effect of Trichoderma fermentation liquid and pyraclostrobin on corn Curvularia leaf spot

[0123]

[0124]

[0125] ④The effect of Trichoderma fermentation liquid on preventing premature aging of leaves

[0126] As shown in Table 8 and Figure 6 As shown, Trichoderma fermentation liquid has a certain anti-premature aging effect on Zhengdan 958, with an anti-premature aging effect of 8.4-13.3%. Trichoderma fermentation liquid also has a certain anti-premature aging effect on Xianyu 335, with an anti-premature aging effect of 3.2-6.4%.

[0127] Table 8 Effect of Trichoderma fermentation liquid on preventing premature aging of leaves

[0128]

[0129] ⑤ The yield-increasing effect of Trichoderma fermentation liquid

[0130] Compared with the control, there were no significant differences in ear length, ear diameter, number of ear rows, number of rows, and bald tip length of Zhengdan 958 after treatment with Trichoderma fermentation solution. Indicators such as ear weight and 100-grain weight were significantly higher than those of the control, with an increase in yield per mu of 2.57%-9.71% (Tables 9-11).

[0131] Compared with the control, there were no significant differences in ear length, number of ear rows, number of rows and bald tip length of Xianyu 335 after treatment with Trichoderma fermentation solution. The indicators such as ear diameter, ear weight and 100-grain weight were significantly higher than those of the control, and the yield per mu increased by 3.78%-14.92% (Table 9-Table 11).

[0132] Table 9

[0133]

[0134] Table 10

[0135]

[0136] Table 11

[0137]

[0138]

[0139] Example 5: Efficacy verification - field trial

[0140] The bio-source synergistic fermentation liquid of Example 2.1 was used alone to test the effects of corn diseases, premature aging, and yield increase in field demonstration areas.

[0141] (1) Experimental materials and methods

[0142] The corn varieties tested are Danyu 405 and Liangyu 99, sown on May 4, 2023.

[0143] Test location: Research base of Dandong Academy of Agricultural Sciences, Fengcheng City, Liaoning Province.

[0144] Test reagent: Bio-source enhanced fermentation liquid of Example 2.1 (hereinafter referred to as Trichoderma fermentation liquid), 100mL / 667m 2 .

[0145] Area: 10 acres.

[0146] Spraying time: On July 20, the DJI drone sprayed the pesticide. The temperature was 28°C and the air humidity was 78%. The highest temperature of the day was 37°C and the lowest temperature was 20°C.

[0147] Survey time: Leaf diseases were surveyed on September 4, 2023, ear rot, stem rot and insect pests were surveyed on September 25, 2003, and the seeds were tested on September 27, 2023.

[0148] (2) Results and analysis

[0149] ① The effect of Trichoderma fermentation liquid on the prevention and control of corn leaf spot

[0150] As shown in Table 12, compared with the control, the prevention effect of corn leaf blight treated with Danyu 405 Trichoderma fermentation liquid reached 85.41%.

[0151] Table 12 Effect of Trichoderma fermentation liquid on the control of corn leaf spot

[0152]

[0153] ②The effect of Trichoderma fermentation liquid on the prevention and control of corn leaf spot

[0154] As shown in Table 13, compared with the control, the control efficacy of Danyu 405 treated with Trichoderma fermentation solution against corn leaf blight was 40.16%, and the control efficacy of Liangyu 99 treated with Trichoderma fermentation solution against corn leaf blight was 47.65%.

[0155] Table 13

[0156]

[0157] ③The effect of Trichoderma fermentation liquid on the prevention and control of corn gray spot

[0158] As shown in Table 14, compared with the control, the control effect of Danyu 405 Trichoderma fermentation liquid on corn gray spot disease was 33.23%, and the control effect of Liangyu 99 Trichoderma fermentation liquid on corn gray spot disease was 35.37%.

[0159] Table 14

[0160]

[0161] ④The effect of Trichoderma fermentation liquid on ear rot

[0162] As shown in Table 15, compared with the control, the control effect of Danyu 405 Trichoderma fermentation liquid on ear rot was 34.04%, and the control effect of Liangyu 99 Trichoderma fermentation liquid on ear rot was 32.88%.

[0163] Table 15

[0164]

[0165]

[0166] ⑤ Effect of Trichoderma fermentation liquid on yield

[0167] As shown in Table 16, compared with the control, the yield of Danyu 405 increased by 6.70% after treatment with Trichoderma fermentation solution, and the average 1000-grain weight increased by 4.87% at 14% water; the yield of Liangyu 99 increased by 10.47% after treatment with Trichoderma fermentation solution, and the average 1000-grain weight increased by 10.64% at 14% water.

[0168] Table 16

[0169]

[0170] In summary, the present invention relates to a Trichoderma aspergillus and a biosource synergistic fermentation liquid containing the metabolites of Trichoderma aspergillus. The biosource fermentation liquid containing the metabolites of Trichoderma aspergillus CTCCSJ-WXH-CF002 can effectively prevent and control corn leaf blight, corn leaf blight and corn curvature leaf spot when used alone or in combination with low-toxic chemical pesticides through plot tests, and the combined use improves the control effect of chemical pesticides and plays a synergistic role. It is further verified through field test demonstration that the biosource fermentation liquid containing the metabolites of Trichoderma aspergillus CTCCSJ-WXH-CF002 has significant preventive effects on corn leaf blight, corn leaf blight, corn gray spot and corn ear rot when used alone. The biosource fermentation liquid containing the metabolites of Trichoderma aspergillus CTCCSJ-WXH-CF002, used alone or in combination with low-toxic chemical pesticides, can effectively prevent corn premature aging and has a significant yield-increasing effect after plot and field tests.

[0171] The above describes the specific embodiments of the present invention. It should be understood that the present invention is not limited to the above specific embodiments, and those skilled in the art may make various modifications or variations within the scope of the claims, which do not affect the essence of the present invention.

Claims

1. A Trichoderma asperellum CTCCSJ-WXH-CF002, whose deposit number is CCTCCNO: M2024435.

2. A bio-source enhanced fermentation liquid, characterized in that: The biosource enhanced fermentation liquid comprises the liquid fermentation liquid of Trichoderma aspergillus CTCCSJ-WXH-CF002 as claimed in claim 1.

3. The bio-source enhanced fermentation liquid according to claim 2, characterized in that: The bio-source enhanced fermentation liquid also contains auxiliary agents; the auxiliary agents include preservatives and surface dispersants; The preservative is xanthan gum, and the surface dispersant is tea saponin; The biosource enhanced fermentation liquid contains 1-3 g / L of xanthan gum and 3-7 g / L of tea saponin.

4. The bio-source enhanced fermentation liquid according to claim 2 or 3, characterized in that: The biosource enhanced fermentation liquid is prepared by a method comprising the following steps: S1. Strain culture: inoculate the activated Trichoderma aspergillus CTCCSJ-WXH-CF002 into liquid PD culture medium for culture to obtain Trichoderma aspergillus CTCCSJ-WXH-CF002 seed liquid; S2. Preparation of fermentation raw materials: Take the following components in parts by weight: 50-60 parts of corn flour, 2-4 parts of potassium dihydrogen phosphate, 1-3 parts of sodium nitrate, 0.5-2 parts of ammonium sulfate, 0.5-2 parts of sodium chloride, 0.1-1 parts of magnesium sulfate, add water to adjust the water content to 10%-40%; add trace element composition and chebulic acid; sterilize and cool; S3, fermentation liquid preparation: taking the fermentation raw material obtained in step S2, adding the Trichoderma aspergillus CTCCSJ-WXH-CF 002 seed liquid, stirring evenly for fermentation; obtaining fermentation liquid.

5. The bio-source enhanced fermentation liquid according to claim 4, characterized in that: In step S1, the characteristic value of the number of live bacteria in the seed liquid of Trichoderma spinulosa CTCCSJ-WXH-CF002 is 1.0×10 8 -6.0×10 8 cfu / mL.

6. The bio-source enhanced fermentation liquid according to claim 4, characterized in that: In step S2, the trace element composition is composed of trace elements Mo, Mn, B, and Cu in a molar ratio of 1-3:1-3:0.5-2:0.5-2.

7. The bio-source enhanced fermentation liquid according to claim 4, characterized in that: In step S2, based on the fermentation raw material adjusted to a water content of 10%-40% by adding water, the trace element composition is added in an amount of 0.05-0.2%; and the chebulic acid is added in an amount of 0.05-0.2%.

8. The bio-source enhanced fermentation liquid according to claim 4, characterized in that: In step S3, the fermentation process of the fermentation is: When the fermentation time is 0-24h, the fermentation temperature is 28°C, the rotation speed is 200rpm, the ventilation volume is 0.6vvm, and the pH is natural; When the fermentation time is 24-96h, the fermentation temperature is 28°C, the rotation speed is 150rpm, the ventilation volume is 0.8vvm, and the pH is natural; When the fermentation time is more than 96h, the fermentation temperature is 28℃, the rotation speed is 120rpm, the ventilation volume is 0.6vvm, and the pH is ≤5.5; When the fermentation exceeds 96 hours and the pH is greater than 5 and less than 5.5, the fermentation is stopped, heated to 65-70℃ and maintained for 15 minutes to kill the Trichoderma, and then put into the tank.

9. A method for using the bio-source enhanced fermentation liquid as claimed in claim 2, characterized in that: Use alone or in combination with chemical pesticides.

10. Use of the Trichoderma aspergillus according to claim 1 or the bio-source enhanced fermentation liquid according to claim 2 in at least one of the following: (1) Promote corn growth, (2) Prevent and control corn leaf spot, corn leaf spot, corn leaf spot, corn gray spot, and corn ear rot; (3) Prevent premature senescence of corn leaves; (4) Increase corn yield.

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