CN112322502A - Microbial inoculum prepared from Trichoderma africanum Ta97 and application thereof in preventing and treating continuous cropping diseases - Google Patents

Microbial inoculum prepared from Trichoderma africanum Ta97 and application thereof in preventing and treating continuous cropping diseases Download PDF

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CN112322502A
CN112322502A CN202011310018.3A CN202011310018A CN112322502A CN 112322502 A CN112322502 A CN 112322502A CN 202011310018 A CN202011310018 A CN 202011310018A CN 112322502 A CN112322502 A CN 112322502A
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microbial inoculum
trichoderma
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吴晓青
张新建
周方园
周红姿
赵晓燕
谢雪迎
王加宁
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Ecology Institute Of Shandong Academy Of Sciences (the Sino-Japanese Friendship Biotechnology Research Center Shandong Academy Of Sciences)
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Abstract

The invention provides a microbial inoculum prepared by using Trichoderma africanum Ta97 and application thereof in preventing and treating continuous cropping diseasesThe preparation process of the microbial inoculum comprises the following steps: the number of viable bacteria contained in the culture medium is 1 × 107～1×108Inoculating cfu/mL seed liquid into a solid fermentation culture medium, uniformly stirring, culturing for 5-8 days under the conditions of constant temperature of 25-28 ℃, light illumination for 12 hours and darkness, naturally drying, sequentially sieving by a 60-100-mesh sieve, and uniformly mixing the sieved part with diatomite according to the weight ratio of 5-7: 1 to obtain trichoderma conidium powder; mixing trichoderma conidium powder with an auxiliary agent to obtain a microbial inoculum; the application of the microbial inoculum in the aspect of preventing and treating continuous cropping diseases. The control effect of the microbial inoculum on continuous cropping diseases is more than or equal to 80 percent, which shows that the microbial inoculum has good control effect on the continuous cropping diseases and is beneficial to continuous cropping planting in fields.

Description

Microbial inoculum prepared from Trichoderma africanum Ta97 and application thereof in preventing and treating continuous cropping diseases
Technical Field
The invention relates to the technical field of biological inoculants, in particular to an inoculant prepared by utilizing Trichoderma africanum Ta97 and application thereof in the aspect of preventing and treating continuous cropping diseases.
Background
Continuous cropping is also continuous cropping and is a planting mode for reasonably utilizing land, wherein corn-wheat multiple cropping is one of typical continuous cropping planting modes. Although the planting mode achieves the maximum utilization of the land, the plant pathogenic bacteria are carried in the growth process of the crops, and the pathogenic bacteria can be gradually accumulated in the soil in the continuous cropping process, so that continuous cropping diseases are caused, and the agricultural production safety is seriously harmed.
The continuous cropping disease mainly comes from pathogenic bacteria parasitized in the overground part and the root system of the crop, the pathogenic bacteria of the overground part naturally fall off along with the epidermis of the crop or enter the soil when the straw is returned to the field, and the pathogenic bacteria and the root system pathogenic bacteria are gradually accumulated in the soil, and after years of continuous cropping, the accumulation of the pathogenic bacteria is increased, and large-area diseases are very easy to develop.
In order to reduce the threat of continuous cropping diseases to crops, pathogenic bacteria are usually killed by chemical drugs, however, the pesticide residue in soil seriously exceeds the standard after long-term use of the chemical drugs, and the chemical drugs enter underground water along with rainwater to pollute surrounding water resources, so that the requirements of environmental protection are not met.
In recent years, intensive research on microorganisms opens a new window for treating continuous cropping diseases, and the search for microorganisms which can solve continuous cropping diseases and do not harm soil is an important research subject in continuous cropping planting.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a microbial inoculum prepared by utilizing Trichoderma africanum Ta97 and application thereof in the aspect of preventing and treating continuous cropping diseases, wherein Trichoderma africanum Ta97 strain for modifying the microbial inoculum is preserved in China general microbiological culture Collection center in 14 months at 07-14 years 2020, the preservation unit is CGMCC for short, and the address is as follows: xilu No.1 Hospital, Beijing, Chaoyang, North Chen, with a deposit number: CGMCC No. 19930. The effective viable count of the microbial inoculum prepared by the trichoderma harzianum Ta97 is more than or equal to 10 hundred million/g; when the microbial inoculum is applied to the aspect of preventing continuous cropping diseases, the microbial inoculum is sprayed on the ground surface for the first time 15-30 days before crop planting, and the field is subjected to rotary tillage; when the germination rate of crops is more than or equal to 80%, the microbial inoculum is sprayed for the second time. The control effect of the microbial inoculum on continuous cropping diseases is more than or equal to 80 percent, which shows that the microbial inoculum has good control effect on the continuous cropping diseases and is beneficial to continuous cropping planting in fields.
The technical scheme of the invention is as follows:
a microbial inoculum prepared by Trichoderma africanum Ta97 is prepared by the following steps: the number of viable bacteria contained in the culture medium is 1 × 107～1×108Inoculating cfu/mL seed liquid into a solid fermentation culture medium, uniformly stirring, culturing for 5-8 days under the conditions of constant temperature of 25-28 ℃, 12 hours of illumination and 12 hours of semi-darkness, naturally drying, sequentially sieving by a 60-100-mesh sieve, and uniformly mixing the sieved part with diatomite according to the weight ratio of 5-7: 1 to obtain trichoderma conidium powder; mixing the trichoderma conidium powder with an auxiliary agent to obtain the microbial inoculum.
Furthermore, the Trichoderma africanum (Trichoderma afroharizianum) Ta97 has been deposited in the common microorganism center of the chinese committee for culture collection of microorganisms at 14/07/2020, the collection unit is abbreviated as CGMCC, address: xilu No.1 Hospital, Beijing, Chaoyang, North Chen, with a deposit number: CGMCC No. 19930.
Further, the trichoderma africanum Ta97 has a bacterial inhibition rate of 100% against Rhizoctonia solani (Rhizoctonia solani), 100% against alternaria alternata (aleernaria sp), 100% against Fusarium oxysporum (Fusarium oxysporum), and 79% against Fusarium moniliforme (Fusarium moniliforme).
Preferably, the seed liquid is prepared as follows:
inoculating Trichoderma africanum Ta97 preserved at-80 ℃ to a PDA solid plate, activating at 25 ℃ for 3 days, taking hyphae at the edge of a bacterial colony, inoculating to the PDA solid plate again, culturing at 25 ℃ for 3 days, taking hyphae at the edge of the bacterial colony, and culturing at 25 ℃ for 3 days to obtain an activated strain; culturing the activated strain on PDA plate at 25 deg.C under 12 hr illumination and 12 hr dark condition for 10 days, collecting spores, and preparing into 107Inoculating the conidium suspension into a PDB culture solution with the volume ratio of 1:100, and performing shake culture at 25 ℃ and 180rpm for 3-5 days to obtain a seed solution with viable count of 1 × 107～1×108cfu/mL。
Preferably, the preparation process of the trichoderma conidium powder is as follows:
inoculating a seed solution into a solid fermentation culture medium, wherein the volume mass ratio of the seed solution to the solid fermentation culture medium is 10-20: 100, uniformly stirring, culturing for 5-8 days under the conditions of constant temperature of 25-28 ℃, 12 hours of illumination and 12 hours of darkness, naturally drying, sequentially sieving by a 60-100-mesh sieve, and uniformly mixing the sieved part with diatomite according to the weight ratio of 5-7: 1 to obtain trichoderma conidium powder; in the trichoderma conidium powder, the content of spores is more than or equal to 50 hundred million/g;
the solid fermentation medium comprises the following components in parts by weight:
40-60 parts of corn flour, 40-60 parts of wheat bran, 20-40 parts of rice hull powder, 10-20 parts of glucose, (NH)4)2SO41-2 parts of KH2PO40.04 to 0.06 portion of MgSO40.005-0.01 part of CaCO30.005-0.01 part of water and 60-100 parts of water; mixing the above materials, packaging in gas-permeable bottle, sealing with sealing film, sterilizing, and cooling.
Preferably, the number of effective viable bacteria in the microbial inoculum is more than or equal to 10 hundred million/g.
Preferably, the microbial inoculum is wettable powder.
Further, the wettable powder comprises the following components in parts by weight:
10-20 parts of trichoderma conidium powder, 30-60 parts of diatomite, 2-4 parts of a wetting agent, 3-6 parts of a dispersing agent and 0.2-0.5 part of an adhesive.
The application of the microbial inoculum prepared by using the Trichoderma africanum Ta97 in the aspect of preventing and treating the continuous cropping diseases has the prevention effect on the continuous cropping diseases more than or equal to 80 percent.
The application of the microbial inoculum prepared by using the Trichoderma africanum Ta97 in the aspect of preventing and treating the continuous cropping diseases comprises the following steps: after the previous crops are harvested, uniformly spraying the microbial inoculum on the ground surface for the first time, immediately carrying out rotary tillage for about 15cm, before the next crops are sown, carrying out deep tillage for about 30cm, and applying base fertilizer and sowing in a conventional mode; when the germination rate of the next crop is more than or equal to 80 percent, the microbial inoculum is sprayed for the second time.
Compared with the prior art, the invention has the beneficial effects that:
1. according to the invention, the high-purity conidial powder with the conidial content of more than or equal to 50 hundred million/g is obtained by controlling the viable count in the seed liquid and controlling the fermentation conditions, and the wettable powder with the effective viable count of more than or equal to 10 hundred million/g is prepared, so that the control effect of the microbial inoculum on the stubble pathogens can be improved; the diatomite in the microbial inoculum not only can be used as an effective carrier, but also has the performance and the adhesion performance of improving soil, so that the microbial inoculum can be better suitable for neutralizing the surface of crops in the soil, and the control time of the microbial inoculum is prolonged.
2. In the confrontation experiment, the bacteriostasis rate of the trichoderma africanum Ta97 in the microbial inoculum is 100 percent on Rhizoctonia solani (Rhizoctonia solani), 100 percent on alternaria alternata (Alernaria sp), 100 percent on Fusarium oxysporum (Fusarium oxysporum) and 79 percent on Fusarium moniliforme (Fusarium moniliforme).
3. By using the microbial inoculum provided by the invention, in the continuous cropping of corn and wheat, the disease of wheat sharp eyespot of succeeding wheat can be reduced by 82.1-83.7%, the wheat leaf blight can be reduced by 91.7-92.7%, and the wheat root rot can be reduced by 82.6-85.2%; can reduce the disease condition of corn sheath blight disease of succeeding corn by 80.6-82.1%, reduce corn leaf blight by 81.1-88.9% and reduce corn root rot by 80.0-80.8%.
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In order to more clearly illustrate the embodiments or technical solutions in the prior art of the present invention, the drawings used in the description of the embodiments or prior art will be briefly described below, and it is obvious for those skilled in the art that other drawings can be obtained based on these drawings without creative efforts.
FIG. 1 is a plate antagonism of T.africana Ta97 against four phytopathogenic fungi of the invention;
in FIG. 1, A is Trichoderma africanum Ta97 antagonistic Rhizoctonia solani, B is Trichoderma africanum Ta97 antagonistic Alternaria alternata, C is Trichoderma africanum Ta97 antagonistic Fusarium oxysporum, and D is Trichoderma africanum Ta97 antagonistic Fusarium moniliforme.
Detailed Description
In order to make those skilled in the art better understand the technical solutions in the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1 determination of antagonism of Trichoderma africanum Ta97 against four pathogenic bacteria
(1) Source of pathogenic bacteria
Pathogenic bacteria are collected from corn-wheat multiple cropping farmlands in Jinan, Shandong province. Through morphological and molecular identification, Rhizoctonia solani (Rhizoctonia solani) is pathogenic bacteria of wheat and corn sheath blight, alternaria (Alernaria sp.) is pathogenic bacteria of wheat leaf blight and corn leaf blight, and Fusarium oxysporum (Fusarium oxysporum) and Fusarium moniliforme (Fusarium moniliforme) are pathogenic bacteria of wheat and corn root rot.
(2) Confrontation test
Respectively inoculating Trichoderma africanum Ta97 and the four pathogenic bacteria into a PDA culture medium, activating twice at 25 ℃, and after the second activation and growth for three days, punching bacterial cakes from the edges of bacterial colonies by using a 5mm puncher; placing Trichoderma africanum Ta97 and Rhizoctonia solani at two ends of the central line of the same 9cm PDA plate, and recording plate number A; placing Trichoderma africanum Ta97 and Alternaria alternata at two ends of the central line of the same 9cm PDA plate, and recording the plate number B; placing Trichoderma africanum Ta97 and Fusarium oxysporum at two ends of the center line of the same 9cm PDA plate, and recording plate number C; placing Trichoderma africanum Ta97 and Fusarium moniliforme at two ends of the center line of the same 9cm PDA plate, and recording plate number D; the plates numbered A, B, C and D were all cultured at 25 ℃ for 6-8 days, and the distance (T, cm) from the growth front of Ta97 to the pathogenic fungus cake was measured. The distance (C, cm) from the front end of the growth of the pathogenic bacteria to the pathogenic bacteria cake is measured by taking a plate cultured by four kinds of pathogenic bacteria alone as a reference. Five biological replicates were set. The formula for calculating the bacteriostasis rate is as follows: (C-T)/Cx 100%.
As can be seen by combining the attached figure 1, the bacteriostasis rate of the Trichoderma africanum Ta97 to Rhizoctonia solani is 100.00% + -0.00%, the bacteriostasis rate to Alternaria alternata is 100.00% + -0.00%, the bacteriostasis rate to Fusarium oxysporum is 100.00% + -0.00%, and the bacteriostasis rate to Fusarium moniliforme is 79.65% + -1.52%. The results show that the Trichoderma africanum Ta97 has obvious antagonistic action on the four pathogenic bacteria.
Example 2 preparation of conidia powder of Trichoderma Africa Ta97 Trichoderma
The fermentation process of the trichoderma conidium powder is as follows:
(1) obtaining a seed solution: inoculating Trichoderma africanum Ta97 preserved at-80 deg.C to PDA solid plate, activating at 25 deg.C for 3 days, and forming colonyTaking hyphae from the edge, inoculating the hyphae into a PDA plate again, culturing at 25 ℃ for 3 days, taking hyphae at the edge of a colony, and culturing at 25 ℃ for 3 days to obtain an activated strain; culturing the activated strain on PDA plate under 12 hr illumination at 25 deg.C for 10 days, collecting spores, and preparing into 107Inoculating conidium suspension into PDB culture solution at a volume ratio of 1:100, shake culturing at 25 deg.C and 180rpm for 4 days to obtain seed solution with viable count of 5 × 107cfu/mL；
(2) Obtaining conidium powder: inoculating the seed solution in the step (1) into a solid fermentation culture medium, wherein the volume mass ratio of the seed solution to the solid fermentation culture medium is 15:100, uniformly stirring, culturing for 7 days under the conditions of constant temperature of 25 ℃, 12 hours of illumination and 12 hours of darkness, naturally air-drying, and sequentially sieving by a 60-mesh sieve and a 100-mesh sieve; uniformly mixing the undersize part with diatomite according to the weight ratio of 6.6:1 to obtain trichoderma conidium powder; in the trichoderma conidium powder, the content of spores is 55 hundred million/g;
wherein the solid fermentation medium comprises the following components in parts by weight:
40 parts of corn flour, 40 parts of wheat bran, 20 parts of rice hull powder, 10 parts of glucose, (NH)4)2SO41 part of KH2PO40.04 part of MgSO 240.005 part of CaCO30.005 part and 60 parts of water; mixing the above materials, packaging in gas-permeable bottle, sealing with sealing film, sterilizing, and cooling.
Example 3 preparation of conidia powder of Trichoderma Africa Ta97 Trichoderma
(1) Obtaining a seed solution: inoculating Trichoderma africanum Ta97 preserved at-80 ℃ to a PDA solid plate, activating at 25 ℃ for 3 days, taking hyphae at the edge of a bacterial colony, inoculating to the PDA solid plate again, culturing at 25 ℃ for 3 days, taking hyphae at the edge of the bacterial colony, and culturing at 25 ℃ for 3 days to obtain an activated strain; culturing the activated strain on PDA plate under 12 hr illumination at 25 deg.C for 10 days, collecting spores, and preparing into 107Inoculating the conidium suspension into PDB culture solutionWherein the volume ratio of conidium suspension to PDB culture solution is 1:100, and shake culturing is carried out at 25 deg.C and 180rpm for 5 days to obtain seed solution with viable count of 1 × 108cfu/mL；
(2) Obtaining conidium powder: inoculating the seed solution obtained in the step (1) into a solid fermentation culture medium, wherein the volume mass ratio of the seed solution to the solid fermentation culture medium is 20:100, uniformly stirring, culturing for 8 days under the conditions of constant temperature of 27 ℃, 12 hours of illumination and 12 hours of darkness, naturally drying, sequentially sieving by 60-100 meshes, and uniformly mixing the sieved part with diatomite according to the weight ratio of 7:1 to obtain trichoderma conidium powder; in the trichoderma conidium powder, the content of spores is 57 hundred million/g;
wherein the solid fermentation medium comprises the following components in parts by weight:
50 parts of corn flour, 50 parts of wheat bran, 30 parts of rice hull powder, 15 parts of glucose, (NH)4)2SO41.5 parts of KH2PO40.05 part of MgSO 240.007 part of CaCO30.006 part of water and 80 parts of water; mixing the above materials, packaging in gas-permeable bottle, sealing with sealing film, sterilizing, and cooling.
Example 4 preparation of conidia powder of Trichoderma Africa Ta97 Trichoderma
(1) Obtaining a seed solution: inoculating Trichoderma africanum Ta97 preserved at-80 ℃ to a PDA solid plate, activating at 25 ℃ for 3 days, taking hyphae at the edge of a bacterial colony, inoculating to the PDA solid plate again, culturing at 25 ℃ for 3 days, taking hyphae at the edge of the bacterial colony, and culturing at 25 ℃ for 3 days to obtain an activated strain; culturing the activated strain on PDA plate under 12 hr illumination at 25 deg.C for 10 days, collecting spores, and preparing into 107Inoculating conidium suspension into PDB culture solution at a volume ratio of 1:100, shake culturing at 25 deg.C and 180rpm for 3 days to obtain seed solution with viable count of 1 × 107cfu/mL；
(2) Obtaining conidium powder: inoculating the seed solution obtained in the step (1) into a solid fermentation culture medium, wherein the volume mass ratio of the seed solution to the solid fermentation culture medium is 10:100, uniformly stirring, culturing for 5 days under the conditions of constant temperature at 28 ℃, 12 hours of illumination and 12 hours of darkness, naturally drying, sequentially sieving by a 60-100-mesh sieve, and uniformly mixing the sieved part with diatomite according to the weight ratio of 5:1 to obtain trichoderma conidium powder; in the trichoderma conidium powder, the content of spores is 54 hundred million/g;
wherein the solid fermentation medium comprises the following components in parts by weight:
60 parts of corn flour, 60 parts of wheat bran, 40 parts of rice hull powder, 20 parts of glucose, (NH)4)2SO42 parts of KH2PO40.06 part of MgSO (MgSO)40.01 part of CaCO30.01 part and 100 parts of water; mixing the above materials, packaging in gas-permeable bottle, sealing with sealing film, sterilizing, and cooling.
Example 5
The wettable powder comprises the following components in parts by weight:
16 parts of trichoderma conidium powder obtained in example 2, 50 parts of diatomite, 3 parts of a wetting agent, 4 parts of a dispersing agent and 0.4 part of an adhesive; the components are uniformly mixed to obtain the wettable powder, wherein the effective viable count is 12 hundred million/g.
Example 6
The wettable powder comprises the following components in parts by weight:
10 parts of trichoderma conidium powder obtained in example 3, 30 parts of diatomite, 2 parts of a wetting agent, 3 parts of a dispersing agent and 0.2 part of an adhesive; the components are uniformly mixed to obtain the wettable powder, wherein the effective viable count is 13 hundred million/g.
Example 7
The wettable powder comprises the following components in parts by weight:
20 parts of trichoderma conidium powder obtained in example 4, 60 parts of diatomite, 4 parts of a wetting agent, 6 parts of a dispersing agent and 0.5 part of a sticking agent; the components are uniformly mixed to obtain the wettable powder, wherein the effective viable count is 12 hundred million/g.
Field test
Determination of disease reduction effect of test (I) wettable powder on succeeding wheat
(1) Test site and soil characteristics
The test site is near the Huangwang farmland in the high and new area of Jinwang, Jining, Shandong province, and the corn-wheat multi-cropping continuous cropping farmland area with flat terrain and few artificial influence factors is selected, and the continuous cropping diseases are monitored every year when the corn-wheat continuous cropping plantation is carried out on the farmland all year round. The soil type is the loess, the organic matter content is about 2 percent, the total nitrogen content is about 800mg/kg, the total phosphorus content is about 700mg/kg, the total potassium content is about 18g/kg, and the pH value is about 7.
(2) Test method
Test area: each processing cell is randomly arranged with an area of 15 square meters, and the processing cells are separated by guard lines and are repeated with 3 cells.
The test set was treatment and control, specifically:
treatment group: treatment 1 (T1): the wettable powder of example 5 was used; treatment 2(T2) use the wettable powder of example 6; treatment 3 (T3): the wettable powder of example 7 was used;
control (CK): and (5) clear water control.
In the test process, the dosage of the treatment group and the control group is as follows:
treatment group: the amount of the wettable powder used per mu is 2.5-5 kg/mu; when spraying, the volume weight ratio of water to the wettable powder is 100: 0.8;
the control was the same as the treatment except that the wettable powder was not added.
The treatment groups and controls were applied separately to the test sites provided in (1) by the following procedure:
after the corn is harvested and 15 days before the wheat is planted, the wettable powder is sprayed on the ground surface according to the dosage of 5 kilograms per mu, and the spraying can be repeated, so that the uniformity of the ground surface microbial inoculum is improved; after spraying, carrying out rotary tillage on the field for 15cm, before planting wheat, carrying out deep tillage for 30cm, applying base fertilizer and sowing in a conventional mode, and carrying out conventional field nursing;
when the germination rate of the wheat is more than or equal to 80 percent, spraying the wettable powder on the overground part and the roots of the malt according to the dosage of 2.5 kilograms per mu, and investigating the occurrence conditions of sheath blight, leaf blight and root rot of each district at the flowering stage of the wheat; the control and treatment groups were in agreement; respectively counting and calculating the prevention and treatment effect, wherein the prevention and treatment effect is that (contrast disease index-treatment disease index)/treatment disease index is multiplied by 100, the disease index is that (the number of each stage of disease is multiplied by the corresponding grade value)/(the number of the total investigated plants is multiplied by the highest grade), the sheath blight disease is graded and referred to as prediction and prevention of main crop diseases and insect pests, the leaf blight disease is graded and referred to as NY/T1443.1-2007 wheat disease resistance evaluation technical specification, and the root rot disease is graded and referred to as NY/T1464.16-2007 pesticide field efficacy test criterion.
(3) Test results
The effect of preventing and controlling continuous cropping diseases is investigated on the aspect of wheat diseases in the flowering period of wheat, the results are shown in table 1,
TABLE 1 wheat flowering phase diseases
Figure BDA0002789456590000101
As can be seen by combining the table 1, in a cell sprayed with the wettable powder, the control effect on the wheat sharp eyespot is 82.1-83.7%, the control effect on the wheat leaf blight is 91.7-92.7%, and the control effect on the wheat root rot is 82.6-85.2%. For continuous cropping fields, the control effect reaches an advanced level.
Determination of disease-reducing effect of wettable powder in test (II) on succeeding corn
(1) Test site and soil characteristics
The second test and the first test are a continuous cropping planting process in one year, and the test places are the same as the first test.
(2) Test method
Test area: each processing cell is 15 square meters in area, all the processing cells are randomly arranged, guard lines are arranged between the processing cells at intervals, and each processing cell is repeated by 3 cells.
The test set was treatment and control, specifically:
treatment group: treatment 1 (T1): the wettable powder of example 5 was used; treatment 2(T2) use the wettable powder of example 6; treatment 3 (T3): the wettable powder of example 7 was used;
control (CK): and (5) clear water control.
In the test process, the dosage of the treatment group and the control group is as follows:
treatment group: 3-4 kg/mu of wettable powder for each mu; when spraying, the volume weight ratio of water to the wettable powder is 100: 1;
the control was the same as the treatment except that the wettable powder was not added.
The treatment groups and controls were applied separately to the test sites provided in (1) by the following procedure:
after the wheat is harvested, the wettable powder is sprayed on the ground surface according to the use amount of 4 kilograms per mu, and the spraying can be repeated, so that the uniformity of the ground surface microbial inoculum is improved; after spraying, carrying out rotary tillage on the field for 15cm, before planting corn, carrying out deep tillage for 30cm, applying base fertilizer and sowing in a conventional mode, and carrying out conventional field nursing;
when the germination rate of the corn is more than or equal to 80 percent, spraying the wettable powder on the corn sprouts according to the using amount of 3 kilograms per mu, and investigating the diseases of the corn in the milk stage of the corn; the control and treatment groups were in agreement; respectively counting and calculating the prevention and treatment effects, wherein the prevention and treatment effects are that the control disease index is that the treatment disease index is multiplied by 100, wherein the disease index is that the disease index is multiplied by 100, the disease number of each grade is multiplied by the corresponding grade value, the total number of the examined plants is multiplied by the highest grade, and the sheath blight disease is graded and referred to 'prediction and prevention of main crop diseases and insect pests', leaf blight and root rot disease grading and referred to 'NY/T1464.16-2007 pesticide field efficacy test criteria'.
(3) Test results
The effect of preventing and controlling continuous cropping diseases is investigated on the aspect of corn diseases in the milk stage of corn, the results are shown in table 2,
TABLE 2 maize diseases at the milk stage
Figure BDA0002789456590000121
As can be seen by combining the table 2, in the plot where the wettable powder is sprayed, the control effect on the corn sheath blight is between 80.6 and 82.1 percent, the control effect on the corn leaf blight is between 81.1 and 88.9 percent, and the control effect on the corn root rot is between 80.0 and 80.8 percent; compared with the control group, the effect is remarkable.
The combination of the test (I) and the test (II) shows that the microbial inoculum prepared by the Trichoderma africanum Ta97 can well prevent and treat continuous cropping diseases in a continuous cropping mode, and has obvious effect compared with a control. The microbial inoculum is a microbial inoculum, and the microorganisms come from soil, so that the secondary pollution to the soil can not be caused, and the microbial inoculum has the advantage of environmental protection.
Although the present invention has been described in detail by referring to the preferred embodiments, the present invention is not limited thereto. Various equivalent modifications or substitutions can be made on the embodiments of the present invention by those skilled in the art without departing from the spirit and scope of the present invention, and these modifications or substitutions are within the scope of the present invention/any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.

Claims (10)

1. A microbial inoculum prepared by using Trichoderma africanum Ta97 is characterized by comprising the following steps: the number of viable bacteria contained in the culture medium is 1 × 107～1×108Inoculating cfu/mL seed liquid into a solid fermentation culture medium, uniformly stirring, culturing for 5-8 days under the conditions of constant temperature of 25-28 ℃, light illumination for 12 hours and darkness, naturally drying, sequentially sieving by a 60-100-mesh sieve, and uniformly mixing the sieved part with diatomite according to the weight ratio of 5-7: 1 to obtain trichoderma conidium powder; mixing the trichoderma conidium powder with an auxiliary agent to obtain the microbial inoculum.
2. The microbial inoculum of claim 1, wherein Trichoderma africanum Ta97 (Trichoderma africanum) Ta97 has been deposited at the China general microbiological culture Collection center (CGMCC) at 14/07/2020, and the accession number is CGMCC: xilu No.1 Hospital, Beijing, Chaoyang, North Chen, with a deposit number: CGMCC No. 19930.
3. The microbial preparation according to claim 2, wherein the Trichoderma africanum Ta97 is used, wherein Trichoderma africanum Ta97 has a rate of 100% inhibition against Rhizoctonia solani (Rhizoctonia solani), a rate of 100% inhibition against Alernaria sp, a rate of 100% inhibition against Fusarium oxysporum (Fusarium oxysporum), and a rate of 79% inhibition against Fusarium moniliforme (Fusarium moniliforme).
4. The microbial inoculum of any of claims 1-3 prepared from Trichoderma africanum Ta97, wherein the seed solution is prepared by:
inoculating Trichoderma africanum Ta97 preserved at-80 ℃ to a PDA solid plate, activating at 25 ℃ for 3 days, taking hyphae at the edge of a bacterial colony, inoculating to the PDA solid plate again, culturing at 25 ℃ for 3 days, taking hyphae at the edge of the bacterial colony, and culturing at 25 ℃ for 3 days to obtain an activated strain; culturing the activated strain on PDA plate at 25 deg.C under 12 hr illumination and 12 hr dark condition for 10 days, collecting spores, and preparing into 107Inoculating the conidium suspension into a PDB culture solution with the volume ratio of 1:100, and performing shake culture at 25 ℃ and 180rpm for 3-5 days to obtain a seed solution with viable count of 1 × 107～1×108cfu/mL。
5. The microbial inoculum of any of claims 1-3 prepared from Trichoderma africanum Ta97, wherein Trichoderma conidium powder is prepared by the following process:
inoculating a seed solution into a solid fermentation culture medium, wherein the volume mass ratio of the seed solution to the solid fermentation culture medium is 10-20: 100, uniformly stirring, culturing for 5-8 days under the conditions of constant temperature of 25-28 ℃, 12 hours of illumination and 12 hours of darkness, naturally drying, sequentially sieving by a 60-100-mesh sieve, and uniformly mixing the sieved part with diatomite according to the weight ratio of 5-7: 1 to obtain trichoderma conidium powder; in the trichoderma conidium powder, the content of spores is more than or equal to 50 hundred million/g;
the solid fermentation medium comprises the following components in parts by weight:
40-60 parts of corn flour, 40-60 parts of wheat bran, 20-40 parts of rice hull powder, 10-20 parts of glucose, (NH)4)2SO41-2 parts of KH2PO40.04 to 0.06 portion of MgSO40.005-0.01 part of CaCO30.005-0.01 part of water and 60-100 parts of water; mixing the above materials, packaging in gas-permeable bottle, sealing with sealing film, sterilizing, and cooling.
6. The microbial inoculum of any one of claims 1-3, prepared from Trichoderma africanum Ta97, wherein the number of viable bacteria available in the microbial inoculum is 10 hundred million/g or more.
7. The microbial inoculum of claim 6 prepared from Trichoderma africanum Ta97, wherein the microbial inoculum is a wettable powder.
8. The microbial inoculum of claim 7 prepared by using Trichoderma africanum Ta97, wherein the wettable powder comprises the following components in parts by weight:
10-20 parts of trichoderma conidium powder, 30-60 parts of diatomite, 2-4 parts of a wetting agent, 3-6 parts of a dispersing agent and 0.2-0.5 part of an adhesive.
9. The application of the microbial inoculum prepared by using Trichoderma africanum Ta97 according to any one of claims 1-8 in the control of continuous cropping diseases, wherein the control effect on continuous cropping diseases is more than or equal to 80%.
10. The use as claimed in claim 9, wherein the microbial inoculum is uniformly sprayed on the ground surface for the first time after the previous crops are harvested, rotary tillage is carried out for about 15cm, deep tillage is carried out for about 30cm before the next crops are sowed, and base fertilizer is applied and sowed in a conventional manner; when the germination rate of the next crop is more than or equal to 80 percent, the microbial inoculum is sprayed for the second time.
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