A special combination agent for Solanaceae plants with disease resistance and its application method

Abstract

The invention discloses a special combination agent for Solanaceae plants with disease resistance and an application method, belonging to the technical field of Solanaceae plant cultivation. The special combination agent comprises an antagonistic composite bacterial agent added to a planting matrix, a moisture and heat resistant microbial agent added to an organic fertilizer, and oyster powder applied to the field, thereby solving the problem that the current microbial agents are difficult to use in high temperature weather.

Description

Technical Field

[0001] The invention relates to the technical field of solanaceous plant cultivation, in particular to a special combination agent for solanaceous plants with disease resistance and an application method. Background Art

[0002] A healthy soil ecosystem is fundamental to maintaining sustainable soil productivity and the foundation for sustainable agricultural development. However, in protected vegetable production systems, the long-term overuse of agrochemicals has disrupted the dynamic balance maintained by diverse soil microbial communities, significantly reducing the number of beneficial microorganisms. Furthermore, irrational production practices in protected areas have severely damaged soil structure, leading to increased soil compaction. Poor soil aeration further encourages the growth of harmful microorganisms and the production of toxins, threatening the survival of beneficial microorganisms. Beneficial microorganisms often produce antagonistic substances, competing with pathogens for ecological niches and nutrients, and inducing plant resistance. These properties help them suppress or eliminate pathogens in the soil or within plants. Therefore, the appropriate use of these microorganisms can, to a certain extent, prevent and control the occurrence of soil-borne crop diseases. Once the diversity of beneficial biological communities that support plant growth is disrupted and eliminated, plants become extremely susceptible to disease, placing significant pressure on biological control.

[0003] Solanaceae plants are one of the most important fruiting vegetables in vegetable production. However, root and stem diseases of Solanaceae vegetables primarily occur during hot seasons and are classified as high-temperature, high-humidity diseases. They primarily affect the stem base or underground main lateral roots. The diseased area begins as a dark brown color and then spreads around the stem base, causing cortical rot, yellowing and wilting of above-ground leaves, and in severe cases, the entire plant to die. The incidence rate in high-temperature, high-humidity environments is as high as 85% or more. Traditional prevention and control measures involve applying chemical root irrigation every 5-7 days for 3-5 times after seedlings are established. Root irrigation with chemical agents also increases surface humidity. While this prevents and controls the disease, it can also cause damage and seedling mortality in approximately 20%, impacting production. Although microbial antagonists can play a role in disease resistance when used to control Solanaceae plants, the use range of microbial antagonists is generally 5-35°C, the optimal use temperature is 18-30°C, and the optimal humidity is 60%. If microbial agents are used under high temperature, high humidity or low temperature conditions, they are difficult to ferment and reproduce. Even when the temperature is too high, the microorganisms will be burned, resulting in a decrease in bacterial activity. Therefore, it is necessary to find an organic compound agent with disease resistance that does not hurt seedlings and is suitable for high temperature and high humidity environments to prevent and control the occurrence of root and stem diseases, while improving the yield and quality of Solanaceae vegetables. Summary of the Invention

[0004] In view of the shortcomings of the existing technology, the purpose of the present invention is to provide a special combination agent for Solanaceae plants with disease resistance and an application method to solve the problem that current microbial agents are difficult to use in high temperature weather.

[0005] The present invention solves the above technical problems through the following technical means:

[0006] A special combination agent for Solanaceae plants with disease resistance comprises an antagonistic composite bacterial agent added to a planting matrix, a moisture and heat resistant microbial agent added to an organic fertilizer, and oyster powder applied to the field.

[0007] Furthermore, the microorganisms in the antagonistic composite bacterial agent are one or more of Bacillus polymyxa, Trichoderma harzianum, Bacillus cereus, and Bacillus subtilis.

[0008] Furthermore, the anti-humidity and heat microbial agent includes the following raw materials: composite fermentation broth, mannitol, arachidonic acid, sodium succinate, cysteine, corn flour, manganese sulfate, bentonite, and glucose. The composite fermentation broth is composed of jelly-like Bacillus fermentation broth and Bacillus licheniformis fermentation broth in a mass ratio of 1:1.

[0009] Antagonistic compound microbial agents are mixed in the planting matrix for use, and the preparation of the planting matrix can avoid use in high temperature and high humidity weather, so the antagonistic compound microbial agents do not need to be protected from moisture and heat. However, the compound microbial agents of Bacillus gelatinosa and Bacillus licheniformis added to organic fertilizers are usually used outdoors and used for the seedling stage of Solanaceae vegetables. This stage often encounters high temperature and high humidity weather, so the agents at this stage are difficult to exert their activity. Therefore, the compound microbial agents are processed to expand the applicable conditions.

[0010] Furthermore, the preparation method of the anti-humidity and heat microbial agent is as follows:

[0011] (1) Primary fermentation: Glucose, cysteine, and manganese sulfate were dissolved in water to obtain a mixed matrix, the composite fermentation broth was added, the pH was adjusted to 6.5-7, and the mixture was fermented at 35-38°C for 1 day to complete the primary fermentation;

[0012] (2) Secondary fermentation: Add mannitol and arachidonic acid to the primary fermentation solution, adjust the pH to 7-7.5, and ferment at 38-42°C for 1 day to complete the secondary fermentation;

[0013] (3) Tertiary fermentation: Mix corn flour and water and add to the secondary fermentation solution, then add sodium succinate and ferment at 42-45°C for 2 days to complete the tertiary fermentation;

[0014] (4) Preparation of microbial agent: The fermentation liquid after three fermentations was mixed with bentonite, stirred thoroughly and then dried to obtain a moisture-heat resistant microbial agent.

[0015] In addition to being related to genes, the heat resistance of Bacillus is also affected by the external environment. The present invention uses three fermentation treatments on Bacillus to improve the heat stress adaptability of microorganisms by affecting the spore flora, and gradually improves its heat resistance and moisture resistance. During the first fermentation process, the environmental pH is adjusted to 6.5-7 and the temperature is 35-38°C, so that it is in a neutral or neutral to acidic environment. Then, raw materials such as glucose, cysteine, and manganese sulfate are added to improve the thermal stability of Bacillus alkaline protein, preventing it from undergoing strong biochemical reactions in the subsequent fermentation environment, thereby causing the death of the strain. During the secondary fermentation process, stress protection substances mannitol and arachidonic acid are added to adjust the pH to a neutral or neutral to alkaline environment, and the fermentation temperature is continued to increase. The added raw materials keep the biofilm of Bacillus stable, protect the normal operation of the internal cell structure, and enable it to adapt to a higher temperature environment during the three fermentations. During the third fermentation process, sodium succinate and cornstarch are added. The cornstarch provides nutrients for the Bacillus spores, while the sodium succinate causes the cortex of the Bacillus spores to swell, increasing its water content and creating a buffer. This mitigates the effects of heat stimulation at higher temperatures, allowing the Bacillus spores to maintain a normal physiological environment even in high temperature and high humidity conditions, thus providing disease resistance. After the fermentation process is complete, the resulting fermentation liquid is mixed with bentonite and dried at low temperature to produce a powdered, heat-resistant microbial inoculant for easy use.

[0016] Furthermore, in the first fermentation step of step (1), the mass ratio of glucose, cysteine, manganese sulfate, water, and composite fermentation liquid is (0.1-0.5): (0.1-0.5): (0.01-0.05): (2-8):

[0017] (1-3).

[0018] Furthermore, the bacterial content of the jelly-like Bacillus fermentation liquid and the Bacillus licheniformis fermentation liquid in the composite fermentation liquid is not less than 1×10 5 cfu / g.

[0019] Furthermore, in the secondary fermentation step of step (2), the mass ratio of the primary fermentation bacterial liquid to mannitol and arachidonic acid is 1: (0.05-0.08): (0.1-0.2).

[0020] Furthermore, in the third fermentation step of step (3), the mass ratio of corn flour, sodium succinate, and secondary fermentation bacterial liquid is (0.5-1): (0.05-0.1):5.

[0021] Furthermore, in the bacterial agent preparation step (4), the mass ratio of the fermentation liquid to the bentonite is 10:(1-5).

[0022] The present invention also discloses a method for applying a special combination agent for Solanaceae plants with disease resistance, and the application steps are as follows:

[0023] (1) Add 100 g / ton of antagonistic compound bacteria to the substrate, mix well, and then sow the seeds of Solanaceae plants in the substrate;

[0024] If the antagonistic compound fungicide is not applied in time during the seedling stage, it can be applied by root irrigation or nest application after transplanting, but it is not suitable to be sprayed on the leaves of the seedbed. When applying, avoid direct mixing with fungicides and strong acid and alkaline substances.

[0025] (2) When turning the soil for planting, spread oyster powder at a rate of 5-100 kg / mu and plow it evenly into the ground as the soil is turned. When applying, avoid direct contact of oyster powder with the roots of the seedlings and keep a distance of more than 5-10 cm from the roots.

[0026] Preferably, when the soil pH value of the planting field is below 5.5, the application rate is 100kg / mu; when the pH value is above 5.5, the application rate is 50kg / mu; generally, when the pH value is around 7.0, the application rate is 5-10kg / mu; then plow the soil evenly;

[0027] (3) After 60 days of floating seedling cultivation in the greenhouse, the Solanaceae seedlings were planted;

[0028] (4) When planting Solanaceae plants, add anti-humidity and heat microbial agents to fully decomposed organic fertilizers, mix well and apply in strips or broadcast at a rate of 2 kg / mu;

[0029] (5) Apply Jinweiguo No. 5 during the vegetative growth period of Solanaceae plants, and apply Jinweiguo No. 7 during the fruit-bearing period of Solanaceae plants. Spray foliar fertilizer evenly on the surface of plant leaves, and the application amount is 50g / mu.

[0030] Furthermore, the solanaceous plants are tobacco, potato, tomato, eggplant, and pepper.

[0031] Furthermore, the special composition for Solanaceae plants disclosed in the present invention can be used in a high temperature climate of 35-55°C.

[0032] The beneficial effects of the present invention are as follows:

[0033] 1. The present invention improves the disease resistance of plants by mixed application of antagonistic composite bacterial agents, anti-humidity and heat microbial agents, and oyster powder, replacing chemical control, greatly reducing the incidence of root and stem diseases of Solanaceae plants, and will not cause damage to the plants, saving the expenditure of chemical agents and labor costs, and has the characteristics of safety and environmental protection.

[0034] 2. The present invention standardizes the field operation measures during the cultivation of Solanaceae plants, facilitates the production of Solanaceae vegetables to save costs and stabilize yields, meets people's living needs, realizes the new concept of green agriculture, and has good social and economic benefits.

[0035] 3. The heat-resistant microbial agent prepared by the present invention can be used in a high-temperature environment, and is particularly suitable for use in areas with high heat and high humidity in summer, such as Chongqing. The activity of the agent will not be reduced in a high-temperature environment, and the yield and quality of Solanaceae vegetables can be improved while preventing and controlling root and stem diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 It is the planting base of Experiment 2 beauty pepper. DETAILED DESCRIPTION

[0037] The present invention will be described in detail below with reference to specific embodiments and accompanying drawings:

[0038] Example 1: Preparation of a special combination agent for Solanaceae plants

[0039] The microorganisms in the antagonistic composite bacterial agent are Bacillus polymyxa, Trichoderma harzianum, and Bacillus cereus, with a mass ratio of 1:1:1.

[0040] Oyster powder: mainly obtained by burning and grinding oyster shells into powder and passing through a 230-325 mesh sieve.

[0041] The anti-humidity and heat microbial agent is prepared by itself. The specific raw materials and preparation steps are as follows:

[0042] (1) Primary fermentation: 0.1 kg of glucose, 0.1 kg of cysteine, and 0.01 kg of manganese sulfate were mixed and dissolved in 2 kg of water to obtain a mixed matrix, 1 kg of the composite fermentation liquid was added, the pH was adjusted to 6.5, and the mixture was fermented at 35-38° C. for 1 day to complete the primary fermentation;

[0043] The composite fermentation liquid used is a mixture of jelly-like Bacillus fermentation liquid and Bacillus licheniformis fermentation liquid in a mass ratio of 1:1, wherein the bacterial content of the jelly-like Bacillus fermentation liquid and the Bacillus licheniformis fermentation liquid is not less than 1×10 5 cfu / g.

[0044] (2) Secondary fermentation: 0.15 kg of mannitol and 0.3 kg of arachidonic acid were added to 3 kg of the primary fermentation solution, the pH was adjusted to 7, and the mixture was fermented at 38-42°C for 1 day to complete the secondary fermentation.

[0045] (3) Tertiary fermentation: 0.3 kg corn flour was mixed with 0.5 kg water and added to 3 kg of the bacterial solution from the secondary fermentation. 0.03 kg of sodium succinate was then added and fermented at 42-45 °C for 2 days to complete the tertiary fermentation.

[0046] (4) Preparation of microbial agent: 3 kg of fermentation liquid after three fermentations was mixed with 1.5 kg of bentonite, stirred thoroughly and then dried at a low temperature of 30-35 ° C to obtain a moisture-resistant microbial agent.

[0047] Example 2: Preparation of a special combination agent for Solanaceae plants

[0048] The microorganisms in the antagonistic composite bacterial agent are Bacillus polymyxa, Trichoderma harzianum, and Bacillus cereus, with a mass ratio of 1:1:1.

[0049] Oyster powder: mainly obtained by burning and grinding oyster shells into powder and passing through a 230-325 mesh sieve.

[0050] The anti-humidity and heat microbial agent is prepared by itself. The specific raw materials and preparation steps are as follows:

[0051] (1) Primary fermentation: 0.5 kg of glucose, 0.5 kg of cysteine, and 0.05 kg of manganese sulfate were mixed and added to 8 kg of water to dissolve to obtain a mixed matrix, 3 kg of the composite fermentation liquid was added, and the mixture was adjusted to pH = 7 and fermented at 35-38° C. for 1 day to complete the primary fermentation;

[0052] The composite fermentation liquid used is a mixture of jelly-like Bacillus fermentation liquid and Bacillus licheniformis fermentation liquid in a mass ratio of 1:1, wherein the bacterial content of the jelly-like Bacillus fermentation liquid and the Bacillus licheniformis fermentation liquid is not less than 1×10 5 cfu / g.

[0053] (2) Secondary fermentation: 0.12 kg of mannitol and 0.4 kg of arachidonic acid were added to 2 kg of the primary fermentation solution, the pH was adjusted to 7.5, and the mixture was fermented at 38-42°C for 1 day to complete the secondary fermentation.

[0054] (3) Tertiary fermentation: 0.4 kg corn flour was mixed with 0.8 kg water and added to 2 kg of the bacterial solution from the secondary fermentation. 0.4 kg sodium succinate was then added and fermented at 42-45°C for 2 days to complete the tertiary fermentation.

[0055] (4) Preparation of microbial agent: 2 kg of fermentation liquid after three fermentations was mixed with 0.5 kg of bentonite, stirred thoroughly and then dried at low temperature at 30-35 °C to obtain a moisture-heat resistant microbial agent.

[0056] Example 3: Preparation of a special combination agent for Solanaceae plants

[0057] The microorganisms in the antagonistic composite bacterial agent are Bacillus polymyxa, Trichoderma harzianum, and Bacillus cereus, with a mass ratio of 1:1:1.

[0058] Oyster powder: mainly obtained by burning and grinding oyster shells into powder and passing through a 230-325 mesh sieve.

[0059] The anti-humidity and heat microbial agent is prepared by itself. The specific raw materials and preparation steps are as follows:

[0060] (1) Primary fermentation: 0.3 kg of glucose, 0.3 kg of cysteine, and 0.03 kg of manganese sulfate were mixed and added to 6 kg of water to dissolve to obtain a mixed matrix, 2 kg of the composite fermentation liquid was added, and the mixture was adjusted to pH = 7 and fermented at 35-38° C. for 1 day to complete the primary fermentation;

[0061] The composite fermentation liquid used is a mixture of jelly-like Bacillus fermentation liquid and Bacillus licheniformis fermentation liquid in a mass ratio of 1:1, wherein the bacterial content of the jelly-like Bacillus fermentation liquid and the Bacillus licheniformis fermentation liquid is not less than 1×10 5 cfu / g.

[0062] (2) Secondary fermentation: 0.16 kg of mannitol and 0.3 kg of arachidonic acid were added to 2 kg of the primary fermentation solution, the pH was adjusted to 7.5, and the mixture was fermented at 38-42°C for 1 day to complete the secondary fermentation.

[0063] (3) Tertiary fermentation: 0.1 kg corn flour was mixed with 0.2 kg water and added to 0.5 kg of the bacterial solution from the secondary fermentation. 0.01 kg of sodium succinate was then added and fermented at 42-45 °C for 2 days to complete the tertiary fermentation.

[0064] (4) Preparation of microbial agent: 0.5 kg of fermentation liquid after three fermentations was mixed with 0.25 kg of bentonite, stirred thoroughly and then dried at a low temperature of 30-35 ° C to obtain a moisture-resistant microbial agent.

[0065] Comparative Example 1:

[0066] The present invention is in contrast to the heat-resistant microbial agent prepared in Example 1. The main difference in the preparation of the microbial agent is that the present comparative example does not undergo two or three fermentations, but only one fermentation. The specific preparation steps are as follows:

[0067] (1) Primary fermentation: 0.1 kg of glucose, 0.1 kg of cysteine, and 0.01 kg of manganese sulfate were mixed and dissolved in 2 kg of water to obtain a mixed matrix, 1 kg of the composite fermentation liquid was added, and the mixture was adjusted to pH = 6.5 and fermented at 35°C for 1 day to complete the primary fermentation;

[0068] (2) Preparation of microbial agent: 3 kg of fermentation liquid after one fermentation was mixed with 1.5 kg of bentonite, stirred thoroughly and then dried at low temperature at 30-35°C to obtain a microbial agent.

[0069] Comparative Example 2:

[0070] The present invention is in contrast to the heat-resistant microbial agent prepared in Example 1. The main difference in the preparation of the microbial agent is that the present comparative example does not perform the first or third fermentation, but only the second fermentation step. The specific preparation steps are as follows:

[0071] (1) Fermentation: 0.15 kg of mannitol and 0.3 kg of arachidonic acid were added to 3 kg of the composite fermentation liquid, the pH was adjusted to 7, and the mixture was fermented at 42° C. for 1 day to complete the primary fermentation.

[0072] (2) Preparation of microbial agent: 3 kg of fermentation liquid after fermentation was mixed with 1.5 kg of bentonite, stirred thoroughly and then dried at low temperature at 30-35 °C to obtain a microbial agent.

[0073] Comparative Example 3:

[0074] The present invention is in contrast to the heat-resistant microbial agent prepared in Example 1. The main difference in the preparation of the microbial agent is that the present comparative example does not perform the first or second fermentation, but only performs the third fermentation step. The specific preparation steps are as follows:

[0075] (1) Fermentation: 0.3 kg corn flour and 0.5 kg water were mixed and added to 3 kg of composite fermentation liquid, and then 0.03 kg of sodium succinate was added. The mixture was fermented at 45° C. for 2 days to complete the fermentation.

[0076] (2) Preparation of microbial agent: 3 kg of fermentation liquid after three fermentations was mixed with 1.5 kg of bentonite, stirred thoroughly and then dried at a low temperature of 30-35°C to obtain a microbial agent.

[0077] Comparative Example 4:

[0078] The present invention is in contrast to the heat-resistant microbial agent prepared in Example 1. The main difference in the preparation of the microbial agent is that the pH is kept neutral during the preparation process of this comparative example. The specific preparation steps are as follows:

[0079] (1) Primary fermentation: 0.1 kg of glucose, 0.1 kg of cysteine, and 0.01 kg of manganese sulfate were mixed and dissolved in 2 kg of water to obtain a mixed matrix, 1 kg of the composite fermentation liquid was added, the pH was adjusted to 7, and the mixture was fermented at 35-38° C. for 1 day to complete the primary fermentation;

[0080] (2) Secondary fermentation: 0.15 kg of mannitol and 0.3 kg of arachidonic acid were added to 3 kg of the primary fermentation solution, the pH was adjusted to 7, and the mixture was fermented at 38-42°C for 1 day to complete the secondary fermentation.

[0081] (3) Tertiary fermentation: 0.3 kg corn flour was mixed with 0.5 kg water and added to 3 kg of the bacterial solution from the secondary fermentation. 0.03 kg of sodium succinate was then added and fermented at 42-45 °C for 2 days to complete the tertiary fermentation.

[0082] (4) Preparation of microbial agent: 3 kg of fermentation liquid after three fermentations was mixed with 1.5 kg of bentonite, stirred thoroughly and then dried at a low temperature of 30-35 ° C to obtain a moisture-resistant microbial agent.

[0083] Comparative Example 5:

[0084] The present invention is in contrast to the heat-resistant microbial agent prepared in Example 1. The main difference in the preparation of the microbial agent is that the temperature is not adjusted during the preparation process of this comparative example, and the temperature is kept constant at 38-42°C during the fermentation process. The specific preparation steps are as follows:

[0085] (1) Primary fermentation: 0.1 kg of glucose, 0.1 kg of cysteine, and 0.01 kg of manganese sulfate were mixed and dissolved in 2 kg of water to obtain a mixed matrix, 1 kg of the composite fermentation liquid was added, and the mixture was adjusted to pH = 6.5 and fermented at 38-42° C. for 1 day to complete the primary fermentation;

[0086] (2) Secondary fermentation: 0.15 kg of mannitol and 0.3 kg of arachidonic acid were added to 3 kg of the primary fermentation solution, the pH was adjusted to 7, and the mixture was fermented at 38-42°C for 1 day to complete the secondary fermentation.

[0087] (3) Tertiary fermentation: 0.3 kg corn flour was mixed with 0.5 kg water and added to 3 kg of the bacterial solution from the secondary fermentation. 0.03 kg of sodium succinate was then added and fermented at 38-42 °C for 2 days to complete the tertiary fermentation.

[0088] (4) Preparation of microbial agent: 3 kg of fermentation liquid after three fermentations was mixed with 1.5 kg of bentonite, stirred thoroughly and then dried at a low temperature of 30-35 ° C to obtain a moisture-resistant microbial agent.

[0089] Comparative Example 6:

[0090] The present invention is in contrast to the heat-resistant microbial agent prepared in Example 1. The main difference in the preparation of the microbial agent is that cysteine ​​is not added during the first fermentation process of this comparative example. The subsequent preparation process is the same. The specific preparation steps are as follows:

[0091] (1) Primary fermentation: 0.1 kg of glucose was added to 2 kg of water to dissolve to obtain a mixed matrix, 1 kg of the composite fermentation liquid was added, the pH was adjusted to 6.5, and the mixture was fermented at 35-38°C for 1 day to complete the primary fermentation;

[0092] Steps (2)-(4) are the same.

[0093] Comparative Example 6:

[0094] The present invention is in contrast to the heat-resistant microbial agent prepared in Example 1. The main difference in the preparation of the microbial agent is that mannitol is not added during the secondary fermentation process of this comparative example. The subsequent preparation process is the same. The specific preparation step (2) secondary fermentation is as follows:

[0095] (2) Secondary fermentation: Add 0.3 kg of arachidonic acid to 3 kg of the primary fermentation solution, adjust the pH to 7, and ferment at 38-42°C for 1 day to complete the secondary fermentation;

[0096] Steps (1), (3), and (4) are the same.

[0097] Comparative Example 7:

[0098] The present invention is in contrast to the heat-resistant microbial agent prepared in Example 1. The main difference in the preparation of the microbial agent is that sodium succinate is not added during the three fermentation processes of this comparative example. The subsequent preparation process is the same. The specific preparation step (3) of the second fermentation is as follows:

[0099] (3) Tertiary fermentation: 0.3 kg corn flour was mixed with 0.5 kg water and added to 3 kg of the bacterial solution from the secondary fermentation. The mixture was fermented at 42-45 °C for 2 days to complete the tertiary fermentation.

[0100] Steps (1), (2) and (4) are the same.

[0101] Experiment 1:

[0102] A heat resistance comparison experiment was conducted on Comparative Examples 1-7 and Example 1. The specific experimental process is as follows:

[0103] (1) Take 10 g each of the anti-heat and humidity microbial agent prepared in Example 1 and the microbial agents prepared in Comparative Examples 1-7;

[0104] (2) Add 0.2g of brown sugar to 100g of clear water (about 30°C) after sun exposure, stir and split, then add the above-mentioned bacterial agents respectively, and oxygenate with an aerator pump. After 2-4 hours, the bacteria will be revived;

[0105] (3) Pick a ring of activated bacteria and inoculate it into 100 ml of liquid culture medium (the components and weight ratios of the liquid culture medium are: glucose 5 g, yeast extract 5 g, peptone 10 g, sodium chloride 5 g, water 1000 mL), shake culture at 30°C for 12 hours to obtain seed liquid, then take 5% volume of the seed liquid and inoculate it into fermentation medium (the components and weight ratios of the fermentation medium are: glucose 20 g, soluble starch 40 g, peptone 1 g, NaNO3 10 g, K2HPO4 3 g, MgSO4·7H2O 1 g, CaCO3 1 g, water 1000 mL), shake culture at 50°C for 48 hours, then take out 10 mL for staining to observe the spore formation, and stop fermentation when the spore formation rate is greater than 90% to obtain fermentation liquid;

[0106] (4) The fermentation broth was diluted in 10-fold gradients to 10 -n Divide the LB culture dish into four sections and apply 20 μL of the dilution to each section. Incubate the dish at 35°C, 45°C, and 55°C for 8 hours. Count the colonies in each section, aiming for a concentration of 10-40 colonies. Calculate the number of spores per mL of fermentation broth and observe the number of surviving spores. The data obtained are as follows:

[0107] Table 1

[0108]

[0109] According to the data in Table 1, the heat-resistant microbial agents prepared in Example 1 and Comparative Examples 1-7 had good activity when cultured at 35°C. However, when the culture temperature was 45°C, the number of spores in Comparative Examples 1-7 decreased, with the decrease in Comparative Examples 1, 2, and 3 being greater, indicating that the composite agent without the first, second, and third fermentation steps had poor heat resistance. When cultured at 55°C, the number of spores in Comparative Examples 1-7 decreased by an order of magnitude, indicating that the activity of Bacillus at 55°C was poor and that it could not reproduce well. This shows that after the treatment of the present invention, Bacillus can obtain better heat resistance and is suitable for preventing and controlling the occurrence of rhizome diseases at high temperatures.

[0110] Example 4: Application method of special combination agent for Solanaceae plants

[0111] (1) Cultivating Solanaceae plant seedlings using a floating seedling cultivation method, adding an antagonistic compound bacterial agent to the substrate at a rate of 100 g / ton, mixing the mixture evenly, and then sowing Solanaceae plant seeds in the substrate;

[0112] If the antagonistic compound fungicide is not applied in time during the seedling stage, it can be applied by root irrigation or nest application after transplanting, but it is not suitable to be sprayed on the leaves of the seedbed. When applying, avoid direct mixing with fungicides and strong acid and alkaline substances.

[0113] (2) When turning the soil for planting, spread oyster powder at a rate of 5-100 kg / mu and plow it evenly into the ground as the soil is turned. When applying, avoid direct contact of oyster powder with the roots of the seedlings and keep a distance of more than 5-10 cm from the roots.

[0114] Preferably, when the soil pH value of the planting field is below 5.5, the application rate is 100kg / mu; when the pH value is above 5.5, the application rate is 50kg / mu; generally, when the pH value is around 7.0, the application rate is 5-10kg / mu; then plow the soil evenly;

[0115] (3) After 60 days of floating seedling cultivation in the greenhouse, plant the Solanaceae seedlings in the afternoon of a sunny day or on a cloudy day. The requirements for planting the seedlings are: shallowly so that the roots are not exposed, deep enough so that the leaves are not buried, and leaves refer to cotyledons.

[0116] (4) When planting Solanaceae plants, add anti-humidity and heat microbial agents to fully decomposed organic fertilizers, mix well and apply in strips or broadcast at a rate of 2 kg / mu;

[0117] (5) Apply Jinweiguo No. 5 during the vegetative growth period of Solanaceae plants, and apply Jinweiguo No. 7 during the fruit-bearing period of Solanaceae plants. Spray foliar fertilizer evenly on the surface of plant leaves, and the application amount is 50g / mu.

[0118] The special combination of the present invention is best used for solanaceous plants, mainly including tobacco, potato, pepper, eggplant, and pepper.

[0119] Experiment 2:

[0120] A planting experiment was conducted on the special combination prepared in Example 1. The specific experimental process is as follows:

[0121] 1. Test variety: Capsicum annuum (Pepper); Location: Beibei District, Chongqing;

[0122] Planting period: March 2022 - October 2022; Maximum temperature: 44.6°C, Extreme temperature difference: 23°C, Maximum ground temperature: 51°C, Relative humidity: 70-80%;

[0123] Second, the experiment was set up with three treatments, randomly arranged, and replicated three times. The area of ​​each plot was 0.1 mu;

[0124] Treatment 1: antagonistic composite bacterial agent + heat-resistant microbial agent prepared in Example 1 + oyster powder;

[0125] Treatment 2: antagonistic compound bacterial agent + commercial rhizome rehabilitation compound bacterial agent (Bacillus amyloliquefaciens, Paenibacillus gelatinosa, Brevibacillus laterosporus, effective viable count ≥ 2 billion / g) + oyster powder;

[0126] Treatment 3: organic fertilizer (application rate is 2 tons / mu).

[0127] 4. Planting process:

[0128] (1) Peppers were grown using a floating seedling method. Antagonistic compound bacteria or farmyard manure was added to a common pepper cultivation substrate at a rate of 100 g / ton, mixed evenly, and pepper seeds were then sown in the substrate.

[0129] (2) When turning the soil for planting, spread oyster powder or farmyard manure at a rate of 10 kg / mu and evenly plow it into the ground as the soil is turned;

[0130] (3) After 60 days of floating seedling cultivation in the greenhouse, pepper seedlings are transplanted in the afternoon of a sunny day or on a cloudy day, with a planting density of about 5,000 plants per mu. When transplanting seedlings, the requirements are: shallowly so that the roots are not exposed, deep enough so that the leaves are not buried, and leaves refer to cotyledons;

[0131] (4) When planting Solanaceae plants, add anti-heat and humidity microbial agents or rhizome rehabilitation synthetic agents or farmyard manure to fully decomposed organic fertilizer, mix well and apply in strips or broadcast at a rate of 2 kg / mu;

[0132] (5) Apply Jinweiguo No. 5 during the growth period of peppers and Jinweiguo No. 7 during the fruiting period of Solanaceae plants. Spray foliar fertilizer evenly on the surface of plant leaves, and the application amount is 50g / mu.

[0133] Field management during the planting period was carried out according to conventional pepper planting management methods.

[0134] 5. Output is shown in Table 2:

[0135] Table 2

[0136]

[0137] analyze:

[0138] Treatment 1 used a combination of antagonistic composite bacteria + the heat-resistant microbial agent prepared in Example 1 + oyster powder to grow peppers. Compared with treatment 2 using an antagonistic composite bacteria + a commercially available rhizome rehabilitation compound bacteria + oyster powder, the yield per mu increased by 188.3 kg, and compared with treatment 3 farmyard manure, it increased by 530.7 kg. Although extremely high temperatures were encountered during the experiment, treatment 1 was still high-yielding, indicating that the heat-resistant microbial agent prepared by the present invention can be used in a high-temperature environment, and is particularly suitable for use in areas with high-temperature and high-humidity environments.

[0139] The above embodiments are intended to illustrate the technical solutions of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art will appreciate that the technical solutions of the present invention may be modified or replaced with equivalents without departing from the spirit and scope of the technical solutions of the present invention, and such modifications or equivalents shall be encompassed by the claims of the present invention. Any techniques, shapes, and structures not described in detail herein are well known.

Claims

1. A special combination agent for Solanaceae plants with disease resistance, characterized in that: The special combination agent is composed of an antagonistic composite bacterial agent added to the planting matrix, a moisture and heat resistant microbial agent added to the organic fertilizer, and oyster powder applied to the field; The preparation method of the anti-humidity and heat microbial agent is as follows: (1) Primary fermentation: Glucose, cysteine, and manganese sulfate were dissolved in water to obtain a mixed matrix, the composite fermentation broth was added, the pH was adjusted to 6.5-7, and the mixture was fermented at 35-38°C for 1 day to complete the primary fermentation. (2) Secondary fermentation: Add mannitol and arachidonic acid to the primary fermentation solution, adjust the pH to 7-7.5, and ferment at 38-42°C for 1 day to complete the secondary fermentation; (3) Tertiary fermentation: Mix corn flour and water and add it to the bacterial solution of the secondary fermentation, then add sodium succinate and ferment at 42-45°C for 2 days to complete the tertiary fermentation; (4) Preparation of microbial agent: The fermentation liquid after three fermentations is mixed with bentonite, stirred thoroughly and then dried to obtain a moisture-heat resistant microbial agent; The composite fermentation liquid consists of a jelly-like Bacillus fermentation liquid and a Bacillus licheniformis fermentation liquid in a mass ratio of 1:

1.

2. The special combination agent for Solanaceae plants with disease resistance according to claim 1, characterized in that: The microorganisms in the antagonistic composite bacterial agent are one or more of Bacillus polymyxa, Trichoderma harzianum, Bacillus cereus and Bacillus subtilis.

3. The special combination agent for Solanaceae plants with disease resistance according to claim 2, characterized in that: In the first fermentation step of step (1), the mass ratio of glucose, cysteine, manganese sulfate, water, and composite fermentation liquid is (0.1-0.5): (0.1-0.5): (0.01-0.05): (2-8): (1-3).

4. The special combination agent for Solanaceae plants with disease resistance according to claim 3, characterized in that: In the secondary fermentation step of step (2), the mass ratio of the primary fermentation bacterial liquid to mannitol and arachidonic acid is 1: (0.05-0.08): (0.1-0.2).

5. The special combination agent for Solanaceae plants with disease resistance according to claim 4, characterized in that: In the third fermentation step of step (3), the mass ratio of corn flour, sodium succinate, and secondary fermentation bacterial liquid is (0.5-1): (0.05-0.1):

5.

6. The special combination agent for Solanaceae plants with disease resistance according to claim 5, characterized in that: In the bacterial agent preparation step (4), the mass ratio of the fermentation liquid to the bentonite is 10:(1-5).

7. The method for applying the disease-resistant combination agent for Solanaceae plants according to claim 1, characterized in that: The application steps are as follows: (1) Add antagonistic compound bacteria to the substrate at 100 g / ton, mix well, and then sow the seeds of Solanaceae plants in the substrate; (2) When turning over the soil for planting, spread oyster powder at a rate of 5-100 kg / mu; (3) After 60 days of floating seedling cultivation in the greenhouse, the Solanaceae seedlings are planted; (4) When planting Solanaceae plants, add anti-humidity and heat microbial agents to fully decomposed organic fertilizers, mix them evenly, and then apply them in strips or broadcast at a rate of 2 kg / mu; (5) Apply Jinweiguo No. 5 during the vegetative growth period of Solanaceae plants, and apply Jinweiguo No. 7 during the fruit-bearing period of Solanaceae plants. Spray foliar fertilizer evenly on the surface of plant leaves, and the application amount is 50g / mu.

8. The method for applying the disease-resistant combination agent for Solanaceae plants according to claim 7, characterized in that: The solanaceous plants are tobacco, potato, tomato, eggplant and pepper.

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