Compound microbial agent and application thereof
Through the application of compound microbial agents, the problem of prevention and control of tobacco root and stem diseases has been solved, the agronomic traits and disease resistance of tobacco have been improved, environmentally friendly and efficient disease control has been achieved, the healthy growth of tobacco has been promoted and environmental pollution has been reduced.
Patent Information
- Application Number
- CN202510613460.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-05-13
- Publication Date
- 2025-09-12
AI Technical Summary
Existing technologies are difficult to effectively prevent and control tobacco root and rhizome diseases, and the long-term use of chemical pesticides has led to increased pathogen resistance and serious environmental pollution. There is an urgent need to develop environmentally friendly and efficient biological control methods.
Composite agent 1 is prepared by combining microbial agents including methylotrophic Bacillus, Streptomyces albus, Bacillus halodurans, Streptomyces oryzae and Bacillus carsonii with carriers such as corn flour, diatomaceous earth, peat, and coffee grounds. Composite agent 2 is prepared by combining seaweed polysaccharide, soluble starch, fulvic acid, Streptomyces oryzae and Bacillus carsonii. These agents are applied to tobacco in the seedbed stage and the field stage, respectively, to promote tobacco growth and prevent and control diseases.
Significantly improve tobacco agronomic traits, enhance disease resistance, effectively control tobacco root and stem diseases, reduce economic losses, and achieve green and sustainable development of tobacco cultivation.
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Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of biological control, and in particular to a composite microbial agent and application thereof in preventing and controlling tobacco root and stem diseases. Background Art
[0002] With global population growth and increasing scarcity of arable land resources, improving crop yield and quality has become a key goal of agricultural production. In this context, tobacco, as a key cash crop, is crucial for the development of the agricultural economy. However, tobacco is susceptible to various diseases, which significantly reduce its yield and quality, resulting in significant economic losses worldwide each year.
[0003] Among various tobacco diseases, rhizome diseases are one of the important factors restricting tobacco development. Common tobacco rhizome diseases include tobacco bacterial wilt, tobacco black shank, tobacco root rot, tobacco anthrax, tobacco sclerotinia, etc. These diseases harm the roots, stems and even leaves of tobacco, causing serious symptoms in tobacco, and posing a serious threat to the healthy growth and yield quality of tobacco.
[0004] Traditional methods for controlling tobacco root and stem diseases primarily rely on chemical pesticides. However, long-term excessive use of chemical pesticides not only leads to increased pathogen resistance and reduced efficacy, but also causes environmental pollution and ecological damage. Therefore, the development of new, environmentally friendly and efficient control methods has become a hot topic in current agricultural science and technology research. In recent years, microbial agents and fertilizers, as a new type of biological control method, have attracted widespread attention due to their environmental friendliness, high efficiency, safety, convenience, and economy. Although microbial agents and fertilizers have shown great potential in the prevention and control of tobacco diseases, tobacco root and stem diseases have not yet been effectively controlled. Their actual application effect is affected by many factors, and further research and development are urgently needed to achieve effective prevention and control of tobacco root and stem diseases, thereby achieving the green and sustainable development of the tobacco planting industry. Summary of the Invention
[0005] The purpose of the present invention is to provide a composite microbial agent for preventing and treating tobacco root and stem diseases and improving the agronomic traits of tobacco. In order to achieve the above-mentioned object, the present invention provides a composite microbial agent, which comprises a composite microbial agent 1 and a composite microbial agent 2, wherein the composite microbial agent 1 comprises a methylotrophic Bacillus with a preservation number of CCTCCNO: M 20221661, a white-flavor Streptomyces with a preservation number of CCTCCNO.M 20231644, and a halotolerant Bacillus with a preservation number of CCTCCNO: M20232276, and the composite microbial agent 2 comprises a blastomyces Streptomyces with a preservation number of CCTCCNO.M 20231643 and a carinii Bacillus with a preservation number of CCTCCNO: M 20221660.
[0006] Preferably, the composite bacterial agent 1 further comprises corn flour, diatoms, peat, and coffee grounds, wherein the contents of methylotrophic Bacillus, Streptomyces albus, and Bacillus halophilus are not less than 1×10 8 cfu / g.
[0007] The present invention also provides a more preferred composite bacterial agent 1, which is solid and comprises, by weight: 10% methylotrophic Bacillus, 10% Streptomyces alboflavin, 10% halodurans, 20% corn flour, 10% diatomaceous earth, 20% peat, and 20% coffee grounds.
[0008] Preferably, the composite bacterial agent 2 further comprises seaweed polysaccharide, soluble starch, and fulvic acid, wherein the content of Streptomyces oryzae and Bacillus carsnerii is not less than 1×10 9 cfu / mL.
[0009] The present invention also provides a more preferred composite bacterial agent 2, which is a liquid and contains: seaweed polysaccharide with a mass concentration of 3 g / L, soluble starch with a mass concentration of 2 g / L, and fulvic acid with a mass concentration of 3 g / L.
[0010] The composite microbial agent provided by the present invention can be used in tobacco cultivation, including improving tobacco agronomic traits and preventing and controlling tobacco root and stem diseases. In particular, it can be used to improve tobacco plant height, stem girth, leaf number, maximum leaf length, and maximum leaf width. It can also be used to prevent and control tobacco root and stem diseases such as tobacco root rot, tobacco black shank disease, tobacco bacterial wilt, and tobacco sclerotinia rot.
[0011] The present invention also provides a method for improving the agronomic traits of tobacco and preventing and controlling tobacco rhizome diseases. By applying the above-mentioned composite microbial agent during the seedbed stage and field stage of tobacco, the agronomic traits of tobacco and the prevention and control of tobacco rhizome diseases can be improved, including tobacco plant height, stem girth, number of leaves, maximum leaf length and maximum leaf width. It can also be used for the prevention and control of tobacco rhizome diseases such as tobacco root rot, tobacco black shank disease, tobacco bacterial wilt, and tobacco sclerotinia disease, and has potential application value in the field of tobacco cultivation.
[0012] The present invention has the following advantages: The present invention provides a composite microbial agent comprising five bacterial strains. Furthermore, a composite agent 1 is prepared by mixing carriers such as corn flour, diatomaceous earth, peat, and coffee grounds with methylotrophic Bacillus, Streptomyces albus, and Bacillus halogenus. The composite agent 1 utilizes the protective and adsorption effects of different carrier combinations on the microorganisms, thereby extending the actual use activity of the microorganisms, enabling the microorganisms to have a wider adaptability in practical applications and fully exert their functions. This microbial fertilizer can promote various agronomic traits of tobacco and enhance soil vitality during both the seedbed and field stages, thereby enhancing the disease resistance of tobacco plants.
[0013] The present invention proposes mixing seaweed polysaccharides, soluble starch, and fulvic acid with Streptomyces oryzae and Bacillus carlesii to form a composite microbial agent 2. The carrier protects the microorganisms, allowing them to function effectively for a longer period of time, enhancing their practical application. Furthermore, the microbial agent can effectively control and prevent the occurrence and invasion of various tobacco root and stem diseases, effectively reducing economic losses in tobacco cultivation.
[0014] The present invention targets the pathogenic species diversity, pathogenesis and crop defense mechanism of various tobacco rhizome diseases, and combines the selected different strains to have a synergistic effect, which helps promote the healthy growth of tobacco and enhance its resistance to diseases, thereby effectively controlling the occurrence and prevalence of rhizome diseases. DETAILED DESCRIPTION
[0015] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.
[0016] Note: Unless otherwise noted, the experimental methods in the following examples are conventional methods, performed according to the techniques and conditions described in literature in the field or according to product specifications. Materials and reagents used in the following examples, unless otherwise noted, are commercially available.
[0017] Example 1 Preparation of composite microbial agent and its use in the prevention and treatment of tobacco diseases This experimental example provides a composite microbial agent, which contains five strains of bacteria, namely: Bacillus halodurans with a preservation number of CCTCCNO: M 20232276, Streptomyces alboflavin with a preservation number of CCTCCNO.M 20231644, Streptomyces oryzae with a preservation number of CCTCCNO.M 20231643, Bacillus carlesii with a preservation number of CCTCCNO: M 20221660, and Bacillus methylotrophicus with a preservation number of CCTCCNO: M 20221661.
[0018] The composite microbial agent comprises composite agent 1 and composite agent 2, which are specifically as follows: The composite bacterial agent 1 is a solid, and contains, by weight, 10% methylotrophic Bacillus, 10% Streptomyces alboflavin, 10% halodurans, 20% corn flour, 10% diatomaceous earth, 20% peat, and 20% coffee grounds, wherein the contents of halodurans, methylotrophic Bacillus, and Streptomyces alboflavin are not less than 1×10 8 cfu / g.
[0019] The composite microbial agent 2 is a liquid containing: seaweed polysaccharide, soluble starch, and fulvic acid at a mass concentration of 3 g / L, respectively; Streptomyces oryzae and Bacillus carlesii; the contents of Streptomyces oryzae and Bacillus carlesii are not less than 1×10 9 cfu / mL.
[0020] This embodiment provides a method for applying a composite microbial agent for preventing and treating tobacco root and stem diseases, as follows: During the tobacco seedling bed stage: add 1.0~2kg of compound fungicide 1 per cubic meter of seedling substrate; and / or, when the 4th leaf of the tobacco seedling grows out, apply 1.0~1.5kg of compound fungicide 1 per 100 square meters; and / or, when the 7th leaf of the tobacco seedling grows out, apply 1.5-2kg of compound fungicide 1 per 100 square meters; and / or, within 15 days before transplanting, apply 1.0~2.0kg of compound fungicide 1 per 100 square meters; "and" indicates that in addition to adding 1.0~2kg of compound fungicide 1 per cubic meter of seedling substrate, it is also applied in other stages; "or" indicates that after adding 1.0~2kg of compound fungicide 1 per cubic meter of seedling substrate, it can be applied or not in other stages.
[0021] Starting from the cross stage of tobacco seedlings, spray compound fungicide 2 at 200mL / mu every 5 days for a total of 3 times.
[0022] During the tobacco field stage: apply compound microbial agent 1 at a rate of 300-400 kg / mu when forming ridges; and / or, apply compound microbial agent 1 at a rate of 150-200 kg / mu 10-25 days after transplanting, and apply 1-2 times after transplanting; Before and after the tobacco clumping stage, spray the compound bacterial agent 2 at a dosage of 200 mL / mu, once; before and after the tobacco budding stage, spray the compound bacterial agent 2 again at a dosage of 200 mL / mu, once.
[0023] Example 2 A method for preventing and controlling tobacco root and stem diseases based on a composite microbial agent is as follows: Apply Compound Fertilizer 1 at 300 kg / mu during ridge formation and 200 kg / mu 25 days after transplanting, for a total of two applications. During the tobacco seedling stage, spray Compound Fertilizer 2 at 200 mL / mu every 5 days, starting from the cross stage, for a total of three applications.
[0024] During the tobacco field period, spray the compound bacterial agent 2 at a dosage of 200 mL / mu once before and after the tobacco cluster stage; spray the compound bacterial agent 2 again before and after the tobacco bud stage at a dosage of 200 mL / mu once.
[0025] Example 3 A method for preventing and controlling tobacco root and stem diseases based on a composite microbial agent is as follows: The compound fungicide 1 was applied during the tobacco seedling bed period. The application method was as follows: 2 kg of compound fungicide 1 was added to each cubic meter of seedling medium; when the fourth leaf of the tobacco seedlings grew out, 1.5 kg of compound fungicide 1 was applied per 100 square meters; when the seventh leaf of the tobacco seedlings grew out, 1.5 kg of compound fungicide 1 was applied per 100 square meters. A total of three fertilizations were applied.
[0026] During the tobacco seedling stage, starting from the cross stage of the tobacco seedlings, spray the compound fungicide 2 at 200mL / mu every 5 days, for a total of 3 applications.
[0027] Example 4 A method for preventing and controlling tobacco root and stem diseases based on a composite microbial agent is as follows: Apply compound fungicide 1 at a rate of 300 kg / mu when forming ridges on the cultivated land; apply compound fungicide 1 at a rate of 200 kg / mu 25 days after transplanting, for a total of two fertilizations.
[0028] During the tobacco field period, spray the compound bacterial agent 2 at a dosage of 200 mL / mu once before and after the tobacco cluster stage; spray the compound bacterial agent 2 again before and after the tobacco bud stage at a dosage of 200 mL / mu once.
[0029] Example 5 A method for preventing and controlling tobacco root and stem diseases based on a composite microbial agent is as follows: During the tobacco seedling stage, starting from the cross stage of the tobacco seedlings, spray the compound fungicide 2 at 200mL / mu every 5 days, for a total of 3 applications.
[0030] Apply compound fungicide 1 during the tobacco field period, and apply compound fungicide 1 at a rate of 300 kg / mu when ridge-forming; apply compound fungicide 1 at a rate of 200 kg / mu 25 days after transplanting, for a total of two fertilizations.
[0031] Example 6 A method for preventing and controlling tobacco root and stem diseases based on a composite microbial agent is as follows: The compound fungicide 1 was applied during the tobacco seedling bed period. The application method was as follows: 2 kg of compound fungicide 1 was added to each cubic meter of seedling medium; when the fourth leaf of the tobacco seedlings grew out, 1.5 kg of compound fungicide 1 was applied per 100 square meters; when the seventh leaf of the tobacco seedlings grew out, 1.5 kg of compound fungicide 1 was applied per 100 square meters. A total of three fertilizations were applied.
[0032] During the tobacco field stage, spray the compound bacterial agent 2 at a dosage of 200 mL / mu once before and after the tobacco clumping stage; spray the compound bacterial agent 2 again before and after the tobacco budding stage at a dosage of 200 mL / mu once.
[0033] Example 7 A method for preventing and controlling tobacco root and stem diseases based on a composite microbial agent is as follows: The compound fungicide 1 was applied during the tobacco seedling bed period. The application method was as follows: 2 kg of compound fungicide 1 was added to each cubic meter of seedling substrate; when the fourth leaf of the tobacco seedlings grew out, 1.5 kg of compound fungicide 1 was applied per 100 square meters; when the seventh leaf of the tobacco seedlings grew out, 1.5 kg of compound fungicide 1 was applied per 100 square meters. A total of three fertilizations were applied.
[0034] Apply compound fungicide 1 during the tobacco field period, and apply compound fungicide 1 at a rate of 300 kg / mu when ridge-forming; apply compound fungicide 1 at a rate of 200 kg / mu 25 days after transplanting, for a total of two fertilizations.
[0035] During the tobacco seedling stage, starting from the cross stage of the tobacco seedlings, spray the compound fungicide 2 at 200mL / mu every 5 days, for a total of 3 applications.
[0036] During the tobacco field period, spray the compound bacterial agent 2 at a dosage of 200 mL / mu once before and after the tobacco cluster stage; spray the compound bacterial agent 2 again before and after the tobacco bud stage at a dosage of 200 mL / mu once.
[0037] Comparative Example During the tobacco planting stage, no compound microbial agents are added to tobacco in the seedbed stage and the field stage, and other planting methods are carried out normally.
[0038] During the tobacco maturity period, a five-point sampling method was used. Five random sampling points were surveyed per treatment group, with 10 plants at each point. Plant height, stem girth, leaf number, maximum leaf length, and maximum leaf width were measured, and average values were calculated. Tobacco disease status was also investigated, and control efficacy was calculated.
[0039] The disease investigation method is as follows: Refer to the national standard "Grading and Investigation Methods of Tobacco Pests and Diseases" (GB / T 23222-2008).
[0040] Grading standards for tobacco root rot: Level 0: Plants grow normally.
[0041] Level 1: The plant grows basically normally or is slightly dwarfed, and the lowest leaves are yellowing or wilting.
[0042] Level 3: Diseased plants are 1 / 4 to 1 / 3 shorter than healthy plants, and 1 / 2 to 2 / 3 of the leaves are yellowing or wilting.
[0043] Level 5: The diseased plants are 1 / 3 to 1 / 2 shorter than the healthy plants, and more than 2 / 3 of the leaves are yellowing or wilting.
[0044] Level 7: The diseased plant is more than 1 / 2 shorter than the healthy plant, and all the leaves of the plant are yellowing or wilting.
[0045] Level 9: The diseased plants are basically dead.
[0046] Grading standards for tobacco black shank: Level 0: The whole plant is disease-free.
[0047] Level 1: The lesions on the stem do not exceed one-third of the stem circumference, or less than one-third of the leaves are withered.
[0048] Level 3: The lesions on the stem surround one-third to one-half of the stem circumference, or one-third to two-half of the leaves are slightly withered, or lesions appear on a few leaves at the bottom.
[0049] Level 5: The lesions on the stem exceed half of the stem circumference, but do not completely surround the stem circumference, or one-half to two-thirds of the leaves are withered.
[0050] Level 7: The stem lesions completely surround the stem, or more than two-thirds of the leaves wilt.
[0051] Level 9: Diseased plants are basically dead Grading standards for tobacco bacterial wilt: Level 0: The whole plant is disease-free.
[0052] Level 1: There are occasional chlorotic spots on the stem, or less than half of the leaves on the diseased side wilt.
[0053] Level 3: There are black streaks on the stem, but they do not exceed half of the stem height, or one-half to two-thirds of the leaves on the diseased side are withered.
[0054] Level 5: The black streaks on the stem exceed half of the stem height but do not reach the top of the stem, or more than two-thirds of the leaves on the diseased side wilt.
[0055] Level 7: Black streaks on the stem reach the top of the stem, or all the leaves of the diseased plant wilt.
[0056] Level 9: Diseased plants are basically dead Grading standards for tobacco sclerotinia rot: Level 0: No symptoms or mild symptoms.
[0057] Level 1: Water-soaked yellow-brown spots with a diameter of 1 to 4 mm are formed.
[0058] Level 2: Water-soaked yellow-brown spots with a diameter of 5 to 9 mm are formed.
[0059] Level 3: Water-soaked yellow-brown spots with a diameter of 10 to 14 mm are formed.
[0060] Level 4: Water-soaked yellow-brown spots with a diameter of 15 to 19 mm are formed.
[0061] Level 5: Water-soaked yellow-brown spots with a diameter of more than 20 mm are formed.
[0062] Disease index = [(Σ (number of diseased plants × representative level)) / (total number of plants × highest representative level)] × 100 Preventive and therapeutic effect (%) = [(disease index of control group - disease index of treatment group) / disease index of control group] × 100% The effects of different treatments on the agronomic traits of tobacco plants are shown in Table 1 below. From the perspective of the effects of different treatments on the agronomic traits of tobacco plant growth, the application of compound microbial agents at different stages of the tobacco seedling bed and field stages can effectively promote the growth of tobacco plants, and promote the increase in plant height, stem girth, number of leaves, maximum leaf length and maximum leaf width. In particular, the combined application of compound microbial agents at the tobacco seedling bed and field stages can significantly promote the growth of tobacco plants.
[0063] Table 1 Effects of different treatments on agronomic traits of tobacco plants
[0064] The control effects of different treatments on tobacco rhizome diseases are shown in Table 2 below. The effects of different treatments on various tobacco rhizome diseases show that applying the composite microbial agent at different tobacco growth stages has a more significant control effect on various tobacco rhizome diseases. In particular, when the composite microbial agent is applied during both the seedbed and field stages, the control of various rhizome diseases is more significant. This indicates that in order to better control rhizome diseases, the composite microbial agent should be applied at different tobacco growth stages to achieve a synergistic effect.
[0065] Table 2 Control effects of different treatments on tobacco rhizome diseases
[0066] From the results in Table 1 and Table 2, it can be seen that the application method adopted in Example 7 is the best. The composite microbial agent is applied in both the seedling stage and the field stage of tobacco, including the combined application of composite agent 1 and composite agent 2, which has the best effect on tobacco growth and rhizome disease prevention and control.
[0067] Although the present invention has been described in detail through the above preferred embodiments, it should be understood that the above description is not intended to limit the present invention. After reading the above description, various modifications and substitutions of the present invention will become apparent to those skilled in the art. Therefore, the scope of protection of the present invention should be defined by the appended claims.
Claims
1. A composite microbial agent, characterized in that: The composite microbial agent comprises a composite agent 1 and a composite agent 2, wherein the composite agent 1 comprises a methylotrophic Bacillus with a preservation number of CCTCCNO: M 20221661, a white-flavor Streptomyces with a preservation number of CCTCCNO.M 20231644, and a halotolerant Bacillus with a preservation number of CCTCCNO: M 20232276, and the composite agent 2 comprises a blastomyces Streptomyces with a preservation number of CCTCCNO.M 20231643 and a carinii Bacillus with a preservation number of CCTCCNO: M 20221660.
2. The composite microbial agent according to claim 1, characterized in that The composite bacterial agent 1 further comprises corn flour, diatoms, peat, and coffee grounds, wherein the contents of methylotrophic Bacillus, Streptomyces albus, and Bacillus halophilus are not less than 1×10 8 cfu / g.
3. The composite microbial agent according to claim 2, characterized in that The composite bacterial agent 1 is solid and contains, by weight, 10% methylotrophic Bacillus, 10% Streptomyces alboflavin, 10% halodurans, 20% corn flour, 10% diatomaceous earth, 20% peat, and 20% coffee grounds.
4. The composite microbial agent according to claim 1, characterized in that The composite bacterial agent 2 further comprises seaweed polysaccharide, soluble starch, and fulvic acid, wherein the content of Streptomyces oryzae and Bacillus carlesii is not less than 1×10 9 cfu / mL.
5. The composite microbial agent according to claim 4, characterized in that The composite bacterial agent 2 is a liquid, comprising: seaweed polysaccharide at a mass concentration of 3 g / L, soluble starch at a mass concentration of 2 g / L, and fulvic acid at a mass concentration of 3 g / L.
6. Use of the composite microbial agent according to any one of claims 1 to 5 in the field of tobacco cultivation.
7. The use according to claim 6, characterized in that The application includes improving the agronomic traits of tobacco and preventing and controlling tobacco root and stem diseases.
8. The use according to claim 7, characterized in that The agronomic traits include tobacco plant height, stem girth, number of leaves, maximum leaf length and maximum leaf width; the tobacco rhizome diseases include tobacco root rot, tobacco black shank disease, tobacco bacterial wilt and tobacco sclerotinia disease.
9. A method for improving agronomic traits of tobacco and preventing and controlling tobacco root and stem diseases, characterized in that: By applying the composite microbial agent according to any one of claims 1 to 5 during the seedbed stage and field stage of tobacco, the agronomic traits of tobacco and the prevention and control of tobacco root and stem diseases can be improved.
10. The method according to claim 9, characterized in that The agronomic traits include tobacco plant height, stem girth, number of leaves, maximum leaf length and maximum leaf width; the tobacco rhizome diseases include tobacco root rot, tobacco black shank disease, tobacco bacterial wilt and tobacco sclerotinia disease.