Microbial agent for retarding phytotoxicity of tobacco herbicide as well as preparation method and application of microbial agent
By using complex microbial agents of Arthurium paraffin, Bacillus cerealis and Bacillus licheniformis, the problem of low mitigation efficiency of herbicide damage in the prior art has been solved, and the healthy development of tobacco roots and the quality of yield has been improved.
Patent Information
- Application Number
- CN202411763785.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-03
- Publication Date
- 2025-03-14
AI Technical Summary
The prior art is difficult to effectively slow down the harm of tobacco herbicides, especially due to the wide variety of herbicides and the complex transformation process in the soil, which makes screening of degraded strains take a long time and low efficiency.
Complex microbial bacterial solution mixed with Arthrobacter paraffin, Bacillus cerealis and Bacillus licheniformis is prepared into microbial bacterial agents through extrusion and granulation technology, and applied to tobacco planting holes or rows to form a physical isolation barrier to reduce the contact between the root system and the residual soil of the herbicide.
This method can effectively reduce the toxicity of herbicides to the roots of tobacco seedlings, improve the load and biological activity of functional microorganisms, promote root development, enhance resistance, and improve tobacco yield and quality, and does not require special screening of different herbicides and intermediate products.
Smart Images
Figure CN119614550A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of microorganisms, and in particular to a microbial agent for alleviating the damage caused by tobacco herbicides, and a preparation method and application thereof. Background Art
[0002] Weeds in tobacco fields hinder the normal growth of tobacco, increase the cost of tobacco field production management, and reduce tobacco yield and quality. In the southern tobacco-growing areas, rotation is the main method, and chemical weed control is mainly used in the field, which saves time, effort and cost. However, tobacco is more sensitive to herbicides, and unscientific use of herbicides can easily lead to hidden dangers of pesticide damage.
[0003] Tobacco field herbicide damage includes seasonal damage caused by improper use of herbicides, residual damage caused by chemical control of previous crops, and droplet drift damage formed during the spraying of liquid herbicides in surrounding fields. Herbicide damage causes tobacco plants to become dwarfed, stunted, deformed, and yellowing of old leaves, and the damage is getting worse year by year. The problem of herbicide damage seriously restricts the healthy growth of tobacco and threatens the production safety and quality safety of the tobacco industry.
[0004] There are three main types of herbicide soil residue remediation technologies: physical, chemical, and biological, including soil tillage, irrigation-drainage, acid-base neutralization, animal plant remediation, use of soil conditioners, spraying of growth regulators, application of herbicide safeners, and use of microbial control methods. Among them, physical methods are time-consuming, labor-intensive, and costly, and improper chemical methods are prone to secondary pollution.
[0005] Microbial methods are currently a popular remediation measure because of their economic, safe, efficient, and low risk of secondary pollution. Microorganisms in nature are huge in number, diverse in variety, reproduce rapidly, and have strong adaptability to the environment. Researchers screen specific degradation microorganisms to degrade herbicides and eliminate herbicide residue damage. However, there are many types of herbicides, and the transformation process in the soil is complex. Some intermediates, like herbicide residues, have an inhibitory effect on crops. Screening for specific herbicide-degrading strains requires special screening for target herbicides and their intermediates, which is time-consuming and inefficient.
[0006] Therefore, how to provide a new, efficient, green and environmentally friendly biological control method for reducing tobacco herbicide damage is a problem that needs to be solved in the field.
[0007] The invention discloses a microbial agent for alleviating the herbicide damage of tobacco and a method for using the microbial agent, which can deal with various herbicide damages without distinction and does not require special means for special screening of target herbicides and intermediates thereof, and is easy to use and low in cost. The invention is characterized in that beneficial microbial strains that have a promoting effect on the root system and plant development of tobacco are preferably selected; a microbial agent is prepared by a special carrier formula using a conventional microbial agent production means-extrusion granulation technology; the agent is used before the tobacco seedlings are transplanted, and the tobacco seedlings are transplanted on the agent by applying the agent particles to the planting holes and planting rows, and the agent and its carrier create a physical isolation barrier between the tobacco seedling root system and the soil, avoiding and greatly reducing the direct contact between the root system and the soil containing herbicide residues, thereby directly reducing the toxic effect of the herbicide on the tobacco seedling root system, and at the same time, the composite microbial liquid is effectively adsorbed around the tobacco seedling root system through the excellent adsorption capacity of the carrier, effectively increasing the load of functional microorganisms, having good biological activity, being stable and not easy to lose, and having a long action time; through the growth-promoting effect of the functional microorganisms, the generation of growth regulators in the plant body is stimulated, the root system development is promoted, the root system vitality is enhanced, the tobacco plant resistance is improved, and the tobacco yield and quality are increased. The results of greenhouse potted plants and field tests show that the application of the present invention can effectively improve the growth of tobacco plants in fields with herbicide residues, achieve early growth and rapid development, and has broad application prospects. Summary of the invention
[0008] Technical problems to be solved: Since the screening time of herbicide-degrading microorganisms is long and the efficiency is low, and different types of herbicides need to be specially screened for target herbicides and their intermediates, the application of herbicide-degrading microorganisms is limited. The present invention provides a microbial agent for reducing tobacco herbicide damage and its preparation method and application, and its technical entry point and approach are completely different from the current research direction and approach for eliminating herbicide damage.
[0009] Technical solution: A microbial agent for alleviating the damage caused by tobacco herbicides, wherein the agent is prepared by compounding a plant growth-promoting bacterial liquid on a carrier, wherein the plant growth-promoting bacteria include Arthrobacter paraffineus, Bacillus cereus and Bacillus licheniformis.
[0010] The above Arthrobacter paraffineus has a collection number of CGMCC No. 1.1464 and was purchased from China General Microbiological Culture Collection Center.
[0011] The above-mentioned Bacillus cereus has a deposit number of ACCC 10611 and was purchased from the China Agricultural Microbiological Culture Collection Center.
[0012] The above-mentioned Bacillus licheniformis has a deposit number of CFCC 1222 and was purchased from the China Forestry Microbial Culture Collection Management Center.
[0013] The carrier raw materials are agar powder, activated carbon and vermiculite.
[0014] The preparation method of the above-mentioned bacterial agent includes the following steps: 1) preparing a composite microbial bacterial solution; 2) preparing a microbial agent carrier; 3) combining the composite microbial bacterial solution, agar powder, activated carbon and vermiculite at a liquid-solid ratio of 1L:5g:25kg:1kg.
[0015] The preparation steps of the composite microbial bacterial liquid are as follows: 1) activating Arthrobacter paraffineus and inoculating it into a nutrient broth medium, and shaking and culturing it at 30-35°C and 150-200rpm until the logarithmic growth phase to obtain Arthrobacter paraffineus seed liquid; expanding the seed liquid and inoculating it into a fermentation medium at an inoculum volume ratio of 10%, and culturing it at 30-35°C and 150-200rpm for 45-50h to obtain a fermentation liquid, wherein the content of Arthrobacter paraffineus in the fermentation liquid is greater than 1.0×10 9 cfu / mL; 2) activating Bacillus cereus and inoculating it into a nutrient broth medium, and shaking and culturing it at 30-35°C and 150-200rpm until the logarithmic growth phase to obtain a Bacillus cereus seed solution; inoculating the above seed solution into a fermentation medium at an inoculum volume ratio of 10%, and culturing it at 30-35°C and 150-200rpm for 45-50h to obtain a fermentation liquid, wherein the content of Bacillus cereus in the fermentation liquid is greater than 1.0×10 9 cfu / mL; 3) activating Bacillus licheniformis and inoculating it into a nutrient broth medium, shaking and culturing it at 30-35°C and 150-200rpm until the logarithmic growth phase to obtain a Bacillus licheniformis seed solution; inoculating the above seed solution into a fermentation medium at an inoculum volume ratio of 10%, culturing it at 30-35°C and 150-200rpm for 45-50h to obtain a fermentation liquid, wherein the content of Bacillus licheniformis in the fermentation liquid is greater than 1.0×10 9 cfu / mL; 4) After adjusting the above three bacterial solutions to the same concentration, mix them in a liquid volume ratio of 1:1:1 to obtain a composite microbial bacterial solution.
[0016] The composition of the nutrient gravy medium is as follows: 10.0 g of peptone, 3.0 g of beef extract, 1.0 L of distilled water, pH adjusted to 7.0, and sterilized at 121° C. with steam for 30 min.
[0017] The operation steps of the above step (3) are: pouring the composite microbial liquid, agar powder, activated carbon and vermiculite into a blender and mixing them evenly, and then extruding them into bacterial agent particles using an extrusion granulator.
[0018] The above-mentioned microbial agent is applied at a dosage of 10 kg per mu. The microbial agent particles are placed in tobacco planting holes or planting rows, and then tobacco seedlings are placed on top of the microbial agent particles. Field cultivation management is carried out in accordance with local production standardization requirements.
[0019] Beneficial effects: (1) The present invention selects paraffinic Bacillus, cereus and Bacillus licheniformis and mixes them in equal proportions to form a composite microbial liquid. The three have a synergistic effect and have a significant growth-promoting effect on tobacco; at the same time, they can effectively improve the resistance of tobacco plants and increase tobacco yield and quality; the three functional microbial strains are all bacteria, the culture method and culture process are simple, the reproduction speed is fast, the tolerance to complex environments is strong, and the adaptability in the actual soil rhizosphere environment is good.
[0020] (2) The raw materials of the microbial agent carrier of the present invention are agar powder, activated carbon and vermiculite. The developed pore structure and large surface area of activated carbon give it good adsorption performance; agar powder is an environmentally friendly polysaccharide substance that can absorb water equivalent to 20 times its own volume; vermiculite is a natural, inorganic, non-toxic mineral with good ion exchange capacity; the combination of the three not only has a good granulation effect, but also can provide effective protection for the survival of microorganisms in the agent particles and the colonization in the soil. After the agent is applied to the soil, on the one hand, a physical isolation barrier is formed to avoid and greatly reduce the direct contact between the roots and the soil with herbicide residues, eliminating the damage of the herbicide to the young roots of the seedlings; on the other hand, the functional microbial agent can be effectively gathered around the root system, promoting the colonization of microorganisms in the soil and their interaction with the root system of crops. It has high biological activity, is stable and not easy to lose, and has a long action time.
[0021] (3) By applying the present invention, the microbial agent forms a dominant bacterial community around the roots of tobacco seedlings, reproduces rapidly, increases the production of growth regulators in the plant, enhances root vitality, indirectly inhibits the effects of herbicide damage, improves the growth of tobacco plants, and facilitates their early growth and rapid development, thus having broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS
[0022] Figure 1 This is a picture of the prevention and control effects of greenhouse tobacco potted plants.
[0023] Figure 2 This is a picture of the tobacco prevention and control effects in the greenhouse Liupanshui field. DETAILED DESCRIPTION
[0024] The technical solutions in the embodiments of the present invention are described clearly and completely below, although the described embodiments are only part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
[0025] For further understanding of the present invention, the present invention is described in detail below in conjunction with the embodiments. Unless otherwise specified, the reagents involved can be purchased through commercial channels, and the methods not mentioned are conventional experimental methods and are not described in detail here.
[0026] Example 1: A microbial agent for reducing the damage caused by tobacco herbicides
[0027] A microbial agent for alleviating the damage caused by tobacco herbicides, the agent comprising a composite microbial liquid and a microbial agent carrier.
[0028] The composite microbial bacterial liquid comprises: Arthrobacter paraffineus, Bacillus cereus and Bacillus licheniformis.
[0029] The Arthrobacter paraffineus has a collection number of CGMCC No. 1.1464, purchased from the China General Microbiological Culture Collection Center; the Bacillus cereus has a collection number of ACCC 10611, purchased from the China Agricultural Microbiological Culture Collection Center; the Bacillus licheniformis has a collection number of CFCC 1222, purchased from the China Forestry Microbiological Culture Collection Center.
[0030] The preparation method of the composite microbial liquid is as follows:
[0031] 1) Activating Arthrobacter paraffinus and inoculating it into a nutrient gravy agar medium, shaking and culturing it at 30-35° C. and 150-200 rpm until the logarithmic growth phase to obtain Arthrobacter paraffinus seed liquid; inoculating the above seed liquid into a fermentation medium at an inoculum amount of 10% (volume ratio), culturing it at 30-35° C. and 150-200 rpm for 45-50 hours to obtain a fermentation liquid, wherein the content of Arthrobacter paraffinus in the fermentation liquid is greater than 1.0×10 9 cfu / mL;
[0032] 2) activating Bacillus cereus and inoculating it into a nutrient gravy agar medium, and culturing it at 30-35° C. and 150-200 rpm with shaking until the logarithmic growth phase to obtain a Bacillus cereus seed solution; inoculating the seed solution into a fermentation medium at an inoculum amount of 10% (volume ratio), and culturing it at 30-35° C. and 150-200 rpm for 45-50 hours to obtain a fermentation liquid, wherein the content of Bacillus cereus in the fermentation liquid is greater than 1.0×10 9 cfu / mL;
[0033] 3) After activating Bacillus licheniformis, inoculate it into PDA medium, and culture it at 30-35° C. and 150-200 rpm with shaking until the logarithmic growth phase to obtain Bacillus licheniformis seed liquid; inoculate the above seed liquid into fermentation medium at an inoculum amount of 10% (volume ratio), and culture it at 30-35° C. and 150-200 rpm for 45-50 hours to obtain fermentation liquid, wherein the content of Bacillus licheniformis in the fermentation liquid is greater than 1.0×10 9 cfu / mL;
[0034] 4) After adjusting the above three bacterial solutions to the same concentration, mix them at a liquid volume ratio of 1:1:1 to obtain a composite microbial bacterial solution.
[0035] The composition of the nutrient broth medium is: 10.0 g of peptone, 3.0 g of beef extract, 1.0 L of distilled water, the pH is adjusted to 7.0, and the medium is sterilized at a high temperature of 121° C. with steam for 30 minutes.
[0036] The preparation of the microbial agent carrier and the agent granules: the composite microbial bacterial solution, agar powder, activated carbon and vermiculite are poured into a blender at a liquid-solid ratio of 1L:5g:25kg:1kg and mixed evenly, and then extruded into agent granules by an extrusion granulator.
[0037] The microbial agent is applied at a dosage of 10 kg per mu, and the microbial agent particles are placed in tobacco planting holes or planting rows, and then tobacco seedlings are placed on the microbial agent particles. Field cultivation management is carried out in accordance with local production standardization requirements.
[0038] Example 2: Tobacco greenhouse potted plant test of a microbial agent for reducing tobacco herbicide damage
[0039] Take the tobacco field soil with serious herbicide damage in Liupanshui, air dry naturally, and pass through a 10-mesh sieve. Each pot is filled with 25kg of soil, and 60kg / mu of local tobacco-specific base fertilizer (10-10-24) is applied. The base fertilizer and soil are evenly mixed for standby use. The tobacco seedling variety used in the experiment is Yunyan 87, and the seedling raising method is the traditional floating seedling raising method. When the tobacco seedlings have 6 true leaves, select the seedlings with consistent growth and relatively strong for transplanting. It is divided into three treatments: control, composite microbial liquid, and microbial agent granules. Among them, the control treatment is transplanted normally. Composite microbial liquid treatment applies the composite microbial liquid around the planting hole, and covers the soil after the tobacco seedlings are transplanted. Microbial agent granules are applied around the planting hole, and the soil is covered after the tobacco seedlings are transplanted. The amount of composite microbial liquid used is 30mL / plant, and the amount of microbial agent granules used is 10g / plant. Each treatment is repeated 4 times. The preparation of composite microbial liquid and microbial agent granules is shown in Example 1.
[0040] The survey results during the clustering stage showed that compared with the control, the plant height, maximum leaf area and stem girth of the microbial agent treatment increased by 21.15%, 11.16% and 5.13% respectively; the plant height, leaf number, maximum leaf area and stem girth of the microbial agent attached to the carrier treatment increased by 42.31%, 18.75%, 92.65% and 39.48% respectively.
[0041] The survey results during the vigorous growth period showed that the plant height, leaf number, maximum leaf area and stem girth of the plants treated with the inoculant increased by 11.12%, 13.04%, 16.64% and 9.98% respectively; the plant height, leaf number, maximum leaf area and stem girth of the plants treated with the inoculant attached to a carrier increased by 87.55%, 39.13%, 133.09% and 49.08% respectively.
[0042] In addition, the number of leaves harvested after the treatment with the microbial agent increased by 30.79%, and the tobacco yield increased by 19.03%; the number of leaves harvested after the treatment with the microbial agent and carrier increased by 50.94%, and the tobacco yield increased by 43.14%. The application of microbial agents can slow down the effects of herbicides on tobacco, and the microbial agent and carrier can further promote the early growth of tobacco seedlings, and effectively improve the growth of tobacco plants in soils with severe herbicide damage.
[0043]
[0044] Example 3: Field test of a microbial agent for reducing tobacco herbicide damage
[0045] The test site is located in Zhudong Village, Zhudong Township, Panzhou City, Liupanshui City. The selected tobacco field has serious herbicide damage, and the field area is 2.2 mu. According to the local fertilization guidance, base fertilizer is applied, and ridges, ponds, transplanting, and topdressing are normal. Among them, the application rate of tobacco-specific base fertilizer (10-10-24) is 60kg / mu; the application rate of seedling fertilizer (22-14-10) is 60kg / mu; the application rate of topdressing (15-0-30) is 18kg / mu. The tobacco seedling variety used in the experiment is Yunyan 87, and the seedling raising method is the traditional floating seedling raising method. It is divided into two treatments: control and microbial agent with carrier. Among them, the control treatment is transplanted normally. The microbial agent treatment requires the microbial agent to be applied around the tobacco seedlings before transplanting, and the soil is covered after transplanting. Each treatment has 3 parallels.
[0046] The survey results show that compared with the control, the plant height and leaf number of tobacco treated with bacterial agents during the bush stage increased by 21.83% and 11.11%; the plant height, leaf number, stem girth and maximum leaf area during the vigorous growth stage increased by 23.53%, 11.59%, 3.30% and 10.85% respectively; the plant height, leaf number, stem girth and maximum leaf area during the topping stage increased by 16.91%, 11.17%, 16.17% and 21.24% respectively.
[0047] The application of bacterial agents can effectively reduce the impact of herbicide residues on tobacco, improve the growth of tobacco plants, and promote early growth and rapid development.
[0048]
[0049] Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the present invention will not be limited to the embodiments shown herein, but will conform to the widest scope consistent with the principles and novel features disclosed herein.
Claims
1. A microbial agent for reducing the damage caused by tobacco herbicides, characterized in that: The bacterial agent is obtained by combining a plant growth-promoting bacterial liquid with a carrier, and the plant growth-promoting bacteria include Arthrobacter paraffinum ( Arthrobacter paraffineus ), Bacillus cereus ( Bacillus cereus ) and Bacillus licheniformis ( Bacillus licheniformis ).
2. The microbial agent for alleviating the herbicide damage to tobacco according to claim 1, characterized in that: The paraffinic bacterium ( Arthrobacter paraffineus ) with a deposit number of CGMCC No.1.1464, purchased from China General Microbiological Culture Collection Center.
3. The microbial agent for alleviating the herbicide damage to tobacco according to claim 1, characterized in that: The Bacillus cereus ( Bacillus cereus ) with the deposit number of ACCC 10611, purchased from China Agricultural Microbiological Culture Collection Center.
4. The microbial agent for alleviating the herbicide damage to tobacco according to claim 1, characterized in that: The Bacillus licheniformis ( Bacillus licheniformis ) with the deposit number of CFCC 1222, purchased from China Forestry Microbial Culture Collection Management Center.
5. The microbial agent for alleviating the herbicide damage to tobacco according to claim 1, characterized in that: The carrier raw materials are agar powder, activated carbon and vermiculite.
6. The method for preparing the bacterial agent according to any one of claims 1 to 5, characterized in that: The following steps are involved: 1) Preparation of composite microbial culture liquid; 2) Preparation of microbial agent carrier; 3) Combining composite microbial culture liquid, agar powder, activated carbon and vermiculite at a liquid-solid ratio of 1L:5g:25kg:1kg.
7. The method for preparing the bacterial agent according to claim 6, characterized in that: The preparation steps of the composite microbial liquid are as follows: 1) Arthrobacter paraffinum ( Arthrobacter paraffineus ) after activation, inoculated into nutrient broth medium, and cultured at 30-35°C and 150-200rpm with shaking until the logarithmic growth phase to obtain paraffinic Arthrobacter seed liquid; the above seed liquid was expanded and inoculated into fermentation medium at an inoculum volume ratio of 10%, and cultured at 30-35°C and 150-200rpm for 45-50h to obtain fermentation liquid, wherein the content of paraffinic Arthrobacter in the fermentation liquid is greater than 1.0×10 9 cfu / mL; 2) Bacillus cereus ( Bacillus cereus ) after activation, inoculated into nutrient broth medium, and cultured at 30-35°C and 150-200rpm to the logarithmic growth phase to obtain Bacillus cereus seed liquid; the seed liquid was inoculated into the fermentation medium at an inoculum volume ratio of 10%, and cultured at 30-35°C and 150-200rpm for 45-50h to obtain a fermentation liquid, wherein the content of Bacillus cereus in the fermentation liquid is greater than 1.0×10 9 cfu / mL; 3) Activating Bacillus licheniformis and inoculating it into a nutrient broth medium, shaking and culturing it at 30-35°C and 150-200rpm until the logarithmic growth phase to obtain a Bacillus licheniformis seed solution; inoculating the above seed solution into a fermentation medium at an inoculum volume ratio of 10%, culturing it at 30-35°C and 150-200rpm for 45-50h to obtain a fermentation liquid, wherein the content of Bacillus licheniformis in the fermentation liquid is greater than 1.0×10 9 cfu / mL; 4) After adjusting the above three bacterial solutions to the same concentration, mix them in a liquid volume ratio of 1:1:1 to obtain a composite microbial bacterial solution.
8. The method for preparing the bacterial agent according to claim 7, characterized in that: The composition of the nutrient broth medium is: 10.0 g of peptone, 3.0 g of beef extract, 1.0 L of distilled water, the pH is adjusted to 7.0, and the medium is sterilized at a high temperature of 121° C. with steam for 30 min.
9. The method for preparing the bacterial agent according to claim 6, characterized in that: The operation steps of step (3) are as follows: pouring the composite microbial liquid, agar powder, activated carbon and vermiculite into a blender and mixing them evenly, and then extruding them into bacterial agent granules using an extrusion granulator.
10. The use of the bacterial agent according to any one of claims 1 to 5, characterized in that: At a dosage of 10 kg per mu, the inoculant particles are placed in tobacco planting holes or rows, and then tobacco seedlings are placed on top of the inoculant particles. Field cultivation management is carried out in accordance with local production standardization requirements.