CN109971680B - Preparation and application of compound microbial agent with disease-resistant and growth-promoting functions - Google Patents
Preparation and application of compound microbial agent with disease-resistant and growth-promoting functions Download PDFInfo
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Abstract
The invention discloses a preparation method and application of a compound microbial agent with disease-resistant and growth-promoting functions, wherein the preparation method comprises the following steps: (1) selecting bacillus subtilis numbered as WSWFJ-2, bacillus subtilis numbered as WSWFJ-7, bacillus amyloliquefaciens numbered as WSWFJ-8 and bacillus amyloliquefaciens numbered as WSWFJ-11, and fermenting respectively; (2) mixing 0-35 parts by weight of a bacillus subtilis strain fermentation liquid numbered as WSWFJ-2, 0-35 parts by weight of a bacillus subtilis strain fermentation liquid numbered as WSWFJ-7, 25-50 parts by weight of a bacillus amyloliquefaciens strain fermentation liquid numbered as WSWFJ-8 and 25-50 parts by weight of a bacillus amyloliquefaciens strain fermentation liquid numbered as WSWFJ-11 to obtain a compound microbial agent with disease resistance and growth promotion functions; the compound microbial agent prepared by the preparation method is applied to agricultural production, can effectively reduce the use amount of chemical pesticides, and promotes the development of green agriculture.
Description
Technical Field
The invention relates to the field of fertilizers, in particular to preparation and application of a compound microbial agent with disease-resistant and growth-promoting functions.
Background
The microbial agent is a viable bacteria preparation prepared by processing fermentation liquor for adsorbing bacteria by using porous substances as adsorbents (such as turf and vermiculite) after industrial production and propagation of target microorganisms (effective bacteria). The bactericide is used for dressing seeds or dipping roots, and has the effects of directly or indirectly improving soil, restoring land capability, preventing soil-borne diseases, maintaining rhizosphere microflora balance, degrading toxic and harmful substances and the like. The agricultural microbial agent can be properly used for improving the yield of agricultural products, improving the quality of the agricultural products, reducing the using amount of chemical fertilizers, reducing the cost, improving the soil and protecting the ecological environment.
The existing microbial inoculum generally adopts a single strain, has poor application effect in the actual use process, and cannot be popularized in a large quantity.
Disclosure of Invention
The invention aims to provide preparation and application of a compound microbial agent with disease-resistant and growth-promoting functions aiming at the defects of the prior art, microbial strains with different functions are compounded, and the microbial agents with different disease-resistant and growth-promoting functions are formed by increasing and decreasing the proportioning strains and adjusting the addition of the strains, so that the compound microbial agent is applied to agricultural production, can effectively reduce the use amount of chemical pesticides and promotes the development of green agriculture.
In order to achieve the above object, the present invention is achieved by the following technical solutions.
A preparation method of a compound microbial agent with disease-resistant and growth-promoting functions comprises the following steps:
(1) selecting Bacillus subtilis numbered WSWFJ-2, Bacillus subtilis numbered WSWFJ-7, Bacillus amyloliquefaciens numbered WSWFJ-8 and Bacillus amyloliquefaciens numbered WSWFJ-11, respectively fermenting, wherein the effective viable bacteria number in the fermentation liquor of each strain is more than 2.0 multiplied by 109cfu/ml；
(2) Mixing 0-35 parts by weight of a bacillus subtilis strain fermentation liquid numbered as WSWFJ-2, 0-35 parts by weight of a bacillus subtilis strain fermentation liquid numbered as WSWFJ-7, 25-50 parts by weight of a bacillus amyloliquefaciens strain fermentation liquid numbered as WSWFJ-8 and 25-50 parts by weight of a bacillus amyloliquefaciens strain fermentation liquid numbered as WSWFJ-11 to obtain a compound microbial agent with disease resistance and growth promotion functions;
the culture medium formula of the two bacillus subtilis strains comprises the following components in parts by weight: 10-16 parts of soybean meal, 7-13 parts of cane sugar, 1-5 parts of peptone, 2-8 parts of corn flour, 3-9 parts of calcium carbonate, 3-7 parts of fish meal, 0.2-0.8 part of calcium chloride, 0.2-0.7 part of potassium dihydrogen phosphate, 0.2-0.8 part of dipotassium hydrogen phosphate, 0.1-0.5 part of manganese sulfate, 0.1-0.5 part of sodium hydroxide, 2-7 parts of sodium chloride and 1-2 parts of defoaming agent;
the inoculation amount (mass percentage) of the bacillus subtilis is 0.5-1.5%,
the aerobic culture temperature of the bacillus subtilis is 35-39 ℃, and the bacillus subtilis is cultured for 1-3 days;
the culture medium formula of the two bacillus amyloliquefaciens strains comprises the following components in parts by weight: 7-13 parts of soybean meal, 5-9 parts of cane sugar, 3-8 parts of peptone, 2-7 parts of corn flour, 3-8 parts of calcium carbonate, 2-8 parts of fish meal, 0.2-0.7 part of calcium chloride, 0.2-0.7 part of potassium dihydrogen phosphate, 0.2-0.8 part of dipotassium hydrogen phosphate, 0.1-0.6 part of manganese sulfate, 0.1-0.4 part of sodium hydroxide, 3-7 parts of sodium chloride and 1-2 parts of defoaming agent;
the inoculation amount (mass percentage) of the bacillus amyloliquefaciens is 0.3 to 1 percent,
the aerobic culture temperature of the bacillus amyloliquefaciens is 33-40 ℃, and the bacillus amyloliquefaciens is cultured for 1-3 days.
The compound microbial agent is diluted to a solution of 700 times of 300 times by weight with water and is used for dipping roots when transplanted after seedling raising of crops.
The compound microbial inoculum is diluted to 200-fold and 1100-fold liquid by water according to the weight ratio and is used for irrigating roots after transplanting crops, and the application amount per mu of the compound microbial inoculum is 0.3-3.0 kg/mu.
Bacillus subtilis (WSWFJ-2) is preserved in China general microbiological culture Collection center (China Committee for culture Collection of microorganisms), and the preservation address is as follows: west road No.1 hospital No. 3, north jing, chaoyang district, preservation date: 6 months and 4 days in 2018, and the accession number is: CGMCC NO. 15842;
bacillus subtilis with the number of WSWFJ-7 is preserved in the China general microbiological culture Collection center, and the preservation address is as follows: west road No.1 hospital No. 3, north jing, chaoyang district, preservation date: 6 months and 4 days in 2018, and the accession number is: CGMCC NO. 15847;
bacillus amyloliquefaciens (Bacillus amyloliquefaciens) with the number of WSWFJ-8 is preserved in the China general microbiological culture Collection center of the China Committee for culture Collection of microorganisms with the preservation address: west road No.1 hospital No. 3, north jing, chaoyang district, preservation date: 6 months and 4 days in 2018, and the accession number is: CGMCC NO. 15848;
bacillus amyloliquefaciens (Bacillus amyloliquefaciens) with the number of WSWFJ-11 is preserved in the China general microbiological culture Collection center of the China Committee for culture Collection of microorganisms with the preservation address: west road No.1 hospital No. 3, north jing, chaoyang district, preservation date: 6 months and 4 days in 2018, and the accession number is: CGMCC NO. 15851.
Detailed Description
The present invention is further illustrated by the following examples, but is not limited to the details of the description.
A preparation method of a compound microbial agent with disease-resistant and growth-promoting functions comprises the following steps:
(1) selecting Bacillus subtilis numbered WSWFJ-2, Bacillus subtilis numbered WSWFJ-7, Bacillus amyloliquefaciens numbered WSWFJ-8 and Bacillus amyloliquefaciens numbered WSWFJ-11, respectively performing separate fermentation, wherein the effective viable count of each strain in the fermentation liquid after fermentation is more than 2.0 × 109cfu/ml；
(2) 0-35 parts of the Bacillus subtilis strain fermentation liquor numbered as WSWFJ-2, 0-35 parts of the Bacillus subtilis strain fermentation liquor numbered as WSWFJ-7, 25-50 parts of the Bacillus amyloliquefaciens strain fermentation liquor numbered as WSWFJ-8 and 25-50 parts of the Bacillus amyloliquefaciens strain fermentation liquor numbered as WSWFJ-11 are mixed according to weight percentage to obtain the compound microbial agent with the functions of resisting diseases and promoting growth.
The culture medium formula of the two bacillus subtilis strains comprises the following components in parts by weight: 10-16 parts of soybean meal, 7-13 parts of cane sugar, 1-5 parts of peptone, 2-8 parts of corn flour, 3-9 parts of calcium carbonate, 3-7 parts of fish meal, 0.2-0.8 part of calcium chloride, 0.2-0.7 part of potassium dihydrogen phosphate, 0.2-0.8 part of dipotassium hydrogen phosphate, 0.1-0.5 part of manganese sulfate, 0.1-0.5 part of sodium hydroxide, 2-7 parts of sodium chloride and 1-2 parts of defoaming agent;
the inoculation amount (mass percentage) of the bacillus subtilis is 0.5-1.5%,
the aerobic culture temperature of the bacillus subtilis is 35-39 ℃, and the bacillus subtilis is cultured for 1-3 days.
The culture medium formula of the two bacillus amyloliquefaciens strains comprises the following components in parts by weight: 7-13 parts of soybean meal, 5-9 parts of cane sugar, 3-8 parts of peptone, 2-7 parts of corn flour, 3-8 parts of calcium carbonate, 2-8 parts of fish meal, 0.2-0.7 part of calcium chloride, 0.2-0.7 part of potassium dihydrogen phosphate, 0.2-0.8 part of dipotassium hydrogen phosphate, 0.1-0.6 part of manganese sulfate, 0.1-0.4 part of sodium hydroxide, 3-7 parts of sodium chloride and 1-2 parts of defoaming agent;
the inoculation amount (mass percentage) of the bacillus amyloliquefaciens is 0.3 to 1 percent,
the aerobic culture temperature of the bacillus amyloliquefaciens is 33-40 ℃, and the bacillus amyloliquefaciens is cultured for 1-3 days.
The defoaming agent is a commercial product which is expensive, such as a THIX-298 product of Nicotine Henxin chemical industry.
The application of the compound microbial agent with the functions of resisting diseases and promoting growth can adopt two methods: firstly, diluting the composite microbial inoculum by water to 300-fold and 700-fold liquid according to the weight ratio, and dipping roots when the composite microbial inoculum is transplanted after seedling raising of crops; secondly, diluting the composite microbial inoculum by water according to the weight ratio to 200-fold liquid of 1100 times, and irrigating roots of the transplanted crops, wherein the application amount per mu of the composite microbial inoculum is 0.3-3.0 kg/mu.
The following description is given in conjunction with specific embodiments.
Example 1
A compound microbial agent with disease-resistant and growth-promoting functions is prepared from 4 strains of Bacillus subtilis (WSWFJ-2), Bacillus subtilis (WSWFJ-7), Bacillus amyloliquefaciens (WSWFJ-8) and Bacillus amyloliquefaciens (WSWFJ-11).
A preparation method of a compound microbial agent with disease-resistant and growth-promoting functions comprises the following steps: fermenting each strain individually, wherein the effective viable bacteria number in the fermentation liquid of each strain is 2.0 × 109More than cfu/ml, and mixing 25 parts of WSWFJ-2 strain fermentation liquor, 25 parts of WSWFJ-7 strain fermentation liquor, 25 parts of WSWFJ-8 strain fermentation liquor and 25 parts of WSWFJ-11 strain fermentation liquor according to weight percentage to obtain the compound microbial agent with the functions of resisting diseases and promoting growth. The compound microbial agent of the formula is suitable for planting leaf vegetables.
The application of the compound microbial agent with the functions of resisting diseases and promoting growth adopts the following method: firstly, diluting the composite microbial inoculum to 400 times of solution by weight with water, and dipping roots when the composite microbial inoculum is used for transplanting after seedling raising of crops; secondly, diluting the compound microbial inoculum by water to 800 times of the solution according to the weight ratio, and irrigating roots of the transplanted crops, wherein the application amount of the compound microbial inoculum per mu is 1.0 kg/mu.
Further, the two bacillus subtilis culture medium formulas are as follows according to parts by weight: 12 parts of soybean meal, 10 parts of cane sugar, 4 parts of peptone, 6 parts of corn flour, 5 parts of calcium carbonate, 5 parts of fish meal, 0.5 part of calcium chloride, 0.3 part of monopotassium phosphate, 0.2 part of dipotassium hydrogen phosphate, 0.5 part of manganese sulfate, 0.3 part of sodium hydroxide, 2 parts of sodium chloride and 1 part of defoaming agent;
the inoculation amount (mass percentage) of the bacillus subtilis is 1.0 percent,
the aerobic culture temperature of the bacillus subtilis is 37 ℃, and the bacillus subtilis is cultured for 2 days.
Further, the culture medium formula of the two bacillus amyloliquefaciens strains is as follows according to parts by weight: 10 parts of soybean meal, 7 parts of cane sugar, 8 parts of peptone, 4 parts of corn flour, 3 parts of calcium carbonate, 6 parts of fish meal, 0.7 part of calcium chloride, 0.2 part of monopotassium phosphate, 0.5 part of dipotassium hydrogen phosphate, 0.3 part of manganese sulfate, 0.2 part of sodium hydroxide, 5 parts of sodium chloride and 2 parts of defoaming agent;
the inoculation amount (mass percentage) of the bacillus amyloliquefaciens is 0.5 percent,
the aerobic culture temperature of the bacillus amyloliquefaciens is 35 ℃, and the bacillus amyloliquefaciens is cultured for 2 days.
Example 2
A composite microbial agent with disease-resistant and growth-promoting functions is prepared from 2 strains of Bacillus amyloliquefaciens (WSWFJ-8) and Bacillus amyloliquefaciens (WSWFJ-11).
A preparation method of a compound microbial agent with disease-resistant and growth-promoting functions comprises the following steps: fermenting each strain individually, wherein the effective viable bacteria number in the fermentation liquid of each strain is 2.0 × 109More than cfu/ml, and mixing 50 parts of WSWFJ-8 strain fermentation liquor and 50 parts of WSWFJ-11 strain fermentation liquor according to the weight percentage to obtain the compound microbial agent with the functions of resisting diseases and promoting growth. The compound microbial agent of the formula is suitable for planting crops such as oil plants, grains, fruit trees and the like.
The application of the compound microbial agent with the functions of resisting diseases and promoting growth adopts the following method: diluting the compound microbial inoculum to 200 times of solution by weight ratio with water, and irrigating roots after transplanting crops, wherein the application amount per mu of the compound microbial inoculum is 3.0 kg/mu.
The culture medium formula of the two bacillus amyloliquefaciens strains comprises the following components in parts by weight: 13 parts of soybean meal, 5 parts of cane sugar, 5 parts of peptone, 3 parts of corn flour, 3 parts of calcium carbonate, 8 parts of fish meal, 0.5 part of calcium chloride, 0.4 part of monopotassium phosphate, 0.6 part of dipotassium hydrogen phosphate, 0.6 part of manganese sulfate, 0.4 part of sodium hydroxide, 5 parts of sodium chloride and 1 part of defoaming agent;
the inoculation amount (mass percentage) of the bacillus amyloliquefaciens is 1 percent,
the aerobic culture temperature of the bacillus amyloliquefaciens is 36 ℃, and the bacillus amyloliquefaciens is cultured for 3 days.
Example 3
A composite microbial agent with disease-resistant and growth-promoting functions is prepared from 3 strains of Bacillus subtilis (WSWFJ-7), Bacillus amyloliquefaciens (WSWFJ-8) and Bacillus amyloliquefaciens (WSWFJ-11),
a preparation method of a compound microbial agent with disease-resistant and growth-promoting functions comprises the following steps: fermenting each strain individually, wherein the effective viable bacteria number in the fermentation liquid of each strain is 2.0 × 109More than cfu/ml, 30 parts of WSWFJ-7 strain fermentation liquor by weight percentage,35 parts of WSWFJ-8 strain fermentation liquor and 35 parts of WSWFJ-11 strain fermentation liquor are mixed to obtain the compound microbial agent with disease resistance and growth promotion functions. The compound microbial agent of the formula is suitable for planting crops such as tobacco, medicinal materials and the like.
The application of the compound microbial agent with the functions of resisting diseases and promoting growth adopts the following method: firstly, diluting the composite microbial inoculum by water to 500 times of solution according to the weight ratio, and dipping the roots when the composite microbial inoculum is transplanted after the seedling of crops. Secondly, diluting the compound microbial inoculum to 300 times of solution by weight with water, and irrigating roots after transplanting crops, wherein the application amount of the compound microbial inoculum per mu is 2.0 kg/mu.
The bacillus subtilis culture medium comprises the following components in parts by weight: 16 parts of soybean meal, 7 parts of cane sugar, 2 parts of peptone, 2 parts of corn flour, 3 parts of calcium carbonate, 7 parts of fish meal, 0.5 part of calcium chloride, 0.7 part of monopotassium phosphate, 0.8 part of dipotassium hydrogen phosphate, 0.1 part of manganese sulfate, 0.5 part of sodium hydroxide, 2 parts of sodium chloride and 2 parts of defoaming agent;
the inoculation amount (mass percentage) of the bacillus subtilis is 1 percent,
the aerobic culture temperature of the bacillus subtilis is 38 ℃, and the bacillus subtilis is cultured for 1 day.
The culture medium formula of the two bacillus amyloliquefaciens strains comprises the following components in parts by weight: 7 parts of soybean meal, 9 parts of cane sugar, 8 parts of peptone, 2 parts of corn flour, 4 parts of calcium carbonate, 5 parts of fish meal, 0.3 part of calcium chloride, 0.5 part of monopotassium phosphate, 0.24 part of dipotassium hydrogen phosphate, 0.1 part of manganese sulfate, 0.1 part of sodium hydroxide, 3 parts of sodium chloride and 2 parts of defoaming agent;
the inoculation amount (mass percentage) of the bacillus amyloliquefaciens is 1 percent,
the aerobic culture temperature of the bacillus amyloliquefaciens is 37 ℃, and the bacillus amyloliquefaciens is cultured for 1 day.
Example 4
A composite microbial agent with disease-resistant and growth-promoting functions is prepared from 3 strains of Bacillus subtilis (WSWFJ-2), Bacillus amyloliquefaciens (WSWFJ-8) and Bacillus amyloliquefaciens (WSWFJ-11),
a preparation method of a compound microbial agent with disease-resistant and growth-promoting functions comprises the following steps: fermenting each strain individually, wherein the effective viable bacteria number in the fermentation liquid of each strain is 2.0 × 109More than cfu/ml, and mixing 25 parts of WSWFJ-2 strain fermentation liquor, 40 parts of WSWFJ-8 strain fermentation liquor and 35 parts of WSWFJ-11 strain fermentation liquor according to weight percentage to obtain the compound microbial agent with the functions of resisting diseases and promoting growth. The compound microbial agent of the formula is suitable for planting crops such as tobacco, medicinal materials and the like.
The application of the compound microbial agent with the functions of resisting diseases and promoting growth adopts the following method: firstly, diluting the composite microbial inoculum to 600 times of solution by weight with water, and dipping roots when the composite microbial inoculum is transplanted after seedling raising of crops. Secondly, diluting the compound microbial inoculum by water to 500 times of the solution according to the weight ratio, and irrigating roots of the transplanted crops, wherein the application amount of the compound microbial inoculum per mu is 1.5 kg/mu.
The bacillus subtilis culture medium comprises the following components in parts by weight: 15 parts of soybean meal, 6 parts of cane sugar, 3 parts of peptone, 4 parts of corn flour, 4 parts of calcium carbonate, 6 parts of fish meal, 0.6 part of calcium chloride, 0.6 part of monopotassium phosphate, 0.7 part of dipotassium hydrogen phosphate, 0.2 part of manganese sulfate, 0.5 part of sodium hydroxide, 3 parts of sodium chloride and 1 part of defoaming agent;
the inoculation amount (mass percentage) of the bacillus subtilis is 0.5 percent,
the aerobic culture temperature of the bacillus subtilis is 37 ℃, and the bacillus subtilis is cultured for 2 days.
The culture medium formula of the two bacillus amyloliquefaciens strains comprises the following components in parts by weight: 8 parts of soybean meal, 10 parts of cane sugar, 6 parts of peptone, 3 parts of corn flour, 3 parts of calcium carbonate, 5 parts of fish meal, 0.3 part of calcium chloride, 0.5 part of monopotassium phosphate, 0.24 part of dipotassium hydrogen phosphate, 0.1 part of manganese sulfate, 0.1 part of sodium hydroxide, 3 parts of sodium chloride and 2 parts of defoaming agent;
the inoculation amount (mass percentage) of the bacillus amyloliquefaciens is 0.8 percent,
the aerobic culture temperature of the bacillus amyloliquefaciens is 37 ℃, and the bacillus amyloliquefaciens is cultured for 2 days.
The test effects of the complex microbial agent of the present invention are as follows
The first embodiment is as follows: influence of compound microbial agent on Chinese cabbage planting
1. Purpose of the experiment
Different functional strains produced by our company are compounded, and the influence of the compound microbial agent on the planting of the Chinese cabbage is determined through a plot experiment, so that a basis is provided for the popularization and the application of products on leaf vegetable crops.
2. Test materials and methods
2.1 test site
A test place outside a greenhouse of a Qinglong town factory area in Anning city.
2.2 test products
The compound microbial agent and the organic fertilizer are prepared by four strains of WSWFJ-2, WSWFJ-7, WSWFJ-8 and WSWFJ-11, and the compound fertilizer has the formula of 16-8-22.
2.3 test article
Chinese cabbage.
2.4 test design and method
The experiment totally designs 2 treatments, wherein 5000 plants are planted per mu, the fertilizing amount of the organic fertilizer applied per mu is 100 kg/mu, the compound fertilizer is 50 kg/mu, and the total fertilizing amount of each vegetable seedling is 0.03 kg. Treating a patient without compound microbial agent as a blank control; in the second treatment, 400 times of liquid of the compound microbial inoculum is dipped in roots when the Chinese cabbage is transplanted, 800 times of liquid is irrigated to the roots after the Chinese cabbage is transplanted for one week, and the application amount per mu of the compound microbial inoculum is 1.0 kg/mu when the roots are irrigated, specifically as follows:
TABLE 1 fertilization situation table
As the test aims at testing the disease prevention and growth promotion characteristics of the compound microbial inoculum, no additional fertilizer is applied.
The test field is planted in the open air, 5 soil moisture is selected for testing, each treatment design is repeated three times, 16 vegetables are planted in each cell, test vegetable seedlings are planted in the middle of 3 soil moisture, 2 soil moisture on two sides is used as a protection row, two plants are reserved before and after the middle of 3 soil moisture respectively and used as protection rows, and the test design is shown as follows.
All the fertilizer is required to be transplanted and is applied into the pond, the fertilizer is uniformly mixed with the bottom soil, 5cm of soil is covered on the fertilizer layer, and then vegetable seedling transplantation is carried out, so that fertilizer seedling burning is avoided. All processing is managed uniformly according to a conventional management mode.
3. Investigation and record
3.1 agronomic trait survey
Agronomic trait surveys were conducted 5 months and 4 days with the following results:
TABLE 2 statistical table of agronomic traits for one week after transplantation
| Treatment of | Plant height (cm) | Crown width (cm) |
| Process one | 18.00 | 57.42 |
| Treatment two | 20.10 | 70.50 |
The table shows that the first treatment can be weaker than the second treatment in terms of plant height and crown width, and the Chinese cabbage of the second treatment has better growth vigor.
Agronomic traits for 6 months and 15 days are shown in the following table:
TABLE 3 agronomic traits statistical Table before harvest
| Treatment of | Plant height (cm) | Crown width (cm) |
| Process one | 26.56 | 57.78 |
| Treatment two | 34.50 | 86.50 |
From the table above, the second treatment is superior to the first treatment in terms of plant height and crown width, and the growth advantage of the second Chinese cabbage is very obvious from the continuous growth of the Chinese cabbage to the harvest period, which shows that the compound microbial agent has very obvious growth promoting effect on the Chinese cabbage.
3.2 mu yield statistics
The acre yields for the different treatments are given in the table below:
TABLE 4 mu yield statistical table
As can be seen from the table above, the yield of the compound microbial agent is improved by 38.52% compared with that of the compound microbial agent when the compound microbial agent is applied to the production of Chinese cabbages, so that the compound microbial agent has good effects of increasing the yield and the income.
3.3 disease investigation statistics
TABLE 5 statistical table for disease investigation
| Treatment of | Soft rot (plant) | Clubroot (Berry) |
| Process one | 5 | 8 |
| Treatment two | 0 | 3 |
From the above table, compared with the first treatment, the second treatment added with the compound microbial agent has no soft rot, and the clubroot is greatly reduced, which shows that the compound microbial agent has obvious effects on enhancing the resistance of the Chinese cabbage, preventing and reducing the soft rot and other diseases.
4. Results and analysis
The compound microbial agent is applied to the planting process of the Chinese cabbage, has good effects of promoting the early growth and the quick growth of the Chinese cabbage, improving the yield and the output value of the Chinese cabbage and preventing and reducing the diseases of the Chinese cabbage, and can be popularized and applied in the field.
Example two: the compound microbial preparation has effect on the planting of Lincang Lingxiang rape
1. Demonstration purpose
The method combines the requirements of lincang soil conservation, adopts a mode of increasing the application of the compound microbial inoculum, supplements the compound microbial inoculum under the condition of consistent fertilization conditions in the rape planting process, improves the soil environment, improves the yield and the quality of the rape, and develops demonstration and popularization of the compound microbial inoculum in the lincang rape planting area.
2. Demonstration site
Lincang Lingxiangxiang rape planting area
3. Materials and methods
The demonstration variety is as follows: local conventional main-cultivated rape
Demonstration area: 50 mu m
Fertilizer to be tested: compound fertilizer (formula is 15-5-), compound microbial inoculum composed of 2 strains of WSWFJ-8 and WSWFJ-11.
An exemplary design: in the Lincang Lingxiangxiangsheng rape planting area, 50 mu are selected as demonstration sites, the compound microbial agent is added as demonstration in a root irrigation mode on the basis of local conventional fertilizers, and compared with the conventional fertilizers, the management mode is carried out according to a local recommended mode.
And (3) applying a base fertilizer: mixed and scattered on the bottom of the pond for transplantation.
4. Design of an exemplary scheme:
table 6 exemplary design fertilizing table
5. Investigation and recording
5.1 growth phase survey
And (4) surveying and recording the development of the growing periods of transplanting, growing, bolting, final flowering, harvesting and the like of the demonstration area and the control area, and understanding the influence of different fertilization modes on the development of the growing period of the rapes. 5.2 agronomic trait survey
And (4) investigating and recording the agronomic traits of the demonstration area and the control area, such as seedling vigor, effective plant number, branch number of each plant, effective bean clamp of each plant, thousand seed weight and the like, and knowing the influence of different fertilization modes on the agronomic trait development of the rapes.
5.3 economic character survey
And (4) investigating and recording economic characters such as fertilizer cost, labor days and the like of the demonstration area and the control area, and knowing the influence of different fertilization modes on the economic characters of the rapes.
5.4 fertilization benefit comparison
And (4) investigating and recording the fertilizing benefits of seed grain wind dry weight, reduced yield per mu and the like of the demonstration area and the control area, and comparing the fertilizing benefits of the rapes in different modes.
6. Data statistics
TABLE 7 survey of agronomic traits at flowering time
TABLE 8 Agronomic traits at knot-pinch stage
TABLE 9 rape test yield data
7. Summary of the invention
7.1 agronomic traits
As can be seen from tables 7 and 8, the growth vigor of the complex microbial inoculum is obviously better than that of the contrast, the plant height is higher, the branches are more, the catkin is long,
7.2 average yield per mu
The rape grows well in the year, and the yield exceeds the expected target. Through field measurement, the rape demonstration yield per mu is 257.7kg, and compared with the conventional yield per mu, the yield per mu is increased by 5 percent. The thousand kernel weight demonstration is higher than the contrast, which shows that the plumpness of rape kernel can be improved by using the microbial inoculum. The oil yield of the demonstration rape is 52kg by pressing 150kg of rapeseed oil respectively for the demonstration and the comparison, the oil yield is 50kg for the comparison, and the oil yield is 1.4 percent higher than that of the comparison.
7.3 demonstration effects
The yield per mu is high by using the compound microbial agent, and the yield per mu in a demonstration area reaches 257.6kg, is increased by 5 percent compared with the contrast and is obviously higher than the contrast from the yield measurement condition.
Secondly, the yield value is higher than that of contrast by using the compound microbial inoculum, and according to the price of the rape in the last year (4.6 yuan/kg), the yield value in the circle is increased by 156.4 yuan/mu, and the yield value of the Boshang is increased by 53.8 yuan/mu.
And thirdly, the compound microbial agent can improve the oil yield of the rapeseed, and the oil yield of the demonstration area is improved by 1.4 percent compared with that of the control area.
Example three: application research of compound microbial agent for preventing and treating tobacco black spots
In the tobacco planting process, a plurality of diseases need to be prevented in the early stage of planting, otherwise, large-area outbreak can be caused, and research shows that the disease-preventing functional microbial inoculum developed by the research center of microbial fermentation engineering of Yunnan province, Limited, is applied to dip and irrigate roots in the early stage of the growth period of a field, so that the tobacco has good prevention and treatment effects on the 'two-black disease'. The research applies the functional microbial inoculum to the tobacco planting process and aims to provide a new idea for preventing and treating the tobacco two-black disease in the wenshan tobacco area.
1. Materials and methods
1.1 test time and location
The test is carried out in a town of Yangtze county of inkstone county of Wenshan, Yunnan province in 2018 in a method of 3-10 months, and the test soil is moderate in fertility and good in irrigation and drainage conditions.
1.2 test materials and methods
The test varieties are as follows: yunyan 87
Test site: in the experiment, a greenhouse of a house of a new village is selected for seedling culture and cultivation according to a method of Yangtian town of inkstone county of Wenshan.
1.3 design of the experiment
1.3.1 growing seedlings
The experiment has 2 treatments, which are respectively as follows:
and (3) treatment: the fertilizer is applied according to a local conventional mode, no chemical agent is adopted for disease prevention and control, 500 times of liquid of the compound microbial inoculum produced by Yunnan province microbial fermentation engineering research center limited company is adopted for dipping roots before transplanting, 300 times of liquid of the compound microbial inoculum is adopted for irrigating the roots 7 days after transplanting, and the application amount of the compound microbial inoculum per mu is 2.0 kg/mu.
Comparison: the fertilizer is applied according to a local conventional mode, a chemical agent is adopted for preventing and controlling diseases, and a compound microbial agent is not applied.
The rest field management measures of each treatment are implemented according to local management schemes.
2. Process survey recording
2.1 investigation of disease and insect pests
And (4) investigating and counting the attack time and the damage degree (incidence rate and disease index, insect plant rate and insect population density) of various pests and diseases in each treatment in the whole field growth period.
2.2 investigation of chemical pesticide usage in conventional control measures
The using times, the prevention and treatment cost and the like of chemical pesticides used in conventional prevention and treatment measures are investigated and counted.
3. Results and analysis
3.1 statistical analysis of disease incidence
It can be seen from table 10 that the morbidity of mosaic disease, black shank, bacterial wilt, climatic spots and aphid treated by the microbial inoculum is lower than that of the control in the colony stage and the vigorous growth stage, indicating that the disease prevention effect of the microbial inoculum is slightly better than that of the chemical pesticide.
TABLE 10 disease statistics
3.2 statistics of the usage of the drugs and the prevention and treatment costs
It can be seen from table 11 that the use cost of the microbial inoculum of the treatment group is 12.5 yuan lower than that of the control group, the control group has various types of medicaments, the use times are more, the labor cost is increased, and the microbial inoculum has certain toxic action on the medicaments for preventing and treating the black shank.
TABLE 11 cost of medication
| Demonstration of | Comparison of | |
| Number of times of administration | 2 | 9 |
| Cost of medication (Yuan) | 32.5 | 20 |
4. Conclusion and discussion
The following conclusions can be drawn by experiments: the functional microbial agent has a certain prevention effect on black shank and bacterial wilt of tobacco plants in the field growth period, and can reduce the incidence of black spots in the colony stage and the vigorous growth stage.
Compared with the conventional planting process, the control cost and the drug cost of farmers by the functional biological agent are increased, and compared with the conventional planting process, the labor frequency is reduced, and the labor cost is reduced.
It should be understood that the above-described embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. Not all embodiments are exhaustive. All obvious changes and modifications which are obvious to the technical scheme of the invention are covered by the protection scope of the invention.
Claims (3)
1. A preparation method of a compound microbial agent with disease-resistant and growth-promoting functions is characterized by comprising the following steps: the method comprises the following steps:
(1) selecting Bacillus subtilis numbered WSWFJ-2, Bacillus subtilis numbered WSWFJ-7, Bacillus amyloliquefaciens numbered WSWFJ-8 and Bacillus amyloliquefaciens numbered WSWFJ-11, respectively fermenting, wherein the effective viable bacteria number in the fermentation liquor of each strain is more than 2.0 multiplied by 109cfu/ml；
(2) Mixing 25-35 parts by weight of a bacillus subtilis strain fermentation liquid numbered as WSWFJ-2, 0-35 parts by weight of a bacillus subtilis strain fermentation liquid numbered as WSWFJ-7, 25-50 parts by weight of a bacillus amyloliquefaciens strain fermentation liquid numbered as WSWFJ-8 and 25-50 parts by weight of a bacillus amyloliquefaciens strain fermentation liquid numbered as WSWFJ-11 to obtain a compound microbial agent with disease resistance and growth promotion functions;
the culture medium formula of the two bacillus subtilis strains comprises the following components in parts by weight: 10-16 parts of soybean meal, 7-13 parts of cane sugar, 1-5 parts of peptone, 2-8 parts of corn flour, 3-9 parts of calcium carbonate, 3-7 parts of fish meal, 0.2-0.8 part of calcium chloride, 0.2-0.7 part of potassium dihydrogen phosphate, 0.2-0.8 part of dipotassium hydrogen phosphate, 0.1-0.5 part of manganese sulfate, 0.1-0.5 part of sodium hydroxide, 2-7 parts of sodium chloride and 1-2 parts of defoaming agent;
the mass percentage of the inoculation amount of the bacillus subtilis is 0.5-1.5%,
the aerobic culture temperature of the bacillus subtilis is 35-39 ℃, and the bacillus subtilis is cultured for 1-3 days;
the culture medium formula of the two bacillus amyloliquefaciens strains comprises the following components in parts by weight: 7-13 parts of soybean meal, 5-9 parts of cane sugar, 3-8 parts of peptone, 2-7 parts of corn flour, 3-8 parts of calcium carbonate, 2-8 parts of fish meal, 0.2-0.7 part of calcium chloride, 0.2-0.7 part of potassium dihydrogen phosphate, 0.2-0.8 part of dipotassium hydrogen phosphate, 0.1-0.6 part of manganese sulfate, 0.1-0.4 part of sodium hydroxide, 3-7 parts of sodium chloride and 1-2 parts of defoaming agent;
the mass percentage of the inoculation amount of the bacillus amyloliquefaciens is 0.3-1 percent,
the aerobic culture temperature of the bacillus amyloliquefaciens is 33-40 ℃, and the bacillus amyloliquefaciens is cultured for 1-3 days;
the preservation registration number of the bacillus subtilis with the number of WSWFJ-2 is CGMCC NO. 15842;
the preservation registration number of the bacillus subtilis with the number of WSWFJ-7 is CGMCC NO. 15847;
the preservation registration number of the bacillus amyloliquefaciens with the number of WSWFJ-8 is CGMCC NO. 15848;
the preservation registration number of the bacillus amyloliquefaciens with the number of WSWFJ-11 is CGMCC NO. 15851.
2. The application of the compound microbial inoculant prepared by the preparation method of the compound microbial inoculant with disease-resistant and growth-promoting functions according to claim 1, wherein the preparation method comprises the following steps: the compound microbial agent is diluted to 700 times of solution by water according to the weight ratio and is used for dipping roots when transplanting after seedling raising of crops.
3. The application of the compound microbial inoculant prepared by the preparation method of the compound microbial inoculant with disease-resistant and growth-promoting functions according to claim 1, wherein the preparation method comprises the following steps: the compound microbial inoculum is diluted to 200-1100 times of liquid by water according to the weight ratio and is used for irrigating roots after transplanting crops, and the application amount per mu of the compound microbial inoculum is 0.3-3.0 kg/mu.
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| CN113249266B (en) * | 2021-06-22 | 2023-02-03 | 云南农业大学 | Bacillus subtilis capable of efficiently dissolving phosphorus and promoting sugarcane germination and application thereof |
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