CN111109026A - Biological control method for diseases in rice growth process and application - Google Patents
Biological control method for diseases in rice growth process and application Download PDFInfo
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- CN111109026A CN111109026A CN201911390335.8A CN201911390335A CN111109026A CN 111109026 A CN111109026 A CN 111109026A CN 201911390335 A CN201911390335 A CN 201911390335A CN 111109026 A CN111109026 A CN 111109026A
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- rice
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/20—Cereals
- A01G22/22—Rice
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C21/00—Methods of fertilising, sowing or planting
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protecting plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
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Abstract
The invention discloses a biological control method for diseases in a rice growth process and application thereof. The bio-control bacteria, a seedling culture substrate and an organic fertilizer are mixed according to a certain proportion on the basis of bio-control bacteria B579 fermentation production, and technologies such as bio-control bacteria liquid spraying are matched, so that the bio-control bacteria can protect the rice plant in the rice growth process, namely the rice seedling stage, the transplanting stage and the crevasse stage, infection of pathogenic bacteria to the rice plant is prevented, and the purpose of preventing and treating diseases is achieved. The invention has a comprehensive control effect of more than 65 percent on main diseases such as rice blast, banded sclerotial blight, damping off, bakanae disease and the like which are frequently generated in rice production.
Description
Technical Field
The invention belongs to the technical field of agricultural plant protection, and relates to a biological control method for diseases in a rice growth process and application thereof.
Background
The rice is the main grain crop in China, has the largest area, the highest yield per unit and the most total yield in the grain crop, and plays a significant role in guaranteeing the grain safety in China. With the breakthrough of rice technologies such as matrix seedling raising, mechanical seedling transplanting, formula fertilization and the like of rice and the transformation and upgrading of the industry, the traditional rice cultivation mode and prevention and control technology cannot meet the pursuit of people on the taste quality and the green health of rice. Green prevention and control is the development direction of plant protection technology in new period. The excessive use of pesticide causes various negative problems of pesticide residue exceeding standard, ecological environment pollution, increase of pesticide resistance of pests and the like. In addition, problems of agricultural product production safety and food quality safety occur.
The biological agent is an environment-friendly and sustainable agricultural biological agent advocated and applied by the current country, and has the following effects: (1) the pest control and the crop growth promotion are integrated, and the economic benefit is obvious; (2) make up for the deficiency of chemical fertilizer, chemical pesticide, reduce the pollution of chemical fertilizer, pesticide to soil and environment; (3) improving the ecological balance of soil microorganisms; (4) increasing the health level of the crop; (5) improve the quality of agricultural products. However, China has a great difference from the world in the aspect of researching agricultural microecological preparations, and the biocontrol bacteria has very good performance in laboratories and unstable effect when used in fields, so that the search for application technology is very important.
Disclosure of Invention
On the basis of the fermentation production of the biocontrol bacteria B579, the biocontrol bacteria, a seedling raising substrate and an organic fertilizer are mixed according to a certain proportion, and technologies such as biocontrol bacteria liquid spraying and the like are matched for preventing and controlling the rice damping-off, bakanae disease, banded sclerotial blight and rice blast. Preventing the infection of pathogenic bacteria to rice plants and achieving the purpose of preventing and treating diseases. Through extensive research, most of the application of the existing biocontrol bacteria is only one measure, or the biocontrol bacteria is directly sprayed before the breach of rice, or the biocontrol bacteria is applied in the form of a biological organic fertilizer, and the three technologies are rarely matched or integrated for use.
The invention uses the microorganism seedling raising matrix to raise rice seedlings, applies the biological organic fertilizer into the field in the form of base fertilizer before transplanting rice seedlings, directly sprays bacillus subtilis liquid 5d-7d before the rice breaks, and the prevention effect of the technology on rice diseases (damping off, bakanae disease, sheath blight and rice blast) reaches more than 65 percent.
To achieve the above object, the present invention discloses the following technical contents
A biological control method for diseases in the growth process of rice is characterized by comprising the following steps:
(1) fermentation production of bacillus subtilis
Fermentation process: adopting a stepwise amplification fermentation culture method, namely, from a test tube inclined plane (culture medium PDA), a 500ml triangular flask (culture medium PD), a 1 ton fermentation tank (culture medium NA) and a 10 ton fermentation tank (screened culture medium), the spore content of the bacillus subtilis liquid per milliliter is 1.05 multiplied by 1010cfu；
The components of the culture medium PDA are potato 200g, glucose (or sucrose) 20g, agar 18g and water 1L, which are subpackaged in test tubes of 1.8 x 20cm, 10ml of each test tube and sterilized for 30 minutes at 121 ℃;
the components of the culture medium PD refer to: potato 200, glucose (or sucrose) 20, and water 1L;
the components of the culture medium NA are as follows: 3g of beef extract, 10g of peptone, 5g of NaCl and 1L of water; the pH value of the mixture is 7.2,
the 10 ton fermenter medium composition was: 27% of corn flour, 3.1% of soybean meal, 0.17% of cane sugar, 0.06% of fish meal, 0.08% of magnesium sulfate, 0.003% of monopotassium phosphate, 0.001% of manganese sulfate, 0.001% of calcium chloride, 0.21% of yeast powder, 0.002% of NaCl0, 4 tons of liquid loading capacity of a 10-ton fermentation tank, 33 ℃, the ventilation volume of 400m3/h and the culture time of 19 hours;
the conditions for the progressive amplification culture of the bacillus subtilis are as follows:
(2) mixing the bacillus subtilis with a seedling culture substrate: mixing 100ml of bacillus subtilis liquid with 50 liters of seedling substrate to obtain a mixture containing 6 multiplied by 10 per gram7A seedling raising substrate of cfu spores;
(3) bacillus subtilis andmixing organic fertilizers: 20L of bacillus subtilis liquid is mixed with 1 ton of organic fertilizer to obtain the organic fertilizer containing 2.1 multiplied by 10 per gram8Spores of cfu;
(4) directly spraying the bacillus subtilis: 500ml of Bacillus subtilis liquid is added with 40kg of water, and each ml contains 1.3 multiplied by 108And (5) cfu spores are sprayed in the break-open period of the rice.
The invention further discloses an application of the biological disease control method in the rice growth process in preventing and controlling rice damping-off, bakanae disease, sheath blight and rice blast. The experimental result shows that the comprehensive control effect on the four diseases reaches more than 65 percent.
The invention is described in more detail below:
a biological control method for diseases in the growth process of rice is characterized in that:
1. fermentation production of bacillus subtilis
1.1 fermentation procedure
Adopts a step-by-step amplification fermentation culture method, namely, a test tube inclined plane is adopted to form a 500ml triangular bottle and a 1 ton fermentation tank to a 10 ton fermentation tank.
1.2 fermentation conditions
The fermentation conditions at each stage are shown in Table 1
TABLE 1 conditions for the stepwise amplification culture of Bacillus subtilis
1.3 spore content
The spore content of the bacillus subtilis liquid per milliliter is 1.05 multiplied by 1010cfu。
Secondly, mixing the bacillus subtilis and the seedling substrate
100ml of bacillus subtilis liquid and 50 liters of seedling substrate are mixed and stirred to obtain the mixture containing 6 multiplied by 10 per gram7A seedling culture substrate of cfu spores.
Thirdly, mixing the bacillus subtilis with the organic fertilizer
20L of bacillus subtilis liquid is mixed with 1 ton of organic fertilizer to obtain the organic fertilizer containing 2.1 multiplied by 10 per gram8And (5) spores of cfu.
Fourthly, directly spraying the bacillus subtilis
500ml of Bacillus subtilis liquid is added with 40kg of water, and each ml contains 1.3 multiplied by 108And (5) spraying cfu spores 5-7 days before the rice breaks.
Application technology and effect of bacillus subtilis
A large amount of bacillus subtilis is arranged around the rhizosphere of rice seedlings bred by a microorganism seedling breeding matrix, then the rice seedlings are transplanted with bacteria in a field applied with a bio-organic fertilizer (the dosage of each mu is 1.5 tons), so that biocontrol bacteria exist around the rhizosphere and soil, the rhizosphere is protected from or less infected by pathogenic bacteria, and in addition, the bacteria liquid is directly sprayed on the rice 5-7 days before the opening of the rice, so that the infection of pathogenic bacteria in the air in the sensitive period of rice disease attack is prevented, and the effect of whole-course prevention is achieved.
5.1 prevention and control effects of microorganism seedling raising matrix on rice damping-off
TABLE 2 prevention and treatment effects of the microorganism seedling substrate on rice damping-off
5.2 prevention and treatment effects of microbial seedling substrate on bakanae disease of rice
TABLE 3 prevention and treatment effects of microorganism seedling raising substrate on bakanae disease of rice
The microbial seedling raising matrix is used for raising rice seedlings, and has a rice damping-off preventing effect of 65.4 percent and a rice bakanae disease preventing effect of 69.3 percent.
5.3 prevention and control effects of Bacillus subtilis on banded sclerotial blight
TABLE 4 prevention and treatment effects of Bacillus subtilis on rice sheath blight disease
5.4 prevention and control effects of Bacillus subtilis on Rice blast
TABLE 5 prevention and control Effect of Bacillus subtilis on Rice blast
From tables 4 and 5, it can be seen that the control effect on the rice sheath blight disease reaches 72.2%, and the control effect on the rice blast reaches 70.3%.
In conclusion, through the application of the technology, diseases such as damping off, bakanae disease, sheath blight and rice blast which are frequently generated in the growth period of rice can be effectively prevented and controlled, and the prevention effect reaches more than 65%.
The invention mainly solves the problems of poor and unstable biocontrol bacterium control effect, mainly inspects the control effect of the biocontrol bacterium application method on rice damping off, bakanae disease, sheath blight and rice blast, and has the main difficulty that how to organically combine and use the biocontrol bacterium application method in a matched manner.
Detailed Description
The invention is described below by means of specific embodiments. Unless otherwise specified, the technical means used in the present invention are well known to those skilled in the art. In addition, the embodiments should be considered illustrative, and not restrictive, of the scope of the invention, which is defined solely by the claims. It will be apparent to those skilled in the art that various changes or modifications in the components and amounts of the materials used in these embodiments can be made without departing from the spirit and scope of the invention. The raw materials and reagents used in the present invention are commercially available. The seedling substrate and the organic fertilizer are sold in the market. The separation, screening and identification of the biocontrol bacterium B579 used in the invention are detailed in Shandong agricultural science 2008, 3: 91-94. The preservation number of the bacillus subtilis strain is as follows: CGMCC 2270.
Example 1
A biological control method for diseases in the growth process of rice comprises the following steps:
fermentation process: adopting a step-by-step amplification fermentation culture method, namely, from a test tube inclined plane (culture medium PDA) to 500ml triangular flask (culture medium PD), 1 ton fermentation tank (culture medium NA) and 10 ton fermentation tank, the fermentation time is 19 hours, and the spore content of bacillus subtilis liquid per milliliter is 1.05 multiplied by 1010cfu；
(1) Fermentation production and fermentation process of bacillus subtilis:
1) slant tube culture: potato 200g, glucose (or sucrose) 20g, agar 18g, water 1L, divided into 1.8 x 20cm tubes, 10ml each. Sterilizing at 121 deg.C for 30 min. B579 was inoculated on a slant and cultured at 30 ℃ for 18 h.
2) Culturing in 500ml triangular flask: 200g of potato, 20g of glucose (or sucrose) and 1L of water are subpackaged in 500ml triangular bottles, each bottle is 100ml, and the sterilization is carried out for 30 minutes at the temperature of 121 ℃. Transferring the bacillus subtilis growing on the inclined plane to a triangular flask for continuous culture at 29 ℃ and the rotating speed of 180r/min for 48 h.
3) Culturing in 1 ton fermentation tank with beef extract 3g, peptone 10g, NaCl 5g, and water 1L; pH 7.2, liquid loading of 1 ton fermenter 400L, 33 deg.C, and air flow of 30m3And h, culturing for 6 h.
4) 10 ton fermentation tank: 27% of corn flour, 3.1% of soybean meal, 0.17% of cane sugar, 0.06% of fish meal, 0.08% of magnesium sulfate, 0.003% of potassium dihydrogen phosphate, 0.001% of manganese sulfate, 0.001% of calcium chloride, 0.21% of yeast powder, 0.002% of NaCl0, 4 tons of liquid loading capacity of a 10 ton fermentation tank, 33 ℃ and ventilation capacity of 400m3H, incubation time 19 hours.
The spore content of the bacillus subtilis liquid per milliliter is 1.05 multiplied by 1010cfu, conditions for stepwise amplification culture of bacillus subtilis are as follows:
(2) mixing the bacillus subtilis with a seedling culture substrate: mixing 100ml of bacillus subtilis liquid with 50 liters of seedling substrate to obtain a mixture containing 6 multiplied by 10 per gram7A seedling raising substrate of cfu spores;
(3) mixing bacillus subtilis with an organic fertilizer: 20L of bacillus subtilis liquid is mixed with 1 ton of organic fertilizer to obtain the organic fertilizer containing per gramHas a size of 2.1 × 108Spores of cfu;
(4) directly spraying the bacillus subtilis: 500ml of Bacillus subtilis liquid is added with 40kg of water, and each ml contains 1.3 multiplied by 108And (5) cfu spores are sprayed in the break-open period of the rice.
The results were: has certain control effect on rice blast and sheath blight of rice in the ear stage.
Claims (2)
1. A biological control method for diseases in the growth process of rice is characterized by comprising the following steps:
(1) fermentation production of bacillus subtilis
Fermentation process: adopting a stepwise amplification fermentation culture method, namely fermenting for 19 hours from a test tube inclined plane (culture medium PDA), a 500ml triangular flask (culture medium PD), a 1 ton fermentation tank (culture medium NA) and a 10 ton fermentation tank, wherein the spore content of bacillus subtilis liquid per milliliter is 1.05 multiplied by 1010cfu；
The components of the culture medium PDA are potato 200g, glucose (or sucrose) 20g, agar 18g and water 1L, which are subpackaged in test tubes of 1.8 x 20cm, 10ml of each test tube and sterilized at 121 ℃ for 30 minutes;
the components of the culture medium PD refer to: potato 200, glucose (or sucrose) 20, and water 1L;
the components of the culture medium NA are as follows: 3g of beef extract, 10g of peptone, 5g of NaCl and 1L of water; the pH value of the mixture is 7.2,
10 ton fermentation tank: 27% of corn flour, 3.1% of soybean meal, 0.17% of cane sugar, 0.06% of fish meal, 0.08% of magnesium sulfate, 0.003% of potassium dihydrogen phosphate, 0.001% of manganese sulfate, 0.001% of calcium chloride, 0.21% of yeast powder, 0.002% of NaCl0, 4 tons of liquid loading capacity of a 10 ton fermentation tank, 33 ℃ and ventilation capacity of 400m3H, culturing for 19 hours; the conditions for the progressive amplification culture of the bacillus subtilis are as follows:
(2) mixing the bacillus subtilis with a seedling culture substrate: 100ml of Bacillus subtilisThe bacterial liquid is mixed with 50 liters of seedling substrate to obtain a mixture containing 6 multiplied by 10 per gram7A seedling raising substrate of cfu spores;
(3) mixing bacillus subtilis with an organic fertilizer: 20L of bacillus subtilis liquid is mixed with 1 ton of organic fertilizer to obtain the organic fertilizer containing 2.1 multiplied by 10 per gram8Spores of cfu;
(4) directly spraying the bacillus subtilis: 500ml of Bacillus subtilis liquid is added with 40kg of water, and each ml contains 1.3 multiplied by 108And (5) spraying the cfu spores 5-7 days before the opening period of the rice.
2. The method for biologically controlling diseases in the growth process of rice according to claim 1, wherein the method is used for controlling rice seedling blight, bakanae disease, sheath blight and rice blast.
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Cited By (2)
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| CN108977366A (en) * | 2018-08-16 | 2018-12-11 | 天津市植物保护研究所 | A kind of method that phytopathogen expands culture |
| CN112759455A (en) * | 2021-01-19 | 2021-05-07 | 甘南县龙头岭有机肥有限公司 | Nutrient solution for rice seedling stage in cold region and preparation method thereof |
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|---|---|---|---|---|
| CN108977366A (en) * | 2018-08-16 | 2018-12-11 | 天津市植物保护研究所 | A kind of method that phytopathogen expands culture |
| CN112759455A (en) * | 2021-01-19 | 2021-05-07 | 甘南县龙头岭有机肥有限公司 | Nutrient solution for rice seedling stage in cold region and preparation method thereof |
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