CN113951279A - Bacteriostatic combination liquid for preventing and treating tobacco bacterial wilt as well as preparation method and application thereof - Google Patents
Bacteriostatic combination liquid for preventing and treating tobacco bacterial wilt as well as preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to a bacteriostatic combined solution for preventing and treating tobacco bacterial wilt, a preparation method and application, belonging to the technical field of plant disease prevention and treatment, wherein the bacteriostatic combined solution is prepared by mixing a microbial liquid and a chemical composition; the microbial solution is obtained by mixing lactobacillus bulgaricus lysate and bacillus pumilus fermentation liquor fermented to logarithmic growth phase in a volume ratio of 1: 1; the chemical composition comprises fulvic acid, sinapine, boron plus selenium, puerarin, ellagic acid and chitosan oligosaccharide; the pH value of the antibacterial combined liquid is 7.2-7.8. The antibacterial combination liquid is prepared by effectively combining a microbial agent and chemical components, has a remarkable inhibition effect on tobacco bacterial wilt bacteria through effective proportioning, has a certain promotion effect on the growth and development of tobacco seedlings, and has great significance on the prevention and treatment of tobacco bacterial wilt.
Description
Technical Field
The invention belongs to the technical field of plant disease control, and particularly relates to a bacteriostatic combination liquid for controlling tobacco bacterial wilt, a preparation method and application.
Background
The tobacco bacterial wilt is also called as myxopathy, tobacco plague and loblolly, and is a disease which is caused by the infection of ralstonia solanacearum and occurs on tobacco. Tobacco bacterial wilt is a typical vascular bundle disease, all parts of roots, stems and leaves can be damaged, and the most typical symptom is blight. The survival time of Ralstonia solanacearum in different environments or soils varies widely. Survives can survive for 7 months on host disease residues, survive for 2-3 years in soil or compost, compete for 8-25 years in soil, but die quickly under dry conditions.
The germs live through the winter mainly on the soil and the diseased plant residues left in the soil, and also on other living hosts. Therefore, the main primary sources of bacterial wilt of tobacco are soil, disease residues and fertilizers. Germs generally invade from the root wound and develop from the bottom to the top. The germs enter host tissues and then divide and propagate, enter vascular bundles, expand to other tissues and generate mucus containing complex polysaccharide, so that the viscosity of the vascular bundle liquid flow is improved, and a catheter is blocked, so that the disease plants wither. The pathogenic bacteria can secrete pectase and cellulolytic enzyme, dissolve the mesocolloid layer of host cell, and cause cell separation and decay and collapse of host cortex and medulla tissue. Cell wall disruption and cell collapse provide a convenient means for lateral movement of bacteria from one cell to another, further hindering water transport and accelerating plant blight. After the diseased tissue is rotten, the germs are left in the soil or in the manure for overwintering and become the initial infection source of the next year.
The tobacco leaf green disease has great influence on the yield and the quality of tobacco, and restricts the continuous and stable development of the tobacco industry in China. The prevention and treatment of tobacco bacterial wilt is a problem which is concerned by the world, researchers at home and abroad do a lot of work in the aspects of disease-resistant variety breeding, agricultural prevention and treatment, chemical agent prevention and treatment, biological prevention and treatment and the like, but the disease-resistant variety breeding cost is high, the agricultural prevention and treatment time is long, and long-term use of chemical agents can cause pathogenic bacteria to generate drug resistance to bactericides and cause imbalance of the ecological system of soil, environmental pollution and drug residue of tobacco leaves, so that the human health is seriously influenced.
Disclosure of Invention
Aiming at the dilemma in the aspect of prevention and control of tobacco bacterial wilt at present, the invention aims at providing the bacteriostatic combination liquid for prevention and control of tobacco bacterial wilt, aims at providing the preparation method of the bacteriostatic combination liquid, and aims at providing the application of the bacteriostatic combination liquid in the aspect of prevention and control of tobacco bacterial wilt. The antibacterial combination liquid prepared by adopting two effective microbial strains and combining six chemical components of fulvic acid, sinapine, boron plus selenium, puerarin, ellagic acid and chitosan oligosaccharide has an obvious inhibiting effect on ralstonia solanacearum, and plays an important role in preventing and controlling tobacco bacterial wilt.
In order to achieve the purpose, the invention adopts the specific scheme that:
a bacteriostatic combination solution for preventing and treating tobacco bacterial wilt is prepared by mixing a microbial solution and a chemical composition; the microbial solution is obtained by mixing lactobacillus bulgaricus lysate and bacillus pumilus fermentation liquor fermented to logarithmic growth phase in a volume ratio of 1: 1; the chemical composition comprises fulvic acid, sinapine, selenone, puerarin, ellagic acid and chitosan oligosaccharide, wherein the concentration of the fulvic acid is 0.5-1.0g/L, the concentration of the sinapine is 0.1-0.15g/L, the concentration of the selenone is 1.0-2.0mL/L, the concentration of the puerarin is 0.05-0.08g/L, the concentration of the ellagic acid is 2.0-2.5g/L, and the concentration of the chitosan oligosaccharide is 3.6-4.5 g/L; the pH value of the antibacterial combined liquid is 7.2-7.8.
The preparation method of the antibacterial combined liquid comprises the following steps:
1) preparation of lactobacillus bulgaricus lysate: weighing 22-25g of lactobacillus bulgaricus strain powder; adding 40-50ml of sterile water, uniformly mixing by vortex, centrifuging, and removing supernatant to obtain bacterial sludge; adding 40-50ml of sterile water and 1-2g of glass beads into 5-10g of the bacterial sludge, carrying out vortex oscillation, and carrying out ice-bath cracking for 15-20min by using an ultrasonic cracker to obtain a cracking stock solution; centrifuging the lysis stock solution, and recovering the supernatant to obtain lactobacillus bulgaricus lysate;
2) preparing a bacillus pumilus fermentation liquid: taking a Bacillus pumilus strain, sequentially carrying out strain activation and amplification culture on the Bacillus pumilus strain with the inoculum size of 10-15%, culturing the Bacillus pumilus strain for 15-18h at 36-38 ℃ and 200-220rpm, and filtering the Bacillus pumilus strain by 6-8 layers of sterilization gauze to obtain a Bacillus pumilus fermentation liquid;
3) mixing the lactobacillus bulgaricus lysate obtained in the step 1) and the bacillus pumilus fermentation liquid obtained in the step 2) according to the volume ratio of 1-3: 1 to obtain a microbial liquid;
4) adding fulvic acid, sinapine, boron selenium, puerarin, ellagic acid and chitosan oligosaccharide into the microbial solution obtained in the step 3), mixing uniformly and dissolving to obtain the antibacterial combined solution.
Preferably, in the preparation method of the bacteriostatic combination solution, the bacillus pumilus is cultured by using a spore-forming culture solution, and the spore-forming culture solution comprises the following components: 1 percent of glucose, 0.5 percent of molasses, 1 percent of protein powder, 0.3 percent of potassium chloride and 0.5 percent of manganese sulfate, and the pH value is 7.0.
The application also requests to protect the application of the bacteriostatic combination solution in the aspect of biological-chemical comprehensive prevention and control of tobacco bacterial wilt.
Preferably, the application refers to a method for biological-chemical integrated control of tobacco leaf disease, the method comprises the following steps: preparing an antibacterial combination solution, and diluting with sterile water by 50-100 times to obtain a diluent for later use; selecting healthy and disease-free tobacco seeds, soaking the seeds in the obtained diluent at room temperature for 15-18h, and then sowing the seeds in a seedling tray; when transplanting, the obtained diluent is adopted for foliage spraying and root irrigation; the spraying amount of the leaf surfaces is 5-10ml, and the root irrigation amount is 30-40 ml; respectively spraying the whole plant with the obtained diluent at 25-30d and 50-55d after transplanting, wherein the spraying amount is 40-50 ml; the rest period is carried out according to the normal cultivation mode of tobacco.
Compared with the prior art, the invention has the following technical effects:
the application provides an antibacterial combined solution, is to carry out effective combination with the chemical composition with microbial inoculum, adopts Bulgaria lactobacillus lysate and Bacillus pumilus zymotic fluid, is equipped with fulvic acid, sinapine, boron and selenium, puerarin, ellagic acid and chitosan oligosaccharide, through effective ratio, makes antibacterial combined solution has played apparent inhibitory effect to tobacco bacterial wilt germ, and its growth and development to the tobacco seedling has certain degree's promotion effect, has great meaning to the prevention and cure of tobacco bacterial wilt.
According to the application, various health-care cultivation technologies such as root irrigation, leaf surface spraying and the like are adopted, so that tobacco plants grow robustly, the disease resistance and the stress resistance of the tobacco plants are enhanced, the incidence rate of bacterial wilt of the tobacco plants is obviously reduced, the tobacco plants grow well, the appearance quality of the baked tobacco leaves is good, and the economic benefit is obviously improved.
Detailed Description
The indicator pathogenic bacteria employed in this application are ralstonia solanacearum ()Ralstonia solanacearum) The strain is provided by the agricultural college of the university of technology in Henan.
The tobacco variety is MS Yunyan 87.
Culture medium: TTC medium formula (g/L): beef extract 3, peptone 10, sodium chloride 5, glucose 6, TTC 0.05, pH 7.2-7.4.
A bacteriostatic combination solution for preventing and treating tobacco bacterial wilt is prepared by mixing a microbial solution and a chemical composition; the microbial solution is obtained by mixing lactobacillus bulgaricus lysate and bacillus pumilus fermentation liquor fermented to logarithmic growth phase in a volume ratio of 1: 1; the chemical composition comprises fulvic acid, sinapine, selenone, puerarin, ellagic acid and chitosan oligosaccharide, wherein the concentration of the fulvic acid is 0.5-1.0g/L, the concentration of the sinapine is 0.1-0.15g/L, the concentration of the selenone is 1.0-2.0mL/L, the concentration of the puerarin is 0.05-0.08g/L, the concentration of the ellagic acid is 2.0-2.5g/L, and the concentration of the chitosan oligosaccharide is 3.6-4.5 g/L; the pH value of the antibacterial combined liquid is 7.2-7.8.
The microbial liquid adopts lactobacillus bulgaricus and bacillus pumilus to respectively carry out cracking and fermentation.
The preparation method of the lactobacillus bulgaricus lysate comprises the following steps:
lactobacillus bulgaricus (Lactobacillus delbrueckii subsp. bulgaricus), Taiwan Suzhou core Biotechnology Limited, the number of lyophilized powder is more than or equal to 100 hundred million/g;
(1) weighing 22-25g of the strain powder, placing the strain powder in a 50ml centrifuge tube, adding sterile water into the centrifuge tube until the scale is about 40-50ml, carrying out vortex oscillation for 1 min, shaking up, centrifuging for 5 min at 3000 rpm by using a centrifuge, removing supernatant, continuously adding 40-50ml of sterile water into the precipitate, carrying out vortex mixing, carrying out high-speed centrifugation again, removing supernatant, and repeating for 4 times to finally obtain toothpaste-like bacterial sludge without culture medium components;
(2) weighing 5-10g of the bacterial sludge, putting the bacterial sludge into a 50ml centrifuge tube, adding 40-50ml of sterile water, simultaneously adding 1-2g of glass beads with the diameter of 0.5 mm, uniformly mixing by vortex oscillation, carrying out ice bath cracking for 30 min by using an ultrasonic cracker, setting the parameters to be 500 watt power, and switching on for 5s and switching off for 5 s;
(3) and (3) centrifuging the lysate after the cracking is finished at a high speed of 4000 rpm for 30 min, recovering the supernatant, and discarding the precipitate to obtain the lactobacillus bulgaricus lysate.
The preparation method of the bacillus pumilus fermentation liquid comprises the following steps:
bacillus pumilus (commercially available) is adopted, strain activation and expanded culture are sequentially carried out, the inoculation amount is 10-15%, the culture is carried out for 15-18h at 36-38 ℃ and 200-220rpm, and then the filtration is carried out through 6-8 layers of sterilization gauze, so as to obtain the Bacillus pumilus fermentation liquor; the bacillus pumilus is cultured by using a spore-producing culture solution, and the spore-producing culture solution comprises the following components: 1 percent of glucose, 0.5 percent of molasses, 1 percent of protein powder, 0.3 percent of potassium chloride and 0.5 percent of manganese sulfate, and the pH value is 7.0.
Example 1
A bacteriostatic combination solution for preventing and treating tobacco bacterial wilt is prepared by mixing a microbial solution and a chemical composition; the microbial solution is obtained by mixing lactobacillus bulgaricus lysate and bacillus pumilus fermentation liquor fermented to logarithmic growth phase in a volume ratio of 1: 1; the chemical composition comprises fulvic acid, sinapine, selenium boronate, puerarin, ellagic acid and chitosan oligosaccharide, wherein the concentration of the fulvic acid is 0.5 g/L, the concentration of the sinapine is 0.15g/L, the concentration of the selenium boronate is 2.0mL/L, the concentration of the puerarin is 0.08g/L, the concentration of the ellagic acid is 2.0 g/L, and the concentration of the chitosan oligosaccharide is 4.5 g/L; the pH value of the bacteriostatic combined solution is 7.5.
Example 2
A bacteriostatic combination solution for preventing and treating tobacco bacterial wilt is prepared by mixing a microbial solution and a chemical composition; the microbial solution is obtained by mixing lactobacillus bulgaricus lysate and bacillus pumilus fermentation liquor fermented to logarithmic growth phase in a volume ratio of 1: 1; the chemical composition comprises fulvic acid, sinapine, selenium boronate, puerarin, ellagic acid and chitosan oligosaccharide, wherein the concentration of the fulvic acid is 0.8 g/L, the concentration of the sinapine is 0.15g/L, the concentration of the selenium boronate is 1.5 mL/L, the concentration of the puerarin is 0.06 g/L, the concentration of the ellagic acid is 2.5g/L, and the concentration of the chitosan oligosaccharide is 4.0 g/L; the pH value of the bacteriostatic combined solution is 7.8.
Example 3
A bacteriostatic combination solution for preventing and treating tobacco bacterial wilt is prepared by mixing a microbial solution and a chemical composition; the microbial solution is obtained by mixing lactobacillus bulgaricus lysate and bacillus pumilus fermentation liquor fermented to logarithmic growth phase in a volume ratio of 1: 1; the chemical composition comprises fulvic acid, sinapine, selenium boronate, puerarin, ellagic acid and chitosan oligosaccharide, wherein the concentration of the fulvic acid is 1.0g/L, the concentration of the sinapine is 0.1 g/L, the concentration of the selenium boronate is 1.0 mL/L, the concentration of the puerarin is 0.05 g/L, the concentration of the ellagic acid is 2.5g/L, and the concentration of the chitosan oligosaccharide is 3.6 g/L; the pH value of the bacteriostatic combined solution is 7.2.
Comparative example 1: preparing a bacteriostatic combined solution, wherein the bacteriostatic combined solution comprises a microbial solution and a chemical composition, the microbial solution is a lactobacillus bulgaricus lysate, and the preparation method is the same as that in example 1; the chemical composition and the concentration are the same as those in example 1, and the pH of the bacteriostatic combined solution is 7.5.
Comparative example 2: preparing a bacteriostatic combined solution, wherein the bacteriostatic combined solution comprises a microbial liquid and a chemical composition, the microbial liquid is a bacillus pumilus fermentation liquid, and the preparation method is the same as that of the embodiment 1; the chemical composition and the concentration are the same as those in example 1, and the pH of the bacteriostatic combined solution is 7.5.
Comparative example 3: the antibacterial combination liquid is prepared by mixing lactobacillus bulgaricus lysate and bacillus pumilus fermentation liquid fermented to logarithmic growth phase in a volume ratio of 1:1, and does not contain a chemical composition, and the pH value of the antibacterial combination liquid is 7.5.
Comparative example 4: preparing a bacteriostatic combined solution, wherein the bacteriostatic combined solution is prepared by mixing a microbial solution and a chemical composition; the microbial solution is obtained by mixing lactobacillus bulgaricus lysate and bacillus pumilus fermentation liquor fermented to logarithmic growth phase in a volume ratio of 1: 1; the chemical composition comprises fulvic acid and chitosan oligosaccharide, wherein the concentration of the fulvic acid is 0.5 g/L, and the concentration of the chitosan oligosaccharide is 4.5 g/L. The pH value of the bacteriostatic combined solution is 7.5.
Comparative example 5: preparing a bacteriostatic combined solution, wherein the bacteriostatic combined solution is prepared by mixing a microbial solution and a chemical composition; the microbial solution is obtained by mixing lactobacillus bulgaricus lysate and bacillus pumilus fermentation liquor fermented to logarithmic growth phase in a volume ratio of 1: 1; the chemical composition comprises fulvic acid, sinapine and boron plus selenium, wherein the concentration of the fulvic acid is 0.5 g/L, the concentration of the sinapine is 0.15g/L, and the concentration of the boron plus selenium is 2.0 mL/L; the pH value of the bacteriostatic combined solution is 7.5.
Control group 6: preparing an antibacterial combined solution, wherein the antibacterial combined solution comprises fulvic acid, sinapine, boron plus selenium, puerarin, ellagic acid and chitosan oligosaccharide, wherein the concentration of the fulvic acid is 1.0g/L, the concentration of the sinapine is 0.1 g/L, the concentration of the boron plus selenium is 1.0 mL/L, the concentration of the puerarin is 0.05 g/L, the concentration of the ellagic acid is 2.5g/L, and the concentration of the chitosan oligosaccharide is 3.6 g/L; the pH value of the bacteriostatic combined solution is 7.2; the pH value of the bacteriostatic combined solution is 7.5.
Comparative example 7: preparing a bacteriostatic combination solution, wherein the bacteriostatic combination solution is obtained by mixing bacillus subtilis fermentation liquor and bacillus pumilus fermentation liquor in a volume ratio of 1:1, and does not contain a chemical composition, and the pH value of the bacteriostatic combination solution is 7.5.
Comparative example 8: the antibacterial combination liquid is prepared by mixing lactobacillus bulgaricus lysate and bacillus subtilis fermentation liquor in a volume ratio of 1:1, and does not contain a chemical composition, and the pH value of the antibacterial combination liquid is 7.5.
Comparative example 9: preparing a bacteriostatic combined solution, wherein the bacteriostatic combined solution is prepared by mixing a microbial solution and a chemical composition; the microbial liquid is bacillus subtilis fermentation liquid; the chemical composition comprises fulvic acid, sinapine, selenium boronate, puerarin, ellagic acid and chitosan oligosaccharide, wherein the concentration of the fulvic acid is 0.5 g/L, the concentration of the sinapine is 0.15g/L, the concentration of the selenium boronate is 2.0mL/L, the concentration of the puerarin is 0.08g/L, the concentration of the ellagic acid is 2.0 g/L, and the concentration of the chitosan oligosaccharide is 4.5 g/L; the pH value of the bacteriostatic combined solution is 7.5.
Comparative example 10: 20% benomyl wettable powder 600 times liquid.
The safety and bacteriostatic activity of the bacteriostatic combination solution for preventing and treating tobacco bacterial wilt, which is disclosed by the application, are analyzed.
Firstly, analyzing the influence on tobacco seedling culture.
Selecting healthy tobacco seeds, diluting the bacteriostatic combination solution obtained in the embodiment 1-3 by 100 times, soaking the seeds for 15 hours at room temperature, sowing the soaked seeds in a seedling tray by taking sterile water soaking (CK) as a control, sowing 1 seed in each grid, culturing for 25 days in a 25 ℃ illumination incubator, taking out the tobacco seedlings, cleaning, then sucking water, and calculating the germination rate; 8 tobacco seedlings with similar growth vigor are selected, the height, weight and root length of the seedlings are respectively measured, and the results are shown in table 1.
Table 1: the effect of the bacteriostatic combined solution on the germination rate, the seedling height, the seedling weight and the root length of tobacco seeds.
Note: the values in the table are mean ± standard error (n = 8).
As can be seen from table 1 above, the tobacco seedlings treated by the bacteriostatic combination solution described in the present application have good growth vigor, and compared with CK, the bacteriostatic combination solution has no adverse effect on the growth and development of tobacco seedlings, but rather has certain promotion effects on germination percentage, seedling weight, root length and seedling height.
And secondly, analyzing the bacterial inhibition activity of ralstonia solanacearum.
Preparing an antibacterial combination solution, using a 96-well plate, respectively adding 50 mu L of the antibacterial combination solution, 50 mu L of bacterial suspension and 100 mu L of culture medium into each well, repeating each group of test for three times, putting the test into an incubator to cultivate for 12 hours at 30 ℃, measuring an OD value by using an enzyme-labeling instrument at 630nm, repeating the steps for three times, and calculating the inhibition rate. Setting a blank group and a control group at the same time; the blank group uses sterile water and a spore-forming culture solution (1: 1) to replace the bacteriostatic combined solution, uses a culture medium to replace the bacterial suspension, and the control group uses sterile water and a spore-forming culture solution (1: 1) to replace the bacteriostatic combined solution.
Inhibition (%) = (control OD value-test OD value)/(control OD value-blank OD value) × 100%.
Table 2: comparison of bacteriostatic Activity
Therefore, the bacteriostatic combination solution has better inhibitory capacity on ralstonia solanacearum.
The method for biologically-chemically and comprehensively preventing and treating the tobacco leaf disease by using the bacteriostatic combination solution comprises the following steps: preparing an antibacterial combination solution, and diluting with sterile water by 50-100 times to obtain a diluent for later use; selecting healthy and disease-free tobacco seeds, soaking the seeds in the diluent obtained in the step one at room temperature for 15-18h, and then sowing the seeds in a seedling tray; when transplanting, the obtained diluent is adopted for foliage spraying and root irrigation; the spraying amount of the leaf surfaces is 5-10ml, and the root irrigation amount is 30-40 ml; respectively spraying the whole plant with the obtained diluent at 25-30d and 50-55d after transplanting, wherein the spraying amount is 40-50 ml; the rest period is carried out according to the normal cultivation mode of tobacco. By adopting the method, the morbidity and disease index of the tobacco bacterial wilt can be effectively reduced through practice, and the relative control effect can reach 88.39 percent after the tobacco bacterial wilt is transplanted for 120 days.
It should be noted that the above-mentioned embodiments illustrate rather than limit the scope of the invention, which is defined by the appended claims. It will be apparent to those skilled in the art that certain insubstantial modifications and adaptations of the present invention can be made without departing from the spirit and scope of the invention.
Claims (5)
1. A bacteriostatic combination liquid for preventing and treating tobacco bacterial wilt is characterized in that: the bacteriostatic combined solution is formed by mixing a microbial solution and a chemical composition; the microbial solution is obtained by mixing lactobacillus bulgaricus lysate and bacillus pumilus fermentation liquor fermented to logarithmic growth phase in a volume ratio of 1: 1; the chemical composition comprises fulvic acid, sinapine, selenone, puerarin, ellagic acid and chitosan oligosaccharide, wherein the concentration of the fulvic acid is 0.5-1.0g/L, the concentration of the sinapine is 0.1-0.15g/L, the concentration of the selenone is 1.0-2.0mL/L, the concentration of the puerarin is 0.05-0.08g/L, the concentration of the ellagic acid is 2.0-2.5g/L, and the concentration of the chitosan oligosaccharide is 3.6-4.5 g/L; the pH value of the antibacterial combined liquid is 7.2-7.8.
2. The preparation method of the bacteriostatic combination solution according to claim 1, characterized in that: the method comprises the following steps:
step one, preparing lactobacillus bulgaricus lysate: weighing 22-25g of lactobacillus bulgaricus strain powder; adding 40-50ml of sterile water, uniformly mixing by vortex, centrifuging, and removing supernatant to obtain bacterial sludge; adding 40-50ml of sterile water and 1-2g of glass beads into 5-10g of the bacterial sludge, carrying out vortex oscillation, and carrying out ice-bath cracking for 15-20min by using an ultrasonic cracker to obtain a cracking stock solution; centrifuging the lysis stock solution, and recovering the supernatant to obtain lactobacillus bulgaricus lysate;
step two, preparing a bacillus pumilus fermentation liquid: taking a Bacillus pumilus strain, sequentially carrying out strain activation and amplification culture on the Bacillus pumilus strain with the inoculum size of 10-15%, culturing the Bacillus pumilus strain for 15-18h at 36-38 ℃ and 200-220rpm, and filtering the Bacillus pumilus strain by 6-8 layers of sterilization gauze to obtain a Bacillus pumilus fermentation liquid;
step three, mixing the lactobacillus bulgaricus lysate obtained in the step one and the bacillus pumilus fermentation liquor obtained in the step two according to the volume ratio of 1-3: 1 to obtain a microbial solution;
4) and (3) adding fulvic acid, sinapine, boron, selenium, puerarin, ellagic acid and chitosan oligosaccharide into the microbial solution obtained in the step three respectively, and uniformly mixing and dissolving to obtain the antibacterial combined solution.
3. The method of claim 2, wherein: in the preparation method of the bacteriostatic combined solution, the bacillus pumilus is cultured by using a spore-forming culture solution, and the spore-forming culture solution comprises the following components: 1 percent of glucose, 0.5 percent of molasses, 1 percent of protein powder, 0.3 percent of potassium chloride and 0.5 percent of manganese sulfate, and the pH value is 7.0.
4. The application of the bacteriostatic combination solution according to claim 1 in the biological-chemical comprehensive control of tobacco bacterial wilt.
5. Use according to claim 4, characterized in that: the application refers to a method for biological-chemical comprehensive control of tobacco leaf disease, which comprises the following steps: preparing an antibacterial combination solution, and diluting with sterile water by 50-100 times to obtain a diluent for later use; selecting healthy and disease-free tobacco seeds, soaking the seeds in the obtained diluent at room temperature for 15-18h, and then sowing the seeds in a seedling tray; when transplanting, the obtained diluent is adopted for foliage spraying and root irrigation; the spraying amount of the leaf surfaces is 5-10ml, and the root irrigation amount is 30-40 ml; respectively spraying the whole plant with the obtained diluent at 25-30d and 50-55d after transplanting, wherein the spraying amount is 40-50 ml; the rest period is carried out according to the normal cultivation mode of tobacco.
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Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102296036A (en) * | 2011-06-01 | 2011-12-28 | 中国农业科学院烟草研究所 | Bacillus pumilus and application in disease prevention and growth promotion thereof |
| CN103931635A (en) * | 2014-05-03 | 2014-07-23 | 海南正业中农高科股份有限公司 | Agricultural composition containing chitosan oligosaccharide and toyocamycin, and application of agricultural composition |
| CN104531559A (en) * | 2014-12-08 | 2015-04-22 | 山西省农业科学院生物技术研究中心 | Bacillusamyloliquefaciens subsp Lh-1 and application thereof |
| CN105052933A (en) * | 2015-07-29 | 2015-11-18 | 山东亿嘉农化有限公司 | Efficient seed coating agent, preparation method and applications thereof, and method using same to process seeds |
| CN105638665A (en) * | 2016-04-08 | 2016-06-08 | 广西南宁益土生物科技有限责任公司 | Composition containing lactose high-temperature treatment product and amino-oligosaccharin |
| CN106222108A (en) * | 2016-08-10 | 2016-12-14 | 青岛科拓恒通乳酸菌产业化开发研究院有限公司 | For promoting compound lactobacillus microbial inoculum and the preparation method of grape quality |
| CN106561717A (en) * | 2015-10-12 | 2017-04-19 | 成都新朝阳作物科学有限公司 | Pesticide for preventing and controlling ginger blast, and preparation method thereof |
| CN109699681A (en) * | 2019-01-28 | 2019-05-03 | 江苏省农业科学院 | One plant of prevention and treatment bacteriosis bacillus pumilus ZR3-3 and its application |
| CN111387209A (en) * | 2020-05-21 | 2020-07-10 | 湖北大学 | Composite antagonistic bacterium agent for preventing and controlling tobacco diseases and application thereof |
| CN113151085A (en) * | 2021-04-23 | 2021-07-23 | 贵州省烟草公司遵义市公司 | Compound microbial agent for preventing and treating soil-borne diseases of tobacco and application thereof |
| CN113455507A (en) * | 2021-07-02 | 2021-10-01 | 中国科学院兰州化学物理研究所 | Application of natural flavonoid compound as antibacterial agent in inhibiting plant pathogenic bacteria |
-
2021
- 2021-11-04 CN CN202111300436.9A patent/CN113951279A/en active Pending
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102296036A (en) * | 2011-06-01 | 2011-12-28 | 中国农业科学院烟草研究所 | Bacillus pumilus and application in disease prevention and growth promotion thereof |
| CN103931635A (en) * | 2014-05-03 | 2014-07-23 | 海南正业中农高科股份有限公司 | Agricultural composition containing chitosan oligosaccharide and toyocamycin, and application of agricultural composition |
| CN104531559A (en) * | 2014-12-08 | 2015-04-22 | 山西省农业科学院生物技术研究中心 | Bacillusamyloliquefaciens subsp Lh-1 and application thereof |
| CN105052933A (en) * | 2015-07-29 | 2015-11-18 | 山东亿嘉农化有限公司 | Efficient seed coating agent, preparation method and applications thereof, and method using same to process seeds |
| CN106561717A (en) * | 2015-10-12 | 2017-04-19 | 成都新朝阳作物科学有限公司 | Pesticide for preventing and controlling ginger blast, and preparation method thereof |
| CN105638665A (en) * | 2016-04-08 | 2016-06-08 | 广西南宁益土生物科技有限责任公司 | Composition containing lactose high-temperature treatment product and amino-oligosaccharin |
| CN106222108A (en) * | 2016-08-10 | 2016-12-14 | 青岛科拓恒通乳酸菌产业化开发研究院有限公司 | For promoting compound lactobacillus microbial inoculum and the preparation method of grape quality |
| CN109699681A (en) * | 2019-01-28 | 2019-05-03 | 江苏省农业科学院 | One plant of prevention and treatment bacteriosis bacillus pumilus ZR3-3 and its application |
| CN111387209A (en) * | 2020-05-21 | 2020-07-10 | 湖北大学 | Composite antagonistic bacterium agent for preventing and controlling tobacco diseases and application thereof |
| CN113151085A (en) * | 2021-04-23 | 2021-07-23 | 贵州省烟草公司遵义市公司 | Compound microbial agent for preventing and treating soil-borne diseases of tobacco and application thereof |
| CN113455507A (en) * | 2021-07-02 | 2021-10-01 | 中国科学院兰州化学物理研究所 | Application of natural flavonoid compound as antibacterial agent in inhibiting plant pathogenic bacteria |
Non-Patent Citations (2)
| Title |
|---|
| 孔德英 等: "氨基寡糖素对番茄青枯病防治作用", 《西南农业大学学报（自然科学版）》 * |
| 施河丽 等: "黄腐酸与微生物菌剂协同对烟草青枯病及根际土壤细菌群落的影响", 《烟草科技》 * |
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