CN110558340A - biological composite plant vaccine and preparation method thereof - Google Patents
biological composite plant vaccine and preparation method thereof Download PDFInfo
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- CN110558340A CN110558340A CN201910711759.3A CN201910711759A CN110558340A CN 110558340 A CN110558340 A CN 110558340A CN 201910711759 A CN201910711759 A CN 201910711759A CN 110558340 A CN110558340 A CN 110558340A
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- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/08—Magnoliopsida [dicotyledons]
- A01N65/12—Asteraceae or Compositae [Aster or Sunflower family], e.g. daisy, pyrethrum, artichoke, lettuce, sunflower, wormwood or tarragon
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Abstract
the invention discloses a biological composite plant vaccine which is prepared from the following raw material components in percentage by mass: acrylic acid: sodium styrene sulfonate: potassium sulfate: b-1, 4-N-amino-glucose ═ 31.5: 3.5: 5.5: 3.5; alternatively, crotonic acid: sodium styrene sulfonate: potassium sulfate: b-1, 4-N-amino-glucose ═ 31.5: 3.5: 5.5: 3.5; the volume of the mushroom extract accounts for 20% of the total solution volume, and the volume of the purified biogas slurry accounts for 40% of the total solution volume. The invention also provides a preparation method of the biological type composite plant vaccine. The vaccine is a novel multifunctional, safe, nontoxic, residue-free and efficient compound biological preparation. The plant vaccine has multiple effects of disease resistance, insect expelling, yield increase, quality improvement, quick absorption, quick response and the like on various main crops such as grain crops, vegetables, fruits and the like, and has excellent diffusivity and permeability.
Description
Technical Field
the invention belongs to the technical field of disease and pest control of crops, and particularly relates to a biological type composite plant vaccine and a preparation method thereof.
Background
plant virus diseases are diseases which have large damage loss and are difficult to control, are also called as plant 'cancer', and become a worldwide problem; because plant virus obligate parasitism is proliferated in a plant body, consumes plant nutrition and causes abnormal growth or leaf yellowing of the plant, the structure of virus particles is difficult to damage by common chemical drugs, and thus no ideal chemical pesticide exists at present; on the basis of plant immunity inducer, the first antiviral protein biological pesticide of China, namely 6 percent oligosaccharide-catenin wettable powder, developed by the plant protection of Chinese academy of agricultural sciences is approved for marketing; the field test result proves that the control effect of the pesticide on the rice stripe disease is 65 percent; the prevention and treatment effect on strawberry yellow leaf curl virus is 68 percent; the prevention and treatment effect on tobacco mosaic virus is 75 percent; in addition, the plant with the trans-resistance gene transfers the resistance gene into a plant body, so that the plant generates resistance protein to resist the invasion of pathogenic bacteria, and the plant immunity inducer has the same important function as the immunity resistance of the resistance gene to plant diseases and insect pests by inducing the plant to generate resistance substances; the plant immune activation protein is a novel plant immune induced resistance pesticide; the protein is a novel structural protein separated from various pathogenic fungi, and a plurality of varieties have good efficacy of stimulating the enhancement of the immunological activity of plants; representative products include "wettable powder of 3% alternaria tenuissima activator protein" which has been temporarily registered for agricultural chemicals in the department of agriculture, "powder of pulegol plant immunity protein" which has been temporarily registered for national fertilizers, and the like; the plant immunity inducer (or called as plant vaccine) is a new field of vaccine engineering technology after human vaccine and animal vaccine, and is also a popular research direction for the creation of biological pesticides in the world; the plant vaccine is a practice of scientifically controlling plant diseases and insect pests by scientists on the theoretical basis of revealing the relationship among plants, plant diseases and insect pests and biological pesticides, and has the capability of improving the resistance of crops and effectively preventing and controlling the crop diseases; the plant vaccines produced on the market at present have single functions, most of the plant vaccines are pesticides essentially, and the plant vaccines have great dependence on equipment during synthesis, for example, most of oligosaccharide products are compounded with chemical components, and are pesticides essentially, and drug residues exist.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a biological type composite plant vaccine suitable for inducing cultivated crops to resist diseases and pests and a preparation method thereof.
In order to achieve the above object, the present invention provides a biotype composite plant vaccine, which is characterized in that: the vaccine is prepared from the following raw material components in percentage by mass: acrylic acid: sodium styrene sulfonate: potassium sulfate: b-1, 4-N-amino-glucose ═ 31.5: 3.5: 5.5: 3.5; alternatively, crotonic acid: sodium styrene sulfonate: potassium sulfate: b-1, 4-N-amino-glucose ═ 31.5: 3.5: 5.5: 3.5; the volume of the mushroom extract accounts for 20% of the total solution volume, and the volume of the purified biogas slurry accounts for 40% of the total solution volume.
the invention also provides a preparation method of the biological compound plant vaccine, which is characterized by comprising the following steps: it comprises the following steps:
(1) Taking 100-120ml of distilled water from a flask, adjusting the pH value to 11-12, adding 5-6 g of titanium dioxide and phosphorus pentoxide respectively, raising the temperature to 50-52 ℃, fully stirring for reaction for 1-2 hours, taking supernate, adding the supernate into another flask, adjusting the pH value to 6-6.5, and taking the supernate as a raw material A;
(2) Mixing acrylic acid, potassium sulfate and sodium styrene sulfonate according to a mass ratio of 31.5: 3.5: 3.5, continuously stirring for 5-6min, adjusting the pH value to 6-6.5 by using sodium hydroxide, adding zeolite and 30ml of a first raw material, simultaneously gradually increasing the temperature of a reaction solution in a flask to 85 ℃, controlling the polymerization process to be finished within 4-5 hours, continuously heating to 95 ℃ after the polymerization is finished, stirring for reaction for 5-7 hours, and naturally cooling at room temperature to prepare a raw material B;
(3) mixing potassium sulfate, B-1, 4-N-amino-glucose and water in a mass ratio of 5.5:3.5:50, fully shaking up, adding the raw material A in a mass ratio of 40%, and continuously stirring for 3.5 to 4 hours at a water bath environment of between 85 and 90 ℃; after the reaction is finished, adjusting the pH value of the solution to 7-8, adding absolute ethyl alcohol according to the volume ratio of 1.2:1, filtering out precipitates after 3-60min, washing with distilled water for 2-3 times, and drying to prepare a raw material C;
weighing pulverized Lentinus edodes powder, adding distilled water, continuously stirring at 90-100 deg.C for 30min, cooling to room temperature, vacuum filtering, discarding residue, and concentrating to obtain Lentinus edodes extract with total volume of 40% of the original volume;
(4) Taking chicory stems and leaves or waste melon, fruit and vegetables, unfermented chicken manure and water according to the mass ratio of 1: 1:3: 4, adding the activated sludge into the container in proportion, taking the activated sludge at the bottom of the septic tank, adding the activated sludge into the container, and sealing and fermenting; fully stirring for 10min after fermentation, filtering with a 200-mesh screen, and standing the filtrate to obtain the prepared purified biogas slurry; the purified biogas slurry is sealed completely to ensure an anoxic environment in the fermentation process; the activated sludge at the bottom of the septic tank is activated sludge at the bottom of the septic tank of a farm; the using amount of the activated sludge at the bottom of the dividing times is 20% of the total fermented materials, the fermentation is required to be carried out for more than 60 days at the temperature of below 20 ℃, and the fermentation is only required to be carried out for 40 days at the temperature of 20 ℃; uniformly mixing the raw material B and the raw material C according to the mass ratio of 1:3, continuously stirring in a water bath at 70-80 ℃ for reacting for 18-20 hours, and cooling to room temperature; and then adding the lentinus edodes extract which accounts for 20% of the total solution volume, and the purified biogas slurry which accounts for 40% of the total solution volume, and uniformly mixing to obtain the biological compound vaccine.
Further, the step (4) is carried out with sealed fermentation: when the temperature is below 20 ℃, fermenting for 60 days; when the temperature is above 20 ℃, the fermentation is only needed for 40 days.
The invention has the following advantages and beneficial effects: the effective components of the plant vaccine comprise K, purified biogas slurry, potassium phosphite, phosphorus, potassium, sodium, calcium, lentinan, humic acid, gibberellin, auxin, antibiotics and B-1, 4-N-acetyl-glucosamine oligomer, can provide organic carbon sources, potash fertilizers, phosphate fertilizers, biological hormones and sterilizing and insect-resisting agents for crops, and is a novel multifunctional, safe, nontoxic, residue-free and efficient compound biological agent. The plant vaccine has multiple effects of disease resistance, insect expelling, yield increase, quality improvement, quick absorption, quick response and the like on various main crops such as grain crops, vegetables, fruits and the like, and has excellent diffusivity and permeability; continuously stirring on a magnetic stirrer; the composite plant vaccine is stored in a dry and dark environment, and is mixed with water for 1: diluting at the ratio of 50-100, and spraying the diluted solution on plant leaves, stalks and the like.
Drawings
FIG. 1 is a graph of the effect of plant vaccines on the growth of strawberry gray mold and early blight strains;
in fig. 1: a is a strawberry gray mold control group, B is a strawberry gray mold treatment group, C is a strawberry early blight control group, and D is a strawberry early blight treatment group.
FIG. 2 is the effect of plant vaccine on the growth of tomato gray mold and early blight strains and the effect of plant vaccine on the growth of green pepper wilt strains;
in fig. 2: e is a tomato gray mold control group, F is a tomato gray mold treatment group, G is a tomato early blight control group, H is a tomato early blight treatment group,
FIG. 3 is a graph showing the effect of plant vaccines on the growth of a bacterial strain of green pepper wilt;
In fig. 3: is a control group of the green pepper wilt disease Y, and is a treatment group of the green pepper wilt disease Z.
Detailed Description
the reagent used in the present invention is not particularly limited, and commercially available conventional reagents can be used, and the technical solution of the present invention will be further specifically described below with reference to examples and drawings.
The preparation method of the biological composite plant vaccine comprises the following specific operation steps:
1. taking 100ml of distilled water from a flask, adjusting the pH value to about 12, adding five grams of titanium dioxide and phosphorus pentoxide respectively, raising the temperature to 50 ℃, fully stirring for reaction for 1-2 hours, taking supernatant liquid by a liquid transfer device, adding the supernatant liquid into another flask, and adjusting the pH value to 6 to be used as a raw material 1;
2. mixing acrylic acid, potassium sulfate and sodium styrene sulfonate according to a mass ratio of 31.5: 5.5:3.5, continuously stirring for 5min, adjusting the pH value to 6-6.5 by using sodium hydroxide, adding zeolite, adding 30ml of the raw material 1, simultaneously gradually increasing the temperature of the reaction liquid in the flask to 85 ℃, controlling the polymerization process to be finished within 4-5 hours, continuously heating to 95 ℃ after finishing the polymerization, then stirring for reaction for 5-7 hours, and naturally cooling at room temperature to prepare a raw material B;
3. mixing potassium sulfate, B-1, 4-N-amino-glucose and water in a mass ratio of 5.5:3.5:50, fully shaking up, adding 40ml of raw material 1, continuously stirring for 3.5 to 4 hours in a water bath environment at 85 to 90 ℃, adjusting the pH of a solution to be within the range of 7 to 8 after the reaction is finished, adding absolute ethyl alcohol according to a volume ratio of 1.2:1, filtering out precipitates after 3 to 60 minutes, washing with distilled water for 2 to 3 times, and drying to prepare a raw material C:
weighing 10g of pulverized shiitake mushroom powder, adding water according to a ratio of 1:15, adding 90-100 ℃, continuously stirring for 30min under the condition, cooling to room temperature, carrying out vacuum filtration, discarding filter residues, reserving filtrate, and concentrating to 40ml to obtain shiitake mushroom extract;
4. taking chicory stems and leaves or waste melon, fruit and vegetables, unfermented chicken manure and water according to the mass ratio of 1: 1:3: 4, adding the activated sludge at the bottom of the septic tank into a container, sealing and fermenting, wherein the fermentation time is over 60 days when the temperature is below 20 ℃, the fermentation time is over 20 ℃, the fermentation time is only over 40 days when the temperature is above 20 ℃, fully stirring for 10min after fermentation, filtering with a 200-mesh screen, and standing the filtrate to obtain the prepared purified biogas slurry; the purified biogas slurry is sealed completely to ensure an anoxic environment in the fermentation process;
uniformly mixing the raw material B and the raw material C according to the mass ratio of 1:3, continuously stirring in a water bath at 70-80 ℃ for 18 hours to react for 20 hours, cooling to room temperature, adding 40ml of mushroom extract, adding 50ml of purified biogas slurry, and uniformly stirring to prepare the biological compound vaccine.
firstly, pathogenic bacteria of gray mold, blight and early blight are taken from diseased plants, wherein the diseased plants are respectively strawberry gray mold and early blight plants, tomato gray mold and early blight plants and green pepper blight plants;
Respectively preparing suspension with spore concentration of 100000/mL by using sterile water, respectively taking 0.1 mL suspension by using a pipette, coating the suspension on a PAD plate, wherein the total number of three pathogenic bacteria is 15 plates, and distributing the pathogenic bacteria taken by each pathogenic plant to three plates; sealing the PAD plate with newspaper, transferring to a constant temperature incubator, separately packaging the three pathogenic bacteria plates in the packaging process, setting the incubator temperature at 25 ℃ and 1 ℃, culturing for 7 days, and inoculating bacterial plaque with the diameter of 0.5mm to the PAD plate containing the plant vaccine (treatment group: the volume content of the plant vaccine is 20%) and the PAD plate not containing the plant vaccine (control group); after culturing for 7 days, measuring the diameter of the circular bacterial plaque, repeating the experiment for 3 times, taking the average value of the bacterial plaque on 10 flat plates each time, and taking the average value of the experiments as the final value to carry out the experiment on pathogenic bacteria under the constant temperature condition;
The results are shown in FIG. 1.
As can be seen from the figure, the bacterial plaque area of the gray mold of strawberries and tomatoes is obviously reduced compared with that of a control group after the plant vaccine is sprayed, the bacterial plaque area in the control group reaches about 5mm, and the bacterial plaque area is only about 1.9mm and 3.1mm after the plant vaccine is treated, so that the plant vaccine can obviously inhibit the growth of the gray mold bacteria; it can be seen from the figure that the bacterial plaque area of strawberry and tomato early blight after the plant vaccine of the invention is sprayed is obviously reduced compared with that of a control group, the bacterial plaque area in the control group reaches about 6mm, and the bacterial plaque area is only about 2.7mm and 1.9mm after the plant vaccine of the invention is treated, which shows that the plant vaccine of the invention obviously inhibits the growth of tomato early blight bacteria. It can also be seen from the figure that the area of bacterial plaque of green pepper wilt disease sprayed by the plant vaccine of the invention is significantly reduced compared with that of the control group, the area of bacterial plaque in the control group reaches about 4.8mm, and after the plant vaccine of the invention is treated, the area of bacterial plaque is only about 1.6mm, which shows that the plant vaccine of the invention significantly inhibits the growth of green pepper wilt disease bacteria by the plant vaccine;
the first embodiment is as follows:
taking 100-120ml of distilled water from a flask, adjusting the pH value to 11-12, adding 5-6 g of titanium dioxide and phosphorus pentoxide respectively, raising the temperature to 50-52 ℃, fully stirring for reaction for 1-2 hours, taking supernate by using a pipette, adding the supernate into another flask, adjusting the pH value to 6-6.5, and taking the supernate as a raw material A; mixing acrylic acid, potassium sulfate and sodium styrene sulfonate according to a mass ratio of 31.5: 3.5: 3.5, continuously stirring for 5-6min, adjusting the pH value to 6-6.5 by using sodium hydroxide, adding zeolite and 30ml of a first raw material, simultaneously gradually increasing the temperature of a reaction solution in a flask to 85 ℃, controlling the polymerization process to be finished within 4-5 hours, continuously heating to 95 ℃ after the polymerization is finished, stirring for reaction for 5-7 hours, and naturally cooling at room temperature to prepare a raw material B; mixing potassium sulfate, B-1, 4-N-amino-glucose and water in a mass ratio of 5.5:3.5:50, fully shaking up, adding 40% of raw material A in a mass ratio, and continuously stirring for 3.5 to 4 hours at a water bath environment of between 85 and 90 ℃; after the reaction is finished, adjusting the pH value of the solution to 7-8, adding absolute ethyl alcohol according to the volume ratio of 1.2:1, filtering out precipitates after 3-60min, washing with distilled water for 2-3 times, and drying to prepare a raw material C;
Uniformly mixing the raw material B and the raw material C according to the mass ratio of 1:3, continuously stirring in a water bath at 70-80 ℃ for 18 hours to react for 20 hours, cooling to room temperature, adding 40ml of mushroom extract, adding 50ml of purified biogas slurry, and uniformly stirring to prepare a biological composite plant vaccine; diluting the plant vaccine with water at a ratio of 1:10, and then carrying out an experiment;
Strawberry is selected as a test crop, and the variety is octopus and red pigment. The test is carried out in a strawberry cultivation base in Wu lake village in a Yang-logical economic development area in a New Zhou area in Wuhan city; each variety is cultivated with 20 plants, wherein 10 plants are used as a reference, 6 fruits are left in each plant through conventional fertilizer and water management, the plant vaccine of the invention is not used, the other 10 plants are used as treatment, 6 fruits are left in each plant through conventional waste water management, the plant vaccine of the invention is sprayed once every day from the seedling stage and diluted by 10 times of water for use until the harvest is finished. Counting the initial flowering phase, the single fruit weight and the total weight of all plants; the statistical results are shown in table 1:
TABLE 1
as can be seen from table 1, after the plant vaccine provided by the present invention is used, the two varieties are superior to the control group of the same variety in three aspects of the initial flowering phase, the weight of the single fruit, the total fruit weight, etc. (i.e. the plant vaccine provided by the present invention is not used), the strawberry variety zhiji is 7 days earlier than the control group, and the strawberry variety jasper is 5 days earlier than the control group; in the aspect of average single fruit weight, the strawberry variety octopus is increased by 23.0 percent compared with a control group, after the plant vaccine provided by the invention is applied to the ruby of the strawberry variety, the strawberry variety is increased by 10.1 percent compared with the control group, and the total yield of the two varieties is relatively increased by 274g and 126 g;
example two:
Taking 100-120ml of distilled water from a flask, adjusting the pH value to 11-12, adding 5-6 g of titanium dioxide and phosphorus pentoxide respectively, raising the temperature to 50-52 ℃, fully stirring for reaction for 1-2 hours, taking supernate by using a pipette, adding the supernate into another flask, adjusting the pH value to 6-6.5, and taking the supernate as a raw material A; mixing acrylic acid, potassium sulfate and sodium styrene sulfonate according to a mass ratio of 31.5: 3.5: 3.5, continuously stirring for 5-6min, adjusting the pH value to 6-6.5 by using sodium hydroxide, adding zeolite and 30ml of a first raw material, simultaneously gradually increasing the temperature of a reaction solution in a flask to 85 ℃, controlling the polymerization process to be finished within 4-5 hours, continuously heating to 95 ℃ after the polymerization is finished, stirring for reaction for 5-7 hours, and naturally cooling at room temperature to prepare a raw material B; mixing potassium sulfate, B-1, 4-N-amino-glucose and water in a mass ratio of 5.5:3.5:50, fully shaking up, adding 40% of raw material A in a mass ratio, and continuously stirring for 3.5 to 4 hours at a water bath environment of between 85 and 90 ℃; after the reaction is finished, adjusting the pH value of the solution to 7-8, adding absolute ethyl alcohol according to the volume ratio of 1.2:1, filtering out precipitates after 3-60min, washing with distilled water for 2-3 times, and drying to prepare a raw material C; uniformly mixing the raw material B and the raw material C according to the mass ratio of 1:3, continuously stirring in a water bath at 70-80 ℃ for 18-20 hours, cooling to room temperature, adding 40ml of mushroom extract, adding 50ml of purified biogas slurry, and uniformly stirring to prepare a biological composite plant vaccine; diluting the plant vaccine with water at a ratio of 1:10, and then carrying out an experiment;
the test is carried out in a cercis 301 laboratory in Wuhan bioengineering college of Hanshi highway in China-Tuohan economic development area of Yang prowl district in Wuhan city; each variety is cultivated with 20 plants, wherein 10 plants are used as a reference, 10 fruits are left in each plant through conventional fertilizer and water management, the plant vaccine of the invention is not used, the other 10 plants are used as treatment, 10 fruits are left in each plant through conventional waste water management, the plant vaccine of the invention is sprayed once every day from the seedling stage and diluted by 10 times of water for use until the harvest is finished. Counting the initial flowering phase, the single fruit weight and the total weight of all plants; the statistical results are shown in table 2:
TABLE 2
As can be seen from table 2, after the plant vaccine provided by the present invention is used, the two varieties are superior to the control group of the same variety in three aspects of the initial flowering phase, the weight of the single fruit, the total fruit weight and the like (i.e. the plant vaccine provided by the present invention is not used), the red cherry tomatoes are 8 days earlier than the control group, and the pearl tomatoes are 6 days earlier than the control group; in the aspect of average fruit weight, the red cherry tomato variety is improved by 22.3 percent compared with the control group, after the plant vaccine provided by the invention is applied to the ruby of the strawberry variety, the plant vaccine is improved by 13.07 percent compared with the control group, and the total yield of the two varieties is relatively improved by 270g and 170 g;
example three:
Taking 100-120ml of distilled water from a flask, adjusting the pH value to 11-12, adding 5-6 g of titanium dioxide and phosphorus pentoxide respectively, raising the temperature to 50-52 ℃, fully stirring for reaction for 1-2 hours, taking supernate by using a pipette, adding the supernate into another flask, adjusting the pH value to 6-6.5, and taking the supernate as a raw material A; mixing acrylic acid, potassium sulfate and sodium styrene sulfonate according to a mass ratio of 31.5: 3.5: 3.5, continuously stirring for 5-6min, adjusting the pH value to 6-6.5 by using sodium hydroxide, adding zeolite and 30ml of a first raw material, simultaneously gradually increasing the temperature of a reaction solution in a flask to 85 ℃, controlling the polymerization process to be finished within 4-5 hours, continuously heating to 95 ℃ after the polymerization is finished, stirring for reaction for 5-7 hours, and naturally cooling at room temperature to prepare a raw material B; mixing potassium sulfate, B-1, 4-N-amino-glucose and water in a mass ratio of 5.5:3.5:50, fully shaking up, adding 40% of raw material A in a mass ratio, and continuously stirring for 3.5 to 4 hours at a water bath environment of between 85 and 90 ℃; after the reaction is finished, adjusting the pH value of the solution to 7-8, adding absolute ethyl alcohol according to the volume ratio of 1.2:1, filtering out precipitates after 3-60min, washing with distilled water for 2-3 times, and drying to prepare a raw material C; uniformly mixing the raw material B and the raw material C according to the mass ratio of 1:3, continuously stirring in a water bath at 70-80 ℃ for 18-20 hours, cooling to room temperature, adding 40ml of mushroom extract, adding 50ml of purified biogas slurry, and uniformly stirring to prepare a biological composite plant vaccine; diluting the plant vaccine with water at a ratio of 1:10, and then carrying out an experiment;
Green peppers are used as experimental crops, the varieties are oxhorn 211, Xiang Min No. 15 and Ji pepper 549, and the experiments are carried out on open-air cultivation lands for vegetable cultivation in Yuanqiao villages in city cities and towns in Bijie, Guizhou province; 50 plants are cultivated in each variety, 25 plants are used as a reference, 5 fruits are left in each plant through conventional fertilizer and water management, the plant vaccine is not used, the other 25 plants are used as treatment, 5 fruits are left in each plant through conventional waste water management, the plant vaccine is sprayed for 2 times from the beginning of the seedling stage and is diluted by water for 10 times for use until the first harvest is finished. Counting the initial flowering phase, the single fruit weight and the total weight of all plants, and meanwhile, counting the number of common insect pests, namely spider mites, aphids, cotton bollworms, oriental tobacco budworms and the like, wherein the counting results are shown in tables 3 and 4:
TABLE 3
as can be seen from Table 3, after the plant vaccine provided by the invention is used, the three varieties are superior to a control group of the same variety in three aspects of the initial flowering phase, the weight of the single fruit, the total fruit weight and the like (namely, the plant vaccine provided by the invention is not used), the horn 211 of the green pepper variety is 7 days earlier than the control group, the Hunan No. 15 of the green pepper variety is 4 days earlier than the control group, and the Guiji pepper variety 549 is 7 days earlier than the control group; in the aspect of average single fruit weight, the ox horn 211 of the green pepper variety is improved by 5.1 percent compared with a control group, after the plant vaccine provided by the invention is applied to the Hunan Ji No. 15 of the green pepper variety, the plant vaccine provided by the invention is improved by 9.8 percent compared with the control group, after the plant vaccine provided by the invention is applied to the Ji pepper variety 549 of the green pepper variety, the plant vaccine is improved by 8.4 percent compared with the control group, and the total output of the three varieties is relatively improved by 512.5g, 262.5g and 237.5 g;
TABLE 4
as can be seen from Table 4, after the plant vaccine provided by the invention is used, the effects of weather, environment, soil fertility and other external factors are eliminated, and the three varieties are superior to a control group of the same variety in the prevention of three insect pests.
Claims (3)
1. a biological type composite plant vaccine is characterized in that: the vaccine is prepared from the following raw material components in percentage by mass: acrylic acid: sodium styrene sulfonate: potassium sulfate: b-1, 4-N-amino-glucose ═ 31.5: 3.5: 5.5: 3.5; alternatively, crotonic acid: sodium styrene sulfonate: potassium sulfate: b-1, 4-N-amino-glucose ═ 31.5: 3.5: 5.5: 3.5; the volume of the mushroom extract accounts for 20% of the total solution volume, and the volume of the purified biogas slurry accounts for 40% of the total solution volume.
2. A method for preparing a biotype composite plant vaccine according to claim 1, wherein: it comprises the following steps:
(1) Taking 100-120ml of distilled water from a flask, adjusting the pH value to 11-12, adding 5-6 g of titanium dioxide and phosphorus pentoxide respectively, raising the temperature to 50-52 ℃, fully stirring for reaction for 1-2 hours, taking supernate, adding the supernate into another flask, adjusting the pH value to 6-6.5, and taking the supernate as a raw material A;
(2) mixing acrylic acid, potassium sulfate and sodium styrene sulfonate according to a mass ratio of 31.5: 3.5: 3.5, continuously stirring for 5-6min, adjusting the pH value to 6-6.5 by using sodium hydroxide, adding zeolite and 30ml of a first raw material, simultaneously gradually increasing the temperature of a reaction solution in a flask to 85 ℃, controlling the polymerization process to be finished within 4-5 hours, continuously heating to 95 ℃ after the polymerization is finished, stirring for reaction for 5-7 hours, and naturally cooling at room temperature to prepare a raw material B;
(3) Mixing potassium sulfate, B-1, 4-N-amino-glucose and water in a mass ratio of 5.5:3.5:50, fully shaking up, adding the raw material A in a mass ratio of 40%, and continuously stirring for 3.5 to 4 hours at a water bath environment of between 85 and 90 ℃; after the reaction is finished, adjusting the pH value of the solution to 7-8, adding absolute ethyl alcohol according to the volume ratio of 1.2:1, filtering out precipitates after 3-60min, washing with distilled water for 2-3 times, and drying to prepare a raw material C;
weighing pulverized Lentinus edodes powder, adding distilled water, continuously stirring at 90-100 deg.C for 30min, cooling to room temperature, vacuum filtering, discarding residue, and concentrating to obtain Lentinus edodes extract with total volume of 40% of the original volume;
(4) Taking chicory stems and leaves or waste melon, fruit and vegetables, unfermented chicken manure and water according to the mass ratio of 1: 1:3: 4, adding the activated sludge into the container in proportion, taking the activated sludge at the bottom of the septic tank, adding the activated sludge into the container, and sealing and fermenting; fully stirring for 10min after fermentation, filtering with a 200-mesh screen, and standing the filtrate to obtain the prepared purified biogas slurry; the purified biogas slurry is sealed completely to ensure an anoxic environment in the fermentation process; the activated sludge at the bottom of the septic tank is activated sludge at the bottom of the septic tank of a farm; the using amount of the activated sludge at the bottom of the dividing times is 20% of the total fermented materials, the fermentation is required to be carried out for more than 60 days at the temperature of below 20 ℃, and the fermentation is only required to be carried out for 40 days at the temperature of 20 ℃; uniformly mixing the raw material B and the raw material C according to the mass ratio of 1:3, continuously stirring in a water bath at 70-80 ℃ for reacting for 18-20 hours, and cooling to room temperature; and then adding the lentinus edodes extract which accounts for 20% of the total solution volume, and the purified biogas slurry which accounts for 40% of the total solution volume, and uniformly mixing to obtain the biological compound vaccine.
3. the method for preparing a biotype composite plant vaccine according to claim 2, wherein the method comprises the following steps: and (4) performing sealed fermentation: when the temperature is below 20 ℃, fermenting for 60 days; when the temperature is above 20 ℃, the fermentation is only needed for 40 days.
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Application publication date: 20191213 |