CN113826644A - Entomopathogenic nematode preparation for preventing and controlling spodoptera frugiperda, preparation method and application thereof - Google Patents
Entomopathogenic nematode preparation for preventing and controlling spodoptera frugiperda, preparation method and application thereof Download PDFInfo
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- CN113826644A CN113826644A CN202111311696.6A CN202111311696A CN113826644A CN 113826644 A CN113826644 A CN 113826644A CN 202111311696 A CN202111311696 A CN 202111311696A CN 113826644 A CN113826644 A CN 113826644A
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Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/10—Animals; Substances produced thereby or obtained therefrom
- A01N63/12—Nematodes
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/002—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing a foodstuff as carrier or diluent, i.e. baits
- A01N25/006—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing a foodstuff as carrier or diluent, i.e. baits insecticidal
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
- A01N43/14—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings
- A01N43/16—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings with oxygen as the ring hetero atom
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N45/00—Biocides, pest repellants or attractants, or plant growth regulators, containing compounds having three or more carbocyclic rings condensed among themselves, at least one ring not being a six-membered ring
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Pest Control & Pesticides (AREA)
- Engineering & Computer Science (AREA)
- Dentistry (AREA)
- Plant Pathology (AREA)
- Agronomy & Crop Science (AREA)
- Wood Science & Technology (AREA)
- Environmental Sciences (AREA)
- Toxicology (AREA)
- Food Science & Technology (AREA)
- Insects & Arthropods (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Virology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses an entomopathogenic nematode preparation for preventing and treating spodoptera frugiperda, a preparation method and application thereof, belongs to the technical field of biological pesticides, and solves the problems of low field biocontrol quick-acting performance and poor control effect persistence when the entomopathogenic nematodes are used for preventing and treating the spodoptera frugiperda in the prior art. The entomopathogenic nematode preparation for preventing and treating spodoptera frugiperda comprises a component A and a component B, wherein the component A contains different active ingredients; wherein the component A comprises entomopathogenic nematodes and an auxiliary agent for maintaining the vitality of the entomopathogenic nematodes; the component B comprises a phagostimulant, ecdysone and a tyrosinase inhibitor. The invention further discloses application of the entomopathogenic nematode preparation in preventing and controlling spodoptera frugiperda, wherein the component B is sprayed in the seedling core of the crops and then sprayed with water to soften the component B, and the component A is applied in the seedling core of the crops after 1 day. According to the invention, entomopathogenic nematodes, insect ecdysone, an insect tyrosinase inhibitor and a phagostimulant are combined for the first time, so that the biocontrol rate of the entomopathogenic nematodes on spodoptera frugiperda larvae is greatly improved.
Description
Technical Field
The invention belongs to the technical field of biological pesticides, and particularly relates to an entomopathogenic nematode preparation for preventing and treating spodoptera frugiperda, and a preparation method and application thereof.
Background
Spodoptera frugiperda (j.e. smith) is an agricultural pest of the moth genus Spodoptera of the family noctuidae. The growth speed of Spodoptera frugiperda becomes faster along with the rise of air temperature, and the Spodoptera frugiperda can be propagated for several generations in one year, so that more than 1000 eggs can be produced by one female moth. Spodoptera frugiperda larvae like to feed on new leaves, can harm over 80 crops such as sorghum, soybean, cotton, barley, rice, peanut, beet, tobacco, tomato, potato, cabbage, wheat and the like, but is the most harmful to gramineous crops and corn.
When Spodoptera frugiperda damages corn, the larvae which hatch initially can dig into the basal part of the leaf sheath, the prevention effect of the chemical agents applied to the leaf surface is not obvious due to the protection of the leaves, and the Spodoptera frugiperda has generated resistance to various chemical agents to different degrees.
Entomopathogenic nematodes (EPNs) are microbial insecticides having the dual characteristics of "natural enemy insects" and "pathogenic microorganisms". The entomopathogenic nematode has the advantages of wide host range, safety to human, livestock and environment, easy mass culture, convenient use and cyclic utilization in environment. However, in the prior art, the research on the use of entomopathogenic nematodes for preventing spodoptera frugiperda is very limited, and a practical field technology with high efficiency and low cost for preventing spodoptera frugiperda by using entomopathogenic nematodes is not formed yet. The applicant finds in earlier researches that the problems of low quick-acting biological control performance and large nematode dosage exist when the entomopathogenic nematodes are independently adopted to control Spodoptera frugiperda.
Therefore, providing an entomopathogenic nematode preparation for controlling spodoptera frugiperda can accelerate the pathogenic process of the entomopathogenic nematodes infecting the spodoptera frugiperda and improve the biocontrol efficiency of pests, which becomes a problem to be solved by technical personnel in the field.
Disclosure of Invention
The invention discloses an entomopathogenic nematode preparation for preventing and treating spodoptera frugiperda, which can accelerate the pathogenic process of the entomopathogenic nematodes infecting the spodoptera frugiperda and improve the biocontrol rate of the entomopathogenic nematodes on spodoptera frugiperda larvae.
The invention also discloses a preparation method of the entomopathogenic nematode preparation.
The invention also discloses application of the entomopathogenic nematode preparation.
The technical scheme adopted by the invention is as follows:
the invention discloses an entomopathogenic nematode preparation for preventing and treating Spodoptera frugiperda, which comprises a component A and a component B, wherein the component A contains different active ingredients; wherein the component A comprises entomopathogenic nematodes and an auxiliary agent for maintaining the vitality of the entomopathogenic nematodes; the component B comprises a phagostimulant, ecdysone and a tyrosinase inhibitor.
The entomopathogenic nematodes in the component A can enter the spodoptera frugiperda body from natural openings such as the epidermis of the spodoptera frugiperda and the like. The entomopathogenic nematodes enter the spodoptera frugiperda body and release symbiotic bacteria, and the symbiotic bacteria resist the immunity of the spodoptera frugiperda by releasing toxins, so that the spodoptera frugiperda becomes septicemic and dead, and the entomopathogenic nematodes achieve the biological control effect of the spodoptera frugiperda.
The ecdysone in the component B of the invention is also called beta ecdysone and has a chemical formula of C27H44O7The chemical name is 20-hydroxy ecdysterone. As an insect growth regulator, ecdysone can block the normal development of Spodoptera frugiperda larvae, disturb the normal metabolic process in the insect body, enable the insect to molt or metamorphose in advance to become a tiny adult or malformed individual, and simultaneously reduce the immunity and weaken the vitality of the insect; in addition, ecdysone can also abnormally accelerate ecdysis of spodoptera frugiperda, which is a fatal injury because abnormal ecdysis causes fragility of the spodoptera frugiperda body surface and thus facilitates invasion by entomopathogenic nematodes.
The tyrosinase inhibitor in the component B is used for inhibiting tyrosinase-mediated immune function in spodoptera frugiperda, so that entomopathogenic nematodes can infect and kill spodoptera frugiperda more quickly and easily.
The phagostimulant in the component B can induce spodoptera frugiperda larvae to selectively feed on the component B, so that ecdysone and tyrosinase inhibitor are ingested, and the infestation of entomopathogenic nematodes on the spodoptera frugiperda larvae is facilitated; in addition, the phagostimulant is used as an alternative food, so that spodoptera frugiperda larvae do not eat tender leaves and tender buds of crops, and leaves are protected from being damaged.
The entomopathogenic nematode preparation of the invention has the function of preventing and controlling Spodoptera frugiperda through the synergistic effect of the four aspects.
In some embodiments of the invention, component A is a suspension, and each 1L A component contains entomopathogenic nematodes 3,000,000 IJs-5,000,000 IJs;
or/and each 1L A component contains the following auxiliary components: 200-400 ml of millet congee filtrate, 5-10 g of humectant, 0.8-1.2 g of preservative, 1.5-2.0 g of nematode symbiotic bacteria nutrient and 0.5-0.8 ml of stabilizer.
The millet congee filtrate is prepared by adding 50g of millet into 500ml of water, boiling for 10-15 min and then filtering with absorbent cotton gauze.
In some embodiments of the invention, the entomopathogenic nematodes include Steinernema carpocapsae;
or/and the humectant comprises trehalose (α, α);
or/and the preservative comprises ascorbic acid;
or/and the nematode symbiotic bacteria nutrient comprises peptone;
or/and the stabilizer comprises tween, preferably tween 20.
The invention selects the high-temperature-resistant commercial strains of entomopathogenic nematodes such as Steinernema carpocapsae and the like, can quickly infect and kill Lepidoptera larvae such as Spodoptera frugiperda and the like, and can be quickly propagated on the corpses of the larvae in a large quantity.
The auxiliary agent is used for maintaining the vitality of the entomopathogenic nematodes. The auxiliary agent provided by the invention has the effects of moisturizing the nematodes, providing nutrient substances for nematode symbiotic bacteria (keeping the activity of the symbiotic bacteria), enabling the nematodes to be dispersed and suspended in liquid without deposition (solving the problem of nematode loss caused by deposition and easy hypoxia death), keeping the preparation stable and preventing corrosion (easy storage), thereby playing a role in maintaining the life of the entomopathogenic nematodes.
In some embodiments of the invention, the component B is a solid granule prepared by drying a semi-finished product of the component B; each 1L B component semi-finished product contains the following phagostimulant components: 2-5 g of agar powder, 40-60 g of corn flour, 5-10 g of wheat flour, 20-40 g of soybean flour, 2-5 g of yeast powder, 3-6 g of trehalose (alpha ), 0.2-0.5 g of ascorbic acid, 2.5-5 g of peptone, 2.5-6 g of beef extract and 1-3 g of soybean oil;
or/and each 1L B component semi-finished product contains 10-15 g of ecdysone;
or/and each 1L B component semi-finished product contains 5-8 mg of tyrosinase inhibitor.
In some embodiments of the invention, the tyrosinase inhibitor is deoxyarbutin. The deoxyarbutin of the invention has the following name: deoxyarbutin, formula C11H14O3。
The invention provides a preparation method of an entomopathogenic nematode preparation for preventing and treating Spodoptera frugiperda, which comprises the following steps of: weighing the auxiliary components according to a proportion, mixing uniformly, sterilizing at high temperature, adding entomopathogenic nematodes, and fixing the volume with distilled water to obtain a component A;
the preparation of the component B comprises the following steps: weighing the phagostimulant components according to a proportion, mixing uniformly, sterilizing at high temperature, adding ecdysone and tyrosinase inhibitor, fixing the volume with distilled water to obtain a semi-finished product of the component B, uniformly stirring the semi-finished product of the component B, drying, and granulating to obtain the component B.
In some embodiments of the invention, the moisture content of the B component is 10 to 12 wt.%.
The invention provides an application of entomopathogenic nematode preparation for preventing and treating spodoptera frugiperda in preventing and treating spodoptera frugiperda.
In some embodiments of the invention, component B is administered first, followed by component A.
In some embodiments of the invention, the B component is sprinkled in the core of the crop seedling, the B component is softened by spraying water after the sprinkling, and the A component is applied in the core of the crop seedling after 1 day;
preferably, the a-component is applied at or after evening.
Compared with the prior art, the invention has the following beneficial effects:
according to the invention, entomopathogenic nematodes, insect ecdysone, an insect tyrosinase inhibitor and a spodoptera frugiperda phagostimulant are combined for the first time, so that the biocontrol rate of the entomopathogenic nematodes on spodoptera frugiperda larvae is greatly improved. Entomopathogenic nematodes can quickly infect and kill Lepidoptera larvae such as Spodoptera frugiperda and the like, and can be rapidly propagated on corpses of the larvae in large quantities; the ecdysone can prevent the normal development of Spodoptera frugiperda larvae, so that the Spodoptera frugiperda larvae can molt or metamorphose in advance to become micro adults or malformed individuals, the immunity of the Spodoptera frugiperda larvae is reduced, and the invasion of entomopathogenic nematodes is facilitated; the tyrosinase inhibitor can inhibit tyrosinase-mediated immune function in Spodoptera frugiperda, and is beneficial to invasion of entomopathogenic nematodes and reproduction of symbiotic bacteria; the phagostimulant can induce spodoptera frugiperda larvae to take in ecdysone and tyrosinase inhibitors, and is beneficial to the invasion of entomopathogenic nematodes on the spodoptera frugiperda larvae. The entomopathogenic nematode, the insect ecdysone, the tyrosinase inhibitor and the phagostimulant of the entomopathogenic nematode preparation disclosed by the invention act synergistically to play a role in preventing and controlling Spodoptera frugiperda.
The preparation and use methods of the invention are simple. By applying the component B and then applying the component A, the entomopathogenic nematodes can infect and kill spodoptera frugiperda faster and more easily.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
Example 1
The embodiment discloses a preparation method of an entomopathogenic nematode preparation for preventing and treating spodoptera frugiperda, which specifically comprises the following steps:
taking 400ml of millet congee filtrate, 8g of trehalose (alpha ), 1.2g of ascorbic acid, 1.5g of peptone and 200.7 g of tween, putting the millet congee filtrate, mixing the mixture uniformly, covering a sealing film, putting the mixture into an autoclave, sterilizing (melting) the mixture for 30min under the conditions that the pressure is 1.0MPa and the temperature is 121 ℃, cooling the obtained product, mixing with 3,000,000IJs of entomopathogenic nematodes, and adding distilled water to a constant volume of 1L to obtain the component A.
Weighing 3g of agar powder, 60g of corn flour, 5g of wheat flour, 30g of soybean meal, 5g of yeast powder, 4g of trehalose (alpha ), 0.4g of ascorbic acid, 2.5g of peptone, 4g of beef extract and 1g of soybean oil, placing the mixture in the same container, uniformly mixing, placing the container in an autoclave with a sealing film, sterilizing (melting) the mixture for 30min under the conditions that the pressure is 1.0MPa and the temperature is 121 ℃, cooling the obtained product, adding 10g of ecdysone and 7mg of tyrosinase inhibitor deoxyarbutin, and fixing the volume to 1L by using distilled water to obtain a B-component semi-finished product.
And (3) uniformly stirring the semi-finished product of the component B into paste, then sending the paste into a dryer, drying the paste at 80 ℃ until the moisture content is 12%, and then putting the paste into a shaft-connected granulator (a particle plate with holes distributed by an insertion and arrangement method and with the aperture of 2.0-3.0 mm, a cutting blade and a grinding disc with the diameter of 4mm are selected to be prepared into small particles to obtain the component B.
The preparation method of the millet congee filtrate in the embodiment comprises the following steps: boiling 50g of semen Setariae in 500ml of water for 15min, and filtering with absorbent cotton gauze.
Example 2
The embodiment discloses a preparation method of an entomopathogenic nematode preparation for preventing and treating spodoptera frugiperda, which specifically comprises the following steps:
taking 300ml of millet congee filtrate, 10g of trehalose (alpha ), 1g of ascorbic acid, 1.7g of peptone and 200.8 g of tween, putting the millet congee filtrate, mixing the mixture uniformly, covering a sealing film, putting the mixture into an autoclave, sterilizing (melting) for 20min under the conditions that the pressure is 1.0MPa and the temperature is 121 ℃, cooling the obtained product, mixing with entomopathogenic nematodes 3,500,000IJs, and adding distilled water to a constant volume of 1L to obtain the component A.
Weighing 4g of agar powder, 45g of corn flour, 9g of wheat flour, 20g of soybean meal, 2g of yeast powder, 3g of trehalose (alpha ), 0.5g of ascorbic acid, 5g of peptone, 2.5g of beef extract and 2g of soybean oil, placing the mixture in the same container, uniformly mixing, placing the container in an autoclave with a sealing film, sterilizing (melting) the mixture for 20min under the conditions that the pressure is 1.0MPa and the temperature is 121 ℃, cooling the obtained product, adding 15g of ecdysone and 8mg of tyrosinase inhibitor deoxyarbutin, and fixing the volume to 1L by using distilled water to obtain a B-component semi-finished product.
And (3) uniformly stirring the semi-finished product of the component B into paste, then sending the paste into a dryer, drying the paste at 80 ℃ until the moisture content is 10%, and then putting the paste into a shaft-connected granulator (a particle plate with holes distributed by an insertion and arrangement method and with the aperture of 2.0-3.0 mm, a cutting blade and a grinding disc with the diameter of 4mm are selected to be prepared into small particles to obtain the component B.
The preparation method of the millet congee filtrate in the embodiment comprises the following steps: boiling 50g of millet in 500ml of water for 10min, and filtering with absorbent cotton gauze.
Example 3
The embodiment discloses a preparation method of an entomopathogenic nematode preparation for preventing and treating spodoptera frugiperda, which specifically comprises the following steps:
taking 250ml of millet congee filtrate, 7g of trehalose (alpha ), 0.9g of ascorbic acid, 1.8g of peptone and 200.6 g of tween, putting the millet congee filtrate, mixing the mixture uniformly, covering a sealing film, putting the mixture into an autoclave, sterilizing (melting) the mixture for 40min under the conditions that the pressure is 1.0MPa and the temperature is 121 ℃, cooling the obtained product, mixing with 4,000,000IJs of entomopathogenic nematodes, and adding distilled water to a constant volume of 1L to obtain the component A.
Weighing 5g of agar powder, 55g of corn flour, 8g of wheat flour, 25g of soybean meal, 3g of yeast powder, 6g of trehalose (alpha ), 0.2g of ascorbic acid, 3g of peptone, 3.5g of beef extract and 1.5g of soybean oil, placing the materials in the same container, uniformly mixing, placing the container in an autoclave after being covered with a sealing film, sterilizing (melting) for 20min under the conditions that the pressure is 1.0MPa and the temperature is 121 ℃, cooling the obtained product, adding 12g of ecdysone and 5mg of tyrosinase inhibitor deoxyarbutin, and fixing the volume to 1L by using distilled water to obtain a B-component semi-finished product.
And (3) uniformly stirring the semi-finished product of the component B into paste, then sending the paste into a dryer, drying the paste at 80 ℃ until the moisture content is 12%, and then putting the paste into a shaft-connected granulator (a particle plate with holes distributed by an insertion and arrangement method and with the aperture of 2.0-3.0 mm, a cutting blade and a grinding disc with the diameter of 4mm are selected to be prepared into small particles to obtain the component B.
The preparation method of the millet congee filtrate in the embodiment comprises the following steps: boiling 50g of semen Setariae in 500ml of water for 15min, and filtering with absorbent cotton gauze.
Example 4
The embodiment discloses a preparation method of an entomopathogenic nematode preparation for preventing and treating spodoptera frugiperda, which specifically comprises the following steps:
taking 200ml of millet congee filtrate, 5g of trehalose (alpha ), 0.8g of ascorbic acid, 1.9g of peptone and 200.5 g of tween, putting the millet congee filtrate, mixing the mixture uniformly, covering a sealing film, putting the mixture into an autoclave, sterilizing (melting) the mixture for 40min under the conditions that the pressure is 1.0MPa and the temperature is 121 ℃, cooling the obtained product, mixing with 4,500,000IJs of entomopathogenic nematodes, and adding distilled water to a constant volume of 1L to obtain the component A.
Weighing 2g of agar powder, 50g of corn flour, 7g of wheat flour, 40g of soybean meal, 4g of yeast powder, 5g of trehalose (alpha ), 0.3g of ascorbic acid, 4g of peptone, 6g of beef extract and 3g of soybean oil, placing the materials in the same container, uniformly mixing, covering a sealing film, placing the container in an autoclave, sterilizing (melting) the mixture for 20min under the conditions that the pressure is 1.0MPa and the temperature is 121 ℃, cooling the obtained product, adding 14g of ecdysone and 6mg of tyrosinase inhibitor deoxyarbutin, and fixing the volume to 1L by using distilled water to obtain a B-component semi-finished product.
And (3) uniformly stirring the semi-finished product of the component B into paste, then sending the paste into a dryer, drying the paste at 80 ℃ until the moisture content is 12%, and then putting the paste into a shaft-connected granulator (a particle plate with holes distributed by an insertion and arrangement method and with the aperture of 2.0-3.0 mm, a cutting blade and a grinding disc with the diameter of 4mm are selected to be prepared into small particles to obtain the component B.
The preparation method of the millet congee filtrate in the embodiment comprises the following steps: boiling 50g of semen Setariae in 500ml of water for 15min, and filtering with absorbent cotton gauze.
Example 5
The embodiment discloses a preparation method of an entomopathogenic nematode preparation for preventing and treating spodoptera frugiperda, which specifically comprises the following steps:
350ml of millet congee filtrate, 9g of trehalose (alpha ), 1.1g of ascorbic acid, 2g of peptone and 200.8 g of tween are put into the same container, mixed uniformly, covered with a sealing film and put into an autoclave for sterilization (melting) for 30min under the conditions of pressure of 1.0MPa and temperature of 121 ℃, the obtained product is cooled and mixed with 5,000,000IJs of entomopathogenic nematodes, and the volume is fixed and supplemented to 1L by distilled water, thus obtaining the component A.
Weighing 2g of agar powder, 40g of corn flour, 10g of wheat flour, 35g of soybean meal, 2g of yeast powder, 4g of trehalose (alpha ), 0.5g of ascorbic acid, 3.5g of peptone, 5g of beef extract and 2.5g of soybean oil, putting the mixture into the same container, uniformly mixing, putting the container into an autoclave after being covered with a sealing film, sterilizing (melting) for 20min under the conditions that the pressure is 1.0MPa and the temperature is 121 ℃, cooling the obtained product, adding 13g of ecdysone and 7.5mg of tyrosinase inhibitor deoxyarbutin, and fixing the volume to 1L by using distilled water to obtain a semi-finished product of the component B.
And (3) uniformly stirring the semi-finished product of the component B into paste, then sending the paste into a dryer, drying the paste at 80 ℃ until the moisture content is 12%, and then putting the paste into a shaft-connected granulator (a particle plate with holes distributed by an insertion and arrangement method and with the aperture of 2.0-3.0 mm, a cutting blade and a grinding disc with the diameter of 4mm are selected to be prepared into small particles to obtain the component B.
The preparation method of the millet congee filtrate in the embodiment comprises the following steps: boiling 50g of semen Setariae in 500ml of water for 15min, and filtering with absorbent cotton gauze.
Example 6
This example provides the use of the formulations of the invention to control spodoptera frugiperda.
The field control test is carried out in a Wu polder corn test base in Guangxi Nanning city, the total number of the field control test is 8, each treatment is set to be 3 times, the total number is 24 cells, and each cell is 32m2(8m 4m), the cells are distributed in random blocks.
In the small horn mouth period of the corn, the component B is sprayed in the corn seedling cores, water is sprayed immediately after the spraying to soften the component B by absorbing water (the water consumption is 50 kg/mu of land), and the component A is injected into the corn seedling cores by a wash bottle (or an injector) in the evening after 1 day. The application amount of the component A is 1 ml-2 ml per corn seedling, and the application amount of the component B is 0.05-0.1 g per corn seedling. Thus, the egg laying characteristic of spodoptera frugiperda is mainly utilized: the larvae of spodoptera frugiperda after egg incubation prefer to be stored in the corncobs to bite tender plant tissues. After the component A and the component B are applied, water is sprayed every 7 days to keep humidity. The survey used 5-point sampling and 20 maize plants were surveyed per test plot. The population base number of insects is investigated before application, the age of insects is mainly 3 years, and part of insects is 4-6 years. The degree of new leaf damage of corn was investigated and classified on the 10 th day after the administration. Grading standard: grade 0, new leaves are not harmed by the insect; grade 1, the new leaves have smaller lesion spots; grade 3, connecting the harmful spots into pieces and forming transparent films; grade 5, which is a defect that causes new leaves to be notched; grade 7, the damage causes the new leaves to have larger and more gaps; grade 9, causing new leaf rupture. And simultaneously, the number of the Spodoptera frugiperda living insects on the corn samples is stripped and recorded.
And calculating the damage index, the leaf protection effect and the insect population prevention effect. The effect of each agent treatment on crops and on non-target organisms was observed during the experiment.
The damage index (%) is [ Σ (number of damaged plants at each stage × relative damaged-stage representative value)/(total number of investigated plants × highest-stage value) ] × 100%
Leaf retention (%) < control area damage index > (control area damage index)/control area damage index >. times.100%
The oral cavity control effect (%) is [1- (number of live insects after drug treatment zone x number of live insects before drug control zone)/(number of live insects before drug treatment zone x number of live insects after drug control zone) ] × 100%
Treatment 1 group: contains only entomopathogenic nematodes;
treatment 2 groups: only the A component (entomopathogenic nematodes and adjuvant);
treatment of 3 groups: only contains the component A (entomopathogenic nematodes and auxiliary agents) and a phagostimulant;
treatment of 4 groups: only contains component A (entomopathogenic nematodes and auxiliary agents), phagostimulant and ecdysone;
treatment of 5 groups: only contains component A (entomopathogenic nematodes and auxiliary agents), phagostimulant and deoxyarbutin;
treatment of 6 groups: contains only entomopathogenic nematodes, and component B (phagostimulant, ecdysone and deoxyarbutin);
treatment of 7 groups: comprises a component A (entomopathogenic nematodes and auxiliaries) and a component B (phagostimulant, ecdysone and deoxyarbutin).
Blank control group: and (5) treating with clear water.
The compositions of the materials treated in groups 1 to 7 are shown in the following table, wherein the semi-finished products of group A and group B are each prepared in 1L, and the preparation method is carried out according to the relevant steps of example 1:
TABLE 1-1
Tables 1 to 2
The results are shown in the following table:
TABLE 2
Note: in the table, the letters a, b, c, d, e and f are different and represent significant differences, and P is less than 0.05.
The results show that the number of live insects in the group 1 treated was reduced compared to the blank group, and there was a significant difference; the insect control effect and the leaf protection effect are improved, and the obvious difference exists; the damage index is reduced and has significant difference, which shows that the application of entomopathogenic nematodes can prevent and control the damage of spodoptera frugiperda larvae to a certain extent.
Compared with the group 1, the group 2 has increased insect population control effect and leaf retention effect, but no significant difference, which indicates that the effect of the entomopathogenic nematodes on controlling spodoptera frugiperda larvae cannot be significantly improved after the addition of the auxiliary agent.
Compared with the group 2, the group 3 has no significant increase in the insect population control effect, but has a decreased damage index and an increased leaf protection effect, and has significant differences, which indicates that the addition of the phagostimulant can improve the effect of the entomopathogenic nematodes on controlling spodoptera frugiperda larvae on the basis of the entomopathogenic nematodes and the auxiliary agent.
Compared with the treatment groups 2 and 3, the treatment group 4 has obviously improved leaf protection effect, reduced damage index and obvious difference; the combination of the phagostimulant and ecdysone is proved to be capable of obviously improving the efficiency of the entomopathogenic nematodes for preventing and controlling Spodoptera frugiperda larvae;
compared with the treatment groups 2 and 3, the treatment group 5 has obviously reduced damage index and obviously improved leaf protection effect; the tyrosinase inhibitor-deoxyarbutin is shown to be capable of remarkably improving the effect of entomopathogenic nematodes on preventing and controlling spodoptera frugiperda larvae;
compared with the treatment groups 6, 5 and 4, the treatment groups 7 have significantly reduced live insect number, significantly improved insect population prevention effect and leaf protection effect, and significantly reduced damage index and significant difference; the auxiliary agent in the component A, the phagostimulant in the component B, the ecdysone and the deoxyarbutin serving as a tyrosinase inhibitor are combined for use, the biological control effect is optimal, and the quick-acting performance of entomopathogenic nematodes on controlling spodoptera frugiperda larvae can be greatly improved.
Example 7
This example discloses the optimal comparison of the application times of the present invention.
The entomopathogenic nematode formulation used in this example was prepared according to the method of example 5.
The test method was the same as example 6 except that the application time was different for each treatment group of this example. The embodiment has 3 processes, each process has 3 repetitions, which is specifically as follows:
treatment 1 group: the component B is applied in the afternoon, and the component A is applied in the evening of the same day;
treatment 2 groups: the component B is applied in the afternoon, and the component A is applied in the evening of the next day;
treatment of 3 groups: the component B is applied in the afternoon, and the component A is applied in the evening of the third day;
wherein treatment 1, treatment 2 and treatment 3 were the same day with component B applied in the afternoon.
Blank control group: spraying clear water in afternoon.
The results are shown in the following table:
TABLE 3
Note: in the table, the letters a, b and c are different and have significant difference, and P is less than 0.05.
Results show that compared with a blank control group, the treatment groups 1-3 have improved insect control effect and leaf retention effect, and have significant difference; the victims decreased exponentially and had significant differences; the three application methods of the embodiment are shown to play a role in preventing and controlling Spodoptera frugiperda larvae.
Compared with the treatment group 1 and the treatment group 3, the treatment group 2 has improved insect control effect and leaf retention effect, and has significant difference; the victims decreased exponentially and had significant differences; the application effect of the component A in the evening of the second day after the component B is applied is shown to be the best, and the application effect of the component A in the control of Spodoptera frugiperda is higher than that of the component B and the component A applied in the same day and is also higher than that of the component A applied in the third day after the component B is applied.
The above-mentioned embodiment is only one of the preferred embodiments of the present invention, and should not be used to limit the scope of the present invention, but all the insubstantial modifications or changes made within the spirit and scope of the main design of the present invention, which still solve the technical problems consistent with the present invention, should be included in the scope of the present invention.
Claims (10)
1. An entomopathogenic nematode preparation for preventing and treating Spodoptera frugiperda is characterized by comprising a component A and a component B which contain different active ingredients; wherein the component A comprises entomopathogenic nematodes and an auxiliary agent for maintaining the vitality of the entomopathogenic nematodes; the component B comprises a phagostimulant, ecdysone and a tyrosinase inhibitor.
2. The entomopathogenic nematode formulation for controlling spodoptera frugiperda of claim 1 wherein component a is a suspension comprising 3,000,000IJs to 5,000,000IJs of entomopathogenic nematodes per 1L A;
or/and each 1L A component contains the following auxiliary components: 200-400 ml of millet congee filtrate, 5-10 g of humectant, 0.8-1.2 g of preservative, 1.5-2.0 g of nematode symbiotic bacteria nutrient and 0.5-0.8 ml of stabilizer.
3. The entomopathogenic nematode formulation for controlling spodoptera frugiperda of claim 2, wherein said entomopathogenic nematodes comprise Steinernema carpocapsae;
or/and the humectant comprises trehalose (α, α);
or/and the preservative comprises ascorbic acid;
or/and the nematode symbiotic bacteria nutrient comprises peptone;
or/and the stabilizer comprises tween, preferably tween 20.
4. The entomopathogenic nematode formulation for controlling spodoptera frugiperda according to any of claims 1 to 3, wherein component B is a solid granule obtained by drying a semifinished product of component B; each 1L B component semi-finished product contains the following phagostimulant components: 2-5 g of agar powder, 40-60 g of corn flour, 5-10 g of wheat flour, 20-40 g of soybean flour, 2-5 g of yeast powder, 3-6 g of trehalose (alpha ), 0.2-0.5 g of ascorbic acid, 2.5-5 g of peptone, 2.5-6 g of beef extract, 1-3 g of soybean oil and the balance of water;
or/and each 1L B component semi-finished product contains 10-15 g of ecdysone;
or/and each 1L B component semi-finished product contains 5-8 mg of tyrosinase inhibitor.
5. The entomopathogenic nematode formulation for controlling spodoptera frugiperda of claim 4, wherein said tyrosinase inhibitor is deoxyarbutin.
6. A process for the preparation of a formulation of entomopathogenic nematodes for the control of Spodoptera frugiperda according to any of claims 1 to 5, characterized in that the preparation of the A-component comprises the following steps: weighing the auxiliary components according to a proportion, mixing uniformly, sterilizing at high temperature, cooling to room temperature, adding entomopathogenic nematodes, and fixing the volume with distilled water to obtain a component A;
the preparation of the component B comprises the following steps: weighing the phagostimulant components according to a proportion, mixing uniformly, sterilizing at high temperature, adding ecdysone and tyrosinase inhibitor, fixing the volume with distilled water to obtain a semi-finished product of the component B, uniformly stirring the semi-finished product of the component B, drying, and granulating to obtain the component B.
7. The method according to claim 6, wherein the moisture content of the component B is 10 to 12 wt.%.
8. Use of an entomopathogenic nematode formulation for the control of spodoptera frugiperda according to any of claims 1 to 5 for the control of spodoptera frugiperda.
9. Use according to claim 8, wherein the B-side is administered first and the A-side is administered second.
10. The use according to claim 9, wherein the B fraction is sprinkled in the core of the crop seedling, the B fraction is softened by spraying water after the sprinkling, and the a fraction is applied in the core of the crop seedling after 1 day;
preferably, the a-component is applied at or after evening.
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| CN116711738B (en) * | 2023-06-14 | 2024-04-05 | 江西省农业科学院植物保护研究所 | Biocontrol agent and application thereof in spodoptera frugiperda control |
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