CN116333895A - Special culture medium for insecticidal fungi without sterilization, production method of insecticidal fungi, and fungi preparation and application - Google Patents
Special culture medium for insecticidal fungi without sterilization, production method of insecticidal fungi, and fungi preparation and application Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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- 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
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P7/00—Arthropodicides
- A01P7/04—Insecticides
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- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
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- C12R2001/645—Fungi ; Processes using fungi
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Abstract
The invention discloses a special culture medium for insecticidal fungi without sterilization, a production method of the insecticidal fungi, a fungus preparation and application. The special culture medium comprises crab shell powder, and can also comprise yellow meal worm powder or/and shrimp powder, is beneficial to the propagation of insecticidal fungi, inhibits the growth of mixed fungi, can be used for the production of the insecticidal fungi without high-temperature or high-pressure sterilization treatment by combining with inoculation of a large amount of insecticidal fungi spores, is convenient to use and simple in production process, can greatly reduce the production cost of the insecticidal fungi, improves the insecticidal control effect and prolongs the control duration. The fungus preparation is obtained after the insecticidal fungus is added into the special culture medium, and can be used for improving soil and preventing and controlling crop diseases and insect pests.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to a special culture medium for insecticidal fungi without sterilization, a method for producing the insecticidal fungi without sterilization, a fungus preparation prepared by using the special culture medium for the insecticidal fungi and application.
Background
Insect pests cause huge losses to agricultural production every year, but chemical pesticides are singly and massively used for a long time, so that environmental pollution is serious, the drug resistance is increased, natural enemies are killed, and ecological balance is destroyed. The insecticidal fungi have the advantages of difficult generation of drug resistance of pests, environmental friendliness and the like, play an important role in reducing the use of chemical pesticides, are agricultural microorganisms widely applied in the agricultural field, and have been registered as insecticidal microbial pesticides, microbial bactericides, microbial fertilizers, microbial seed treatment agents and other agricultural application directions. However, in the production and use of insecticidal fungi, a number of limiting factors have limited the broad use of insecticidal fungi.
Firstly, the production cost is high in the aspect of insecticidal fungus fermentation production. At present, grains (rice and wheat) or combined agricultural and sideline products (wheat bran and rice bran) are mainly used as raw materials, and the raw materials often carry various miscellaneous bacteria (such as various aspergillus), and are also suitable for the growth of the miscellaneous bacteria. Therefore, it is necessary to sterilize raw materials in fermentation production and maintain a sterile environment for the fermentation process, which greatly increases production costs and difficulty. Therefore, the types of the insecticidal fungi products such as the metarhizium anisopliae, the beauveria bassiana and the like are more, but the mass production is extremely small and the cost is high.
Secondly, the application technology of the insecticidal fungi is difficult and the effect is unstable. Fungi are controlled by the way living spores contact the pests, and need to survive and grow and reproduce in the environment. However, the insecticidal fungal spores such as metarhizium anisopliae, beauveria bassiana and the like need to be contacted with plant surfaces and rhizosphere, seed surfaces, soil, organic fertilizers and chemical fertilizers and insect surfaces in the application process, and a large number of other microorganisms exist in the scenes, so that the survival, growth and propagation of the insecticidal fungal spores can be influenced through mechanisms such as quantity advantage, predominance, bacteriostasis and fungi killing function and the like, and the control and lasting effects of the insecticidal fungi on insects are influenced.
In summary, in the production and application of the insecticidal fungi, the promotion of the growth and propagation of the insecticidal fungi while the inhibition of the growth and propagation of the mixed fungi is a technical key for reducing the production cost of the insecticidal fungi, improving the insecticidal control effect and prolonging the control duration.
The body wall is an important barrier of insects against the invasion of foreign microorganisms and mainly consists of substances such as shell proteins, chitin, quinones, phenols and the like. Wherein, the content of the shell and white matter is the highest, a special complex is formed by wrapping a small amount of chitin wires and crosslinking the chitin wires, and the hydrolytic enzyme of most microorganisms is difficult to degrade the shell and white matter; in addition, substances such as quinones, phenols and the like in the insect body wall have the effect of obviously inhibiting the growth of microorganisms. These physical and chemical barriers of the insect body wall effectively prevent the invasion of other microorganisms into the insect. The insecticidal fungi have obtained the ability to overcome physical and chemical barriers of insect body walls in the long-term evolution process, and can produce a series of insect cuticle hydrolases, including rich protease systems, chitinase, peroxidase and other hydrolases, which can efficiently degrade main components such as capsid proteins, chitin, quinones, phenols and the like in the cuticle, and utilize the nutrients for growth and propagation. Therefore, only a very small number of fungi evolve into entomopathogenic fungi (insecticidal fungi) in nature, but non-entomopathogenic fungi such as aspergillus, penicillium, rhizoctonia solani, saprolegnia, root rot and fusarium oxysporum cannot secrete corresponding high-efficiency hydrolytic enzymes, substances such as shell proteins, chitin, quinones and phenols of insect epidermis cannot be effectively utilized, and the substances such as quinones and phenols in the fungi cannot be inhibited, so that the fungi are difficult to grow and reproduce on the insect epidermis. The shell protein accounts for more than half of the dry weight of the insect epidermis, can provide rich nitrogen source and carbon source for the growth and propagation of insecticidal fungi, and is difficult to be utilized by other microorganisms. Therefore, the raw materials containing the shell proteins and the like can effectively promote the growth and the reproduction of the insecticidal fungi after being mixed with the spores of the insecticidal fungi, and inhibit the growth and the reproduction of other fungi and other microorganisms, so that the production cost can be reduced, the insecticidal control effect can be improved, and the control duration can be prolonged in the production and the application of the insecticidal fungi.
The content of the capsid protein in the skin component of arthropods is often 50% or more, and it is difficult to use it by many microorganisms. The present invention thus proposes a technique of using arthropod powder as a medium for pesticidal fungi, in combination with a larger amount of inoculated pesticidal fungi, thereby realizing the production of pesticidal fungi and pesticidal fungi preparations without sterilization, the technique comprising a medium dedicated to pesticidal fungi without sterilization, a method of producing pesticidal fungi without sterilization, and the preparation of fungi preparations and applications using the medium dedicated to pesticidal fungi.
Disclosure of Invention
In view of this, it is an object of the present invention to provide a special medium for pesticidal fungi, which comprises crab shell powder, without sterilization.
Preferably, the special culture medium further comprises flour weevil powder or/and shrimp powder. The crab shell powder and the yellow meal worm powder or/and shrimp powder are used together to be more beneficial to the proliferation of insecticidal fungi.
Preferably, the mass fraction of crab shell powder in the special culture medium is more than 60%.
Preferably, the mass fractions of the crab shell powder, the yellow meal worm powder and the shrimp powder in the special culture medium are respectively 60-80%, 10-40% and 10-40%.
Preferably, the mass fraction of crab shell meal, yellow meal worm meal and shrimp meal in the special culture medium is 60%, 10% and 30%, respectively, or 60%, 30% and 10%, respectively, or 60%, 20% and 20%, respectively. The culture medium comprises crab shell powder, yellow meal worm powder and shrimp powder according to the mass fraction, is optimal for the proliferation of insecticidal fungi and is unfavorable for the proliferation of mixed bacteria.
The second object of the present invention is to provide a method for producing a pesticidal fungus without sterilizing, comprising directly inoculating a pesticidal fungus into any one of the above-mentioned special culture media for fermentation, inoculating a larger amount of spores of the pesticidal fungus without sterilizing the special culture media before inoculation, the inoculation amount being 1X 10 or more 6 Each of the insecticidal fungus spores/gram of the special culture medium. The special culture medium can be used without high temperature or high pressure sterilization, when the inoculation amount is more than or equal to 1 multiplied by 10 6 When the special culture medium is used for killing the spores of the fungi/gram, the mixed bacteria amount is low, the propagation effect of the fungi is good, and compared with the traditional fermentation after the high-temperature sterilization of grains, the production and use cost of the fungi is greatly reduced.
It is a further object of the present invention to provide a pesticidal fungal preparation comprising a pesticidal fungus and any of the aforementioned special culture media, wherein the formulation is pesticidalThe content of insect and fungus is 1×10 or more 6 Each of the insecticidal fungus spores/gram of the special culture medium.
The invention provides a preparation method of a pesticide fungus preparation, which is obtained by adding the pesticide fungus into any special culture medium, wherein the special culture medium is not sterilized before the pesticide fungus is added, and the adding amount of the pesticide fungus is more than or equal to 1 multiplied by 10 6 The special culture medium is used for the spores/gram of the insecticidal fungi, and the insecticidal fungi can grow and reproduce by using the special culture medium after the preparation is applied.
It is a fifth object of the present invention to provide the use of the above fungal formulation for soil improvement and for controlling crop pests. The fungus preparation can be diluted and irrigated to soil for use; the fungus preparation can also be used for sowing after the plant seeds are treated by seed dressing; the fungus preparation can be combined with organic fertilizer or compound fertilizer, namely, the fungus preparation and the fertilizer are mixed and then applied to soil, and the fertilizer comprises the organic fertilizer and the compound fertilizer. The insecticidal fungi have good propagation effect in the special culture medium, and can better protect crops so as to reduce crop diseases and insect pests.
The special culture medium without sterilizing comprises crab shell powder, yellow meal worm powder or/and shrimp powder, and the special culture medium is directly inoculated with a large dose of insecticidal fungi without sterilizing treatment, even if the content is more than or equal to 1 multiplied by 10 6 The special culture medium is used for cultivating the spores/gram of the insecticidal fungi, and then fermentation is carried out, so that the insecticidal fungi are produced, the proliferation effect is good, the mixed bacteria rate is low, the process is simple, and the production cost of the insecticidal fungi produced by high-temperature sterilization at present is greatly reduced; the invention adds the insecticidal fungi into the fungus preparation prepared by the special culture medium, has simple and convenient production, is beneficial to the multiplication of the insecticidal fungi in the field, has convenient use and good effect, and can be used for improving soil and preventing and controlling crop diseases and insect pests.
Detailed Description
The present invention will be described in detail with reference to examples, which are illustrative only and are not limiting the scope of application of the present invention. The present invention is not limited to the following embodiments or examples, and modifications and variations made without departing from the spirit of the present invention are intended to be included in the scope of the present invention.
Material preparation: the insecticidal fungi are selected from Metarhizium anisopliae (Metarhizium anisopliae) and Beauveria bassiana (Beauveria bassiana), and are purchased from Chongqing Polymerise biological engineering Co. Aspergillus flavus and Escherichia coli were given by the university of Chongqing research center. Arthropod powder, wherein crab shell is purchased from Bo Yide pharmaceutical Co., ltd; silkworm chrysalis is purchased by Bo De Yi pharmaceutical sales limited; yellow meal worm is purchased from Ningbo vonic Shannon Biotechnology Co., ltd; shrimp meal was purchased from Shandong Lu Hua ocean technologies Co., ltd; maggot meal is purchased from He-Ji-Sheng Yuan biotechnology Co., ltd; cockroach meal was purchased from Bozhou-city, carrier pharmaceutical Co., ltd; periostracum Cicadae coarse powder is purchased from Biotechnology Inc. of Imperial canteen in Bozhou. Grinding the purchased arthropod dry powder and the purchased arthropod coarse powder by a machine through a 100-mesh sieve to obtain crab shell powder, silkworm chrysalis powder, maggot powder, yellow meal worm powder, cockroach powder, cicada slough powder and shrimp powder for later use. Of course, arthropod powders can also be prepared without sieving.
Experimental example 1 determination of compatibility of arthropod powder with insecticidal fungi
The method comprises the following steps: crab shell powder, silkworm chrysalis powder, maggot powder, yellow meal worm powder, cockroach powder, cicada slough powder and shrimp powder are respectively mixed with metarhizium anisopliae or beauveria bassiana spore powder according to the mass of 100:1, mixing evenly, taking the destruxin or beauveria bassiana spore powder without adding arthropod powder as a control, culturing in a 37 ℃ incubator, detecting spore germination rates respectively for 3 days and 14 days, and determining whether the arthropod powder has an influence on the germination of insecticidal fungus spores. Diluting each composition after culture with sterile water to spore content of 1×10 7 spore/mL, 50. Mu.L of spore suspension is uniformly coated on a 1/4SDAY plate, and the germination rate is detected after culturing for 14-16h at 26 ℃.
Results: as shown in Table 1, the germination rates of the green muscardine fungi and the beauveria bassiana mixed with the crab shell powder, the silkworm chrysalis powder, the yellow mealworm powder, the cicada slough powder and the shrimp powder are about 63% and are equivalent to those of the control, and the germination rates of the green muscardine fungi and the beauveria bassiana mixed with the crab shell powder, the silkworm chrysalis powder, the yellow mealworm powder, the cicada slough powder and the shrimp powder are not negatively influenced. Therefore, the crab shell powder, the silkworm chrysalis powder, the yellow meal worm powder, the cicada slough powder and the shrimp powder have better compatibility with the metarhizium anisopliae and the beauveria bassiana, and are expected to be applied to the production, the preparation and the field use of the insecticidal fungi.
TABLE 1 Effect of arthropod powder on the spore germination Rate of Metarrhizium anisopliae and Beauveria bassiana (37 ℃ C.)
Experimental example 2 Effect of arthropod powder on growth and propagation of insecticidal fungi and other fungi
The method comprises the following steps: preparing a PDA plate and an agar plate containing 10% arthropod powder (crab shell powder, silkworm chrysalis powder, yellow meal worm powder, periostracum Cicadae powder or shrimp powder); selecting a flat plate for growing the metarhizium anisopliae, the beauveria bassiana and the aspergillus flavus for 7 days, punching by a puncher with the diameter of 9mm, inoculating metarhizium anisopliae, beauveria bassiana and aspergillus flavus cakes to the centers of the agar flat plates with different arthropod powders respectively, culturing for 10 days at the temperature of 27+/-1 ℃, observing the growth conditions of the metarhizium anisopliae, the beauveria bassiana and the aspergillus flavus on the flat plate, measuring the size of a colony, and calculating the growth rate of the colony by taking a PDA flat plate as a control, wherein the growth rate=100% × (the diameter of the colony-9 mm)/9 mm.
LB liquid medium and liquid medium containing 10% arthropod powder (crab shell powder, silkworm chrysalis powder, yellow meal worm powder, cicada slough powder or shrimp powder) were prepared. The preparation method of the liquid culture medium containing 10% of arthropod powder comprises the following steps: different arthropod powders sterilized at high temperature were dispersed in sterile water to prepare 10% by mass of arthropod powder liquid medium. 1mL of coliform suspension with the same concentration is inoculated into 100mL of each liquid culture medium (the coliform suspension is obtained by culturing coliform in LB liquid at 36+/-1 ℃ overnight), the temperature is 36+/-1 ℃, the speed is 200r/min, the shaking culture is 16h, the LB culture medium is used as a control, the concentration of the coliform in the culture medium before shaking culture and the concentration of the coliform in the culture medium after shaking culture are respectively determined by using a gradient dilution plating method, and the growth multiple of the coliform before shaking culture and the growth multiple of the coliform after shaking culture relative to each culture medium are calculated, wherein the growth multiple is = (the number of the coliform after shaking culture-the number of the coliform before shaking culture)/the number of the coliform before shaking culture.
Results: as shown in Table 2, it is seen that the culture medium containing crab shell powder, yellow meal worm powder and shrimp powder has growth promoting effect on both green muscardine fungus and white muscardine fungus, and has no obvious growth promoting effect on Aspergillus flavus and Escherichia coli.
TABLE 2 Effect of arthropod powder on growth and reproduction of insecticidal fungi and other fungi
Experimental example 3 Effect of different inoculum sizes of insecticidal fungi on propagation of Mixed fungi
Respectively mixing crab shell powder, yellow meal worm powder or shrimp powder as culture medium with appropriate amount of destruxin or beauveria bassiana spore powder to make spore content in each gram of culture medium be 1×10 respectively 5 Spores, 1×10 6 Spores, 1×10 7 Adding 50% water into the spores according to the weight ratio, uniformly mixing, carrying out constant-temperature and moisture-preserving culture at 26-28 ℃ for 10 days, measuring the total fungus and bacteria number and the metarhizium anisopliae or beauveria bassiana number in each gram of substances after fermentation by adopting a CFU method, calculating the mixed fungus rate,
as can be seen from Table 3, the content of spores of the insecticidal fungi in the culture medium is 1X 10 5 When spores/g, crab shell powder, yellow meal worm powder and shrimp powder are greenThe mixed bacteria rates of the beauveria bassiana and the beauveria bassiana are respectively
And->
Between them; when the content is increased to 1X 10 6 Spores/g, 1X 10 7 The spore/g is low in the mixed bacteria of Metarhizium anisopliae and Beauveria bassiana, and the mixed bacteria are +.>
And
the content of mixed bacteria is far lower than that of the existing production fermentation of the insecticidal fungus raw material.
As can be seen from the above, the inoculation content in the culture medium is higher than 1×10 6 When spores/g, the arthropod powder does not need high-temperature high-pressure sterilization, beauveria bassiana and metarhizium anisopliae can be quickly propagated to form dominant bacterial groups, inhibit the propagation of mixed bacteria, meet the control requirement of the mixed bacteria in production, and have good application prospect.
TABLE 3 Effect of different inoculum sizes on the proliferation of infectious agents
Note that: the difference analysis was performed between the same arthropod powders.
Experimental example 4 determination of conditions of different proportions of culture media for promoting propagation of insecticidal fungi
1. Preparation of culture mediums with different proportions
The components of the culture mediums with different proportions are prepared according to the mass percentage, 0-100% of crab shell powder, 0-100% of yellow meal worm powder and 0-100% of shrimp powder are weighed and fully mixed to obtain the culture mediums with different proportions, 13 culture mediums with different proportions (namely 13 formulas) are prepared in the future experiment, and the specific content of the crab shell powder, the yellow meal worm powder and the shrimp powder in each formula is shown in Table 4.
Table 4 culture medium formulations of different ratios
| Formulation of | Crab shell powder content (%) | Tenebrio molitor powder content (%) | Shrimp powder content (%) |
| Formulation 1 | 100 | - | -- |
| Formulation 2 | - | 100 | - |
| Formulation 3 | - | - | 100 |
| Formulation 4 | 0 | 50 | 50 |
| Formulation 5 | 0 | 75 | 25 |
| Formulation 6 | 0 | 25 | 75 |
| Formulation 7 | 80 | 10 | 10 |
| Formulation 8 | 60 | - | 40 |
| Formulation 9 | 60 | 10 | 30 |
| Formulation 10 | 60 | 20 | 20 |
| Formulation 11 | 60 | 30 | 10 |
| Formulation 12 | 60 | 40 | - |
| Formulation 13 | 40 | 30 | 30 |
2. Effect of culture Medium dedicated to insecticidal fungi on proliferation of insecticidal fungi
According to 13 formulas of the culture medium in Table 4, the culture medium is respectively and uniformly mixed with appropriate amount of destruxin spores or beauveria bassiana spore powder, so that the spore content in each gram of the culture medium is 1×10 6 Adding 50% water according to weight ratio, uniformly mixing, constant-temperature moisturizing and culturing at 26-28deg.C for 7 days, measuring total fungus content and different fungus spore content in each gram of fermented product by CFU method, calculating proliferation multiple and mixed fungus rate, proliferation multiple= (fungus spore content after culturing-fungus spore content before culturing)/fungus spore content before culturing,
the results are shown in Table 5: the culture medium compounded by three arthropod powders of the formula 9-11 (the crab shell powder content is 60%, the yellow meal worm powder and the shrimp powder) has the most obvious effect of promoting the propagation of the metarhizium anisopliae and the beauveria bassiana, and the propagation multiples of the metarhizium anisopliae and the beauveria bassiana are about 1.3 ten thousand times and about 2.4 ten thousand times respectively; formula 7 (crab shell powder content 80%), formula 8 (crab shell powder content 60%), formula 12 (crab shell powder content 60%) have inferior effects of promoting propagation of metarhizium anisopliae and beauveria bassiana, the propagation multiples of metarhizium anisopliae and beauveria bassiana are about 1.1 ten thousand times and about 2 ten thousand times respectively, and the propagation promoting effects of metarhizium anisopliae and beauveria bassiana are poorer than those of formulas 9-11; formula 13 (crab shell powder content 40%) is compounded by three arthropod protein powder, the proliferation times of the metarhizium anisopliae and the beauveria bassiana are about 8000 times and 1.6 ten thousand times respectively, and the proliferation promoting effect of formula 13 on the metarhizium anisopliae and the beauveria bassiana is poorer than that of formulas 7, 8 and 12; formulas 1 to 6 are compounded with 1 arthropod powder or 2 arthropod powdersThe proliferation times of the metarhizium anisopliae and the beauveria bassiana are about 5800 times and about 1.1 ten thousand times respectively, wherein the proliferation times of the metarhizium anisopliae and the beauveria bassiana which are culture mediums compounded by 2 or 3 kinds of powder are higher than that of the culture medium compounded by 1 kind of powder; the content of mixed bacteria in the metarhizium anisopliae culture mediums with different proportions is as follows
The content of mixed bacteria in beauveria bassiana is +.>
The content of the mixed bacteria is low. Therefore, the formula 7-12 (the crab shell powder content is 60-80%) can well promote the propagation of beauveria bassiana and metarhizium anisopliae and is unfavorable for the propagation of mixed bacteria, wherein the formula 9-11 has the best effect.
TABLE 5 determination of propagation conditions of Metarrhizium anisopliae, beauveria bassiana and other miscellaneous fungi in culture medium special for insecticidal fungi
Experimental example 5 production of insecticidal fungi
Mixing appropriate amount of Metarrhizium anisopliae with formula 9 culture medium and Beauveria bassiana with formula 11 culture medium to obtain a total content of 1×10 insecticidal fungus spores per gram of culture medium 6 Adding 50% water into the spores according to the weight ratio, mixing uniformly, putting into a square tray with the thickness of 10-12cm and the square tray at the temperature of 26-30 ℃ for culturing, wherein the square tray is 40cm multiplied by 60cm multiplied by 20 cm.
The traditional solid state fermentation method is as follows: weighing 500g of rice, soaking in water at 20-30deg.C for 1.5h, draining, packaging into fermentation bag (mushroom bag with air permeable membrane at two ends), sterilizing for 30min, cooling to room temperature, inoculating 10% of liquid bacteria of destruxin or PDA of destruxin for 3d at 26-30deg.C, stirring, fermenting in fermentation chamber, and opening bag for 10 days for drying. Controlling the temperature of the fermentation chamber at 26-30deg.C, controlling the humidity at 0-10d at 85-95%, and suction drying for 11-15 d. And (3) measuring the numbers of the insecticidal fungi and the mixed fungi after fermentation by adopting a CFU method at 5d, 10d and 15d respectively, and calculating the spore yield and the mixed fungi rate.
Results: as shown in Table 6, the culture medium of formula 9 without high temperature sterilization is adopted for fermentation to produce the metarhizium anisopliae, the culture medium of formula 11 is adopted for fermentation to produce the beauveria bassiana, the spore yield of the metarhizium anisopliae reaches more than 40 hundred million spores/g in 10 days and 15 days, and the spore yield of the metarhizium anisopliae reaches more than 70 hundred million spores/g in 10 days and 15 days, which is higher than the spore yield of the metarhizium anisopliae (34 hundred million spores/g and 58 hundred million spores/g) in the traditional production method in 15 days. The production amount of the spores produced by the fermentation of the culture mediums of the formulas 9 and 11 is not obviously different from that produced by the fermentation of the culture mediums of the formula 15 in 10 days, and the production amount of the spores is maximum in 15 days of the traditional fermentation, which indicates that the fermentation production of the special culture medium is 5 days earlier than the traditional fermentation. The unsterilized culture medium of formulas 9 and 11 is used for fermenting and producing the metarhizium anisopliae and the beauveria bassiana, the mixed bacteria rate is kept at a low level all the time, and particularly, the mixed bacteria rate of the traditional fermentation is rapidly increased in the drying process. The special culture medium is unfavorable for the growth of mixed bacteria and is inhibited by insecticidal fungi. The invention is characterized by simple production process, no need of sterilization, easy obtaining of raw materials, low spore yield and less mixed bacteria, low comprehensive production cost and important application value in the production of the insecticidal fungal spores.
Table 6 application of culture medium special for insecticidal fungi in fermentation production
Experimental example 6 Effect of culture Medium dedicated to insecticidal fungi on propagation of insecticidal fungi in soil treatment
Mixing the culture medium 9 with appropriate amount of Metarrhizium anisopliae oil suspension (obtained from Chongqing Polymerise biological engineering Co., ltd.) uniformly, mixing the culture medium 11 with appropriate amount of Bearrhizium anisopliae oil suspension (obtained from Chongqing Polymerise biological engineering Co., ltd.) uniformly, and making into 5×10 per gram of culture medium 8 Fungal preparation of spores, which is diluted 500 times to obtain 1×10 6 spore/mL spore nutrient liquid, irrigated soil, 10L per square meter, 1×10 without adding culture medium 6 spore/mL spore suspension is irrigated and used as a control, a proper amount of surface layer (5-10 cm) soil is taken after 10d on the same day, the bacterial colony numbers of the metarhizium anisopliae and the beauveria bassiana are measured by adopting a CFU method after the soil is diluted by sterile water in a gradient way, the number of the insecticidal fungi in each gram of dry weight soil is calculated according to the corresponding dilution multiple, and the multiplication multiple of the bacterial amount of the same day is calculated relative to inoculation.
The results are shown in Table 7, the number of Metarrhizium anisopliae was 34.7X10 after 10 days of the dilution of the soil with the culture medium special for the insecticidal fungi and the spores of the insecticidal fungi 4 Spores/gram proliferate to 4230×10 4 Spores/gram, proliferation of 120.9 times, and number of control metarhizium anisopliae is 32.6X10 4 Spores/gram was reduced to 12.4X10 4 Spores/gram, 0.6 fold reduction; the number of beauveria bassiana is 32.1×10 4 Spores/gram proliferate to 4260×10 4 Spore/g, proliferation 131.7 times, and control beauveria count from 34.5X10 4 Spores/gram reduction to 21 x 10 4 Spores/gram, 0.4-fold reduction. Therefore, the special culture medium can well promote the propagation of the metarhizium anisopliae and the beauveria bassiana by combining the insecticidal fungi and the irrigation soil treatment.
TABLE 7 conditions of the pesticidal fungus-dedicated Medium for promoting propagation of beauveria bassiana and Metarrhizium anisopliae in soil treatment
Experimental example 7 Effect of culture Medium dedicated to insecticidal fungi on propagation of insecticidal fungi in seed treatment
Selecting a formula 9 culture medium, respectively and uniformly mixing the culture medium with a proper amount of a metarhizium anisopliae oil suspending agent and a beauveria bassiana oil suspending agent to prepare a fungus preparation containing 1 hundred million spores per gram of the culture medium, mixing the fungus preparation and cucumber seeds according to a seed proportion of 100g/100kg, air-drying, sowing the seeds in sterilized soil sterilized at a high temperature (200 ℃ for 2 hours) in advance, placing the seeds in an illumination incubator, controlling the temperature to be 25+/-2 ℃, and periodically irrigating sterile water. Seed dressing treatment is carried out according to the proportion of 100mL/100kg seed of the spore suspension and the spore suspension of 1 hundred megaspore/mL of the metarhizium anisopliae spore suspension and 1 hundred megaspore/mL of the beauveria bassiana as a control. And (3) taking seeds of the seed dressing before sowing, respectively measuring the quantity of the green muscardine fungus and the beauveria bassiana which are treated differently by using a CFU method, and calculating the quantity of each seed of the green muscardine fungus and the beauveria bassiana. After 10d of sowing, respectively taking a proper amount of root and rhizosphere soil of cucumber seedlings, respectively measuring the quantity of the green muscardine fungus and the beauveria bassiana which are treated differently by adopting a CFU method, calculating the quantity of the green muscardine fungus and the beauveria bassiana on the root of each plant according to the diluted corresponding multiple, and calculating the multiplication multiple of the green muscardine fungus and the beauveria bassiana on each plant root and rhizosphere soil relative to the green muscardine fungus and the beauveria bassiana on each seed.
As a result, as shown in Table 8, after 10 days of sowing, the amounts of the green muscardine fungus and the beauveria bassiana used in each plant of cucumber root and rhizosphere soil by the special culture medium combined with the insecticidal fungi are significantly increased, which are higher than those of the control, the proliferation of the green muscardine fungus is 48.3 times and the proliferation of the beauveria bassiana is 70.4 times, and the amount of the insecticidal fungi treated by only using the insecticidal fungi for seed dressing is significantly reduced. Therefore, the special culture medium is added into the seed dressing of the insecticidal fungi, so that the propagation of the insecticidal fungi can be effectively promoted, the quantity of the insecticidal fungi in the microenvironment is increased, and the control effect is ensured; meanwhile, the use of the special culture medium can also reduce the dosage of the insecticidal fungal spores, and the fungal spores with lower concentration can also be propagated in a large quantity, so that the use cost is reduced.
TABLE 8 conditions of the pesticidal fungal Medium promoting propagation of beauveria bassiana and Metarrhizium anisopliae in seed treatment
Experimental example 8 Effect of the Special Medium on the propagation of the insecticidal fungi in the Combined use of the insecticidal fungi and the Fertilizer
Special purposeMixing the culture medium 9 with appropriate amount of destruxin spore powder, and mixing the special culture medium 10 with appropriate amount of destruxin spore powder to obtain a mixture containing 1×10 per gram of culture medium 6 Fungal preparation of individual spores. Wherein, the fungus preparation containing the metarhizium anisopliae and the organic granular fertilizer are mixed according to the weight ratio of 1:99 mixing, stirring to obtain the product containing 1×10 Metarrhizium anisopliae 4 Individual spores/gram of organic fertilizer particles; the fungus preparation containing beauveria bassiana and the compound fertilizer particles are mixed according to the weight ratio of 1:99 mixing, stirring to obtain 1×10 spore containing beauveria bassiana 4 Compound fertilizer granules per gram. Then, respectively mixing the organic fertilizer particles or the compound fertilizer particles mixed with the fungus preparation with sandy soil with the water content of 25 percent according to the weight ratio of 1:10 to be mixed with the same spore content (the content of the metarhizium anisopliae is 1 multiplied by 10) 4 Spores per gram, beauveria bassiana content of 1×10 4 Individual spores/gram) of the organic fertilizer particles or the compound fertilizer particles and sandy soil with the water content of 25 percent according to the weight ratio of 1:10 as test controls, each treatment was repeated 3 times. After the treatment is placed at 25 ℃ for 10 days, sampling is carried out, a CFU method is adopted after the sampling is diluted by sterile water in a gradient way, the quantity of the metarhizium anisopliae or the beauveria bassiana in sandy soil is respectively measured, and the quantity of the insecticidal fungi in each gram of fertilizer and the multiple of the fungi proliferation relative to the inoculation are calculated according to the quantity.
As can be seen from Table 9, after the green muscardine fungus alone or the white muscardine fungus in the control was mixed with the fertilizer and left in the sandy soil for 10 days, the number of spores of the green muscardine fungus per gram of the fertilizer was proliferated to 840×10 4 Proliferation is 0.84×10 3 The beauveria spores cannot be detected in the compound fertilizer; after the fungus preparation is mixed with the organic fertilizer or the compound fertilizer and placed in sandy soil for 10 days, the spore numbers of the metarhizium anisopliae and the beauveria bassiana in each gram of the fertilizer respectively reach 13200 multiplied by 10 4 Sum 220×10 4 Respectively proliferate by 13.20×10 3 Double sum 0.22×10 3 The times are all much higher than the control. In conclusion, in the use of the metarhizium anisopliae or beauveria bassiana combined organic fertilizer or compound fertilizer, the metarhizium anisopliae or beauveria bassiana is added with a special culture medium to prepare a fungus preparation and then is combined with the organic fertilizer or compound fertilizer for use, and the growth and propagation of the insecticidal fungi metarhizium anisopliae and beauveria bassiana can be promoted after the application, so that the development of the insecticidal fungi and fertilizer combined product is promoted.
TABLE 9 conditions of the propagation of insecticidal fungi in the combined use of the insecticidal fungi and the fertiliser with the special Medium
Experimental example 9 Effect of fungal formulations in field applications
The insecticidal fungi are added into a special culture medium to prepare a fungi preparation for field experiments, and the preparation method comprises the following steps: mixing formula 9 with appropriate amount of destruxin spore powder, and making into culture medium containing 10 hundred million spores, 5 hundred million spores and 1×10 spores per gram 6 The metarhizium anisopliae fungus preparation of the individual spores is ready for use; with 1X 10 of the preparation 6 The weight ratio of the metarhizium anisopliae fungus preparation to the organic fertilizer is 1:99 are stirred evenly for standby. Mixing formula 10 with appropriate amount of spore powder of Beauveria bassiana, and making into culture medium containing 10 hundred million spores, 5 hundred million spores, and 1×10 spores 6 The beauveria bassiana fungus preparation of the individual spores is reserved; with 1X 10 of the preparation 6 The weight ratio of the beauveria bassiana fungus preparation to the compound fertilizer of the individual spores/gram is 1:99 are stirred evenly for standby.
Wheat and tomato field planting: and selecting land plots with serious soil hardening and serious occurrence of soil-borne diseases in the past year and underground insect pests for testing. (1) seed dressing. Seed dressing of 10 hundred million spores/g of the metarhizium anisopliae fungus preparation is selected, the dosage is 100g of the metarhizium anisopliae fungus preparation/100 kg of seeds, 3.3g of metarhizium anisopliae spore powder/100 kg of seeds are used for seed dressing, 96.7g of special culture medium for formula 9/100 kg of seeds are used for seed dressing, and the treatment of fresh water seed dressing is used as blank control. Investigation of the situation that wheat seedlings are gnawed by cutworms in the period from the time of turning green to the time of jointing the wheat, investigation of the situation of basal rot of wheat stems in the period of grouting, and calculation of prevention effect respectively; and (5) investigating the situation that the tomato seedlings are gnawed by grubs and the situation that the tomato root rot occurs in the tomato seedling stage, and respectively calculating the control effect. (2) soil irrigation. Respectively selecting 5 hundred million spores/g of metarhizium fungus preparation and diluting the special culture medium of formula 9 by 500 times1×10 of the liquid of (2) 6 The spores/g of the metarhizium anisopliae suspension irrigated soil, respectively irrigated after wheat sowing and tomato planting, and the treatment of water irrigation is used as a blank control. Investigation of the biting condition of wheat seedlings by cutworms and investigation of the basal rot of wheat stems in the grouting period from the beginning of the wheat turning green to the jointing period, and calculation of the prevention effect respectively; and (5) investigating the situation that the tomato seedlings are gnawed by grubs and the situation that the tomato root rot occurs in the tomato seedling stage, and respectively calculating the control effect. (3) mixing with organic fertilizer. Uniformly spreading the prepared fertilizer containing the metarhizium anisopliae fungus preparation into the land, wherein the dosage per mu is 400-500kg, uniformly ploughing, planting wheat and tomatoes, and setting an organic fertilizer (1X 10 metarhizium anisopliae spores in each gram of fertilizer) mixed with the metarhizium anisopliae spore powder 4 And (b) mixing the organic fertilizer with the culture medium of the formula 9 (the weight of the culture medium of the formula 9 and the fertilizer is 1:99 mix well) with the treatment of applying only organic fertilizer as a blank. Investigation of the biting condition of wheat seedlings by cutworms and investigation of the basal rot of wheat stems in the grouting period from the beginning of the wheat turning green to the jointing period, and calculation of the prevention effect respectively; and (5) investigating the situation that the tomato seedlings are gnawed by grubs and the situation that the tomato root rot occurs in the tomato seedling stage, and respectively calculating the control effect.
Planting potatoes in the field: and selecting land plots with serious soil hardening and serious occurrence of soil-borne diseases in the past year and underground insect pests for testing. (1) seed dressing. Seed dressing of the beauveria fungus preparation containing 10 hundred million spores/g is selected, the dosage is 100g of the beauveria fungus preparation/100 kg of seeds, 1.25g of beauveria spore powder/100 kg of seeds are used for seed dressing, 98.75g of the culture medium 10 per 100kg of seeds are used for seed dressing respectively, and the treatment of fresh water seed dressing is used as a blank control. And respectively counting the occurrence of insect pests and scab diseases of potato blocks in the potato harvesting period, and calculating the corresponding insect fruit rate and disease control effect. (2) soil irrigation. Selecting soil of potato irrigated with 5 hundred million spores/g beauveria fungus preparation diluted 500 times, respectively diluting 500 times with special culture medium of formula 10, and 1×10 6 The spores/g beauveria bassiana suspension irrigates soil with a clear water irrigated treatment as a blank. Respectively counting the occurrence of insect pests and scab diseases of potato blocks during the harvesting period of the potato, and calculating corresponding insectsFruit rate and disease control effect. (3) mixing with compound fertilizer. Uniformly spreading the prepared fertilizer containing the beauveria bassiana fungus preparation into the land, uniformly ploughing and sowing potatoes with the amount of 30-40kg per mu, and setting a compound fertilizer mixed with beauveria bassiana spore powder (1X 10 beauveria bassiana spores in each gram of fertilizer) 4 And b) treating and mixing the compound fertilizer with a formula 10 culture medium only (the formula 10 culture medium and the fertilizer are 1 by weight: 99 mix well) with the treatment of the applied compound fertilizer alone as a blank. And in the potato harvesting period, counting the occurrence of insect pests and scab diseases of potato blocks respectively, and calculating the corresponding insect fruit rate and disease control effect.
From table 10, it can be seen that the fungus preparation added with the culture medium of formula 9 has better protection effect on wheat in the wheat field application, the prevention effect on the stem rot of wheat is 86%, 91% and 87%, the prevention effect on the tiger is 84%, 88% and 90%, and the prevention effect on the stem rot and the tiger is obviously higher than that of the treatment of the independent green muscardine fungus and the special culture medium; the effect on tomatoes is the same; meanwhile, the observation shows that the improvement effect of the soil hardening condition after the fungus preparation containing the culture medium of the formula 9 and the metarhizium anisopliae is most obvious, and the soil is looser.
Table 10 effects of the Special Medium on Metarrhizium anisopliae field applications
From Table 11, it can be seen that the beauveria bassiana fungus preparation added with the culture medium of formula 10 has better protection effect on potatoes in potato field application, and has the prevention effects on potato scab of 84%, 80% and 77% respectively, and the grubs have the rates of 7%, 8% and 5% respectively, and the prevention effects on scab and grubs are obviously higher than those of the beauveria bassiana alone and the special culture medium. Meanwhile, the observation shows that the fungus preparation containing the culture medium of the formula 10 and beauveria bassiana has the most obvious effect of improving the soil hardening condition and loose soil. In the field application of the insecticidal fungi, the fungus preparation added with the special culture medium for the insecticidal fungi can obviously improve the control effect of the insecticidal fungi on plant diseases and insect pests and improve the soil. Therefore, the fungus preparation has great application prospect in plant disease and pest control.
Table 11 effects of the Special Medium on Beauveria Bicoloris field applications
It should be noted that, the experimental operations related to the above experimental examples have a certain universality, so that the detailed description is not given, and the detailed description is omitted for the content of the related operations in other experimental examples or for the reference to the prior art.
Claims (10)
1. A special culture medium for insecticidal fungi without sterilization, which is characterized in that the special culture medium comprises crab shell powder.
2. The special culture medium according to claim 1, wherein the special culture medium further comprises flour weevil powder or/and shrimp powder.
3. The special culture medium according to claim 1, wherein the crab shell powder accounts for more than 60% of the mass of the special culture medium.
4. A special culture medium according to claim 3, wherein the mass fraction of crab shell meal, yellow meal worm meal and shrimp meal in the special culture medium is 60-80%, 10-40% and 10-40%, respectively.
5. The special culture medium according to claim 4, wherein the mass fraction of crab shell meal, yellow meal worm meal and shrimp meal in the special culture medium is 60%, 10% and 30%, respectively, or 60%, 30% and 10%, respectively, or 60%, 20% and 20%, respectively.
6. A process for producing a pesticidal fungus without sterilization, which comprises inoculating a pesticidal fungus to the specific medium of any one of claims 1 to 5, fermenting, wherein the specific medium is not sterilized before the inoculation, and the inoculum size of the pesticidal fungus is 1X 10 or more 6 Individual spores/gram of dedicated medium.
7. A pesticidal fungal preparation comprising a pesticidal fungus and the medium according to any one of claims 1 to 5, wherein the pesticidal fungus is contained in an amount of 1X 10 or more 6 Individual spores/gram of dedicated medium.
8. A process for producing a pesticidal fungal preparation, comprising adding a pesticidal fungus to the specific medium of any one of claims 1 to 5, wherein the pesticidal fungus is added in an amount of 1X 10 or more without subjecting the specific medium to sterilization treatment before the addition 6 Individual spores/gram of dedicated medium.
9. Use of the pesticidal fungal formulation of claim 7 for soil improvement and control of crop diseases and insect pests.
10. The use according to claim 9, wherein the fungal formulation is used in combination with a fertiliser.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103392942A (en) * | 2013-08-09 | 2013-11-20 | 陈冬强 | Preparation method for ornamental carp feed |
| KR20140117221A (en) * | 2013-03-26 | 2014-10-07 | 주식회사 푸르네 | Plant growth substance and anti-root knot nematode compound and its composition using Lysobacter capsici YS1215 from crab shell amended soil and manufacturing method thereof |
| WO2020226482A1 (en) * | 2019-05-09 | 2020-11-12 | Hong Heng Lai | Synergistic compositions for controlling plant pathogens with chitinolytic microorganisms / organic acid combinations and method of use thereof |
-
2023
- 2023-04-04 CN CN202310348840.6A patent/CN116333895B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20140117221A (en) * | 2013-03-26 | 2014-10-07 | 주식회사 푸르네 | Plant growth substance and anti-root knot nematode compound and its composition using Lysobacter capsici YS1215 from crab shell amended soil and manufacturing method thereof |
| CN103392942A (en) * | 2013-08-09 | 2013-11-20 | 陈冬强 | Preparation method for ornamental carp feed |
| WO2020226482A1 (en) * | 2019-05-09 | 2020-11-12 | Hong Heng Lai | Synergistic compositions for controlling plant pathogens with chitinolytic microorganisms / organic acid combinations and method of use thereof |
Non-Patent Citations (4)
| Title |
|---|
| A OETARI等: "Crustaceous wastes as growth substrates for insect-pathogenic fungus Metarhizium majus UICC 295", 《PROCEEDING OF THE 4TH INTERNATIONAL SYMPOSIUM ON GREEN TECHNOLOGY FOR VALUE CHAINS 2019》, pages 1 - 7 * |
| 徐阿妹等: "应用二次通用旋转组合设计优化黄绿绿僵菌固相培养基配方", 《安徽农业大学学报》, vol. 40, no. 4, pages 649 - 655 * |
| 王福祥等: "黄粉虫虫粉对白僵菌生长的影响", 《安徽农业科学》, pages 201 - 202 * |
| 陶淑霞等: "黄粉虫虫粉对球孢白僵菌孢子粉产量、质量及其毒力的影响", 《江苏农业科学》, pages 122 - 125 * |
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