CN110122509B - Application of shiitake mushrooms in insect pest expelling, insect repellent and preparation method of insect repellent - Google Patents
Application of shiitake mushrooms in insect pest expelling, insect repellent and preparation method of insect repellent Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
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- 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
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
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Abstract
The invention relates to the technical field of edible fungi, and particularly discloses application of shiitake mushrooms in insect pest expelling, an insect repellent and a preparation method of the insect repellent. The preparation method comprises the following steps: the mushroom strains are fermented and cultured to obtain fermentation liquor, bubbles are generated in the fermentation liquor by adopting a physical or chemical method, and the volatile substances are taken out. The preparation method of the insect repellant provided by the invention is simple in process, convenient to operate, green, environment-friendly, safe and reliable, the obtained insect repellant is free of toxic and side effects, free of irritation, safe and efficient, and capable of effectively preventing and treating mushroom flies and mushroom mosquitoes, protecting edible fungi such as oyster mushrooms from insect pests and guaranteeing food safety.
Description
Technical Field
The invention relates to the technical field of edible fungi, and particularly relates to application of mushrooms in insect pest expelling, an insect repellent and a preparation method of the insect repellent.
Background
In agricultural production, particularly edible fungus production, a chemical insecticide is generally adopted for surface spraying to achieve an insect prevention effect, so that pesticide residues are serious, particularly, the adsorptive capacity of the fruiting bodies of the edible fungi is strong, and the serious food safety problem can be caused by the surface spraying of the insecticide. For example, edible fungi such as oyster mushroom and agaricus bisporus have serious insect pests, are generally used for chemical insecticidal pesticides, and cause great harm to food safety.
At present, the odor insect repellent is widely applied to production and life. The odor insect repellant is a preparation for repelling space insects by utilizing volatile substances, does not contact human bodies or objects in use, and reduces the use danger. The existing odor insect repellents are mainly chemical insect repellents, such as camphor balls, mosquito-repellent incense and the like; plant-derived odor insect repellent such as eucalyptus oil, citrus oil, etc. However, the chemical odor insect repellent is often high in toxicity and has a lot of contraindications in use; the plant-derived odor insect repellent is mainly volatile oil substances and has strong irritation.
Therefore, the search for new safe and effective odor repellents is an important topic in the field of biopesticide research today.
Disclosure of Invention
Aiming at the problems of high toxicity, high irritation, influence on food safety and the like of the existing insect repellent, the invention provides an application of shiitake in insect pest repelling, the insect repellent and a preparation method thereof.
In order to achieve the purpose of the invention, the embodiment of the invention adopts the following technical scheme:
an application of Lentinus Edodes in repelling insect pest is provided, wherein the Lentinus Edodes is spore, mycelium or fruiting body.
Further, the application of the mushroom in pest control is that volatile substances generated by the mushroom are used for pest control, and the pest control is agricultural pest control, such as field control of oyster mushroom flies and mushroom mosquitoes.
Further, the shiitake mushroom includes at least one of shiitake mushroom 808, shiitake mushroom 0912, shiitake mushroom L18, shiitake mushroom 868, or shiitake celebration 20.
The invention also provides an insect repellent comprising volatile substances produced by shiitake mushrooms.
The invention also provides a preparation method of the insect repellent derived from the shiitake mushrooms, which comprises the following steps:
the mushroom strains are fermented and cultured to obtain fermentation liquor, bubbles are generated in the fermentation liquor by adopting a physical or chemical method, and the volatile substances are taken out.
Further, the volatile substance is dried and condensed to obtain the liquid insect repellent.
Further, the method for the fermentation culture of the mushroom strains comprises the following steps: activating test tube species of Lentinus Edodes, transferring into PDA culture medium, standing for 2-4 days, shake culturing for 6-8 days, transferring into fermentation culture medium, and shake culturing for 13-17 days to obtain fermentation broth.
Further, the temperature of the static culture and the shaking culture is 23-27 ℃.
Further, the fermentation medium is a PDA (potato dextrose agar) medium.
Further, the fermentation medium comprises the following components in parts by weight: 180 and 220 parts of potato cooking juice; 17-23 parts of glucose; 1.5-2.5 parts of monopotassium phosphate; 0.8-1.2 parts of magnesium sulfate; 0.0018-0.0022 parts of vitamin B1; 800 portions of water, and the pH value of the culture medium system is 7.0.
Further, the fermentation medium comprises the following components in parts by weight: 4.5-5.5 parts of pear wood chips; 18-22 parts of glucose; 1.5-2.5 parts of monopotassium phosphate; 0.8-1.2 parts of magnesium sulfate; 0.0018-0.0022 parts of vitamin B1; 800 portions of water, and the pH value of the culture medium system is 7.0.
Further, the fermentation medium comprises the following components in parts by weight: 4.5-5.5 parts of pear wood chips; 0.01-0.13 parts of guaiacol; 17-23 parts of glucose; 1.5-2.5 parts of monopotassium phosphate; 0.8-1.2 parts of magnesium sulfate; 0.0018-0.0022 parts of vitamin B1; 800 portions of water, and the pH value of the culture medium system is 7.0.
Further, the physical method is that clean air or inert gas source is introduced into the fermentation liquor through the sand core filter plate to generate bubbles.
Further, the chemical method comprises the following steps: and adding sodium chloride and sodium bicarbonate into the fermentation liquor, wherein the addition amount of the sodium chloride is 6-8% of the mass of the fermentation liquor, and the addition amount of the sodium bicarbonate is 4-6% of the mass of the fermentation liquor.
Compared with the prior art, the technical scheme of the invention is adopted to apply the shiitake mushrooms to repelling insect pests and pests such as mosquitoes, flies and the like, and is safe and efficient. The method is characterized in that fermentation liquor is obtained by culturing and fermenting test tube species of the mushrooms, volatile substances in the fermentation liquor are collected to obtain the insect repellent, the process is simple, the operation is convenient, the method is green and environment-friendly, safe and reliable, the obtained insect repellent is free of toxic and side effects, free of irritation, safe and efficient, mushroom flies and mushroom mosquitoes can be effectively prevented and treated, edible fungi such as oyster mushrooms are protected from insect damage, the use of the oyster mushroom insecticide is reduced, and the food safety is guaranteed.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1
The application of the mushrooms in insect pest expelling is used for expelling flies and mosquitoes of oyster mushrooms, and the specific experimental process is as follows:
mushroom fly and mushroom mosquito culture medium: 20g of starch, 20g of corn flour and 5g of bran, adding 55ml of water, uniformly stirring, filling into a wide-mouth glass bottle, and sterilizing for later use.
Obtaining the mushroom flies: 20 mushroom flies are captured in a flat mushroom shed, put into a wide-mouth glass bottle containing a mushroom fly and mushroom mosquito culture medium, cultured for 12 days at 28 ℃, narcotized by ether after mass propagation of the mushroom flies, and transferred into a new culture medium for culture. Obtaining mushroom mosquitoes: as above.
Mushroom stick: lentinus edodes 808, Lentinus edodes 0912, Lentinus edodes L18, Lentinus edodes 868, and Lentinus edodes of 20 varieties of Gengzaceae, with the size of the mushroom bag of 15cm × 58 cm.
And (3) oyster mushroom sticks: clean and no plant diseases and insect pests, and has a mushroom 2026 stick with fruiting body, and the size of the mushroom bag is 17cm × 33 cm.
Insect repelling test: preparing a 30L plastic barrel with a small hole with the diameter of 1cm on the barrel cover, putting the plastic barrel into a clean and closed room, putting a fungus stick into the plastic barrel, and then releasing mushroom flies or mushroom mosquitoes in the room. Putting 1 mushroom stick of shiitake mushroom 808, shiitake mushroom 0912, shiitake mushroom L18, shiitake mushroom 868 and shiitake celeriaceae 20 and 1 oyster mushroom stick into different plastic barrels, setting a contrast for only putting 1 oyster mushroom stick, culturing at 28 ℃ overnight, and counting the number of mushroom flies or mushroom mosquitoes in the barrels. The experiment was repeated 3 times. In the case of the mushroom and the oyster mushroom sticks existing at the same time, the insect repelling effect of the mushroom sticks is shown in table 1.
TABLE 1 comparison of the indoor insect-repelling effect of the mushroom sticks
As can be seen from Table 1, the plastic barrels of the mushroom sticks contain 3.7 to 6.7 mushroom flies and 4.0 to 5.7 mushroom mosquitoes, while the plastic barrels of the mushroom sticks contain 28.3 mushroom flies and 20.0 mushroom mosquitoes, and further cultivation shows that the mushroom flies and the mushroom mosquitoes propagate on the oyster mushrooms. Tests show that the mushroom has a certain insect expelling effect, particularly has a good effect of expelling mushroom flies and mushroom mosquitoes, and has a high application value in the aspect of protecting oyster mushrooms from insect pests.
Example 2
A method for preparing an insect repellent comprising the steps of:
(1) preparation of the culture Medium
PDA liquid medium (L): 200g of potato cooking juice, 20g of glucose and 1000ml of water, wherein the pH value is 7.0;
fermentation medium (L):
firstly, PDA culture medium; 200g of potato cooking juice, 20g of glucose and 1000ml of water, wherein the pH value is 7.0;
② 200g of potato cooking juice, 20g of glucose, 2.0g of monopotassium phosphate, 1.0g of magnesium sulfate, 2mg of vitamin B1, 1000ml of water, the pH value of the culture medium system is 7.0,
③ 5g of 100 meshes peeled pear wood chip, 20g of glucose, 2.0g of monopotassium phosphate, 1.0g of magnesium sulfate, 2mg of vitamin B1, 1000ml of water and 7.0 of pH value of a culture medium system,
5g of peeled 100-mesh pear wood chips, 0.1g of guaiacol, 20g of glucose, 2.0g of monopotassium phosphate, 1.0g of magnesium sulfate, 12 mg of vitamin B, 1000ml of water and the pH value of a culture medium system being 7.0;
(2) activating the 808 test-tube strains of the shiitake mushrooms at 25 ℃ for 7d, transferring the shiitake mushrooms into a 500mL triangular flask containing 100mL of PDA culture medium, standing and culturing the shiitake mushrooms at 25 ℃ for 3 days, culturing the shiitake mushrooms at 25 ℃ for 7 days by a shaker at 100rpm, respectively transferring the shiitake mushrooms into 500mL triangular flasks containing 150mL of the (first), (second), (third) and (fourth) fermentation culture medium according to 20 percent of inoculation quantity after hypha grows thick, and culturing the shiitake mushrooms for 15 days by a shaker at 100rpm under the condition of 25 ℃ to obtain fermentation liquor of a series of different fermentation culture media;
(3) taking a plurality of 50ml plastic bottles with small holes with the diameter of 2mm at the upper part and the middle part, filling 20g of sodium bicarbonate to obtain a plurality of gas production bottles, respectively transferring fermentation liquor of different fermentation culture mediums to wide-mouth bottles, adding sodium chloride accounting for 7% of the mass of the fermentation liquor, sinking one gas production bottle into each wide-mouth bottle to produce gas, and taking out volatile substance insect repellents.
To better illustrate the effect of the insect repellent provided in the examples, experiments were conducted to repel mushroom flies and mushroom mosquitoes. Preparing a plastic barrel with a small hole with the diameter of 1cm on the barrel cover, putting the plastic barrel into a clean and closed room, and then releasing mushroom flies or mushroom mosquitoes in the room. The processing method comprises the following steps: the insect repellents of different fermentation media and the oyster mushroom sticks are respectively put into different plastic barrels, and the oyster mushroom sticks and 0.01 percent of guaiacol and gas production bottle control CK1 and the oyster mushroom sticks and clear water and gas production bottle control CK2 are additionally added, each treatment is repeated for 3 times, the temperature is 28 ℃ overnight, the number of mushroom flies or mushroom mosquitoes in the barrels is counted, and the results are shown in Table 2.
TABLE 2 insect repellant test with insect repellant
As can be seen from the results in Table 2, the insect repellent provided in the examples has a good insect repellent effect, and the insect repellent effects are respectively culture media (c), (c) and (c) from large to small. CK1 added with 0.01% guaiacol, the numbers of mushroom fly and mushroom mosquito are respectively 19.0 per square meter and 14.0 per square meter, which is slightly lower than that of CK2 of gas producing bottle with clear water. The mushroom 808 fermentation liquor can generate insect-repellent substances, and the wood chips and the guaiacol are added to promote the generation of the insect-repellent substances.
Example 3
A method for preparing an insect repellent comprising the steps of:
(1) preparation of the culture Medium
PDA liquid medium (L): 200g of potato cooking juice, 20g of glucose and 1000ml of water, wherein the pH value is 7.0;
fermentation medium (L): 5.5g of peeled 100-mesh pear wood chips, 0.13g of guaiacol, 22g of glucose, 2.5g of monopotassium phosphate, 1.2g of magnesium sulfate, 2.2mg of vitamin B1, 1000ml of water and 7.0 of pH value of a culture medium system;
(2) activating mushroom L18 test tube seeds for 7d at 25 ℃, transferring the mushroom L18 test tube seeds into a 500mL triangular flask containing 100mL PDA culture medium, standing and culturing for 2 days at 27 ℃, culturing for 6 days at 27 ℃ by a shaking table at 100rpm, transferring the mushroom L18 test tube seeds into a fermentation culture medium according to 20% inoculation amount after hypha grows thick, and performing shaking table culture at 100rpm for 13 days at 27 ℃ to obtain fermentation liquor;
(3) taking 50ml of plastic bottles with small holes with the diameters of 2mm at the upper part and the middle part, filling 30g of sodium bicarbonate to obtain a gas production bottle, transferring fermentation liquor to a wide-mouth bottle, adding sodium chloride accounting for 7% of the mass of the fermentation liquor, sinking the gas production bottle into each wide-mouth bottle to generate gas, and taking out volatile substance insect repellents.
To better illustrate the efficacy of the insect repellents provided by the examples, a field insect repellent test was conducted. The fruiting shed for oyster mushroom 2026 is divided into 6 areas, each area is 60 square meters, and the areas are separated by plastic films. Wherein, 12 jars (treatments) capable of generating the shiitake mushroom L18 insect repellant are uniformly placed in 3 areas, the other 3 areas are set as blank Controls (CK), the number of the shiitake mushroom flies and the shiitake mushroom mosquitoes are counted in 1 day, 8 days and 15 days respectively by 5 random points in each area and 4 square meters in each point, and the number of the shiitake mushroom flies and the shiitake mushroom mosquitoes are calculated, and the results are shown in the table 3.
TABLE 3 insect repellant field insect repellant test
The data in table 3 show that when the area capable of producing the mushroom L18 insect repellant jar is placed, the number of mushroom flies and mushroom mosquitoes is greatly reduced after 1 day, after 15 days, the average number of the mushroom flies and the average number of the mushroom mosquitoes are respectively 1.2/square meter and 0.5/square meter, and the average number of the control group mushroom flies and the average number of the control group mushroom mosquitoes are respectively 19.0/square meter and 9.3/square meter, which indicates that the mushroom L18 insect repellant provided by the embodiment of the invention has obvious insect repelling effect and can effectively protect edible mushrooms from insect damage.
Example 4
A method for preparing an insect repellent comprising the steps of:
(1) preparation of the culture Medium
PDA liquid medium (L): 200g of potato cooking juice, 20g of glucose and 1000ml of water, wherein the pH value is 7.0;
fermentation medium (L): 4.5g of peeled pear sawdust of 100 meshes, 0.07g of guaiacol, 18g of glucose, 1.5g of monopotassium phosphate, 0.8g of magnesium sulfate, 1.8mg of vitamin B1, 1000ml of water and 7.0 of pH value of a culture medium system;
(2) activating 20 test tube strains of Lentinus edodes of Generaceae at 25 deg.C for 7d, transferring into 500mL triangular flask containing 100mL PDA culture medium, standing at 23 deg.C for 4 days, shaking at 23 deg.C for 8 days at 100rpm, transferring mycelia into fermentation culture medium according to 20% inoculum size after mycelia grow to be viscous, and shaking at 100rpm at 23 deg.C for 17 days to obtain fermentation broth;
(3) putting 1000mL of fermentation liquor into a 5L suction bottle, introducing an inert gas source into the fermentation liquor through a sand core filter plate to generate bubbles, connecting the suction bottle to a vacuum pump to continuously pump air for 2h, pumping out volatile substances under the action of the bubbles and the vacuum, adsorbing and dehumidifying through a silica gel column and anhydrous copper sulfate, passing through a liquid nitrogen tank provided with a U-shaped stainless steel tube, condensing the volatile substances in the U-shaped tube due to the low-temperature action of the liquid nitrogen when the volatile substances meet the condensation in the U-shaped tube, taking out the condensate (liquid insect repellent) in the U-shaped tube, putting the condensate into a 5mL small glass bottle, repeating for 3 times, wherein the content of the condensate in the fermentation liquor is 23.4mg/L, 24.6mg/L and 25.7mg/L respectively, putting the condensate into a freezer at the temperature of-45 ℃ for refrigeration for standby, and putting the extracted residual liquor into the freezer at the temperature of-.
Example 5
A method for preparing an insect repellent comprising the steps of:
(1) preparation of the culture Medium
PDA liquid medium (L): 200g of potato cooking juice, 20g of glucose and 1000ml of water, wherein the pH value is 7.0;
fermentation medium (L): 4.5g of peeled pear sawdust of 100 meshes, 0.07g of guaiacol, 18g of glucose, 1.5g of monopotassium phosphate, 0.8g of magnesium sulfate, 1.8mg of vitamin B1, 1000ml of water and 7.0 of pH value of a culture medium system;
(2) activating 20 test tube strains of Lentinus edodes of Generaceae at 25 deg.C for 7d, transferring into 500mL triangular flask containing 100mL PDA culture medium, standing at 23 deg.C for 4 days, shaking at 23 deg.C for 8 days at 100rpm, transferring mycelia into fermentation culture medium according to 20% inoculum size after mycelia grow to be viscous, and shaking at 100rpm at 23 deg.C for 17 days to obtain fermentation broth;
(3) putting 1000mL fermentation liquor into a 5L suction flask, adding 50g of sodium bicarbonate, stirring at 80rpm by using a magnetic stirrer, continuously pumping air for 2h by connecting a vacuum pump to a bottle mouth, pumping out volatile substances under the stirring and vacuumizing effects, adsorbing and dehumidifying by a silica gel column and anhydrous copper sulfate, passing through a liquid nitrogen tank provided with a U-shaped stainless steel tube, condensing the volatile substances in the U-shaped tube due to the low temperature effect of the liquid nitrogen when the volatile substances meet the condensation in the U-shaped tube, taking out the condensate (liquid insect repellent) in the U-shaped tube, putting the condensate into a 5mL small glass bottle, repeating for 3 times, wherein the content of the condensate in the fermentation liquor is 22.3mg/L, 23.5mg/L and 26.7mg/L respectively, putting the condensate into a freezer at the temperature of-45 ℃ for cold storage, and putting the residual liquid after extraction into the freezer at the temperature of-45 ℃ for cold storage.
To better illustrate the characteristics of the liquid-like insect repellent provided in the examples of the present invention, the insect repellent obtained in example 5 was subjected to an insect repellent test. Preparing a 20L plastic barrel with a small hole with the diameter of 1cm on the top, putting 20 mushroom flies which are lost by diethyl ether, putting a small glass bottle which is recovered to the room temperature of 25 ℃ and contains coagulates into the plastic barrel, opening the cover, setting a control group of extraction residual liquid, fermentation liquid and clear water, counting the number of the mushroom flies after 8 hours, and obtaining the results shown in Table 4.
Table 4 volatile substance condensate insect repellent test
As can be seen from the data in table 4, the residual liquid after the fermentation broth is subjected to the extraction of the volatile substances has no significant difference in the insect repelling effect from the clear water control, and the volatile substance condensate and the fermentation broth have significant insect repelling effect. The volatile insect repellent obtained by the invention has a remarkable insect repelling effect.
According to the data, the obtained insect repellent has no toxic or side effect, no irritation, safety and high efficiency, can effectively prevent and control mushroom flies and mushroom mosquitoes, protects oyster mushrooms from insect pests, reduces the use of oyster mushroom insecticides, and ensures food safety.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents or improvements made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (6)
1. The application of the shiitake mushrooms in insect pest expelling is characterized in that volatile substances generated by the shiitake mushrooms are used for expelling insect pests, the insect pests are agricultural insect pests, and the preparation method of the volatile substances comprises the following steps: the mushroom strains are fermented and cultured to obtain fermentation liquor, bubbles are generated in the fermentation liquor by adopting a physical or chemical method, and the volatile substances are taken out.
2. The use of shiitake mushrooms according to claim 1 for combating insect pests, wherein: the method for the fermentation culture of the mushroom strains comprises the following steps: activating test tube species of Lentinus Edodes, transferring into PDA culture medium, standing for 2-4 days, shake culturing for 6-8 days, transferring into fermentation culture medium, and shake culturing for 13-17 days to obtain fermentation broth.
3. The use of shiitake mushrooms according to claim 2 for repelling insect pests, wherein: the temperature of the static culture and the shaking culture is 23-27 ℃.
4. The application of the shiitake mushroom in pest repelling according to claim 2, wherein the fermentation medium comprises the following components in parts by mass: 180 and 220 parts of potato cooking juice; 17-23 parts of glucose; 1.5-2.5 parts of monopotassium phosphate; 0.8-1.2 parts of magnesium sulfate; 0.0018-0.0022 parts of vitamin B1; 800 portions of water, and the pH value of the culture medium system is 7.0.
5. The application of the shiitake mushroom in pest repelling according to claim 2, wherein the fermentation medium comprises the following components in parts by mass: 4.5-5.5 parts of pear wood chips; 18-22 parts of glucose; 1.5-2.5 parts of monopotassium phosphate; 0.8-1.2 parts of magnesium sulfate; 0.0018-0.0022 parts of vitamin B1; 800 portions of water, and the pH value of the culture medium system is 7.0.
6. The application of the shiitake mushroom in pest repelling according to claim 2, wherein the fermentation medium comprises the following components in parts by mass: 4.5-5.5 parts of pear wood chips; 0.01-0.13 parts of guaiacol; 17-23 parts of glucose; 1.5-2.5 parts of monopotassium phosphate; 0.8-1.2 parts of magnesium sulfate; 0.0018-0.0022 parts of vitamin B1; 800 portions of water, and the pH value of the culture medium system is 7.0.
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