CN111374129A - Nematicide containing 1-octen-3-ol and preparation method and application thereof - Google Patents
Nematicide containing 1-octen-3-ol and preparation method and application thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of preparation of insecticide, and particularly relates to a nematicide containing 1-octen-3-ol and a preparation method and application thereof. The pesticide mainly contains 1-octen-3-ol as an active ingredient, and the balance of the pesticide can also comprise paecilomyces lilacinus and induced nutrients thereof. The medicament for preventing and treating the root-knot nematodes is safe to use, easy to degrade, harmless to human bodies and environment, free of hormone residue, free of safety problems, good in effect of preventing and treating the root-knot nematodes, good in compatibility with other active ingredients for preventing and treating the root-knot nematodes, and capable of being used in a compatible manner so as to improve the nematode preventing and treating effect. The medicament has strong compatibility of each component and obvious synergistic effect, can obviously improve the nutritional status of crops, promote the growth and development of the crops and improve the immunity of the crops while preventing and treating the root-knot nematodes.
Description
Technical Field
The invention belongs to the technical field of preparation of insecticide, and particularly relates to a nematicide containing 1-octen-3-ol and a preparation method and application thereof.
Background
At present, root knot nematode diseases commonly occur in crops such as cucumbers, tomatoes and the like in China, the diseases are more serious particularly in facility cultivation, and the annual loss of various crops reaches billions of dollars. After root-knot nematodes are infected, root knots are formed at the roots of crops, the absorption function of the roots is damaged, and the overground parts are dwarfed and yellowed and even leaves fall off. Among 29 kinds of root-knot nematodes reported in China, the most serious are meloidogyne incognita (M.incognita), meloidogyne hapla (M.hapla), meloidogyne javanica (M.vanica) and meloidogyne arachidis (M.arenaria), and the most serious is the most common and serious in a facility environment especially meloidogyne incognita.
Because germplasm resources for resisting the root-knot nematode disease are scarce, important measures for preventing the root-knot nematode in the related technology are chemical prevention, such as fenamiphos, potassium bromide, promecarb and the like. The medicaments can seriously reduce the quality of vegetables, pollute the environment and destroy the ecological balance, such as 1, 3-dichloropropene and methyl bromide have extremely strong volatility and large odor irritation, and special matched medicament application instruments or devices are required during use; the effect of dazomet and metam is influenced by soil texture, moisture and temperature. The toxicity of the existing agent or methyl isothiocyanate generated by decomposition in water is generally high, the toxicity to human bodies is high, the time for air dissipation is long, and the growth cycle of the next-stubble crops is influenced.
Therefore, more green control measures are increasingly receiving attention. At home and abroad, paecilomyces lilacinus, verticillium chlamydosporia, arthrobotrys oligosporus, pasteurella and other microbial agents are used for preventing and treating meloidogyne incognita, the predatory fungi and the parasitic bacteria can effectively prevent and treat the nematodes after being propagated in soil, but the predatory fungi are nonselective to the nematode predation, and the pasteurella is obligately parasitic and other characteristics limit the efficient development of the nematicidal compound; more importantly, the soil environment factors are complex, and whether the nematode-killing microorganisms can be colonized and become dominant flora is the key to influence the nematode control effect. In the actual agricultural planting process, most of microbial agents for preventing and controlling nematodes have unstable effects and slow response. Therefore, a green and environment-friendly chemical source nematocide is urgently needed to be found for quickly killing the nematodes, and on the basis, the microbial agent is long-acting in prevention and control, so that the problem of the meloidogyne can be effectively solved.
1-octen-3-ol (1-octen-3-ol), Mushroom alcohol (Mushroom alcohol), Vinyl amyl methanol (Vinyl amyl carbinol), 1-Vinyl hexanol (1-Vinyl hexanol), formula C8H16O, structural formula is CH2＝CHCH(OH)CH2CH2CH2CH2CH3Is aliphatic unsaturated alcohol, is colorless to light yellow oily liquid in appearance, and has fragrance of mushroom, lavender, rose and hay. In nature, it is mainly found in mints, thyme and fresh mushrooms, and belongs to natural flavors. Fruits such as bananas and berries, beer, pork, shrimp, soybeans, and many other foods are also contained in trace amounts. Is insoluble in water and soluble in organic solvent such as ethanol.
1-octen-3-ol is widely used for preparing flavor and medicinal raw materials, and Chinese patent publication No. CN105104377A discloses that 1-octen-3-ol has herbicidal effect on some weeds at a use concentration of 2.5-25 ml/L. Seedling establishment is strong, and the biological activity of five volatile compounds on soil-borne pathogenic fungi and nematodes, plant protection bulletin, volume 39, stage 6, and month 12 2012 disclose that 1-octen-3-ol can inhibit conidium germination of penicillium cerealis; 1-octene-3-ol and linalool have antibacterial and anti-nematode activities as active volatile components of Bacillus polymyxa BM P-11, the lethal activity of 1-octene-3-ol on nematodes is studied, the study shows that the lethal activity of 1-octene-3-ol on 2-instar larvae of heterodera avenae wollenweber is higher than that on southern root nematodes, the literature abstract describes that the mortality rate of southern root-knot nematodes is 72.3% at a concentration of 5.24 muL/L, but the literature lacks rigidness, and the mortality rate of southern root-knot nematodes after being sealed for 24h at a concentration of 10 muL/L is 72.3% and the mortality rate of southern root-knot nematodes at a concentration of 5.24 muL/L is about 20% can be seen by reading in detail and combining with a drawing 2 of the literature, in addition, the experiment is treated in a fumigating mode in a sealed environment, the use concentration of the experiment has poor field practical guiding significance, and the experimental procedure was not set to correct mortality to exclude the mortality of meloidogyne incognita from organic solvents that dissolve 1-octen-3-ol.
Chitarra G S et al, 1-Octen-3-ol inhibition of conidium germination of Penicillium paneum of fine effects on membrane metabolism, precipitation, intercellular pH, and changes the protein composition. FEMSM biology Ecology biology, 2005, 54(1):67-75, reported that 1-Octen-3-ol inhibits conidium germination of Penicillium paneum. The biocompatibility of 1-octen-3-ol with a nematicidal microorganism is not known.
Therefore, the real lethal condition of the 1-octen-3-ol to the root-knot nematodes (such as the meloidogyne incognita) needs to be further scientifically verified, and the 1-octen-3-ol and the nematicidal microorganisms need to be screened in a biocompatible manner and compatible with long-acting prevention and control microorganisms to form a stable product, so that the prevention and control effects of the root-knot nematodes are improved in an environment-friendly manner.
Disclosure of Invention
The invention aims to provide a new application of a nematode medicament containing 1-octen-3-ol in the production of organic agriculture and green agriculture, and the new application is a safe and efficient nematicide taking 1-octen-3-ol and paecilomyces lilacinus as active ingredients. In the nematicide, each active ingredient is synergistic and is taken as the active ingredient together, thereby playing a remarkable role in the aspect of controlling nematodes.
The invention relates to a nematicide containing 1-octen-3-ol, in which the 1-octen-3-ol can be used as an active ingredient alone or together with other active ingredients for preventing and treating root-knot nematodes. The using concentration of the 1-octen-3-ol is 4-24 mmol/L.
Further, the nematicide containing the 1-octen-3-ol also comprises paecilomyces lilacinus and an induced nutrient component mixture thereof, wherein the mass ratio of the paecilomyces lilacinus and the induced nutrient component mixture thereof to the 1-octen-3-ol is 1-1.5: 2-3.
Furthermore, the 1-octen-3-ol-containing nematicide has the effective viable count of the paecilomyces lilacinus in the paecilomyces lilacinus and the induced nutrient component mixture thereof being more than or equal to 10 hundred million cfu.g-1The mixture comprises 325-mesh paecilomyces lilacinus powder, 325-mesh chitin powder, glucose and peptone, and the mass ratio of the paecilomyces lilacinus powder to the chitin powder to the peptone is 0.5-1: 2-3: 0.5-1: 1-2.
Further, the nematicide containing the 1-octen-3-ol further comprises glycerol, volatile silicone oil, alkyl betaine, malonic acid and an organic silicon surfactant, wherein the mass ratio of the glycerol, the volatile silicone oil, the alkyl betaine, the malonic acid and the organic silicon surfactant to the 1-octen-3-ol is 1-1.5: 1-1.75: 1-1.75: 0.25-0.4: 0.05-0.1: 2 to 3.
The invention also provides a preparation method of the nematicide containing the 1-octen-3-ol, which comprises the steps of fully mixing the induced nutrient component mixture of the paecilomyces lilacinus with the same amount of glycerol, adding the 1-octen-3-ol, the volatile silicone oil, the lauryl betaine, the malonic acid and the organic silicon surfactant in proportion, fully mixing and emulsifying uniformly; the paecilomyces lilacinus is characterized in that the weight ratio of an induced nutritional component mixture of the paecilomyces lilacinus to an induced nutritional component mixture of the paecilomyces lilacinus, glycerol, 1-octen-3-ol, volatile silicone oil, alkyl betaine, malonic acid and an organic silicon surfactant is 1-1.5: 1-1.5: 2-3: 1-1.75: 1-1.75: 0.25-0.4: 0.05 to 0.1.
The invention also provides application of the nematicide containing the 1-octen-3-ol in preventing and treating tomato root-knot nematodes.
In the induced nutrient component mixture of the paecilomyces lilacinus, glucose and peptone provide nutrients for rapid proliferation of the paecilomyces lilacinus, and the ultramicro chitin powder can induce the paecilomyces lilacinus to generate a large amount of chitinase, so that nematode epidermis is favorably cracked. The volatile silicone oil is nontoxic, odorless, oilless and greasy, has good compatibility with other components, low surface tension and good spreadability, and can better suspend the paecilomyces lilacinus mixture. The alkyl betaine, the organic silicon surfactant and the paecilomyces lilacinus have good biocompatibility and strong stability and expansibility. Malonic acid maintains faintly acid pH, which is beneficial to strain propagation.
Compared with the prior art, the invention has the following beneficial effects:
1. the medicament for preventing and treating the root-knot nematode is safe to use, easy to degrade, harmless to human bodies and environment, free of hormone residue and free of safety problems.
2. The medicament for preventing and treating the root-knot nematodes has good effect on preventing and treating the root-knot nematodes, has good compatibility with other active ingredients for preventing and treating the root-knot nematodes, and can be used in combination so as to improve the nematode preventing and treating effect.
3. The active ingredient 1-octen-3-ol of the invention has good biocompatibility with nematicidal microorganisms, and has no influence on the preservation period and germination of microbial spores or spores.
4. The medicament for preventing and treating the root-knot nematode can also obviously improve the nutritional status of crops, promote the growth and development of the crops and improve the immunity of the crops.
5. The preparation method of the medicament for preventing and treating the root-knot nematode is simple, and large-scale special equipment is not required.
Drawings
FIG. 1 shows the results of biocompatibility studies on 1-octen-3-ol and Paecilomyces lilacinus;
FIG. 2 is a product appearance diagram of a nematicide containing 1-octen-3-ol;
FIG. 3 is a graph showing the rapid growth in soil of Paecilomyces lilacinus in a nematicidal formulation containing 1-octen-3-ol;
FIG. 4 shows the effect of treatment of T on the control of tomato root-knot nematode by a nematicidal preparation containing 1-octen-3-ol and Paecilomyces lilacinus and a single Paecilomyces lilacinus preparation.
Detailed Description
The present invention will be described in further detail with reference to specific embodiments, but it should not be construed that the scope of the present invention is limited to the following examples. Various substitutions and alterations can be made by those skilled in the art and by conventional means without departing from the spirit of the method of the invention described above.
Example 1
Liquid immersion method for detecting drug effect test of 1-octen-3-ol on plant root-knot nematode
1. Test agent
Dissolving 1-octene-3-alcohol in methanol to prepare 4, 8, 12, 16, 20 and 24 mmol.L respectively-1The test solution was concentrated, and methanol was used as a control.
2. Culture and preparation of nematodes
The nematodes isolated from the root knot of tomato were inoculated in Potato Dextrose Agar (PDA) medium of Fusarium oxysporum (Fusarium solani) and cultured at 28 ℃ for 5-7 days. And (3) after the nematodes are propagated and grow on the whole culture dish, inverting the culture dish, adding 3-5mL of sterile water into the culture dish cover, and standing for more than 6 hours, wherein the nematodes enter the sterile water. The nematodes were diluted with sterile distilled water to about 50 nematodes per 10. mu.L of the nematode suspension according to the nematode density in each field under a microscope.
3. Test method
Transferring 500 mu L of the test solution into a 1.5ml PCR tube, adding 10 mu L of nematode suspension, opening a cover, placing in a constant-temperature incubator at 28 ℃ for dark culture for 3h, pouring into a plate, and checking and calculating the nematode death rate under an optical microscope.
Method for identifying death: dead nematodes are stiff or J-shaped, while live nematodes are twisted. After the stiff nematodes are transferred to sterile water, the nematodes are identified as dead if they have not reacted any more.
Mortality rate (number of dead nematode/total number of nematode) × 100%;
corrected mortality-control mortality.
The whole experiment was repeated three times with three replicates per experiment using methanol without added sample as a control, and the average and corrected mortality was calculated by taking the average.
4. Results of the experiment
1-Octen-3-ol has a good insecticidal effect on nematodes, as shown in Table 1-1.
TABLE 1-11-Octen-3-ol poisoning results for Meloidogyne (%)
From Table 1-1, it can be derived: 1-octen-3-ol can rapidly poison nematodes in an open environment for 3 hours, the nematode death rate is positively correlated with the use concentration of 1-octen-3-ol, and 24 mmol.L-1The mortality rate of the nematodes reaches more than 90 percent under the concentration of the organic solvent, and the organic solvent methanol also has lethal effect on the nematodes.
Example 2
Biocompatibility test of 1-octen-3-ol with Paecilomyces lilacinus
1. Test method
Preparing a solid Martin culture medium for culturing paecilomyces lilacinus, and sucking methanol solutions of 50% by mass fraction 1-octen-3-ol with different volumes by using a pipette when pouring a flat plate to ensure that the concentration of the 1-octen-3-ol in the PDA culture medium is 4-24 mmol.L-1. After the culture medium was solidified, 100. mu.L of a Paecilomyces lilacinus spore suspension (10. mu.L) was aspirated by the method of the confrontation test5cfu·mL-1) On PDA plate, using sterile coated glass rod to fully and uniformly coat spore suspension, placing the plate at 28 deg.C, culturing for 5-7d, taking out, and observing test phenomenon, using PDA plate without 1-octen-3-ol as control. The results are shown in Table 2-1.
2. Test results
The results show that the 1-octen-3-ol and the paecilomyces lilacinus have good biocompatibility and no influence on the germination rate of the paecilomyces lilacinus, and are shown in figure 1.
TABLE 2-11-Octen-3-ol and Paecilomyces lilacinus biocompatibility profiles
| Concentration of drug (mmol. L)-1) | Growth of colonies |
| 0 | The colony grows over the whole plate |
| 4 | The colony grows over the whole plate |
| 8 | The colony grows over the whole plate |
| 12 | The colony grows over the whole plate |
| 16 | The colony grows over the whole plate |
| 20 | The colony grows over the whole plate |
| 24 | The colony grows over the whole plate |
Example 3
Preparation of nematocides containing 1-octen-3-ol
1. Medicament
1-octen-3-ol, 325 mesh Paecilomyces lilacinus powder (effective viable count is more than or equal to 10 hundred million cfu g)-1) Chitin powder of 325 mesh, glucose, peptone, glycerol, cyclopentadimethylsiloxane, alkylbetaine, malonic acid, silicone surfactant, all of which are commercially available.
2. Preparation process
(1) Preparation of Paecilomyces lilacinus and its nutrient inducing mixture:
weighing 1 part of paecilomyces lilacinus powder passing through 325 meshes, 2 parts of chitin powder passing through 325 meshes, 0.5 part of glucose and 1 part of peptone, and fully and uniformly mixing the components to prepare paecilomyces lilacinus and a nutrition inducing mixture thereof for later use.
(2) Preparation of nematocides
Weighing 1 part of paecilomyces lilacinus and a nutrition inducing mixture thereof and 1 part of glycerol, shearing and mixing uniformly, then adding 3 parts of 1-octen-3-ol, 1.5 parts of cyclopentadimethylsiloxane, 1.5 parts of alkyl betaine, 0.25 part of malonic acid and 0.1 part of organic silicon, adding, homogenizing and mixing.
3. Product appearance
The product is uniform and has good suspension property. As shown in picture 2.
Example 4
Liquid immersion method for detecting drug effect test of nematocide containing 1-octen-3-ol on plant root-knot nematodes
1. Preparation of pharmaceutical agents
The preparation method is the same as example 3, and the 1-octene-3-ol is respectively prepared by diluting with water and the mass fractions of the 1-octene-3-ol are 4, 8, 12 and 16 mmol.L-1The concentration of the test solution is 4, 8, 12, 16 mmol.L respectively prepared by dissolving 1-octene-3-alcohol in methanol-1The test liquid medicine with the concentration is used as a positive control, and methanol is used as a negative control.
2. Culture and preparation of nematodes in the same manner as in example 1
3. Test method
Transferring 500 mu L of the test solution into a 1.5ml PCR tube, adding 10 mu L of nematode suspension, opening a tube cover, placing the tube cover in a constant-temperature incubator at 28 ℃ for dark culture for 3h, pouring the tube cover into a flat dish, and checking and calculating the mortality of the nematodes under an optical microscope.
Method for identifying death: dead nematodes are stiff or J-shaped, while live nematodes are twisted. After the stiff nematodes are transferred to sterile water, the nematodes are identified as dead if they have not reacted any more.
Mortality rate (number of dead nematode/total number of nematode) × 100%;
taking 1-octen-3-ol dissolved in methanol as a positive control, taking methanol without a sample as a negative control, repeating the whole experiment three times, taking an average value of three parallels of each experiment, and calculating the average mortality.
4. Results of the experiment
The concentration of the prepared nematicide containing 1-octen-3-ol in 1-octen-3-ol is 4-16 mmol.L-1When the nematode is treated for 3 hours, the lethality of the nematode is 39-90%, and the lethality of the nematode is only 16-52% in the methanol solution only containing 1-octen-3-ol under the same concentration and treatment time, as shown in the table 4-1.
TABLE 4-1 poisoning results of 1-octen-3-ol-containing agent and 1-octen-3-ol alone for Meloidogyne
From Table 4-1, it can be derived: the prepared medicament containing 1-octen-3-ol has good permeability expansibility, remarkably improves the killing rate of nematodes, and can improve the prevention and treatment effect of the nematodes.
Example 5
Nematicide stability containing 1-octen-3-ol and pot culture control effect of nematicide on plant root-knot nematodes
1. Test agent
The nematicide containing 1-octen-3-ol was prepared in the same manner as in example 3. A glycerol suspension containing Paecilomyces lilacinus and its nutrient inducing mixture was used as a control.
2. Spore viability assay for Paecilomyces lilacinus in pharmaceutical Agents
And respectively measuring the effective viable count of the paecilomyces lilacinus in the medicament and the glycerin suspending agent of the paecilomyces lilacinus and the nutrition inducing mixture thereof at 0, 4, 8 and 12 months, wherein the effective viable count is measured by a plate counting method after gradient dilution.
4. Nematicidal agent containing 1-octen-3-ol has effect of preventing and controlling tomato root-knot nematode
The nematode control effect of the medicament is measured according to the device and the method (CN110275001A) for measuring and evaluating the medicament effect of the medicament on the root-knot nematodes.
(1) Seed disinfection: sterilizing tomato seeds with 70% ethanol for 5 minutes, sterilizing the tomato seeds with 4% sodium hypochlorite for 15 minutes, and accelerating germination with sterile water after the tomato seeds are sterilized;
(2) seedling culture: after the seeds germinate, putting the seeds into a nutrition pot filled with a sterile matrix (perlite and vermiculite), keeping moisture after covering the seeds with a film, putting the seeds into a biochemical incubator for seedling culture, and irrigating the seeds with sterile Hoagland nutrient solution; after the third true leaf grows out, cutting off the main root of the seedling, dipping the root in a rooting solution containing 0.5mg/kg of indolebutyric acid and 0.5mg/L of sodium naphthaleneacetate, and continuously placing the root in a matrix for culturing to induce a large amount of lateral roots to grow; in the seedling growth process, 20mg/kg of chlormequat chloride is sprayed to avoid vigorous growth in a biochemical incubator; removing the surface matrix along with the growth of the lateral roots to expose the lateral roots at the base parts of the stems, promoting lignification and facilitating subsequent transplanting treatment;
(3) soil treatment: the method comprises the following steps of (1) filling soil seriously infected by nematodes in the previous crop in a device disclosed in the patent of the invention (CN110275001A), carrying out spray treatment on the soil on one side of a partition plate by using an agent diluted by 100 times, and carrying out spray treatment on the soil on the other side of the partition plate by using a control spray, wherein the amount of the soil on the other side of the partition plate is 1kg per mu; balancing for 3-5 days, and transplanting the seedlings;
(4) and (3) transplanting seedlings: transplanting the tomato seedlings into medicament-treated soil (CK +) and contrast-treated soil (CK-), fixing the tomato seedlings on a baffle fixing column, dividing root systems into 2 parts, respectively entering the soil on two sides of a baffle, wherein the root system entering the medicament-treated soil is (R +), and the root system entering the contrast-treated soil is (R-); after transplanting, fertilizing and watering according to the conventional method, and periodically measuring the plant height, the growth vigor and the yield;
(5) evaluation of the effects: after the tomatoes are picked, the baffle parallel to the middle hydrophobic partition plate in the device is opened, the soil is kept soft by watering, the tomato is kept intact when the tomato root system is pulled out, and the number of the tomato root system processed in different ways is counted.
5. Results of the experiment
(1) The paecilomyces lilacinus in the preparation has good biocompatibility and no influence on the preservation period and germination rate of paecilomyces lilacinus spores, and is shown in table 5-1.
TABLE 5 Effect of 11-octen-3-ol on the shelf life and germination Rate of Paecilomyces lilacinus spores
(2) Paecilomyces lilacinus in nematicidal formulations containing 1-octen-3-ol rapidly propagated in soil as shown in FIG. 3. The control effect of the medicament on the tomato root-knot nematodes is good and is improved by more than 1 time compared with the control effect of single paecilomyces lilacinus, the data statistics are shown in tables 5-2, and the control effect is shown in figure 4.
TABLE 5-2 nematicidal formulations containing 1-octen-3-ol and Paecilomyces lilacinus for control of meloidogyne lycopersicae
Claims (6)
1. The nematicide containing 1-octen-3-ol is characterized by comprising 1-octen-3-ol, wherein the use concentration of the 1-octen-3-ol is 4-24 mmol/L.
2. The nematicide containing 1-octen-3-ol according to claim 1, further comprising paecilomyces lilacinus and its induced nutrient mixture, wherein the mass ratio of the paecilomyces lilacinus and its induced nutrient mixture to 1-octen-3-ol is 1-1.5: 2-3.
3. The nematicide containing 1-octen-3-ol according to claim 2, wherein the number of active bacteria in paecilomyces lilacinus in the paecilomyces lilacinus and its induced nutrient composition mixture is not less than 10 hundred million cfu g-1The mixture comprises 325-mesh paecilomyces lilacinus powder, 325-mesh chitin powder, glucose and peptone, and the mass ratio of the paecilomyces lilacinus powder to the chitin powder to the peptone is 0.5-1: 2-3: 0.5-1: 1-2.
4. The nematicide containing 1-octen-3-ol according to claim 1, further comprising glycerol, volatile silicone oil, alkyl betaine, malonic acid, and a silicone surfactant, wherein the mass ratio of the glycerol, volatile silicone oil, alkyl betaine, malonic acid, and silicone surfactant to 1-octen-3-ol is 1 to 1.5: 1-1.75: 1-1.75: 0.25-0.4: 0.05-0.1: 2 to 3.
5. The method for preparing a nematicide containing 1-octen-3-ol according to any one of claims 1 to 4, wherein the mixture of induced nutritional components of Paecilomyces lilacinus is fully mixed with an equal amount of glycerol, and then the 1-octen-3-ol, volatile silicone oil, alkyl betaine, malonic acid and silicone surfactant are added in proportion and fully mixed and emulsified uniformly.
6. The application of the nematicide containing 1-octen-3-ol in the prevention and treatment of tomato root-knot nematodes, which is characterized in that the nematicide containing 1-octen-3-ol is the nematicide containing 1-octen-3-ol according to any one of claims 1 to 4 or the nematicide containing 1-octen-3-ol prepared by the preparation method according to claim 5.
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| CN114342737A (en) * | 2022-01-26 | 2022-04-15 | 四川省食用菌研究所 | Paecilomyces varioti disease prevention and control method for morchella esculenta |
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