CN117441718A - Application of tetraethylene glycol dimethyl ether in preparation of preparation for preventing and controlling plant root knot nematode - Google Patents
Application of tetraethylene glycol dimethyl ether in preparation of preparation for preventing and controlling plant root knot nematode Download PDFInfo
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- CN117441718A CN117441718A CN202311412875.8A CN202311412875A CN117441718A CN 117441718 A CN117441718 A CN 117441718A CN 202311412875 A CN202311412875 A CN 202311412875A CN 117441718 A CN117441718 A CN 117441718A
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- root
- dimethyl ether
- controlling plant
- knot nematode
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- 235000019319 peptone Nutrition 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- NGFMICBWJRZIBI-UJPOAAIJSA-N salicin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=CC=C1CO NGFMICBWJRZIBI-UJPOAAIJSA-N 0.000 description 1
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- 239000005720 sucrose Substances 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N31/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic oxygen or sulfur compounds
- A01N31/02—Acyclic compounds
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/34—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom
- A01N43/40—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom six-membered rings
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/90—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having two or more relevant hetero rings, condensed among themselves or with a common carbocyclic ring system
-
- 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
- A01N57/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds
- A01N57/26—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds having phosphorus-to-nitrogen bonds
- A01N57/32—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds having phosphorus-to-nitrogen bonds containing heterocyclic radicals
-
- 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
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/22—Bacillus
-
- 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
- A01P5/00—Nematocides
Abstract
The invention discloses application of tetraethylene glycol dimethyl ether in preparation of a preparation for preventing and controlling plant root-knot nematode, and the compound can prevent second-instar larvae of the root-knot nematode, and can remarkably improve the prevention effect of a B16 microbial inoculum and a common chemical nematicide on crop root-knot nematode. The tetraglycol dimethyl ether is used together with various nematicides, and has the characteristics of high control efficiency, low use cost, no residue and safety to people and livestock.
Description
Technical Field
The invention relates to a pesticide and the application field thereof, in particular to the application of tetraethylene glycol dimethyl ether in preparing a preparation for preventing and controlling plant root knot nematode, and the strain and an active compound thereof have the activity of avoiding root knot nematode second-instar larvae (J2) and have great application value in preventing and controlling plant root knot nematode.
Background
The nematicidal bacillus B16 strain is a new species of nematode biocontrol bacteria published by university of yunnan 2005 (Huang et al Systematic andAppliedMicrobiology,2005, 28:323-327), which was isolated from the forest soil of yunnan. Taxonomically, the morphological features of the B16 strain are: gram positive bacteria, aerobic, cell rod shape, length of 2.0-3.5 μm, width of 0.8-1.2 μm. Culturing in LB culture medium at 37deg.C for 3d, and the colony is round, 1.0-2.0mm in diameter, rough and matt, similar to fine powder, and white in lawn; few endospores or spores released outside the cells are seen in the cells, and after 5 days, the cells almost fully produce spores. Physiological and biochemical characteristics of strain B16: can grow in the range of 4-50 ℃ and the optimal growth temperature is 20-37 ℃; the pH range of the growth is between 6.0 and 8.5, and the optimal growth is about 7.0 to 8.0; the bacterial has positive catalase and oxidase reactions, and positive ornithine decarboxylase, beta-glucosidase, lysine decarboxylase, urease, alpha-glucosidase, alpha-galactosidase, alpha-maltosidase, L-aspartyl-alanyl-amidase and lipase; and negative for N-acetyl-beta-aminoglucosidase, arginine ditolylase, beta-galactosidase and beta-glucuronidase. Strain B16 can utilize glucose, maltose, ribose, mannose, mannitol, sucrose, xylose, fructose, salicin, dextrin, galactose, starch, arabinose, cellobiose, amygdalin, sorbitol, and rhamnose. Acid can only be produced from N-acetylglucosamine, lactose and escin. The bacterium can coagulate milk or peptone, liquefy gelatin, decompose Tween-80, hydrolyze starch, but cannot utilize cellulose acetate, H2S and melanin. The prominent menaquinone of this bacterium is MK-7. The major cellular fatty acid components are anti-eiso-C15:0 (48.67%), iso-C15:0 (13.45%), C16:0 (9.06%) and anti-eiso-C17:0 (8.29%). The molecular biological characteristics of the B16 strain are as follows: the genome DNA of the B16 strain is used as a template, the 16SrDNA gene sequence of the strain is amplified by using universal primers 27F and 1541R, and the sequence number of the 16S rDNA sequence of the B16 strain in GenBank is AY820954.
B16 strain possesses a unique set of "Trojan horse" nematicidal mechanisms: the strain B16 can recognize morpholine molecules released by nematodes to sense the presence of the nematodes in the environment, and simultaneously, the strain B16 generates spores in advance, when the spores of the strain B16 reach a certain amount, benzaldehyde, benzyl benzoate, acetophenone, 2-heptanone, indole, naphthalene, 2, 5-dimethyl anisole and various aromatic organic volatile matters attract the nematodes, and the olfactory chemotactic system of the nematodes is destroyed, and the formed spores are used for protecting the nematodes and enter the nematodes, so that the nematodes are positioned in intestinal tracts; after the B16 strain enters the nematode gut, the spores regrind and secrete extracellular proteases (serine proteases and neutral proteases) to destroy the nematode gut structure, thereby killing the nematodes, and the B16 strain is propagated in the nematode by using the worms as nutrients, starting the next round of nematode infestation (Niu et al, PNAS,2010, 107:16631-16636). Currently, the biological pesticides "bacillus nematicides B16 parent drug (PD 20211349), bacillus nematicides B16 powder (PD 20211362)" for root knot nematode disease have been registered with bacillus nematicides B16 strain in the university of yunnan and assigned to shandong sharp biotechnology limited.
At present, the application of tetraethylene glycol dimethyl ether with high control efficiency in preparing a preparation for controlling plant root knot nematodes is lacking.
Disclosure of Invention
The invention aims to provide an application of tetraethylene glycol dimethyl ether with high control efficiency in preparing a preparation for controlling plant root knot nematode.
In order to achieve the purposes of the application, the technical scheme adopted by the application is as follows: in a first aspect, the present application provides the use of tetraethylene glycol dimethyl ether in the preparation of a formulation for controlling plant root knot nematodes.
In a second aspect, the present application provides a formulation for controlling plant root knot nematodes.
The invention relates to application of tetraethylene glycol dimethyl ether in preparation of a preparation for preventing and controlling plant root knot nematodes.
Further, the root-knot nematode is a root-knot nematode second-instar larva.
The preparation for preventing and treating plant root knot nematode comprises tetraethylene glycol dimethyl ether with the final concentration of 3-5% and a chemical nematicide.
Further, the chemical nematicide is a common chemical nematicide or a bacterial agent of bacillus nematicide B16 strain, and the common chemical nematicide is any one of fosthiazate, abamectin or fluopyram.
Further, the pesticide for preventing and controlling plant root knot nematodes is a bacillus nematicide B16 microbial inoculum, and tetraethyleneglycol dimethyl ether with the final concentration of 3-5% is added into the microbial inoculum. The effect of the B16 microbial inoculum for preventing and treating the root knot nematode disease can be improved by more than 10 percent. The compound is tetraethylene glycol dimethyl ether, and can be applied to the prevention and treatment of plant root knot nematode disease.
Furthermore, the chemical for preventing and controlling plant root knot nematode is a common chemical nematicide added with tetraethyleneglycol dimethyl ether with the final concentration of 3-5%. Can improve the effect of preventing and treating root-knot nematode by more than 5% by using the common chemical nematicide.
Further, the agent for preventing and controlling plant root-knot nematodes is nematicidal bacillus B16 strain fermentation liquor capable of avoiding root-knot nematodes and 3-5% of tetraethyleneglycol dimethyl ether.
Further, the preparation method of the nematicidal bacillus B16 strain fermentation liquor capable of repelling root knot nematodes comprises the following steps: the B16 strain is inoculated into a conventional LB liquid medium and cultured for 48 hours at 30-37 ℃ and 150rpm to prepare a fermentation broth of the B16 strain.
The beneficial effects are that: the compound can avoid the root-knot nematode J2, and can obviously improve the control effect of the B16 microbial inoculum and the common chemical nematicide on the root-knot nematode disease of crops. The tetraglycol dimethyl ether is matched with various nematicides, so that the tetraglycol dimethyl ether has the characteristics of high control efficiency, low use cost, no residue and safety to human and livestock, and can meet the control requirements of various crop root knot nematode diseases in agricultural production.
Compared with the prior art, the invention has the following advantages: (1) By using the tetraethylene glycol dimethyl ether as an auxiliary ingredient of the B16 microbial inoculum, the control effect of the B16 microbial inoculum on the root-knot nematode disease can be improved by more than 10 percent, and the control effect of the B16 microbial inoculum on the root-knot nematode disease can be up to more than 70 percent.
(2) The prevention effect can be improved by more than 5% by using the tetraglycol dimethyl ether and the common chemical nematicide in a matched way.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present application, the following description will briefly introduce the drawings that are needed in the embodiments or the description of the prior art, it is obvious that the drawings in the following description are only some embodiments of the present application, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a graph showing the effect of B16 strain fermentation broth on the avoidance of root-knot nematode J2; the bars in the graph represent the results of the analysis of the significance of the differences between the treatments, if the number of bars in two of the graphs is the same, the two treatments are not significantly different, otherwise the differences are significant.
FIG. 2 is a GC-MS detection and comparison chart of volatile components of B16 fermentation broth and LB medium provided in the example of the present application.
Fig. 3 is a graph showing the repellent effect of tetraethylene glycol dimethyl ether on root-knot nematode J2 according to an embodiment of the present application; the bars in the graph represent the results of the analysis of the significance of the differences between the treatments, if the number of bars in two of the graphs is the same, the two treatments are not significantly different, otherwise the differences are significant.
Detailed Description
In order to make the technical problems, technical schemes and beneficial effects to be solved by the present application more clear, the present application is further described in detail below with reference to the embodiments. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the present application.
Example 1
The invention relates to application of tetraethylene glycol dimethyl ether in preparation of a preparation for preventing and controlling plant root knot nematodes.
The root-knot nematode is a root-knot nematode second-instar larva.
The preparation for preventing and treating plant root knot nematode comprises tetraethylene glycol dimethyl ether with the final concentration of 3-5% and a chemical nematicide. The chemical nematicide is a bacillus nematicide B16 strain fungicide.
The pesticide for preventing and controlling plant root knot nematode is tetraethyleneglycol dimethyl ether with the final concentration of 3-5% added into a bacillus nematicide B16 microbial inoculum. The effect of the B16 microbial inoculum for preventing and treating the root knot nematode disease can be improved by more than 10 percent. The compound is tetraethylene glycol dimethyl ether, and can be applied to the prevention and treatment of plant root knot nematode disease.
The chemical for preventing and controlling plant root-knot nematode is a common chemical nematicide added with tetraethyleneglycol dimethyl ether with the final concentration of 3-5%. Can improve the effect of preventing and treating root-knot nematode by more than 5% by using the common chemical nematicide.
The agent for preventing and controlling plant root-knot nematodes is nematicidal bacillus B16 strain fermentation liquor capable of repelling root-knot nematodes and 3-5% of tetraethyleneglycol dimethyl ether.
The preparation method of the nematicidal bacillus B16 strain fermentation liquor capable of avoiding root-knot nematodes comprises the following steps: the B16 strain is inoculated into a conventional LB liquid medium and cultured for 48 hours at 30-37 ℃ and 150rpm to prepare a fermentation broth of the B16 strain.
After intellectual property rights of the B16 strain are obtained by Shandong Sirui biotechnology limited company, the strain is deeply researched and developed, and the fact that the B16 strain can generate a compound tetraethyleneglycol dimethyl ether, the compound has stronger repellent activity on the root-knot nematode J2, and the compound is added into the B16 microbial inoculum and the common chemical nematicide, so that the control effect of each agent on the root-knot nematode can be greatly improved. Although the B16 strain has been reported to trap nematodes and its use in controlling nematode diseases, the active compounds for repelling nematodes produced by the strain and their use in controlling nematode pests have not been reported. The strain is nematicidal bacillus B16 strain, and the strain is B16-w1; the preservation name is nematicidal bacillus; the microbial strain is preserved in China general microbiological culture Collection center (CGMCC), and the preservation address is 1 # 3 of North West Lu in the Chaoyang area of Beijing city; preservation date: 2023, 08, 2; the preservation number is CGMCC No.28088.
Example 2
The preparation for preventing and treating plant root knot nematode comprises tetraethylene glycol dimethyl ether with the final concentration of 3-5% and a common chemical nematicide. The common chemical nematicide is any one of fosthiazate, abamectin or fluopyram.
Test example 1
Determination of effect of B16 strain fermentation liquor in avoiding root-knot nematode J2
The invention adopts LB culture medium to prepare B16 fermentation liquor according to the conventional microbial fermentation method, and the effective viable count of B16 strain in the fermentation liquor is about 10 8 And each ml. The repellent effect of the fermentation broth on the root-knot nematode J2 is determined according to a conventional Pluronic F-127 gel method. The specific operation steps are as follows.
(1) Preparing a flat plate: a23% solution of Pluronic F-127 gel was prepared at 4deg.C and spread on the bottom of a 60mm diameter plate and then placed in a 25℃incubator until Pluronic F-127 was semi-solid.
(2) Sample adding: mu.L of B16 bacterial liquid is added at the position (A) 10mm away from the left edge of the bottom of the flat plate, 5 mu.L of LB liquid culture medium is used as a control and is added at the position (B) 10mm away from the right edge of the bottom of the flat plate, 5 mu.L of nematode suspension containing 100-200 root-knot nematodes J2 is added at the center of the middle of the flat plate, and the mixture is kept stand for 6h, 12h and 24h in a culture box at 25 ℃.
(3) Counting: observed under an optical microscope, the number of J2 on both left and right sides at 6h, 12h and 24h were counted, respectively, and an evasion index (AI) was calculated according to the following formula: ai= (B-ase:Sub>A)/(a+b). When AI is more than or equal to 0.2 and less than or equal to 0.3, the evasion is regarded as medium; when AI is more than or equal to 0.3, the method is regarded as strong evasion.
As shown in FIG. 1, the AI values of the fermentation broth B16 for J2 at 6, 12 and 24 hours are respectively 0.41, 0.44 and 0.38, and the fermentation broth B16 has strong evasion effects; the AI value of the LB liquid culture medium is between-0.08 and 0.09, and has no evasion effect on J2. Indicating that the repellent effect of the B16 fermentation liquor on J2 comes from the fermentation product. FIG. 1B16 shows the repellent effect of strain fermentation broth on root-knot nematode J2.
Test example 2
Identification of active compound of B16 strain for avoiding root-knot nematode J2 and determination of avoiding effect thereof
During the previous experiment, the root-knot nematode J2 and the B16 are not in direct contact, and it is deduced that the B16 strain generates certain volatile substances to avoid the root-knot nematode J2. To verify this conclusion, volatile components in the fermentation broth of strain B16 were tested by GC-MS and the repellent effect of each volatile component on the root-knot nematode J2 was determined one by one using the Pluronic F-127 gel method described in example 1 above with pure products of each volatile component including tetraethyleneglycol dimethyl ether. Each compound was used as a solvent at a concentration of 3% with aniline (pc 1) and n-octanol (pc 2) reported to have a repellent effect on the root-knot nematode J2 as positive controls and deionized water as Negative Controls (NC) to check the accuracy of the experimental results.
Analysis of results: the volatile components of the B16 strain broth were analyzed by GC-MS using LB liquid medium for culturing the B16 strain as a control, and the total of 27 kinds of volatile components unique to the B16 strain broth, including tetraethyleneglycol dimethyl ether (FIG. 2), were subtracted from the volatile components in LB. 24 volatiles were purchased and their repellent activity against J2 was measured one by one, and the results showed that only tetraethyleneglycol dimethyl ether had repellent activity against J2. The AI value of the Negative Control (NC) with deionized water is 0.084, and the repellent activity is not generated; the positive controls had AI values of 0.38 and 0.29 for 3% aniline (pc 1) and 3% n-octanol (pc 2), respectively; and the AI value of 3% of tetraethyleneglycol dimethyl ether is 0.39, which is obviously higher than that of two positive control compounds, thus showing that the tetraglyme has strong repellent activity on the root-knot nematode J2. FIG. 2B16 GC-MS detection comparison patterns of volatile components of fermentation broth and LB medium. Figure 3 the repellent effect of tetraethylene glycol dimethyl ether on root-knot nematode J2.
Test example 3
Field test for improving control effect of B16 microbial inoculum on tomato root knot nematode disease by tetraethylene glycol dimethyl ether
Test site: the tomato planted throughout the year is endangered by root knot nematodes at the vegetable planting base of the Yuxi city of Yunnan province.
Test crop: tomato (variety: pink-80).
Test agent: (1) b16 microbial inoculum: the content of viable bacteria is 5.5 hundred million CFU/g, which is prepared by Shandong Sirui biotechnology Co., ltd, and meets the enterprise standard of biological pesticide ' nematicide bacillus B16 powder (PD 20211362 ') '; (2) "B16 microbial inoculum+1% tetraethyleneglycol dimethyl ether" microbial inoculum: adding tetraethylene glycol dimethyl ether into the B16 microbial inoculum to make the content of the tetraethylene glycol dimethyl ether account for 1 percent (W: W); (3) "B16 microbial inoculum+3% tetraethyleneglycol dimethyl ether": adding tetraethylene glycol dimethyl ether into the B16 microbial inoculum to make the content of the tetraethylene glycol dimethyl ether be 3% (W: W); (4) "B16 microbial inoculum+5% tetraethyleneglycol dimethyl ether": adding tetraethylene glycol dimethyl ether into the B16 microbial inoculum to make the content of the tetraethylene glycol dimethyl ether account for 5 percent (W: W); (5) 3% tetraethyleneglycol dimethyl ether: sterilizing the B16 microbial inoculum at 121 ℃ for 2 hours to inactivate the B16 bacterial strain, and then adding tetraethylene glycol dimethyl ether to make the final concentration reach 3%; (6) 10% fosthiazate granules produced by Shandong province United agricultural chemical industry Co.
The test method comprises the following steps: the dosage of each medicament is 2 kg/mu, and the application method is hole application during transplanting; treatment without any nematicide was used as a blank; each of the four (3) replicates, and 120 tomatoes were each replicated.
Investigation and statistical methods: stopping the test after 180 days of application, pulling out the plants, grading the root nodules according to the following standard, and counting the disease index and the control effect according to the formula.
The nodule grading criteria are as follows:
level 0: no root nodule; stage 1: 1-20% of root systems have root nodules; 2 stages: 21-40% of the root systems have root nodules; 3 stages: 41-60% of the root systems have root nodules; 4 stages: 61-80% of the root systems have root nodules; 5 stages: 81-100% of the root systems have root nodules.
Disease index (%) = (n1×1+n2×2+n3×3+n4×4+n5×5)/(s× 5) ×100; n1-n5 represent the total number of plants corresponding to the root nodule disease grade index of 1-5, respectively, and S represents the total number of plants investigated.
Control (%) = 100 (1-x/y), x, y represent disease index of treatment and blank control, respectively.
Identification of the species of the root-knot nematodes in the field: after the test is finished, 30 tomato plants with root nodules are randomly selected from the test, 5 female insects are selected from the root nodules of each plant root system, perineum pattern sections of each female insect are prepared according to a conventional method, and the perineum pattern sections are observed under a microscope; and finally, identifying the nematode species according to the perineum pattern morphology by a conventional method.
Test results: the control effect of each medicament on tomato root knot nematode disease is shown in table 1:
TABLE 1
Note that: the same letters following the same column of data in the table indicate that the differences are not significant, and the different letters indicate that the differences are significant.
As shown in Table 1, the invention shows the control effect of various tested medicaments on tomato root knot nematode disease, and the control effect reaches 65.11% when the B16 microbial inoculum is singly used. When 3% of tetraethylene glycol dimethyl ether is singly used, the prevention effect is 17.12%, which shows that the tetraethylene glycol dimethyl ether has the effect of preventing and controlling nematodes, but the prevention effect is poor. When 1%, 3% and 5% of tetraethyleneglycol dimethyl ether are respectively added into the B16 microbial inoculum, the control effect is respectively 70.68%, 76.51% and 77.87%, and compared with the control effect of the B16 microbial inoculum, the control effect is respectively improved by 5.56%, 11.4% and 12.76%, which indicates that the control effect of the B16 microbial inoculum can be obviously improved by adding the tetraethyleneglycol dimethyl ether; the content of tetraethylene glycol dimethyl ether is best in effect when being 3% and 5%, the effect is obviously higher than the control effect (68.04%) of the fosthiazate granule which is 10% of the control chemical pesticide, and when the content of glycol dimethyl ether is 3% and 5%, the control effect is not obviously different, so that the proper addition amount of the glycol dimethyl ether in the B16 microbial inoculum is 3%. The species of root-knot nematodes were identified based on 150 nematode perineal pattern morphology, three species of root-knot nematodes infesting tomatoes in this test field: the amounts of the three nematodes are 47.3%, 31.6% and 21.1% respectively, which shows that the addition of tetraethyleneglycol dimethyl ether to the B16 microbial inoculum has control effect on the three root-knot nematodes.
Test example 4
Field test for improving control effect of B16 microbial inoculum on cucumber root knot nematode disease by tetraethyleneglycol dimethyl ether
Test site: the plant base of the Shandong Shou vegetables is damaged by root knot nematodes for perennial tomato and cucumber.
Test crop: cucumber (variety: zhongnong No. 8).
Test agent: (1) b16 microbial inoculum: the content of viable bacteria is 5.5 hundred million CFU/g, which is prepared by Shandong Sirui biotechnology Co., ltd, and meets the enterprise standard of biological pesticide ' nematicide bacillus B16 powder (PD 20211362 ') '; (2) "B16 microbial inoculum+1% tetraethyleneglycol dimethyl ether" microbial inoculum: adding tetraethylene glycol dimethyl ether into the B16 microbial inoculum to make the content of the tetraethylene glycol dimethyl ether account for 1 percent (W: W); (3) "B16 microbial inoculum+3% tetraethyleneglycol dimethyl ether": adding tetraethylene glycol dimethyl ether into the B16 microbial inoculum to make the content of the tetraethylene glycol dimethyl ether be 3% (W: W); (4) "B16 microbial inoculum+5% tetraethyleneglycol dimethyl ether": adding tetraethylene glycol dimethyl ether into the B16 microbial inoculum to make the content of the tetraethylene glycol dimethyl ether account for 5 percent (W: W); (5) 3% tetraethyleneglycol dimethyl ether: sterilizing the B16 microbial inoculum at 121 ℃ for 2 hours to inactivate the B16 bacterial strain, and then adding tetraethylene glycol dimethyl ether to make the final concentration reach 3%; (6) 10% fosthiazate granules produced by Shandong province United agricultural chemical industry Co.
The test method comprises the following steps: the dosage of each medicament is 2 kg/mu, and the application method is hole application during transplanting; treatment without any nematicide was used as a blank; each of the four (3) replicates, and 120 tomatoes were each replicated.
Investigation and statistical methods: stopping the test after 180 days of application, pulling out the plants, grading the root nodules according to the following standard, and counting the disease index and the control effect according to the formula.
The nodule grading criteria are as follows:
level 0: no root nodule; stage 1: 1-20% of root systems have root nodules; 2 stages: 21-40% of the root systems have root nodules; 3 stages: 41-60% of the root systems have root nodules; 4 stages: 61-80% of the root systems have root nodules; 5 stages: 81-100% of the root systems have root nodules.
Disease index (%) = (n1×1+n2×2+n3×3+n4×4+n5×5)/(s× 5) ×100; n1-n5 represent the total number of plants corresponding to the root nodule disease grade index of 1-5, respectively, and S represents the total number of plants investigated.
Control (%) = 100 (1-x/y), x, y represent disease index of treatment and blank control, respectively.
Identification of the species of the root-knot nematodes in the field: after the test is finished, 30 tomato plants with root nodules are randomly selected from the test, 5 female insects are selected from the root nodules of each plant root system, perineum pattern sections of each female insect are prepared according to a conventional method, and the perineum pattern sections are observed under a microscope; and finally, identifying the nematode species according to the perineum pattern morphology by a conventional method.
Test results: the control effect of each medicament on cucumber root knot nematode disease is shown in table 2:
TABLE 2
Note that: the same letters following the same column of data in the table indicate that the differences are not significant, and the different letters indicate that the differences are significant.
As shown in Table 2, the invention shows the control effect of various tested medicaments on tomato root knot nematode disease, and the control effect reaches 63.02% when the B16 microbial inoculum is singly used. When 3% of tetraethylene glycol dimethyl ether is singly used, the prevention effect is 18.35%, which shows that the tetraethylene glycol dimethyl ether has the effect of preventing and controlling nematodes, but the prevention effect is poor. When 1%, 3% and 5% of tetraethyleneglycol dimethyl ether are respectively added into the B16 microbial inoculum, the control effect is 68.89%, 74.46% and 75.7%, and compared with the control effect of the B16 microbial inoculum, the control effect is respectively improved by 5.87%, 11.44% and 12.68%, which indicates that the control effect of the B16 microbial inoculum can be obviously improved by adding the tetraethyleneglycol dimethyl ether; the content of tetraethylene glycol dimethyl ether is 3% and 5%, the effect is best, the effect is equivalent to the control effect (68.04%) of the fosthiazate granule with the content of 10% of the control chemical pesticide, and when the content of glycol dimethyl ether is 3% and 5%, the control effect is not obviously different, so that the proper addition amount of the glycol dimethyl ether in the B16 microbial inoculum is 3%. The species of root-knot nematodes were identified based on 150 nematode perineal pattern morphology, three species of root-knot nematodes infesting tomatoes in this test field: the quantity of the southern root-knot nematodes (Meloidogyne incognita) and the northern root-knot nematodes (M.hapla) are 68.3 percent and 31.7 percent respectively, which shows that the addition of tetraethyleneglycol dimethyl ether into the B16 microbial inoculum has control effects on the two root-knot nematodes.
Test example 5
Field test for improving control effect of common chemical nematicide by tetraethyleneglycol dimethyl ether
Test site: the plant base of the Shandong Shou vegetables is damaged by root knot nematodes for perennial tomato and cucumber.
Test crop: cucumber (variety: zhongnong No. 8).
Test agent: (1) 10% fosthiazate granules produced by Shandong province United pesticide industry Co., ltd; (2) 5% avermectin emulsifiable concentrate produced by Hebei Wilkinsonii chemical industry Co.Ltd; (3) 41.7% fluopyram suspoemulsion, produced by bayer crop science company; (4) 10% fosthiazate granules +3% tetraethyleneglycol dimethyl ether; (5) 5% avermectin emulsifiable solution and 3% tetraethylene glycol dimethyl ether; (6) 41.7% fluopyram suspoemulsion +3% tetraethyleneglycol dimethyl ether; (7) 3% tetraethylene glycol dimethyl ether aqueous solution, the invention is prepared. For the medicaments needing to be added with the tetraethylene glycol dimethyl ether, the tetraethylene glycol dimethyl ether is added into the original package of each medicament to ensure that the final concentration reaches 3 percent.
The test method comprises the following steps: the usage and dosage of each medicament are carried out according to the description of the original product; the application method of the 3% tetraethylene glycol dimethyl ether aqua is to dilute 500 times and irrigate roots; treatment without any nematicide was used as a blank; each of the four (3) replicates, and 120 tomatoes were each replicated.
Investigation and statistical methods: stopping the test after 180 days of application, pulling out the plants, grading the root nodules according to the following standard, and counting the disease index and the control effect according to the formula.
The nodule grading criteria are as follows:
level 0: no root nodule; stage 1: 1-20% of root systems have root nodules; 2 stages: 21-40% of the root systems have root nodules; 3 stages: 41-60% of the root systems have root nodules; 4 stages: 61-80% of the root systems have root nodules; 5 stages: 81-100% of the root systems have root nodules.
Disease index (%) = (n1×1+n2×2+n3×3+n4×4+n5×5)/(s× 5) ×100; n1-n5 represent the total number of plants corresponding to the root nodule disease grade index of 1-5, respectively, and S represents the total number of plants investigated.
Control (%) = 100 (1-x/y), x, y represent disease index of treatment and blank control, respectively.
Test results: the synergistic effect of tetraethylene glycol dimethyl ether on the control of three nematicides is shown in table 3:
TABLE 3 Table 3
As shown in Table 3, the invention shows the synergistic effect of tetraethylene glycol dimethyl ether on three common chemical nematicides for preventing and treating cucumber root knot nematode disease. When the chemical nematicide 10% fosthiazate granule, 5% avermectin emulsifiable solution and 41.7% fluopyram suspension emulsion are independently used, the control effect on cucumber root knot nematode disease is 68.83%, 62.89% and 74.2% respectively; when tetraethyleneglycol dimethyl ether with the final concentration of 3% is added into the three chemical nematicides, the control effects are 74.71%, 69.96% and 79.61%, and the control effects of the three chemical nematicides are improved by 5.88%, 7.07% and 5.41%, respectively. The result shows that the prevention effect is improved by more than 5% when tetraethyleneglycol dimethyl ether with the final concentration of 3% is added into the common nematicide.
The foregoing has shown and described the basic principles, principal features and advantages of the invention. It will be understood by those skilled in the art that the foregoing test examples and descriptions are merely illustrative of the principles of the present invention and that various changes and modifications may be made therein without departing from the spirit and scope of the invention, the scope of which is defined in the appended claims, specification and their equivalents.
Claims (8)
1. The application of tetraethylene glycol dimethyl ether in preparing a preparation for preventing and controlling plant root knot nematode.
2. The use according to claim 1, characterized in that: the root-knot nematode is a root-knot nematode second-instar larva.
3. A preparation for preventing and controlling plant root knot nematodes, which is characterized in that: the agent for preventing and controlling plant root-knot nematodes comprises tetraethylene glycol dimethyl ether with a final concentration of 3-5% in claim 1 and a chemical nematicide.
4. A formulation for controlling plant root knot nematodes as claimed in claim 3 in which: the chemical nematicide is a common chemical nematicide or a nematicide bacillus B16 strain fungicide, and the common chemical nematicide is any one of fosthiazate, abamectin or fluopyram.
5. The preparation for controlling plant root knot nematode according to claim 4, wherein: the pesticide for preventing and controlling plant root knot nematode is tetraethyleneglycol dimethyl ether with the final concentration of 3-5% added into a bacillus nematicide B16 microbial inoculum.
6. The preparation for controlling plant root knot nematode according to claim 4, wherein: the chemical for preventing and controlling plant root-knot nematode is a common chemical nematicide added with tetraethyleneglycol dimethyl ether with the final concentration of 3-5%.
7. A formulation for controlling plant root knot nematodes as claimed in claim 3 in which: the agent for preventing and controlling plant root-knot nematodes is nematicidal bacillus B16 strain fermentation liquor capable of repelling root-knot nematodes and 3-5% of tetraethyleneglycol dimethyl ether.
8. The formulation for controlling plant root knot nematodes as defined in claim 7, in which: the preparation method of the nematicidal bacillus B16 strain fermentation liquor capable of avoiding root-knot nematodes comprises the following steps: the B16 strain is inoculated into a conventional LB liquid medium and cultured for 48 hours at 30-37 ℃ and 150rpm to prepare a fermentation broth of the B16 strain.
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| CN1690189A (en) * | 2004-04-21 | 2005-11-02 | 云南大学 | Bacillus for killing plant parasitic nematode and its preparation process and use thereof |
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| CN1690189A (en) * | 2004-04-21 | 2005-11-02 | 云南大学 | Bacillus for killing plant parasitic nematode and its preparation process and use thereof |
| CN101861880A (en) * | 2010-06-24 | 2010-10-20 | 云南大学 | Microbial agent and application thereof |
| CN105638740A (en) * | 2016-01-25 | 2016-06-08 | 莫明和 | Root-knot nematode lure attraction agent and application thereof |
| CN114600896A (en) * | 2022-01-14 | 2022-06-10 | 杭州益森键生物科技有限公司 | Efficient permeation and conduction type pine trunk injection agent and preparation method thereof |
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