CN105557758A - Locust micro-microsporidia spore water suspension and preparation method thereof - Google Patents
Locust micro-microsporidia spore water suspension and preparation method thereof Download PDFInfo
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- CN105557758A CN105557758A CN201511021621.9A CN201511021621A CN105557758A CN 105557758 A CN105557758 A CN 105557758A CN 201511021621 A CN201511021621 A CN 201511021621A CN 105557758 A CN105557758 A CN 105557758A
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- spore
- locust
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/10—Animals; Substances produced thereby or obtained therefrom
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/02—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing liquids as carriers, diluents or solvents
- A01N25/04—Dispersions, emulsions, suspoemulsions, suspension concentrates or gels
Abstract
The invention relates to a micro-microsporidia spore water suspension used for preventing and controlling locusts, and a preparation method thereof. The active component of the locust micro-microsporidia spore water suspension is locust micro-microsporidia spores, and proper amounts of Morwet D425 and xanthan gum are added. The locust micro-microsporidia spore water suspension has the advantages of simple preparation method, good leaf surface adhesiveness, high heat resistance, substantially improved dispersion and uniformity, normal temperature storage resistance, substantial prolongation of the shelf life, good locust killing effect, and cost reduction, is safe to chemical pesticides, and is important for improving the Chinese locust prevention and treatment technology level.
Description
Technical field
The present invention relates to biological pesticide technical field, specifically, relate to a kind of microsporidian spore aqueous suspension agent.
Background technology
Locust disaster is the natural calamity of serious threat China agricultural production always.Nearly 1000 kinds of the locust that China has recorded, wherein to about more than 60 kinds that agriculture, woods, animal husbandry can work the mischief.The method of current control locust mainly adopts chemical pesticide control, the advantage of chemical pesticide control to force down rapidly the insect density of locust, but its shortcoming also clearly, because it simultaneously often brings contaminated environment, kills and wounds a large amount of useful animals, person poultry poisoning and insect and to develop immunity to drugs etc. adverse consequences.
Microbial pesticide is eliminated locusts not only effectively can control locust disaster, and not killed natural enemies, environmentally friendly, to person poultry safety, there is very wide application prospect.At present, the existing technology about utilizing green muscardine fungus to prevent and treat locust, but its production cost is high, storage life is short, the requirement for environmental conditions of use is harsh, some biological uneasy congruent defect be also limit to the scope of application of this technology.
Nosema locustae is a kind of parasitic protozoa of locust obligate, only have pathogenic to orthopterans such as locusts, to the complete no pathogenicity of vertebrate, experimental study proves also not infect honeybee, safety is good, and this pathogen less by the restriction of environmental condition in field use procedure, be produced into low, therefore very competitive in control locust disaster.Existing formulation and application process are usually adopt low concentration nosema locustae aqua, mainly water and microsporidian spore are formed by directly mixing, suspensibility is low, poor stability, be easy to evaporate and be not suitable for aircraft ultra-low volume and the outstanding problem such as spray.For improving locust effect of going out, the following problem of nosema locustae aqueous suspension agent is studied: the plate-out ability on blade face, preparation evaporation rate, warm tolerance, dispersiveness, storage endurance, uniformity, the stability etc. of liquid in sprinkling process.
The solution of the problems referred to above must consider the cooperation of each component, and this invention have developed a kind of nosema locustae aqueous spore suspension for the problems referred to above, efficiently solves the problems referred to above.
Summary of the invention
The object of this invention is to provide one and can be used for nosema locustae spore fluid suspending agent and preparation method thereof.
Nosema locustae spore fluid suspending agent of the present invention, is except containing except active component nosema locustae spore, with 100 parts of listed as parts by weight, also comprises the component of following weight portion: MorwetD425:10-50 part, xanthans: 1-5 part; Sodium lactate: 1-5 part, and the water of surplus.
Preferably, described nosema locustae spore fluid suspending agent, is except containing except active component nosema locustae spore, with 100 parts of listed as parts by weight, also comprises the component of following weight portion: MorwetD425:20-30 part, xanthans: 3-5 part; Sodium lactate: 3-5 part, and the water of surplus.
Preferred, described nosema locustae spore fluid suspending agent, is except containing except active component nosema locustae spore, with 100 parts of listed as parts by weight, also comprises the component of following weight portion: MorwetD425:30 part, xanthans: 4 parts; Sodium lactate: 5 parts, and the water of surplus.
Wherein, the concentration range of described active component nosema locustae spore is not limit, preferred 100-5000 ten thousand spores/mL; More preferably 1000-2000 ten thousand spores/mL.
Wherein, described D425 is a kind of alkylnaphthalene sulfonate condensation polymer, is the one in the Morwet Series Dispersant described in this area; For preventing nosema locustae from assembling in suspension, the effect of dispersal spore;
Described xanthans, for increasing the proportion of suspending agent, improves the suspensibility of spore;
Described sodium lactate, for suppressing varied bacteria growing, keeps the purity of suspension.
Said components interacts, and ensures that suspending agent can be stablized, preserve for a long time.And the blade face plate-out ability of liquid is good, after spraying, the evaporation rate of liquid is slow, good heat resistance, uniformity are good.
The present invention also provides the preparation method of above-mentioned nosema locustae spore fluid suspending agent.
The preparation method of nosema locustae spore fluid suspending agent of the present invention, comprises the following steps:
1) take xanthans, add appropriate water, be stirred to xanthans and dissolve completely;
2) in xanthan gum solution, add D425 and sodium lactate, mix, obtain mixed solution;
3) the nosema locustae mother liquor of freezen protective is taken out, thaws, shake up, measure mother liquor by allocation ratio and add in mixed solution, then supply water, mix.
Wherein step 1) in, the time of described stirring is 2 hours;
Described step 2) and 3) in, the hybrid mode that described mixing adopts this area conventional, preferably stirs, described mixing time preferably 2 hours.
Experimental studies have found that, nosema locustae suspending agent control uncultivated area migratory locusts control efficiency of the present invention is good, and from control efficiency, recommended dose is amount of formulation 65-80 milliliter/mu.Described uncultivated area includes but not limited to field, grassland, mountain forest etc.
Nosema locustae spore fluid suspending agent preparation method of the present invention is simple, liquid to blade face plate-out ability is good, warm tolerance is high, dispersed and uniformity significantly improves, more resistance to normal temperature is stored, is significantly improved shelf life, maintain the activity of nosema locustae, locust of going out is effective, reduce cost, more safer than chemical pesticide, significant for raising China locust control technical merit.
Embodiment
Further describe the present invention below in conjunction with specific embodiment, but embodiment is only exemplary, does not form any restriction to scope of the present invention.It will be understood by those skilled in the art that and can modify to the details of technical solution of the present invention and form or replace down without departing from the spirit and scope of the present invention, but these amendments and replacement all fall within the scope of protection of the present invention.
As being not particularly illustrated, various reagent of the present invention is commercial reagent.
Embodiment 1
Processing formulation constituent is D425 (alkylnaphthalene sulfonate condensation polymer Morwet), xanthans, sodium lactate and water.Prepare nosema locustae spore fluid suspending agent in the steps below:
1) take the xanthans of 0.04% by configuration sum-rate, add appropriate water, stir 2 hours, dissolve completely to xanthans;
2) by configuration sum-rate, in xanthan gum solution, add the D425 of 0.3%, the sodium lactate of 0.05%, mixes, and obtains mixed solution;
3) 10,000,000,000 nosema locustae spore mother liquors of freezen protective are taken out, thaw, shake up, measuring mother liquor by configuration sum-rate adds in mixed solution, then supplies water, uniform stirring 2 hours, mix, namely can be made into 2,000 ten thousand spores/mL nosema locustae spore suspension agent.
Embodiment 2
Processing formulation constituent is D425 (alkylnaphthalene sulfonate condensation polymer Morwet), xanthans, sodium lactate and water.Prepare nosema locustae spore fluid suspending agent in the steps below:
1) take the xanthans of 0.05% by configuration sum-rate, add appropriate water, stir 2 hours, dissolve completely to xanthans;
2) by configuration sum-rate, in xanthan gum solution, add the D425 of 0.1%, the sodium lactate of 0.03%, mixes, and obtains mixed solution;
3) 10,000,000,000 nosema locustae spore mother liquors of freezen protective are taken out, thaw, shake up, measuring mother liquor by configuration sum-rate adds in mixed solution, then supplies water, uniform stirring 2 hours, mix, namely can be made into 1,000 ten thousand spores/mL nosema locustae spore suspension agent.
Embodiment 3
Processing formulation constituent is D425 (alkylnaphthalene sulfonate condensation polymer Morwet), xanthans, sodium lactate and water.Prepare nosema locustae spore fluid suspending agent in the steps below:
1) take the xanthans of 0.01% by configuration sum-rate, add appropriate water, stir 2 hours, dissolve completely to xanthans;
2) by configuration sum-rate, in xanthan gum solution, add the D425 of 0.5%, the sodium lactate of 0.01%, mixes, and obtains mixed solution;
3) 10,000,000,000 nosema locustae spore mother liquors of freezen protective are taken out, thaw, shake up, measuring mother liquor by configuration sum-rate adds in mixed solution, then supplies water, uniform stirring 2 hours, mix, namely can be made into 5,000 ten thousand spores/mL nosema locustae spore suspension agent.
Embodiment 4
Processing formulation constituent is D425 (alkylnaphthalene sulfonate condensation polymer Morwet), xanthans, sodium lactate and water.Prepare nosema locustae spore fluid suspending agent in the steps below:
1) take the xanthans of 0.02% by configuration sum-rate, add appropriate water, stir 2 hours, dissolve completely to xanthans;
2) by configuration sum-rate, in xanthan gum solution, add the D425 of 0.3%, the sodium lactate of 0.03%, mixes, and obtains mixed solution;
3) 10,000,000,000 nosema locustae spore mother liquors of freezen protective are taken out, thaw, shake up, measuring mother liquor by configuration sum-rate adds in mixed solution, then supplies water, uniform stirring 2 hours, mix, namely can be made into 1,000,000 spores/mL nosema locustae spore suspension agent.
Active determination test in embodiment 5
1. test objective
With Asiatic migratory locust 3 nymph in age (nymph of a locust) for supplying examination target, pressing from both sides malicious blade method according to NY/T1154.2-2006, measuring the activity of nosema locustae spore fluid suspending agent.
2. experimental condition
For examination target: with Asiatic migratory locust 3 nymph in age (nymph of a locust) for supplying examination target.
Instrument and equipment:
Electronic balance (minimum weighing value 0.1mg)
Card punch, capillary dropper
250ml transparent plastic cup
Diameter is the culture dish of 9cm
Beaker
Pipettor
Tweezers
Gauze, rubber band etc.
3. experimental scheme
Examination material prepares:
Asiatic migratory locust is provided by China Agricultural University.
Raising temperature is 28 ± 1 DEG C, and humidity is 50 ± 10%, 24h illumination, feeding maize leaves.
Medicament:
Test medicine: nosema locustae spore mother liquor (1 × 10
10spore/ml), provided by China Agricultural University.
Contrast medicament: the former medicine of 95% ethiprole, is produced by Yancheng City Rui Jie Chemical Co., Ltd. of Jiangsu Province.
4. test procedure
Medicament is prepared:
With nosema locustae spore fluid suspending agent formula (embodiment 1) dilution nosema locustae spore mother liquor of the present invention, be mixed with 1 × 10 respectively
5, 1 × 10
6, 1 × 10
7, 1 × 10
8, 1 × 10
9individual spore/ml spore fluid suspending agent.
Contrast medicament: by the former medicinal acetone solution of 95% ethiprole, make 5mg/L solution, for subsequent use.
Blank: with not having the aqueous suspension agent of nosema locustae spore as CK.
Worm prepares in examination:
Select the length of time consistent 3 age Locusta migratoria manilensis locust nymph, hungry 8h, chooses 50 nymphs of a locust, with scales/electronic balance weighing, calculates every average quality.
Press from both sides malicious leaf production:
Beat with the card punch that diameter is 1cm and get fresh corn blade, with pipettor from low concentration, drip the liquid to be measured of 1ul on blade, involutory the making of leaf butterfly scribbling gelatinized corn starch with another sheet presss from both sides malicious leaf butterfly.After making, fix leaf butterfly with the pin being inserted with hard triangular plate, be positioned in transparent plastic cup, take food for the nymph of a locust.Each process 4 repetition, each repeat examination borer population be 20.Contrast medicament and blank also method process like this.
Chemicals treatment:
Each transparent plastic cup places 1 nymph of a locust, cultivates under being placed in normal condition.After 4h, after examination worm has taken food leaf butterfly completely, (raising temperature is 28 ± 1 DEG C, and humidity is 50 ± 10%, 24 h light, feeding fresh corn leaf) is normally fed to investigation to examination worm.The nymph of a locust not taking food whole Zhang Yedie is completely removed.
5. investigate
According to the action character of nosema locustae, 7d, 14d, 21d, 28d investigate after treatment respectively, record total borer population and dead borer population.
6. data statistics and analysis
Computational methods:
According to 5.1 methods in NY/T1154.2-2006, the examination worm calculating each process eats dose and corrected mortality.
Statistical analysis:
The LD of each medicament is calculated with DAS (SPSS)
50and LD
90, b value and 95% confidence interval thereof.
7. result
After nosema locustae process 3 Locusta migratoria manilensis locust nymph in age of variable concentrations, examination worm eats the death condition (as table 1) of dose and the 28th day, calculates 3 Asiatic migratory locust in the age LD of 28 days after inoculation
50be 2.87 × 10
4spore/g, LD
90be 1.56 × 10
6spore/g.
After table 1. processes 28 days, locust micro-spore spore fluid suspending agent and contrast medicament are to the lethality of Asiatic migratory locust
Note: 3 age Locusta migratoria manilensis locust nymph average weight be 7 × 10
-2g/ head; The corrected mortality of variable concentrations process adopts duncan's new multiple range method to compare, and level of difference is P<0.05.
By toxicity determination test, 1 × 10
9spore/mL and l × l0
8the nymph of a locust corrected mortality of spore/mL nosema locustae spore fluid suspending agent process all reaches more than 90%, suitable with the nymph of a locust corrected mortality 96.94% contrasting the process of medicament ethiprole, illustrate with nosema locustae spore fluid suspending agent effective, and illustrate that the activity of the additive of nosema locustae aqueous suspension agent on nosema locustae does not affect.
Embodiment 6 field control effectiveness test
1. test objective
According to NY/T1464.1-2007 " pesticide field efficacy medicine test criterion part 1; Insecticide control migratory locusts " method that specifies; be intended to the 2000 ten thousand spores/milliliter nosema locustae spore fluid suspending agent of clear and definite various dose to the control efficiency of uncultivated area locust and the safety to uncultivated area plant and non-target mark beneficial insect etc.; its suitable dispenser period clear and definite, consumption and method, lay the foundation for applying this agricultural chemicals.
2. experimental condition
The selection of subjects, crop and kind
Subjects: be Asiatic migratory locust LocustamigratoriamanilensisMeyen.
Trial crops: be uncultivated area, main grass seeds is little reed and Herba Aeluropi sinensis etc., based on little reed.
Environment or Protected Cultivation
Experimental field be selected in Zhong Jie farm, Huanghua City, Hebei province to border on the sea barren beach meadow, beach area is large, and mixed have multiple weeds, and vegetation height is at 50-70cm, and physical features is smooth.Be experimental field saline land of bordering on the sea, test site migratory locusts moderate occurs.
3. experimental scheme and arrangement
Medicament:
Test medicine: 2,000 ten thousand spores/milliliter nosema locustae spore fluid suspending agent (China Agricultural University provides); Contrast medicament: 10,000,000,000 spores/gram Metarhizium anisopliae wetting powder (Jiangxi Tianren Eco-Industrial Co., Ltd).
Dosing and numbering:
Table 2. reagent agent experimental scheme
Reagent agent and the process of contrast medicament 4, add blank, amount to 5 process, each process minizone randomized arrangement (table 2,3).
Table 3. is process field, community arrangement pattern respectively
| CK | A | D | C |
| B | B | B | D |
| A | C | C | B |
| D | D | A | CK |
| C | CK | CK | A |
Plot area and repetition:
Plot area: 666.6 square metres.
Number of repetition: 4 times.
Application method:
Using method: spraying, medicament is evenly sprayed on weeds surface.
Spraying equipment: sprayer is the knapsack sprayer (JACTOHD400 type) that Li Nong private limited partnership of Singapore produces.
Spraying time and number of times: afternoon on July 20th, 2013 sprays medicine, altogether dispenser 1 time.
Use capacity: mu liquid 50 kilograms of (750Kg/hm
2).
Prevent and treat the medicament data of other damage by disease and insect: this experimental field before the test and duration of test do not spray other medicament.
4. investigate, record and measuring method
Meteorology and soil information:
Meteorological data: dispenser was cloudy for the same day, mean temperature of air 25 DEG C, the maximum air temperature 31 DEG C, the minimum air temperature 2019 DEG C, relative moisture 30%, 3 grades, southwester.Duration of test is cloudy weather.
Soil information: experimental field soil is strand moisture soil, and meta-alkalescence, soil fertility is poor.
Investigation method, time and number of times:
Control time and number of times: investigation radix before dispenser, after dispenser 7,14,21 days (July 27, August 3, August 10) investigation prevention effect, investigate 4 times altogether.
Investigation method: each community adopts the method for five point samplings, orientation, 5, all directions is distributed.Often many 30 square metres, paste ground with clap net and catch comprehensively.Investigation borer population alive, and record.
Drug effect calculates:
Method represents control efficiency to correct Revision insect recluced rate, and carry out statistical computation according to " pesticide field efficacy medicine test criterion (one) " prescriptive procedure, computing formula is:
Revision insect recluced rate (%)=[borer population of living before (borer population of living after borer population-dispenser of living before dispenser)/dispenser] × 100
Check plot insect population increasing rate (%)=[borer population of living after (borer population of living before borer population-dispenser of living after dispenser)/dispenser] × 100
Control efficiency (%)=[(treatment region Revision insect recluced rate-check plot insect population increasing rate)/(1-check plot insect population increasing rate)] × 100
Direct impact on crop:
Duration of test is observed, and test medicine and contrast medicament respectively process various vegetation growth without obvious impact.
The Quality and yield of product:
The Quality and yield of failed call observation crop.
Other biological is affected
Impact on other damage by disease and insect: do not find the impact of reagent agent test dose on other damage by disease and insect.
Impact on other non-target organisms: do not find that reagent agent test dose has harmful effect to other non-target organisms.
5. results and analysis
Table 4.2000 ten thousand spores/milliliter nosema locustae spore fluid suspending agent control uncultivated area migratory locusts result of the test
Note: the preventive effect (%) in upper table is each repetition mean value.
By Deng Kenshi new multipole difference (DMRT) method to result of the test data analysis.
Evaluating drug: comprehensive above-mentioned result of the test can be found out, 2000 ten thousand spores/milliliter nosema locustae spore fluid suspending agent control uncultivated area migratory locusts control efficiency is better, when using preparation consumption to be 80 milliliters/mu, after dispenser, 21 days control efficiency can reach 83.06%, respectively process higher than other, process with 50 milliliters/mu and the equal significant difference of control efficiency of control treatment, the equal difference of the control efficiency processed with 65 milliliters/mu is not remarkable.
Recommended dose: recommended dose is amount of formulation 65-80 milliliter/mu.
Safety: field observation shows, under dosage used, 2,000 ten thousand spores/milliliter nosema locustae spore fluid suspending agent has no adverse effects to meadow growth, useful Natural Enemies and quantity etc.
Claims (7)
1. a nosema locustae spore fluid suspending agent, is characterized in that, is except containing except active component nosema locustae spore, with 100 parts of listed as parts by weight, also comprises the component of following weight portion: MorwetD425:10-50 part, xanthans: 1-5 part; Sodium lactate: 1-5 part, and the water of surplus.
2. nosema locustae spore fluid suspending agent as claimed in claim 1, it is characterized in that, described nosema locustae spore fluid suspending agent, except containing except active component nosema locustae spore, with 100 parts of listed as parts by weight, also comprise the component of following weight portion: MorwetD425:20-30 part, xanthans: 3-5 part; Sodium lactate: 3-5 part, and the water of surplus.
3. nosema locustae spore fluid suspending agent as claimed in claim 1 or 2, it is characterized in that, the agent of described nosema locustae spore suspension, except containing except active component nosema locustae spore, with 100 parts of listed as parts by weight, also comprise the component of following weight portion: MorwetD425:30 part, xanthans: 4 parts; Sodium lactate: 5 parts, and the water of surplus.
4. the application of the nosema locustae spore fluid suspending agent as described in claim 1-3 any one, is characterized in that, for preventing and treating locust.
5. the application of nosema locustae spore fluid suspending agent as claimed in claim 4, is characterized in that, for preventing and treating the locust of field, grassland, mountain forest.
6. the application of nosema locustae spore fluid suspending agent as claimed in claim 4, is characterized in that, the dosage of described control locust is amount of formulation 65-80 milliliter/mu.
7. the preparation method of the nosema locustae spore fluid suspending agent as described in claim 1-3 any one, is characterized in that, comprise the following steps:
1) take xanthans, add appropriate water, be stirred to xanthans and dissolve completely;
2) in xanthan gum solution, add D425 and sodium lactate, mix, obtain mixed solution;
3) the nosema locustae spore mother liquor of freezen protective is taken out, thaws, shake up, measure mother liquor by allocation ratio and add in mixed solution, then supply water, mix.
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Cited By (3)
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|---|---|---|---|---|
| CN109169645A (en) * | 2018-09-26 | 2019-01-11 | 中国农业大学 | A kind of composite assistant for the micro- spore biological agent of locust |
| CN111657275A (en) * | 2020-07-20 | 2020-09-15 | 北京嘉景生物科技有限责任公司 | Insecticidal fungal spore aqueous suspension and preparation method thereof |
| CN113749091A (en) * | 2021-10-15 | 2021-12-07 | 张昕然 | Locust microsporidia spore water suspending agent and preparation method and application thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109169645A (en) * | 2018-09-26 | 2019-01-11 | 中国农业大学 | A kind of composite assistant for the micro- spore biological agent of locust |
| CN111657275A (en) * | 2020-07-20 | 2020-09-15 | 北京嘉景生物科技有限责任公司 | Insecticidal fungal spore aqueous suspension and preparation method thereof |
| CN113749091A (en) * | 2021-10-15 | 2021-12-07 | 张昕然 | Locust microsporidia spore water suspending agent and preparation method and application thereof |
| CN113749091B (en) * | 2021-10-15 | 2022-10-11 | 张欣杨 | Locust microsporidia spore water suspending agent and preparation method and application thereof |
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