CN103749547B - A kind of entomophthora water floats type granule and application thereof - Google Patents
A kind of entomophthora water floats type granule and application thereof Download PDFInfo
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- CN103749547B CN103749547B CN201310725551.XA CN201310725551A CN103749547B CN 103749547 B CN103749547 B CN 103749547B CN 201310725551 A CN201310725551 A CN 201310725551A CN 103749547 B CN103749547 B CN 103749547B
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Abstract
A kind of entomophthora water floats type granule and application thereof, belongs to technical field of pesticide.It is characterized in that being prepared using following methods：Mixed to Entomophthorales mycelial concentration to add sodium alginate and corn flour in 10 30 mg/mL zymocyte liquid, then mixed solution is inflated 0.5 0.8 hours with air pump, the mixed liquor of bubbles is added dropwise in the calcium chloride solution of concentration 1 1.5% with pipettor again, according to zymocyte liquid：Calcium chloride solution volume ratio 1:The consumption of 10 20 ratio-dependent calcium chloride solution, obtained shaped particles ball sterile water wash produces entomophthora water and floats type granule.A kind of above-mentioned entomophthora water floats type granule, with good spores release ability, spores release amount is big, the production spore duration is long, and production cost is low, to economic insects and people and animals without threat, it is environmentally safe, high lethal effect is produced to rice grub planthopper, the planthopper preventing and treating that can be applied in rice terrace, prevention effect is good.
Description
Technical field
The invention belongs to technical field of pesticide, specially a kind of entomophthora water floats type granule and application thereof.
Background technology
Planthopper is the agricultural pests that a class seriously endangers Rice Production, and main species have brown paddy plant hopper, white backed planthopper and ash
Plant hopper etc., sucks plant juice by sucking mouth parts and can propagate plant virus, causes the paddy rice underproduction or even total crop failure.Planthopper
Migration is strong, can migrate over long distances, and damaging range is wide, and breeding is fast, is also easy to produce the resistance to the action of a drug, is generally acknowledged to be most difficult to having for preventing and treating
One of evil biological group.Such as the nineties in last century once successfully controlled the imidacloprid of planthopper, because abuse causes the resistance to the action of a drug to go out throughout the year
Now ultimate failure, causes insect pest in recent years to break out repeatedly.Meanwhile, the triggered environment of chemical pesticide abuse and food security are asked
Topic becomes increasingly conspicuous, and has aggravated the difficulty of the export of farm produce the problems such as residues of pesticides again.Therefore, non-harmful insect day is developed
Enemy and its obligate pathogenic microorganism prevention rice grub, the need for meeting safety in production and Organic Farming.
Biological control is had been reported for preventing and treating planthopper, but supper toxic strain in the early time with the muscardine and green muscardine fungus in bacterium
Scarcity, is seldom higher than 70% to the fatal rate of planthopper, and lethal more slow, and incubation period is long.On the other hand, such biological and ecological methods to prevent plant disease, pests, and erosion is true
Bacterium host range is relatively wide so that it has the possibility of the economic insects such as silkworm of harm neighborhood cultivation.Entomophthorales is true
Bacterium is that a class host speciality is higher（Host range is narrow）Biocontrol microorganisms, Pandora Delphacis as used in this application only infects plant hopper
With the insect such as aphid, do not report that refer to it has threat to economic insects and people and animals so far.Meanwhile, such fungi has virulence high,
Lethal fast the advantages of, possess potentiality of the exploitation for biological pesticide.
From unlike chemical pesticide, Entomophthorales are not the chemical substances to host by producing with toxic action
Carry out lethal insect, but host is died of hunger by exhausting in pin main body nutrient.Therefore, it does not have residues of pesticides and environmental disruption etc.
Problem.Entomophthora is generally directed through body wall into host's haemocoele internal breeding by conidium, to preventing and treating sucking pest
Such as planthopper has obvious advantage.The conidium that susceptible lethal worm corpse can actively discharge infectivity again is gone around invasion and attack
Host, can induce pest population is suppressed in host's epidemic disease, short time under economic damage level under given conditions.Rice flies
Lice breeding is strong, and a queen can produce up to a hundred ovum, therefore, and insect density is generally higher in field, beneficial to Pandora Delphacis in rice
It is popular in plant hopper population, play the effect for controlling its population density.
Formulation compared to hyphomycetale biocontrol microorganisms such as muscardine is mainly fungal spore formulation, and the conidium of entomophthora is uncomfortable
Close and carry out formulation.At present, the research and development of entomophthora formulation are made as granule generally using mycelia as principal component.Just there are report in foreign countries in the early time
Road develops the pure mycelia formulation of entomophthora and has applied for patent, but pure mycelia agent cost is high, it is impossible to large-scale application.Several years ago,
There are research natural nutrition matrix such as millet or artificial composite particles in the country as mycelia carrier, and corresponding mycelia particle is made
Agent, cost declines to a great extent, and the production spore duration is long, but this kind of granule is not suitable for paddy field environment, because it is sunken under water, nothing
Method produces spore.Based on this, it is necessary to which exploitation is applied to the entomophthora biological control formulation of paddy field environment, to give full play to such biological and ecological methods to prevent plant disease, pests, and erosion
Function of the bacterium in terms of control paddy field insect and reduction agricultural losses.
The content of the invention
For the above-mentioned problems in the prior art, it is an object of the invention to design a kind of entomophthora water to float type granule
And application thereof technical scheme, the granule can be applied in paddy field, gives full play to its biology to rice grub planthopper and prevents
Control function.
A kind of described entomophthora water floats type granule, it is characterised in that prepared using following methods：To Entomophthorales bacterium
Silk concentration is 10-30 mg/mL（Dry weight/v, active ingredient）Zymocyte liquid in add sodium alginate and corn flour and mix, marine alga
The consumption of sour sodium is 10-20 mg/mL（w/v）, the consumption of corn flour is 10-50 mg/mL（w/v）, then with air pump to mixed
Aeration 0.5-0.8 hours is closed, then the mixed liquor of bubbles is added dropwise in concentration 1-1.5% calcium chloride solutions with pipettor,
According to zymocyte liquid：Calcium chloride solution volume ratio 1:The consumption of 10-20 ratio-dependent calcium chloride solution, obtained shaped particles
After ball sterile water wash, surface moisture is dried, entomophthora water is produced and floats type granule.
A kind of described entomophthora water floats type granule, it is characterised in that zymocyte liquid, sodium alginate and corn flour mixing are molten
Liquid is slowly added dropwise in calcium chloride solution by bore 2-3mm pipettor, by the round bead shape particle that calcification is diameter 3-5 mm.
A kind of described entomophthora water floats type granule, it is characterised in that described Entomophthorales mycelial concentration is 15-25
Mg/mL, preferably 18-20mg/mL.
A kind of described entomophthora water floats type granule, it is characterised in that the consumption of sodium alginate is 12-18mg/mL, preferably
14-16mg/mL。
A kind of described entomophthora water floats type granule, it is characterised in that the consumption of corn flour is 20-40 mg/mL, preferably
25-35mg/mL。
A kind of described entomophthora water floats type granule, it is characterised in that the concentration of calcium chloride solution is 1.2-1.4%.
A kind of described entomophthora water floats type granule, it is characterised in that zymocyte liquid：Calcium chloride solution volume ratio 1:12-
18, preferably 1:14-16.
A kind of described entomophthora water floats type granule, it is characterised in that described Entomophthorales are Pandora Delphacis.
A kind of described entomophthora water floats application of the type granule in the preparation for killing rice hoppers is prepared.
The Pandora Delphacis（Pandora delphacis）For the obligate disease fungus of rice grub planthopper, extensively
It is distributed in the Rice Cropping region in East Asia, Southeast Asia, South Asia and America.
Above-mentioned entomophthora water floats type granule, and its composition is liquid phase fermented hypha, sodium alginate, corn flour, wherein zymophyte
Silk is main active ingredient, and corn flour plays nutrition synergistic effect and improves the spores release ability of formulation, sodium alginate tool
There is the ability that particulate matter is formed in calcium chloride solution, above-mentioned entomophthora mycelia and corn flour can be wrapped in particle, sea
Contain a large amount of vacuole structures in the particle that mosanom is formed, buoyancy can be produced in water, particle is bubbled through the water column release spore,
To infect surrounding host planthopper, insecticidal effect is played.Liquid phase fermented hypha, sodium alginate, corn flour are mixed with each other
Synergistic function, and production cost is reduced, apply also in the environment of paddy field.
To make particle to bubble through the water column, the sodium alginate soln inflated air to by using air pump before calcification makes to mix
Close and a large amount of micro-bubbles are full of in liquid, bubble stays in formation vacuole in particle and produces buoyancy after calcification.
A kind of above-mentioned entomophthora water floats type granule, with good spores release ability, and spores release amount is big, and production spore continues
Time is long, and production cost is low, environmentally safe to economic insects and people and animals without threat, rice grub planthopper is produced high
Lethal effect, the planthopper preventing and treating that can be applied in rice terrace, prevention effect is good.
Embodiment
In conjunction with embodiments of the invention, beneficial effects of the present invention are further illustrated.
Need to meet：Fermented hypha concentration is 10-30 mg/mL（W dry weights/v）, sodium alginate（w/v）For 10-20 mg/
ML and 10-50mg/mL（w/v）Corn flour.
Embodiment 1
In the zymocyte liquid 100mL for being 10mg dry weights/mL to mycelial concentration, 1 gram of sodium alginate and 1 gram of commercially available corn are added
Powder.Mix, after air pump inflation half an hour, the mixed liquor of bubbles is added dropwise to 1 L1% with pipettor（w/v）Calcium chloride solution
In, shaped particles ball sterile water wash is standby, obtains 10 mg/mL entomophthora mycelia and adds 10 mg/mL corn flour synergy water to float type
Bead dosage form.
Embodiment 2
In the zymocyte liquid 100mL for being 10 mg dry weights/mL to mycelial concentration, add 1.5 grams of sodium alginates and 5 grams commercially available
Corn flour.Mix, after air pump is inflated 0.6 hour, the mixed liquor of bubbles is added dropwise to 1 L1% with pipettor（w/v）Chlorination
In calcium solution, shaped particles ball sterile water wash is standby, obtains 10mg/mL entomophthora mycelia and adds 50mg/mL corn flour synergy water
Floating type bead dosage form.
Embodiment 3
In the zymocyte liquid 100mL for being 20mg dry weights/mL to mycelial concentration, 1.5 grams of sodium alginates and 1 gram of commercially available jade are added
Ground rice.Mix, after air pump is inflated 0.8 hour, the mixed liquor of bubbles is added dropwise to 1 L 1% with pipettor（w/v）Calcium chloride
In solution, shaped particles ball sterile water wash is standby, obtains 20 mg/mL entomophthora mycelia and adds 10mg/mL corn flour synergy water
Floating type bead dosage form.
Embodiment 4
In the zymocyte liquid 100mL for being 20 mg dry weights/mL to mycelial concentration, 2 grams of sodium alginates and 5 grams of commercially available jade are added
Ground rice.Mix, after air pump inflation half an hour, the mixed liquor of bubbles is added dropwise to 1 L1% with pipettor（w/v）Calcium chloride is molten
In liquid, shaped particles ball sterile water wash is standby, obtains 20 mg/mL entomophthora mycelia and adds 50 mg/mL corn flour synergy water to float
Type bead dosage form.
Embodiment 5
In the mL of zymocyte liquid 100 for being 30 mg dry weights/mL to mycelial concentration, add 1.5 grams of sodium alginates and 1 gram commercially available
Corn flour.Mix, after air pump inflation half an hour, the mixed liquor of bubbles is added dropwise to 1 L 1% with 50mL pipettors（w/v）
In calcium chloride solution, shaped particles ball sterile water wash is standby, obtains 30mg/mL entomophthora mycelia and adds 10 mg/mL corn flour
Synergy water floats type bead dosage form.
Embodiment 6
In the mL of zymocyte liquid 100 for being 30 mg dry weights/mL to mycelial concentration, 2 grams of sodium alginates and 5 grams of commercially available jade are added
Ground rice.Mix, after air pump inflation half an hour, the mixed liquor of bubbles is added dropwise to 1L1% with 50mL pipettors（w/v）Chlorination
In calcium solution, shaped particles ball sterile water wash is standby, obtains 30 mg/mL entomophthora mycelia and adds 50 mg/mL corn flour synergy
Water floats type bead dosage form.
Beneficial effects of the present invention are further illustrated below in conjunction with corresponding experiment.
Experiment one：Production spore determines and killed planthopper experiment and its result
Bead dosage form unit sporulation quantity result of the test
Embodiment 1-6 bead dosage forms, in self-control fungal spore collector（Structure is like 1.8 mL cryopreservation tubes）Interior observation spore
Release conditions.
Material and method
Material to be tested
Embodiment 1-6 kind granules
Odd-numbered day sporulation quantity is determined with production spore total amount：
By granule, one is pasted at the top of spore collectors, and each collector bottom adds the % dodecyls of 1mL 0.5
Sodium sulphate（SDS）Solution.Observation in 24 degrees Celsius of constant incubators is put into, the particle that top is pasted uninterruptedly discharges mitogenetic spore
Son, is fallen into the SDS solution of bottom by Action of Gravity Field spore, is inactivated and is kept profile complete, is easy to count.It is daily to replace bottom
SDS solution.The daily spore suspension collected blood counting chamber counting day sporulation quantity under the microscope.Observation is to replacing
Untill no longer finding spore in the SDS solution changed, the daily output spore amount that adds up calculates the production spore total amount of embodiment 1-6 granules.Each
Embodiment respectively observation 10（Repeat）.Not spores release amount difference unary variance analysis of be the same as Example granule.
Kill planthopper experimental design and method：
Preparation for trying planthopper：The brown paddy plant hopper occurred to rice field then is caught for examination planthopper.Raise in indoor cage
On cover rice seedlings.During insecticidal test, adult 15-20 is moved into there are 10 plants in the high 15cm of diameter 8cm transparent plastic bottle, in bottle
Rice seedlings（Seedling age 2 weeks）Use is taken food as planthopper, while adding 20mL rice nutrition liquid.
Granule is placed：In the plastic bottle that embodiment 1-6 granules are respectively put into access planthopper, 3 every bottle（Equidistantly
Interval）, plastic bottle seals with ventilated membrane.It is put into and daily work borer population and dead borer population is observed under 24 degrees Celsius.Each embodiment is each
Do 3 bottles of repetitions.Another 3 bottles of preparation is not put into granule（CK is compareed）, observe its natural mortality rate.
Insecticidal effect computational methods：Observation was put into after particle the death condition of planthopper in bottle to the 8th day, respectively at the 4th,
5th, 6,7,8 days, according to borer population living and dead borer population in bottle, killing rate is calculated.And school is carried out to the accumulative killing rate of observation the 8th day
Just, corrected mortality is calculated, and carries out significance difference analysis.
Insecticidal effect presses formula（1）、（2）Calculate：
Killing rate or natural mortality rate（%）= （Originate borer population-residue work borer population）/ starting borer population * 100（1）
Corrected mortality（%）=（Killing rate-CK）/（100-CK）*100 （2）
In formula：CK--- planthopper natural mortality rate in control bottle.
Result of the test is with discussing（It is shown in Table 1）
From the point of view of sporulation quantity：Embodiment 1-6 sporulation quantities are in 7.4-15.6 ' 104Individual spore/, the production spore duration is 8-
15 days, production spore was mostly focused on first three day, and first three day sporulation quantity accounts for the 47.3-68.2% of total sporulation quantity.Wherein, the production of embodiment 6 spore
Amount is maximum, and the duration is at most.Killed from this experiment in terms of planthopper effect：Embodiment 1-6 granules have desinsection to planthopper
Effect, susceptible worm corpse occurs from the 4th day, and peak mortality concentrates on the 5th day and the 6th day, accumulative death in the 8th day
Rate is 43.4-84.1%, and corrected mortality is 40.2-83.2%.
Security, the bacterium only infects a small number of arthropods such as planthopper and aphid from previous literature report, other to field
Beneficial insect or people and animals' non-hazardous.And entomophthora does not produce any toxicant during lethal host.
Every group 3 bottles, initial borer population 15-20 heads in each plastic bottle；CK is control group, is not added with granule；Corrected mortality
For the correction result of the 8th day killing rate.
Table 1 shows that mycelial concentration and corn powder content discharge spore to granule in embodiment 1-6（Burst size and continue
On time）Have a significant impact；Table 2 shows that Pandora Delphacis granule has higher insecticidal effect to planthopper, and mycelia is dense
Degree influence is notable, but corn flour influence is not obvious, and its insecticidal effect is only lifted a little；The corrected mortality of embodiment 6 is significantly higher than
Other embodiments.
Claims (10)
1. a kind of entomophthora water floats type granule, it is characterised in that prepared using following methods：It is to Entomophthorales mycelial concentration
Sodium alginate is added in 10-30 mg/mL zymocyte liquid and corn flour is mixed, the consumption of sodium alginate is 10-20 mg/mL,
The consumption of corn flour is 10-50 mg/mL, and then mixed solution is inflated 0.5-0.8 hours with air pump, then will with pipettor
The mixed liquor of bubbles is added dropwise in concentration 1-1.5% calcium chloride solutions, according to zymocyte liquid：Calcium chloride solution volume ratio 1:10-
After the consumption of 20 ratio-dependent calcium chloride solution, obtained shaped particles ball sterile water wash, surface moisture is dried, is produced
Entomophthora water floats type granule；Zymocyte liquid, sodium alginate and corn flour mixed solution are slowly dripped by bore 2-3mm pipettor
Add in calcium chloride solution, by the round bead shape particle that calcification is the mm of diameter 3 ~ 5；Described Entomophthorales are Pandora Delphacis.
2. a kind of entomophthora water as claimed in claim 1 floats type granule, it is characterised in that described Entomophthorales mycelia is dense
Spend for 15-25 mg/mL.
3. a kind of entomophthora water as claimed in claim 1 floats type granule, it is characterised in that described Entomophthorales mycelia is dense
Spend for 18-20mg/mL.
4. a kind of entomophthora water as claimed in claim 1 floats type granule, it is characterised in that the consumption of sodium alginate is 12-
18mg/mL。
5. a kind of entomophthora water as claimed in claim 1 floats type granule, it is characterised in that the consumption of sodium alginate is 14-
16mg/mL。
6. a kind of entomophthora water as claimed in claim 1 floats type granule, it is characterised in that the consumption of corn flour is 20-40 mg/
mL。
7. a kind of entomophthora water as claimed in claim 1 floats type granule, it is characterised in that the consumption of corn flour is 25-35mg/
mL。
8. a kind of entomophthora water as claimed in claim 1 floats type granule, it is characterised in that the concentration of calcium chloride solution is 1.2-
1.4%。
9. a kind of entomophthora water as claimed in claim 1 floats type granule, it is characterised in that zymocyte liquid：Calcium chloride solution volume
Than 1:12-18.
10. a kind of entomophthora water as claimed in claim 1 floats type granule, it is characterised in that zymocyte liquid：Calcium chloride solution body
Product compares 1:14-16.
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