CN109938046A - Downy mildew of garpe biochemistry Collaborative Control medicament and its application - Google Patents
Downy mildew of garpe biochemistry Collaborative Control medicament and its application Download PDFInfo
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- CN109938046A CN109938046A CN201910312379.2A CN201910312379A CN109938046A CN 109938046 A CN109938046 A CN 109938046A CN 201910312379 A CN201910312379 A CN 201910312379A CN 109938046 A CN109938046 A CN 109938046A
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Abstract
The invention discloses downy mildew of garpe biochemistry Collaborative Control medicament and its applications.The medicament contains Propamocarb and Oudemansiella radicata protein extract；The preparation method of Oudemansiella radicata protein extract includes: to be collected water-soluble substances with flooding after crushing long root mushroom fructification, obtained long root mushroom water-soluble extractive；The ammonium sulfate precipitation that saturation degree is 80% is carried out to long root mushroom water-soluble extractive and collects precipitating, after precipitating deionized water dialysis, obtains Oudemansiella radicata protein extract.Propamocarb and Oudemansiella radicata protein extract are respectively 0.21 and 0.07mg/L to the EC50 of Plasmopara viticola, Propamocarb and Oudemansiella radicata protein extract produce synergistic effect according to the mass ratio mixture of 21:490 and 21:90, and the EC50 to Plasmopara viticola is 0.05mg/L.The advantages of present invention is reduced chemical agent usage amount in terms for the treatment of and preventing downy mildew of garpe.
Description
Technical field
The present invention relates to the prevention and treatment of downy mildew of garpe in field of biotechnology, in particular to downy mildew of garpe biochemistry is assisted
With control medicament and its application.
Background technique
Downy mildew of garpe is a kind of global grape disease.Downy mildew of garpe main harm blade, can also infect young sprout
The tender tissues such as young fruit.Blade is killed, and the clear fleck of translucent, water stain shape, blur margin is initially generated on blade face, after by
Flaring is greatly faint yellow to yellowish-brown polygonal scab, and size shape is different, and several scabs connect together sometimes, forms yellowish-brown
Withered large-scale scab.The scab back side generates white mustiness object (sporophore and sporangium of pathogen) when there was dampness in the air.It is sick afterwards
Spot dry up it is brown, sick leaf easily ahead of time fall off.Tender tip, tendril, petiole, flower spike stalk are susceptible, and scab is just translucent water stain shape spot
Point, after be gradually expanded, scab in yellowish-brown to brown, it is slightly concave fall into, it is mould that sparse white is generated when air humidity is big, on scab
Shape object, sick tip growth stop, and distortion is withered when serious.Young fruit is susceptible, and scab subcircular is in celadon, and surface has white mould
Shape object, rear shrinkage fall off, and fruit is susceptible after growing up, and are generally not formed mustiness object.Cob is susceptible, can cause part fruit ear or entire
Fruit ear falls off.
Downy mildew of garpe is by oomycota, Oomycete, Peronosporales Plasmopara viticola (Berk.dt
Curtis caused by) Berl.Et de Toni. infects, which is obligate parasite.The bacterium mycelium without every multicore,
It is grown between host cell, gos deep into drawing nutrient in host cell with spherical haustorium.Germ is mainly got in fallen leaves with egg spore
In the winter, in warm winter area, the mycelium being attached on bud and in the blade hung on the tree also can be overwintering.
The control method of downy mildew of garpe mainly has physical control and chemical prevention method at present.Physical control is to remove
Cause of disease and reinforcement orchard management.Chemical prevention method is using absorbabilities such as flumorph, aliette, dimethomorph, Propamocarb, cymoxanils
The prevention and treatment of fungicide progress downy mildew.Long-term a large amount uses chemical pesticide in grape production, causes a series of problems, such as cause of disease
Bacterium/disease pest drug resistance increases, and orchard worker has to the usage amount for increasing pesticide, and pesticide cost increases；Fruit agriculture is residual exceeded, environment
Pollution risk increases；Non-target organism is aggrieved and the problems such as chronic toxicity, therefore highly-safe, residual is badly in need of in grape production
Measure low, non-harmful pest and disease damage Control Technology.
Summary of the invention
The technical problem to be solved by the present invention is to how reduce prevention and/or treat the chemical agent in downy mildew of garpe
Usage amount.
In order to solve the above technical problems, the present invention provides a kind of preparations.
Preparation provided by the present invention contains Propamocarb and Oudemansiella radicata protein extract；The long root mushroom Protein Extraction
Object is prepared according to the method included the following steps:
1) flooding is used after crushing long root mushroom fructification, is collected water-soluble substances, is obtained long root mushroom water-soluble extractive；
2) ammonium sulfate precipitation that saturation degree is 80% is carried out to the long root mushroom water-soluble extractive, precipitating is collected, to institute
After stating precipitating deionized water dialysis, the Oudemansiella radicata protein extract is obtained；
The preparation is downy mildew of garpe bacteria inhibitor, or to prevent and/or treating the medicament of downy mildew of garpe.
In above-mentioned preparation, the mass ratio of Propamocarb and the Oudemansiella radicata protein extract can be 21:490, the long root
The quality of mushroom protein extract is in terms of the quality of protein.
In above-mentioned preparation, the mass ratio of Propamocarb and the Oudemansiella radicata protein extract can be 21:90, the long root mushroom
The quality of protein extract is in terms of the quality of protein.
The application of Propamocarb and long root mushroom in preparation prevention and/or treatment downy mildew of garpe medicament also belongs to of the invention
Protection scope.
The application of Propamocarb and the Oudemansiella radicata protein extract in prevention and/or treatment downy mildew of garpe also belongs to
Protection scope of the present invention.
The application of Propamocarb and long root mushroom in preparation downy mildew of garpe bacteria inhibitor also belongs to protection scope of the present invention.
Propamocarb and the Oudemansiella radicata protein extract are inhibiting the application in Plasmopara viticola to also belong to the present invention
Protection scope.
Following any purposes of the Oudemansiella radicata protein extract also belong to protection scope of the present invention:
M1. application of the Oudemansiella radicata protein extract in preparation prevention and/or treatment downy mildew of garpe medicament；
M2. the application in downy mildew of garpe is being prevented and/or treated to the Oudemansiella radicata protein extract；
M3. the application in the Oudemansiella radicata protein extract preparation downy mildew of garpe bacteria inhibitor；
M4. the Oudemansiella radicata protein extract is inhibiting the application in Plasmopara viticola.
Following any purposes of long root mushroom also belong to protection scope of the present invention:
N1. application of the long root mushroom in preparation prevention and/or treatment downy mildew of garpe medicament；
N2. long root mushroom is preventing and/or is treating the application in downy mildew of garpe；
N3. application of the long root mushroom in preparation downy mildew of garpe bacteria inhibitor；
N4. long root mushroom is inhibiting the application in Plasmopara viticola.
Above, the work of the medicament and the downy mildew of garpe bacteria inhibitor of the prevention and/or treatment downy mildew of garpe
Property ingredient can be Propamocarb and the Oudemansiella radicata protein extract.The medicament of the prevention and/or treatment downy mildew of garpe
Active constituent can also contain other components, and described other component those skilled in the art can be according to prevention and/or treatment oenin
The effect of mildew determines.The active constituent of the downy mildew of garpe bacteria inhibitor can also contain other components, other components
Those skilled in the art can determine according to the inhibitory effect to Plasmopara viticola.
The medicament and the downy mildew of garpe bacteria inhibitor of the prevention and/or treatment downy mildew of garpe may also include load
Body.The carrier can be micro mist CaCO3, turf, clay, talcum, kaolin, montmorillonite, white carbon, zeolite, silica, diatomite and
At least one of water.The agent of the medicament and the downy mildew of garpe bacteria inhibitor of above-mentioned prevention and/or treatment downy mildew of garpe
Type can be a variety of dosage forms, such as liquor, emulsion, suspending agent, pulvis, granule, wettable powder or water dispersible granules.
Above-mentioned steps 1) in, the use flooding can be at 2-6 DEG C with flooding 10-14 hours.The water can be from
Sub- water.
The long root mushroom fructification also can be dry fructification for fresh fructification.The fructification of the drying is
Fresh long root mushroom fructification is dried to obtain under room temperature (such as 20-25 DEG C).
The fresh long root mushroom fructification and the volume ratio of water can be 1:40-60, such as 1:40.
Above-mentioned steps 1) in, it can be used and the water-soluble substances are collected by centrifugation.Be collected by centrifugation the water-soluble substances from
Mental and physical efforts can be 6000-15000g (such as 12000g), and centrifugation time can be 10-20 minutes (such as 15 minutes).Above-mentioned steps 2) in,
It can be used and the precipitating is collected by centrifugation.The centrifugal force that the precipitating is collected by centrifugation can be 6000-15000g (such as 12000g), centrifugation
Time can be 10-20 minutes (such as 15 minutes).
Above-mentioned steps 2) in, the sulfuric acid ammonia-sinking that saturation degree is 80% is carried out to the long root mushroom water-soluble extractive at 4 DEG C
It forms sediment.
Above-mentioned steps 2) in, it is described to use molecular cut off to carry out for 3kDa semi-permeable membrane with deionized water dialysis.
Above-mentioned preparation method further includes liquid in the semi-permeable membrane after dialysing in 3000-9000g (such as 6000g), centrifugation
10-20 minutes (such as 15 minutes), supernatant is collected, which is freeze-dried, long root mushroom Protein Extraction is prepared into
The step of object dry powder.
Experiments have shown that Propamocarb is 0.21mg/L, Oudemansiella radicata protein extract oenin to the EC50 of Plasmopara viticola
The EC50 of mildew bacterium is 0.07mg/L, and Propamocarb has significantly Plasmopara viticola with after Oudemansiella radicata protein extract mixture
Synergistic effect: the mass ratio of Propamocarb and protein is the Oudemansiella radicata protein extract of 21:490 and the Mixed Pharmacy of Propamocarb
Poison is altogether with the Mixed Pharmacy of Oudemansiella radicata protein extract and Propamocarb that the mass ratio of Propamocarb and protein is 21:90
Number is respectively 145.83 and 164.06.Illustrate Propamocarb and Oudemansiella radicata protein extract according to Propamocarb and long root mushroom protein
Protein quality in extract is than producing synergistic effect for the ratio mixture of 21:490 and 21:90.Propamocarb and protein
Mass ratio be 21:490 Oudemansiella radicata protein extract and Propamocarb Mixed Pharmacy and Propamocarb and protein quality
There is significant inhibit to Plasmopara viticola than the Mixed Pharmacy of Oudemansiella radicata protein extract and Propamocarb for 21:90
Effect, the EC50 to Plasmopara viticola is 0.05mg/L, much smaller than Propamocarb to the EC50 of Plasmopara viticola.
The present invention is reduced chemical agent usage amount in terms for the treatment of and preventing downy mildew of garpe and guarantees fruit security quality
The advantages of.
Detailed description of the invention
Fig. 1 is the downy mildew of garpe disease leaf photo of field acquisition.
Fig. 2 is Plasmopara viticola to the quick of Oudemansiella radicata protein extract, Oudemansiella radicata protein extract and Propamocarb
Perceptual photo.
Specific embodiment
The present invention is further described in detail With reference to embodiment, and the embodiment provided is only for explaining
The bright present invention, the range being not intended to be limiting of the invention.Examples provided below can be used as the art ordinary skill
The guide that personnel are further improved, is not construed as limiting the invention in any way.
Experimental method in following embodiments is unless otherwise specified conventional method.Material as used in the following examples
Material, reagent etc., are commercially available unless otherwise specified.
Long root mushroom in following embodiments is long root mushroom (Oudemansiella radicata) (also known as long root mushroom
(Oudemansiella radicata (Relhan) Singer)) Chinese microorganism strain has been concealed on April 26th, 2013
Preservation administration committee forestry microorganism center (also known as China Forest Microbiological Culture Collection administrative center, China
Forestry Culture Collection Center, english abbreviation CFCC, abbreviation forestry microorganism center), deposit number is
CFCC 89567, the public can obtain the bacterial strain from CFCC from the collection day.
Clitocybe Maxima in following embodiments is Clitocybe Maxima (Clitocybe maxima) (also known as Clitocybe maxima
(P.Gaertn., G.Mey.&Scherb.) P.Kumm.) Chinese microorganism strain preservation pipe has been concealed on April 26th, 2013
Reason committee forestry microorganism center (also known as China Forest Microbiological Culture Collection administrative center, China Forestry
Culture Collection Center, english abbreviation CFCC, abbreviation forestry microorganism center), deposit number CFCC
89534, the public can obtain the bacterial strain from CFCC from the collection day.
Embodiment 1, Oudemansiella radicata protein extract and Propamocarb and Oudemansiella radicata protein extract mixture are to oenin
The inhibiting effect of mildew bacterium
1, the preparation of edible fungus protein matter extract
1.1, the preparation of entitled Oudemansiella radicata protein extract
It is impregnated long root mushroom fructification 2 hours of freeze-drying with the deionized water of 40 times of volumes, it will using tissue mashing machine
Mixture carries out historrhexis to paste, and after 4 DEG C of extractions (stand) 12h, 12000g is centrifuged 15min, collects supernatant, should
Supernatant is long root mushroom water-soluble extractive.
(NH is added into long root mushroom water-soluble extractive at 4 DEG C4)2SO4To (NH4)2SO4Saturation degree be 80%, in 4 DEG C
Under the conditions of stand 4 hours, 12000g be centrifuged 15min, collect precipitating, dialyse to the precipitating.Wherein, it dialyses the semi-transparent of use
The molecular cut off of film is 3kDa, is deposited in semi-permeable membrane in the tap water of flowing the 5h that dialyses, then dialyse in deionized water
12h.Liquid in semi-permeable membrane after dialysis is centrifuged 15min in 6000g, supernatant is collected, which is placed in liquid nitrogen
Freezing, and be freeze-dried 36 hours, protein powder is obtained, is ground into powder, which is Oudemansiella radicata protein extract.
The protein content of the Oudemansiella radicata protein extract is measured with BCA method, the results showed that the egg of the Oudemansiella radicata protein extract
White matter content is 80.15% (mass percentage).
1.2, the preparation of entitled Clitocybe Maxima protein extract
It is impregnated mug destroying angel 2 hours of freeze-drying with the deionized water of 40 times of volumes, it will using tissue mashing machine
Mixture carries out historrhexis to paste, and after 4 DEG C of extractions (stand) 12h, 12000g is centrifuged 15min, collects supernatant, should
Supernatant is Clitocybe Maxima water-soluble extractive.
(NH is added into Clitocybe Maxima water-soluble extractive at 4 DEG C4)2SO4To (NH4)2SO4Saturation degree be 80%, in 4 DEG C
Under the conditions of stand 4 hours, 12000g be centrifuged 15min, collect precipitating, dialyse to the precipitating.Wherein, it dialyses the semi-transparent of use
The molecular cut off of film is 3kDa, is deposited in semi-permeable membrane in the tap water of flowing the 5h that dialyses, then dialyse in deionized water
12h.Liquid in semi-permeable membrane after dialysis is centrifuged 15min in 6000g, supernatant is collected, which is placed in liquid nitrogen
Freezing, and be freeze-dried 36 hours, protein powder is obtained, is ground into powder, which is Clitocybe Maxima protein extract.
The effective component (active constituent) of the Clitocybe Maxima protein extract is protein.The Clitocybe Maxima protein is measured with BCA method to mention
Take the protein content of object, the results showed that the protein content of the Clitocybe Maxima protein extract is that 73.17% (quality percentage contains
Amount).
2, the configuration of edible fungus protein matter solution
2.1, the Oudemansiella radicata protein extract solution that protein concentration is respectively 5,50,500 μ g/mL is prepared
Take 1.1 Oudemansiella radicata protein extract (protein content 80.15%) 18.71mg in centrifuge tube, every pipe
30mL sterile water is added, concussion several times, is placed in 25 DEG C of water-bath 3min, centrifuge tube is taken out after proteolytic, obtains protein
(Oudemansiella radicata protein extract of referred to as 500 μ g/mL is molten for the Oudemansiella radicata protein extract solution that concentration is 500 μ g/mL
Liquid).
Taking protein concentration is that the Oudemansiella radicata protein extract solution 3mL of 500 μ g/mL adds 27mL sterile water to be made into 30mL
Protein concentration is Oudemansiella radicata protein extract solution (the referred to as Oudemansiella radicata protein extract of 50 μ g/mL of 50 μ g/mL
Solution).
Taking protein concentration is that the Oudemansiella radicata protein extract solution 3mL of 50 μ g/mL adds 27mL sterile water to be made into 30mL
(Oudemansiella radicata protein extract of referred to as 5 μ g/mL is molten for the Oudemansiella radicata protein extract solution that protein concentration is 5 μ g/mL
Liquid).
2.2, the Clitocybe Maxima protein extract solution that protein concentration is respectively 5,50,500 μ g/mL is prepared
Take 1.2 Clitocybe Maxima protein extract (protein percentage 73.17%) 20.5mg in centrifuge tube, often
30mL sterile water is added in pipe, and concussion several times, is placed in 25 DEG C of water-bath 3min, centrifuge tube is taken out after proteolytic, obtains albumen
(the Clitocybe Maxima protein extract of referred to as 500 μ g/mL is molten for the Clitocybe Maxima protein extract solution that matter concentration is 500 μ g/mL
Liquid).
Taking protein concentration is that the Clitocybe Maxima protein extract solution 3mL of 500 μ g/mL adds 27mL sterile water to be made into 30mL
(the Clitocybe Maxima protein extract of referred to as 50 μ g/mL is molten for the Clitocybe Maxima protein extract solution that protein concentration is 50 μ g/mL
Liquid).
Taking protein concentration is that the Clitocybe Maxima protein extract solution 3mL of 50 μ g/mL adds 27mL sterile water to be made into 30mL
(the Clitocybe Maxima protein extract of referred to as 5 μ g/mL is molten for the Clitocybe Maxima protein extract solution that protein concentration is 5 μ g/mL
Liquid).
3, the Propamocarb solution that Propamocarb concentration is 1,5,10 μ g/mL is prepared
(1) preparation of mother liquor: 103 μ L Propamocarb stostes is taken to be configured to 97% solution plus the acetone of 9.897mL；The mother
The concentration of Propamocarb is 1 × 10 in liquid4μg/mL。
(2) it the preparation of dilution: takes the mother liquor of 0.5mL to add the acetone of 4.5mL, obtains the frost of the 1000 μ g/mL of 5mL
Mould prestige solution.
(3) the Propamocarb solution that Propamocarb concentration is 1,5,10 μ g/mL is prepared
Taking the Propamocarb solution 0.4mL of 1000 μ g/mL to be made into 40mL concentration plus the sterile water of 39.6mL is 10 μ g/mL's
Propamocarb solution；
The Propamocarb solution 20mL of 10 μ g/mL is taken to be made into the Propamocarb that 40mL concentration is 5 μ g/mL plus the sterile water of 20mL
Solution；
The Propamocarb solution 8mL of 5 μ g/mL is taken plus the sterile water of 32mL to be made into the Propamocarb that 40mL concentration is 1 μ g/mL molten
Liquid.
4, host and cause of disease
The acquisition of 4.1 grape healthy leaves
In in the vineyard in Yunnan Province, the county Xun Dian, Kunming Yunnan Prov Agriculture University's modern education base, acquisition kind is red
The grape healthy leaves of the earth are used for the correlative study of downy mildew of garpe.
The acquisition and separation of 4.2 P. viticolas
8~September in 2013 is in the vineyard in the area such as Yunnan Prov Agriculture University's modern education base, base periphery vineyard
Middle acquisition downy mildew of garpe disease leaf.Known to field observation result: blade catch an illness just be translucent, the pistac that blur margin is clear
Oil immersion shape spot, the rear yellowly that extends is to brown polygonal spot.When humidity is big, the back side generates the mould layer of white, the i.e. sporangiocyst of germ
Stalk and sporangium, the last browning of scab, blade dry up (Fig. 1).Each vineyard use the sampling of 5 methods with reduce collect it is identical
The probability of bacterial strain.The downy mildew of garpe leaf freshness protection package moisturizing of acquisition, being put in temperature is 21 DEG C, relative humidity 90-
100 ﹪, moisturizing culture in the incubator of illumination 16h/ dark 8h are spare after growing fresh mycelia again after blade.
It is 1 × 10 that downy mildew fresh after culture, which is configured to concentration in micro- sem observation, with 4 DEG C of sterile waters4A spore
Spore suspension can be used for follow-up test by son/ml downy mildew spore suspension.
5, Plasmopara viticola is sensitive to Oudemansiella radicata protein extract, Clitocybe Maxima protein extract and Propamocarb respectively
The foundation of baseline
5.1 chemicals treatment
3-4 leaf phase leaf age, the size red earth blade of consistent healthy grape as far as possible are selected, after aseptic water washing, by 3-5
The overlapping of piece leaf, the leaf dish of ф 15mm is produced with punch, and all leaf dishes is mixed at random spare.It tests in triplicate, often
It is secondary that leaf dish is divided into four groups: Oudemansiella radicata protein extract group, Clitocybe Maxima protein extract group, Propamocarb group and control group.
Oudemansiella radicata protein extract group sets the long root mushroom egg of the Oudemansiella radicata protein extract solution processing of 500 μ g/mL, 50 μ g/mL
The Oudemansiella radicata protein extract solution of the processing of white matter extract solution and 5 μ g/mL handle these three processing, each processing 15
Leaf dish；Clitocybe Maxima protein extract group sets the mug of the Clitocybe Maxima protein extract solution processing of 500 μ g/mL, 50 μ g/mL
The Clitocybe Maxima protein extract solution of the processing of umbrella protein extract solution and 5 μ g/mL handle these three processing, each processing
15 leaf dishes；Propamocarb group sets the frost of the Propamocarb solution processing of 10 μ g/mL, Propamocarb the solution processing and 1 μ g/mL of 5 μ g/mL
Mould prestige solution handles these three processing, 15 leaf dishes of each processing.
It is molten to the Oudemansiella radicata protein extract solution of 500 μ g/mL, the Oudemansiella radicata protein extract of 50 μ g/mL respectively
Polysorbas20 is added in the Oudemansiella radicata protein extract solution of liquid and 5 μ g/mL, makes the 0.1 μ g/mL of concentration of polysorbas20, obtains
Oudemansiella radicata protein extract group treatment fluid；
It is molten to the Clitocybe Maxima protein extract solution of 500 μ g/mL, the Clitocybe Maxima protein extract of 50 μ g/mL respectively
Polysorbas20 is added in the Clitocybe Maxima protein extract solution of liquid and 5 μ g/mL, makes the 0.1 μ g/mL of concentration of polysorbas20, obtains
Clitocybe Maxima extract group treatment fluid；
Add respectively into the Propamocarb solution of the Propamocarb solution of 10 μ g/mL, the Propamocarb solution of 5 μ g/mL and 1 μ g/mL
Enter polysorbas20, makes the 0.1 μ g/mL of concentration of polysorbas20, obtain Propamocarb group treatment fluid.
Control treatment liquid is to use sterile water as solvent, and polysorbas20 is 0.1 μ g/mL as the concentration that solute prepares polysorbas20
Solution.
It is put into 15 leaf dishes in each culture dish lid, the medicine of 10mL corresponding concentration is separately added into according to the concentration of label
Agent carries out blade immersion treatment 20 minutes, completes chemicals treatment.It is face-up when immersion, guarantee that medicament immerses blade back.
The inoculation and culture of 5.2 Plasmopara viticolas
The medical fluid in the leaf dish by 5.1 chemicals treatment is blotted after immersion, blade back is placed in upward with identical medicine
It is then 1 × 10 by 4.2 concentration with sessile drop method on the blotting paper of agent concentration wetting4The downy mildew spore suspension of a spore/ml
Liquid is inoculated into the excised leaf back side of health, and each leaf dish is inoculated with 20 μ L downy mildew spore suspensions (1 × 104A spore/mL),
Leaf dish after inoculation is placed in 19 DEG C of illumination boxs dark culturing for 24 hours, to ensure the intrusion of pathogen, then 19
DEG C, moisturizing culture in the illumination box of 12h alternation of light and darkness, until starting to fall ill.Then the morbidity face in leaf dish is continuously monitored
Product calculates control efficiency and EC50.With this method measurement Plasmopara viticola to Oudemansiella radicata protein extract, Clitocybe Maxima albumen
The sensitivity testing of matter extract and Propamocarb, and establish base-line sensitivities.
The investigation of 5.3 results and statistical analysis
5.3.1 investigation method
Using square method after the onset of culture, (each grid area is 4mm2) measurement leaf dish on onset area, according to following
Formula calculates Oudemansiella radicata protein extract, Clitocybe Maxima protein extract and Propamocarb to the inhibiting rate of Plasmopara viticola.
5.3.2 data statistic analysis
Inhibiting rate is converted to probit value (y), concentration is converted to logarithm (x), finds out each medicament to Plasmopara viticola
Toxicity regression curve equation y=a+bx, and effectively inhibit in concentration EC50 value and correlation coefficient r.
The result shows that Plasmopara viticola has Propamocarb, Oudemansiella radicata protein extract and Clitocybe Maxima protein extract
There is significant sensibility, be mainly shown as that disease incidence significantly reduces, disease index significantly reduces, and disease resistance enhances (Fig. 2).Frost
Mould prestige is 0.21mg/L to the EC50 of Plasmopara viticola, and Oudemansiella radicata protein extract is to the EC50 of Plasmopara viticola
0.07mg/L, Clitocybe Maxima protein extract are 0.05mg/L (table 1) to the EC50 of Plasmopara viticola.Plasmopara viticola pair
Oudemansiella radicata protein extract and the sensibility of Clitocybe Maxima protein extract are above Propamocarb.
Table 1, Plasmopara viticola are quick to Oudemansiella radicata protein extract, Clitocybe Maxima protein extract and Propamocarb respectively
Perceptual measurement result
6. Propamocarb is respectively with Oudemansiella radicata protein extract, Clitocybe Maxima protein extract mixture to Plasmopara viticola
Indoor joint toxicity measuring
Referring to the method for Schepers (1984) and Ishii (2001) etc., Plasmopara viticola is carried out to frost with leaf disk method
The toxicity test of mould prestige and Oudemansiella radicata protein extract, Clitocybe Maxima protein extract.
6.1, the Oudemansiella radicata protein extract solution that protein concentration is respectively 5,50,500 μ g/mL is prepared
With 2.1.
6.2, the Clitocybe Maxima protein extract solution that protein concentration is respectively 5,50,500 μ g/mL is prepared
With 2.2.
6.3, the Propamocarb solution that Propamocarb concentration is 1,5,10 μ g/mL is prepared
With 3.
6.4, the Mixed Pharmacy solution of Propamocarb and Oudemansiella radicata protein extract is prepared
6.4.1 the concentration that the mass ratio for preparing Propamocarb and protein is 21:490 be respectively 3.5mg/L, 35mg/L and
The Oudemansiella radicata protein extract of 350mg/L and the Mixed Pharmacy solution of Propamocarb
The Propamocarb solution of the Oudemansiella radicata protein extract solution of 5 μ g/mL and 1 μ g/mL, which is mixed to get concentration, is
The Oudemansiella radicata protein extract of 3.5mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the long root mushroom albumen of 3.5mg/L
In the Mixed Pharmacy solution of matter extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:490.
The Propamocarb solution of the Oudemansiella radicata protein extract solution of 50 μ g/mL and 5 μ g/mL, which is mixed to get concentration, is
The Oudemansiella radicata protein extract of 35mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the long root mushroom protein of 35mg/L
In the Mixed Pharmacy solution of extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:490.
The Propamocarb solution of the Oudemansiella radicata protein extract solution of 500 μ g/mL and 10 μ g/mL, which is mixed to get concentration, is
The Oudemansiella radicata protein extract of 350mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the long root mushroom albumen of 350mg/L
In the Mixed Pharmacy solution of matter extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:490.
Wherein, the effective component (active constituent) of each Mixed Pharmacy is protein and Propamocarb, each Mixed Pharmacy solution
Concentration be contained protein in Oudemansiella radicata protein extract concentration and the sum of concentration of Propamocarb.
6.4.2 the concentration that the mass ratio for preparing Propamocarb and protein is 21:315 be respectively 3.0mg/L, 30mg/L and
The Oudemansiella radicata protein extract of 300mg/L and the Mixed Pharmacy solution of Propamocarb
The Propamocarb solution of the Oudemansiella radicata protein extract solution of 5 μ g/mL and 1 μ g/mL, which is mixed to get concentration, is
The Oudemansiella radicata protein extract of 3.0mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the long root mushroom albumen of 3.0mg/L
In the Mixed Pharmacy solution of matter extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:315.
The Propamocarb solution of the Oudemansiella radicata protein extract solution of 50 μ g/mL and 5 μ g/mL, which is mixed to get concentration, is
The Oudemansiella radicata protein extract of 30mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the long root mushroom protein of 30mg/L
In the Mixed Pharmacy solution of extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:315.
The Propamocarb solution of the Oudemansiella radicata protein extract solution of 500 μ g/mL and 10 μ g/mL, which is mixed to get concentration, is
The Oudemansiella radicata protein extract of 300mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the long root mushroom albumen of 300mg/L
In the Mixed Pharmacy solution of matter extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:315.
Wherein, the effective component (active constituent) of each Mixed Pharmacy is protein and Propamocarb, each Mixed Pharmacy solution
Concentration be contained protein in Oudemansiella radicata protein extract concentration and the sum of concentration of Propamocarb.
6.4.3 the concentration that the mass ratio for preparing Propamocarb and protein is 21:210 be respectively 2.5mg/L, 25mg/L and
The Oudemansiella radicata protein extract of 250mg/L and the Mixed Pharmacy solution of Propamocarb
The Propamocarb solution of the Oudemansiella radicata protein extract solution of 5 μ g/mL and 1 μ g/mL, which is mixed to get concentration, is
The Oudemansiella radicata protein extract of 2.5mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the long root mushroom albumen of 2.5mg/L
In the Mixed Pharmacy solution of matter extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:210.
The Propamocarb solution of the Oudemansiella radicata protein extract solution of 50 μ g/mL and 5 μ g/mL, which is mixed to get concentration, is
The Oudemansiella radicata protein extract of 25mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the long root mushroom protein of 25mg/L
In the Mixed Pharmacy solution of extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:210.
The Propamocarb solution of the Oudemansiella radicata protein extract solution of 500 μ g/mL and 10 μ g/mL, which is mixed to get concentration, is
The Oudemansiella radicata protein extract of 250mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the long root mushroom albumen of 250mg/L
In the Mixed Pharmacy solution of matter extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:210.
Wherein, the effective component (active constituent) of each Mixed Pharmacy is protein and Propamocarb, each Mixed Pharmacy solution
Concentration be contained protein in Oudemansiella radicata protein extract concentration and the sum of concentration of Propamocarb.
6.4.4 the concentration that the mass ratio for preparing Propamocarb and protein is 21:140 be respectively 2.0mg/L, 20mg/L and
The Oudemansiella radicata protein extract of 200mg/L and the Mixed Pharmacy solution of Propamocarb
The Propamocarb solution of the Oudemansiella radicata protein extract solution of 5 μ g/mL and 1 μ g/mL, which is mixed to get concentration, is
The Oudemansiella radicata protein extract of 2.0mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the long root mushroom albumen of 2.0mg/L
In the Mixed Pharmacy solution of matter extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:140.
The Propamocarb solution of the Oudemansiella radicata protein extract solution of 50 μ g/mL and 5 μ g/mL, which is mixed to get concentration, is
The Oudemansiella radicata protein extract of 20mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the long root mushroom protein of 20mg/L
In the Mixed Pharmacy solution of extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:140.
The Propamocarb solution of the Oudemansiella radicata protein extract solution of 500 μ g/mL and 10 μ g/mL, which is mixed to get concentration, is
The Oudemansiella radicata protein extract of 200mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the long root mushroom albumen of 200mg/L
In the Mixed Pharmacy solution of matter extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:140.
Wherein, the effective component (active constituent) of each Mixed Pharmacy is protein and Propamocarb, each Mixed Pharmacy solution
Concentration be contained protein in Oudemansiella radicata protein extract concentration and the sum of concentration of Propamocarb.
6.4.5 the concentration that the mass ratio for preparing Propamocarb and protein is 21:90 be respectively 1.5mg/L, 15mg/L and
The Oudemansiella radicata protein extract of 150mg/L and the Mixed Pharmacy solution of Propamocarb
The Propamocarb solution of the Oudemansiella radicata protein extract solution of 5 μ g/mL and 1 μ g/mL, which is mixed to get concentration, is
The Oudemansiella radicata protein extract of 1.5mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the long root mushroom albumen of 1.5mg/L
In the Mixed Pharmacy solution of matter extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:90.
The Propamocarb solution of the Oudemansiella radicata protein extract solution of 50 μ g/mL and 5 μ g/mL, which is mixed to get concentration, is
The Oudemansiella radicata protein extract of 15mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the long root mushroom protein of 15mg/L
In the Mixed Pharmacy solution of extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:90.
The Propamocarb solution of the Oudemansiella radicata protein extract solution of 500 μ g/mL and 10 μ g/mL, which is mixed to get concentration, is
The Oudemansiella radicata protein extract of 150mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the long root mushroom albumen of 150mg/L
In the Mixed Pharmacy solution of matter extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:90.
Wherein, the effective component (active constituent) of each Mixed Pharmacy is protein and Propamocarb, each Mixed Pharmacy solution
Concentration be contained protein in Oudemansiella radicata protein extract concentration and the sum of concentration of Propamocarb.
6.5, the Mixed Pharmacy solution of Propamocarb and Clitocybe Maxima protein extract is prepared
6.5.1 the concentration that the mass ratio for preparing Propamocarb and protein is 21:490 be respectively 3.5mg/L, 35mg/L and
The Clitocybe Maxima protein extract of 350mg/L and the Mixed Pharmacy solution of Propamocarb
The Propamocarb solution of the Clitocybe Maxima protein extract solution of 5 μ g/mL and 1 μ g/mL, which is mixed to get concentration, is
The Clitocybe Maxima protein extract of 3.5mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the Clitocybe Maxima albumen of 3.5mg/L
In the Mixed Pharmacy solution of matter extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:490.
The Propamocarb solution of the Clitocybe Maxima protein extract solution of 50 μ g/mL and 5 μ g/mL, which is mixed to get concentration, is
The Clitocybe Maxima protein extract of 35mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the Clitocybe Maxima protein of 35mg/L
In the Mixed Pharmacy solution of extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:490.
The Propamocarb solution of the Clitocybe Maxima protein extract solution of 500 μ g/mL and 10 μ g/mL, which is mixed to get concentration, is
The Clitocybe Maxima protein extract of 350mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the Clitocybe Maxima albumen of 350mg/L
In the Mixed Pharmacy solution of matter extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:490.
Wherein, the effective component (active constituent) of each Mixed Pharmacy is protein and Propamocarb, each Mixed Pharmacy solution
Concentration be contained protein in Clitocybe Maxima protein extract concentration and the sum of concentration of Propamocarb.
6.5.2 the concentration that the mass ratio for preparing Propamocarb and protein is 21:315 be respectively 3.0mg/L, 30mg/L and
The Clitocybe Maxima protein extract of 300mg/L and the Mixed Pharmacy solution of Propamocarb
The Propamocarb solution of the Clitocybe Maxima protein extract solution of 5 μ g/mL and 1 μ g/mL, which is mixed to get concentration, is
The Clitocybe Maxima protein extract of 3.0mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the Clitocybe Maxima albumen of 3.0mg/L
In the Mixed Pharmacy solution of matter extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:315.
The Propamocarb solution of the Clitocybe Maxima protein extract solution of 50 μ g/mL and 5 μ g/mL, which is mixed to get concentration, is
The Clitocybe Maxima protein extract of 30mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the Clitocybe Maxima protein of 30mg/L
In the Mixed Pharmacy solution of extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:315.
The Propamocarb solution of the Clitocybe Maxima protein extract solution of 500 μ g/mL and 10 μ g/mL, which is mixed to get concentration, is
The Clitocybe Maxima protein extract of 300mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the Clitocybe Maxima albumen of 300mg/L
In the Mixed Pharmacy solution of matter extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:315.
Wherein, the effective component (active constituent) of each Mixed Pharmacy is protein and Propamocarb, each Mixed Pharmacy solution
Concentration be contained protein in Clitocybe Maxima protein extract concentration and the sum of concentration of Propamocarb.
6.5.3 the concentration that the mass ratio for preparing Propamocarb and protein is 21:210 be respectively 2.5mg/L, 25mg/L and
The Clitocybe Maxima protein extract of 250mg/L and the Mixed Pharmacy solution of Propamocarb
The Propamocarb solution of the Clitocybe Maxima protein extract solution of 5 μ g/mL and 1 μ g/mL, which is mixed to get concentration, is
The Clitocybe Maxima protein extract of 2.5mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the Clitocybe Maxima albumen of 2.5mg/L
In the Mixed Pharmacy solution of matter extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:210.
The Propamocarb solution of the Clitocybe Maxima protein extract solution of 50 μ g/mL and 5 μ g/mL, which is mixed to get concentration, is
The Clitocybe Maxima protein extract of 25mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the Clitocybe Maxima protein of 25mg/L
In the Mixed Pharmacy solution of extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:210.
The Propamocarb solution of the Clitocybe Maxima protein extract solution of 500 μ g/mL and 10 μ g/mL, which is mixed to get concentration, is
The Clitocybe Maxima protein extract of 250mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the Clitocybe Maxima albumen of 250mg/L
In the Mixed Pharmacy solution of matter extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:210.
Wherein, the effective component (active constituent) of each Mixed Pharmacy is protein and Propamocarb, each Mixed Pharmacy solution
Concentration be contained protein in Clitocybe Maxima protein extract concentration and the sum of concentration of Propamocarb.
6.5.4 the concentration that the mass ratio for preparing Propamocarb and protein is 21:140 be respectively 2.0mg/L, 20mg/L and
The Clitocybe Maxima protein extract of 200mg/L and the Mixed Pharmacy solution of Propamocarb
The Propamocarb solution of the Clitocybe Maxima protein extract solution of 5 μ g/mL and 1 μ g/mL, which is mixed to get concentration, is
The Clitocybe Maxima protein extract of 2.0mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the Clitocybe Maxima albumen of 2.0mg/L
In the Mixed Pharmacy solution of matter extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:140.
The Propamocarb solution of the Clitocybe Maxima protein extract solution of 50 μ g/mL and 5 μ g/mL, which is mixed to get concentration, is
The Clitocybe Maxima protein extract of 20mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the Clitocybe Maxima protein of 20mg/L
In the Mixed Pharmacy solution of extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:140.
The Propamocarb solution of the Clitocybe Maxima protein extract solution of 500 μ g/mL and 10 μ g/mL, which is mixed to get concentration, is
The Clitocybe Maxima protein extract of 200mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the Clitocybe Maxima albumen of 200mg/L
In the Mixed Pharmacy solution of matter extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:140.
Wherein, the effective component (active constituent) of each Mixed Pharmacy is protein and Propamocarb, each Mixed Pharmacy solution
Concentration be contained protein in Clitocybe Maxima protein extract concentration and the sum of concentration of Propamocarb.
6.5.5 the concentration that the mass ratio for preparing Propamocarb and protein is 21:90 be respectively 1.5mg/L, 15mg/L and
The Clitocybe Maxima protein extract of 150mg/L and the Mixed Pharmacy solution of Propamocarb
The Propamocarb solution of the Clitocybe Maxima protein extract solution of 5 μ g/mL and 1 μ g/mL, which is mixed to get concentration, is
The Clitocybe Maxima protein extract of 1.5mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the Clitocybe Maxima albumen of 1.5mg/L
In the Mixed Pharmacy solution of matter extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:90.
The Propamocarb solution of the Clitocybe Maxima protein extract solution of 50 μ g/mL and 5 μ g/mL, which is mixed to get concentration, is
The Clitocybe Maxima protein extract of 15mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the Clitocybe Maxima protein of 15mg/L
In the Mixed Pharmacy solution of extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:90.
The Propamocarb solution of the Clitocybe Maxima protein extract solution of 500 μ g/mL and 10 μ g/mL, which is mixed to get concentration, is
The Clitocybe Maxima protein extract of 150mg/L and the Mixed Pharmacy solution of Propamocarb, the concentration are the Clitocybe Maxima albumen of 150mg/L
In the Mixed Pharmacy solution of matter extract and Propamocarb, the mass ratio of Propamocarb and protein is 21:90.
Wherein, the effective component (active constituent) of each Mixed Pharmacy is protein and Propamocarb, each Mixed Pharmacy solution
Concentration be contained protein in Clitocybe Maxima protein extract concentration and the sum of concentration of Propamocarb.
6.6 chemicals treatment
3-4 leaf phase leaf age, the size healthy red earth blade of grape consistent as far as possible are selected, with after rinsing with ruinning water, by 3-5
The overlapping of piece leaf, the leaf dish of ф 15mm is produced with punch, and all leaf dishes is mixed at random spare.It tests in triplicate, often
It is secondary that leaf dish is divided into 14 groups: Oudemansiella radicata protein extract group, Clitocybe Maxima protein extract group, Propamocarb group, Propamocarb and
The Mixed Pharmacy solution group of Oudemansiella radicata protein extract and Propamocarb that the mass ratio of protein is 21:490 is (referred to as white: long
=21:490 group), the mass ratio of Propamocarb and protein be the Oudemansiella radicata protein extract of 21:315 and the mixture of Propamocarb
The long root mushroom protein that the mass ratio of liquid medicine group (referred to as white: length=21:315 group), Propamocarb and protein is 21:210
The mass ratio of the Mixed Pharmacy solution group (referred to as frost: length=21:210 group) of extract and Propamocarb, Propamocarb and protein is
The Oudemansiella radicata protein extract and the Mixed Pharmacy solution group (referred to as white: length=21:140 group) of Propamocarb, downy mildew of 21:140
The mass ratio of prestige and protein be 21:90 Oudemansiella radicata protein extract and Propamocarb Mixed Pharmacy solution group (referred to as frost:
Length=21:90) group, Propamocarb and protein mass ratio be 21:490 Clitocybe Maxima protein extract and Propamocarb mixture
The Clitocybe Maxima protein that the mass ratio of liquid medicine group (referred to as white: big=21:490 group), Propamocarb and protein is 21:315
The mass ratio of the Mixed Pharmacy solution group (referred to as frost: big=21:315 group) of extract and Propamocarb, Propamocarb and protein is
The Clitocybe Maxima protein extract and the Mixed Pharmacy solution group (referred to as white: big=21:210 group) of Propamocarb, downy mildew of 21:210
The Mixed Pharmacy solution group of Clitocybe Maxima protein extract and Propamocarb that the mass ratio of prestige and protein is 21:140 is (referred to as
Frost: big=21:140 group), the mass ratio of Propamocarb and protein be 21:90 Clitocybe Maxima protein extract and Propamocarb
Mixed Pharmacy solution group (referred to as white: big=21:90 group) and control group.
Oudemansiella radicata protein extract group sets the Oudemansiella radicata protein extract solution processing of 500 μ g/mL, 50 μ g/mL
The Oudemansiella radicata protein extract solution of the processing of Oudemansiella radicata protein extract solution and 5 μ g/mL handle these three processing, each
Handle 15 leaf dishes；Clitocybe Maxima protein extract group sets the Clitocybe Maxima protein extract solution processing of 500 μ g/mL, 50 μ g/
The Clitocybe Maxima protein extract solution processing of mL and the Clitocybe Maxima protein extract solution of 5 μ g/mL handle these three processing,
15 leaf dishes of each processing；Propamocarb group sets the Propamocarb solution processing of 10 μ g/mL, the Propamocarb solution processing of 5 μ g/mL and 1 μ
The Propamocarb solution of g/mL handles these three processing, 15 leaf dishes of each processing.Frost: length=21:490 group sets the length of 3.5mg/L
The processing of the Mixed Pharmacy solution of root mushroom protein extract and Propamocarb, the Oudemansiella radicata protein extract of 35mg/L and Propamocarb
The processing of Mixed Pharmacy solution, the Oudemansiella radicata protein extract of 350mg/L and Propamocarb Mixed Pharmacy solution handle this three
A processing, 15 leaf dishes of each processing；Frost: length=21:315 group sets the Oudemansiella radicata protein extract and Propamocarb of 3.0mg/L
The processing of Mixed Pharmacy solution, the Oudemansiella radicata protein extract of 30mg/L and Propamocarb the processing of Mixed Pharmacy solution,
The Oudemansiella radicata protein extract of 300mg/L and the Mixed Pharmacy solution of Propamocarb handle these three processing, each processing 15
Leaf dish；Frost: length=21:210 group set 2.5mg/L Oudemansiella radicata protein extract and Propamocarb Mixed Pharmacy solution handle,
The Oudemansiella radicata protein extract of 25mg/L and the Mixed Pharmacy solution of Propamocarb are handled, the long root mushroom protein of 250mg/L mentions
The Mixed Pharmacy solution of object and Propamocarb is taken to handle these three processing, 15 leaf dishes of each processing；Frost: length=21:140 group is set
The Oudemansiella radicata protein extract of 2.0mg/L and the Mixed Pharmacy solution of Propamocarb are handled, the long root mushroom protein of 20mg/L mentions
Take the Mixed Pharmacy of the processing of the Mixed Pharmacy solution of object and Propamocarb, the Oudemansiella radicata protein extract of 200mg/L and Propamocarb
Solution handles these three processing, 15 leaf dishes of each processing；Frost: length=21:90 group sets the long root mushroom albumen of 1.5mg/L respectively
The Mixed Pharmacy solution of matter extract and Propamocarb processing, the Oudemansiella radicata protein extract of 15mg/L and Propamocarb mixed medicine
The Mixed Pharmacy solution of agent solution processing, the Oudemansiella radicata protein extract of 150mg/L and Propamocarb handles these three processing, often
15 leaf dishes of a processing.Frost: big=21:490 group sets the Clitocybe Maxima protein extract of 3.5mg/L and the Mixed Pharmacy of Propamocarb
Solution processing, the Mixed Pharmacy solution processing of the Clitocybe Maxima protein extract of 35mg/L and Propamocarb, 350mg/L Clitocybe Maxima
The Mixed Pharmacy solution of protein extract and Propamocarb handles these three processing, 15 leaf dishes of each processing；Frost: big=21:
The Mixed Pharmacy solution of 315 groups of Clitocybe Maxima protein extracts for setting 3.0mg/L and Propamocarb processing, 30mg/L Clitocybe Maxima egg
The processing of the Mixed Pharmacy solution of white matter extract and Propamocarb, the Clitocybe Maxima protein extract of 300mg/L and Propamocarb it is mixed
The agent solution that makes up a prescription handles these three processing, 15 leaf dishes of each processing；Frost: big=21:210 group sets the Clitocybe Maxima egg of 2.5mg/L
The processing of the Mixed Pharmacy solution of white matter extract and Propamocarb, the Clitocybe Maxima protein extract of 25mg/L and the mixture of Propamocarb
The Mixed Pharmacy solution of liquid medicine processing, the Clitocybe Maxima protein extract of 250mg/L and Propamocarb handles these three processing,
15 leaf dishes of each processing；Frost: big=21:140 group sets the Clitocybe Maxima protein extract of 2.0mg/L and the mixed medicine of Propamocarb
Agent solution processing, the Mixed Pharmacy solution processing of the Clitocybe Maxima protein extract of 20mg/L and Propamocarb, 200mg/L mug
The Mixed Pharmacy solution of umbrella protein extract and Propamocarb handles these three processing, 15 leaf dishes of each processing；Frost: big=
21:90 group sets Clitocybe Maxima protein extract and the Mixed Pharmacy solution processing of Propamocarb, 15mg/L of 1.5mg/L respectively
The Mixed Pharmacy solution of Clitocybe Maxima protein extract and Propamocarb processing, 150mg/L Clitocybe Maxima protein extract and frost
The Mixed Pharmacy solution of mould prestige handles these three processing, 15 leaf dishes of each processing.Control treatment liquid is to use sterile water as molten
Agent, the solution that polysorbas20 is 0.1 μ g/mL as the concentration that solute prepares polysorbas20.
15 leaf dishes are put into each culture dish lid, the various processing of remaining each group in addition to control group, according to label
Concentration be separately added into the medicament of 10mL corresponding concentration and polysorbas20 be added simultaneously and make the 0.1 μ g/mL of content of polysorbas20, to leaf
Piece carries out immersion treatment 20 minutes, completes chemicals treatment.It is put into 15 leaf dishes in the culture dish lid of control group, is added 10mL pairs
According to treatment fluid, blade is carried out immersion treatment 20 minutes, completes chemicals treatment.It is face-up when immersion, guarantee that medicament immerses leaf
Back.
The inoculation and culture of 6.7 Plasmopara viticolas
The medical fluid in the leaf dish by 6.6 chemicals treatment is blotted after immersion, blade back is placed in upward with identical medicine
It is then 1 × 10 by 4.2 concentration with sessile drop method on the blotting paper of agent wetting4The downy mildew spore suspension of a spore/ml connects
Kind arrives vacuum side of blade, and each leaf dish is inoculated with 20 μ L downy mildew spore suspensions (1 × 104A spore/mL), by the leaf dish after inoculation
Dark culturing is placed in 19 DEG C of illumination boxs for 24 hours, to ensure the intrusion of pathogen, then in 19 DEG C, 12h alternation of light and darkness
Moisturizing culture in illumination box, until starting to fall ill.Using square method after the onset of culture, (each grid area is 4mm2) survey
The onset area in leaf dish is measured, calculates Oudemansiella radicata protein extract, Clitocybe Maxima protein extract, downy mildew according to following formula
The Mixed Pharmacy of Oudemansiella radicata protein extract and Propamocarb that the mass ratio of prestige, Propamocarb and protein is 21:490 is (referred to as
Frost: length=21:490), the mass ratio of Propamocarb and protein be 21:315 Oudemansiella radicata protein extract and Propamocarb mix
The Oudemansiella radicata protein extract that the mass ratio of dose out powders (abbreviation frost: length=21:315), Propamocarb and protein is 21:210
It is the long root mushroom of 21:140 with the mass ratio of the Mixed Pharmacy of Propamocarb (referred to as frost: length=21:210), Propamocarb and protein
The mass ratio of the Mixed Pharmacy (abbreviation frost: length=21:140) of protein extract and Propamocarb, Propamocarb and protein is 21:
The matter of the Mixed Pharmacy (referred to as frost: length=21:90) of 90 Oudemansiella radicata protein extract and Propamocarb, Propamocarb and protein
Mixed Pharmacy (referred to as white: big=21:490), Propamocarb of the amount than the Clitocybe Maxima protein extract and Propamocarb that are 21:490
With the Mixed Pharmacy of Clitocybe Maxima protein extract and Propamocarb that the mass ratio of protein is 21:315 (referred to as frost: it is big=21:
315) Mixed Pharmacy (letter of Clitocybe Maxima protein extract and Propamocarb that, the mass ratio of Propamocarb and protein is 21:210
Frost: big=21:210), the mass ratio of Propamocarb and protein be referred to as the Clitocybe Maxima protein extract and Propamocarb of 21:140
The Clitocybe Maxima protein extract that the mass ratio of Mixed Pharmacy (referred to as white: big=21:140), Propamocarb and protein is 21:90
Inhibiting rate and effectively inhibition of Mixed Pharmacy (abbreviation frost: big=21:90) the various processing to Plasmopara viticola with Propamocarb
Middle concentration EC50.
Inhibiting rate is converted to probit value (y), concentration is converted to logarithm (x), finds out each medicament to Plasmopara viticola
Toxicity regression curve equation y=a+bx, and effectively inhibit in concentration EC50 value and correlation coefficient r.
The co-toxicity of Mixed Pharmacy is indicated using the co-toxicity coefficient method of Sun Yunpei:
Toxicity index=(standard agent EC50Value/reagent agent EC50Value) × 100
Mixed Pharmacy actual toxicity index ATI=(standard agent EC50Value/reagent agent (mixture) EC50Value) × 100
Mixed Pharmacy theoretical toxicity index TTI=∑ (pharmaceutically active ingredient of the toxicity index of certain medicine × in Mixed Pharmacy
Percentage).
It is synergistic effect when CTC≤120；It is antagonism when CTC≤80；It is addition when 80 < CTC < 120.
The results are shown in Table 2, after showing Propamocarb and Oudemansiella radicata protein extract mixture, has to Plasmopara viticola aobvious
Write synergistic effect: the mass ratio of Propamocarb and protein is the Oudemansiella radicata protein extract of 21:490 and the mixed medicine of Propamocarb
The co-toxicity coefficient of agent is 145.83, and the mass ratio of Propamocarb and protein is the Oudemansiella radicata protein extract and downy mildew of 21:90
The co-toxicity coefficient of the Mixed Pharmacy of prestige is 164.06.Illustrate Propamocarb and Oudemansiella radicata protein extract according to Propamocarb and long root
Protein quality in mushroom protein extract is than producing synergistic effect for the ratio mixture of 21:490 and 21:90.Propamocarb
The Mixed Pharmacy of Oudemansiella radicata protein extract and Propamocarb that mass ratio with protein is 21:490 and 21:90 is to oenin
Mildew bacterium has significant inhibiting effect, and the EC50 to Plasmopara viticola is 0.05mg/L and 0.05mg/L, much smaller than frost
EC50 (0.21mg/L) of the mould prestige to Plasmopara viticola.
The Oudemansiella radicata protein extract and Propamocarb that the mass ratio of Propamocarb and protein is 21:315 and 21:210
The co-toxicity coefficient of Mixed Pharmacy is respectively 84.50 and 91.86 with addition；The mass ratio of Propamocarb and protein is 21:
The co-toxicity coefficient of the Mixed Pharmacy of 210 Oudemansiella radicata protein extract and Propamocarb is 61.76, has antagonism.
The indoor joint toxicity measuring result of 2 Propamocarb of table and Oudemansiella radicata protein extract mixture to Plasmopara viticola
| Medicament | EC50(mg/L) | Toxicity index | Actual toxicity index | Theoretical toxicity index | Co-toxicity coefficient |
| Propamocarb | 0.21 | 100.00 | |||
| Oudemansiella radicata protein extract | 0.07 | 300.00 | |||
| Frost: length=21:490 | 0.05 | 425.50 | 291.78 | 145.83 | |
| Frost: length=21:315 | 0.09 | 242.94 | 287.50 | 84.50 | |
| Frost: length=21:210 | 0.12 | 174.05 | 281.82 | 61.76 | |
| Frost: length=21:140 | 0.08 | 251.62 | 273.91 | 91.86 | |
| Frost: length=21:90 | 0.05 | 430.10 | 262.16 | 164.06 |
Note: data are the average value of three repeated experiments in table.
The results are shown in Table 3, after showing Propamocarb and Clitocybe Maxima protein extract mixture, Propamocarb and Clitocybe Maxima albumen
Matter extract is to Plasmopara viticola without significant synergistic effect, and co-toxicity coefficient is below 120.
The indoor joint toxicity measuring result of 3 Propamocarb of table and Clitocybe Maxima protein extract mixture to Plasmopara viticola
| Medicament | EC50(mg/L) | Toxicity index | Actual toxicity index | Theoretical toxicity index | Co-toxicity coefficient |
| Propamocarb | 0.21 | 100 | |||
| Clitocybe Maxima protein extract | 0.05 | 420 | |||
| Frost: big=21:490 | 0.05 | 420.00 | 406.85 | 103.23 | |
| Frost: big=21:315 | 0.05 | 388.89 | 370.83 | 104.87 | |
| Frost: big=21:210 | 0.04 | 488.37 | 453.06 | 107.79 | |
| Frost: big=21:140 | 0.05 | 396.23 | 371.10 | 106.77 | |
| Frost: big=21:90 | 0.06 | 350.00 | 352.78 | 99.21 |
Note: data are the average value of three repeated experiments in table.
The present invention is had been described in detail above.To those skilled in the art, do not depart from spirit of the invention and
Range, and without carrying out under unnecessary experimental conditions, can synchronization parameters, concentration and under the conditions of, it is real in a wider range
Apply the present invention.Although The present invention gives particular embodiments, it is understood that, the present invention can be improved further.
In short, pressing the principle of the present invention, the application is intended to include any change, purposes or improvement of the present invention, including departing from this Shen
Please in the open scope, and the change carried out with routine techniques known in the art.By the range of following attached claims,
It can carry out the application of some essential characteristics.
Claims (10)
1. preparation, the preparation contains Propamocarb and Oudemansiella radicata protein extract；The Oudemansiella radicata protein extract according to
The method preparation included the following steps:
1) flooding is used after crushing long root mushroom fructification, is collected water-soluble substances, is obtained long root mushroom water-soluble extractive；
2) ammonium sulfate precipitation that saturation degree is 80% is carried out to the long root mushroom water-soluble extractive, collects precipitating, to described heavy
After the deionized water dialysis of shallow lake, the Oudemansiella radicata protein extract is obtained；
The preparation is downy mildew of garpe bacteria inhibitor, or to prevent and/or treating the medicament of downy mildew of garpe.
2. preparation according to claim 1, it is characterised in that: the active constituent of the medicament is Propamocarb and the long root
Mushroom protein extract.
3. preparation according to claim 1 or 2, it is characterised in that: Propamocarb and the Oudemansiella radicata protein extract
Mass ratio is 21:490, and the quality of the Oudemansiella radicata protein extract is in terms of the quality of protein.
4. preparation according to claim 1 or 2, it is characterised in that: Propamocarb and the Oudemansiella radicata protein extract
Mass ratio is 21:90, and the quality of the Oudemansiella radicata protein extract is in terms of the quality of protein.
5. the application of Propamocarb and long root mushroom in preparation prevention and/or treatment downy mildew of garpe medicament.
6. Oudemansiella radicata protein extract described in any claim is being prevented and/or is being controlled in Propamocarb and claim 1-4
Treat the application in downy mildew of garpe.
7. the application of Propamocarb and long root mushroom in preparation downy mildew of garpe bacteria inhibitor.
8. Oudemansiella radicata protein extract described in any claim is inhibiting grape downy mildew in Propamocarb and claim 1-4
Application in germ.
9. following any purposes of Oudemansiella radicata protein extract described in any claim in claim 1-4:
M1. Oudemansiella radicata protein extract described in any claim in preparation prevention and/or treats Portugal in claim 1-4
Application in grape downy mildew medicament；
M2. oenin is being prevented and/or treated to Oudemansiella radicata protein extract described in any claim in claim 1-4
Application in mildew；
M3. Oudemansiella radicata protein extract described in any claim prepares Plasmopara viticola inhibition in claim 1-4
Application in agent；
M4. in claim 1-4 Oudemansiella radicata protein extract described in any claim inhibit Plasmopara viticola in
Application.
10. following any purposes of long root mushroom:
N1. application of the long root mushroom in preparation prevention and/or treatment downy mildew of garpe medicament；
N2. long root mushroom is preventing and/or is treating the application in downy mildew of garpe；
N3. application of the long root mushroom in preparation downy mildew of garpe bacteria inhibitor；
N4. long root mushroom is inhibiting the application in Plasmopara viticola.
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