CN106942301B - A kind of botanical pesticide preparation of prevention and control rice blast and preparation method thereof - Google Patents

A kind of botanical pesticide preparation of prevention and control rice blast and preparation method thereof Download PDF

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CN106942301B
CN106942301B CN201710194026.8A CN201710194026A CN106942301B CN 106942301 B CN106942301 B CN 106942301B CN 201710194026 A CN201710194026 A CN 201710194026A CN 106942301 B CN106942301 B CN 106942301B
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actinomyces
rice blast
chinese medicine
ethanol
botanical pesticide
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钟积东
谭志奇
周艳
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Hunan Geling Technology Co., Ltd
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    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract

The present invention relates to microbial technology field, discloses a kind of botanical pesticide preparation of prevention and control rice blast and preparation method thereof.Botanical pesticide preparation of the present invention by resisting rice blast bacteria actinomyces spore suspension and the Chinese medicine extract form.The Chinese medicine extract is the coptis, the wind-weed, moutan bark, river cassia bark, the root of purple-flowered peucedanum, radix ranunculi ternati, the mixed extract of wilsonii.The invention provides a kind of Chinese medicine extract for the actinomyces preventive effect for strengthening resisting rice blast bacteria, the actinomyces of itself and resisting rice blast bacteria are engaged collaboration prevention and control Pyricularia oryzae, it not only compensate for the deficiency in Antagonistic Actinomycetes preventive effect, and avoid using chemical pesticide, still it ensure that environmentally friendly, have no toxic side effect, be more not easy to make pathogen develop immunity to drugs compared to actinomyces.

Description

A kind of botanical pesticide preparation of prevention and control rice blast and preparation method thereof
Technical field
The present invention relates to microbial technology field, particularly relates to a kind of botanical pesticide of prevention and control rice blast Preparation and preparation method thereof.
Background technology
Rice blast (Blast Fungus) is that whole world rice region endangers one of rice disease of most serious, also known as rice heat Disease, it is commonly called as burning pest, hangs a pest, pinches neck pest etc..The disease can be damaged to rice throughout the year, and it is endangered throughout rice Each position, there are seedling rice blast, leaf pest, pulvinus pest, section rice blast, panicle blast, branch stalk pest and grain pest etc..Typically result in the rice underproduction 10%~20%.By taking China as an example, since the 1990s, the sick year occurring area is in 3,800,000 hectares of hm2More than, Year loss paddy reaches several hundred million kilograms.
At present, chemical pesticide is a kind of major measure for preventing and treating rice blast, but a large amount of and long-term use of chemical pesticide, both Increase production cost, be also easily caused pathogen and develop immunity to drugs and reduce prevention effect, while cause environmental pollution and destroy to give birth to State is balanced, and hidden trouble is brought to human health and Environmental security.And because rice blast pathogen heredity is complicated, it is various to cause a disease and The features such as easily making a variation, the disease-resistant variety newly cultivated need to work hard cultivation, and often lose resistance promoting in the several years, reduce the party Method controls the effect of rice blast.Therefore, development is safe and free of toxic and side effects, free from environmental pollution, is not easy to make pathogen develop immunity to drugs Microbial pesticide it is most important.
Endophyte be it is a kind of can be survived in plant without having the microorganism that substantially endangers to host plant, and by It is grown in factors such as field operation, climate changes, bred and the influence of prevention effect is smaller, be a kind of natural microorganism Live body agricultural chemicals.Current separated obtained disease resistance endophyte is the bacterium and fungi monoid easily cultivated mostly, and is used as difficult point From endogeny rayungus then report it is less.Actinomyces are the main source of antibiotic again, it has now been found that antibiosis be known as closely 6000 kinds, wherein more than 4000 kinds is produced by actinomyces.The microorganism of multiple beneficial metabolite, actinomyces can be produced as one kind Obviously there is very high economic value.
Chinese patent CN104195064A discloses a kind of rice endogeny rayungus of external efficiently antagonism Pyricularia oryzae, its Screening obtains one plant of rice endogeny rayungus from long-grain nonglutinous rice plum No. 4, and it can be mould to the pears spores of at least five kinds of different biological strains Bacterium (pathomycete of rice blast) has antagonism；Chinese patent CN105567596A discloses a kind of actinomyces and its application, It discloses a kind of Fuyang streptomycete, detects that the bacterial strain is to the inhibiting rate of Pyricularia oryzae by flat board dual test 48.6%；Document《Rice blast biocontrol actinomycetes A11The Fermentation Conditions of bacterial strain》(space etc., Chinese agronomy circular, volume 21 5th phase, in May, 2005) also disclose that actinomyces A11To the antagonistic ability of Pyricularia oryzae, bacteriostasis rate average out under given conditions To 57.4%.
Using the actinomyces live body of resisting rice blast bacteria as pesticidal preparations, although can be avoided with its safer biological metabolite Hidden danger caused by chemical pesticide, the antagonism corresponding relation between different actinomyces and pathogen can also be utilized, reduces rice blast Germ is various, the easy influence brought that makes a variation, but the actinomyces screened ability height itself differs, and single use still has It is short of, also needs to further strengthen in preventive effect.
The content of the invention
In view of this, it is used for it is an object of the invention to provide a kind of in prevention and control rice blast botanical pesticide preparation Medicament extracting solution so that the Chinese medicine extract can significantly improve the actinomyces of resisting rice blast bacteria in botanical pesticide preparation to rice The inhibiting rate of seasonal febrile diseases bacterium；
It is another object of the present invention to provide a kind of botanical pesticide preparation of prevention and control rice blast and its preparation Method so that the botanical pesticide preparation has preferable inhibiting rate to Pyricularia oryzae, can be applied in the preventing and treating of rice blast.
To achieve these goals, the present invention provides following technical scheme：
A kind of Chinese medicine extract for prevention and control rice blast botanical pesticide preparation, be the coptis, the wind-weed, moutan bark, The mixed extract of river cassia bark, the root of purple-flowered peucedanum, radix ranunculi ternati, wilsonii.
It is used alone for existing rice endogeny rayungus insufficient in rice blast prevention effect the defects of, including the present invention Plurality of Chinese extract is with the addition of on the basis of raw actinomyces, the suppression that each component is improved to Pyricularia oryzae by synergy is made With.
Preferably, the coptis, the wind-weed, moutan bark, river cassia bark, the root of purple-flowered peucedanum, radix ranunculi ternati, the weight ratio of wilsonii are 1:1: 1:1:1:1:1。
Preferably, the extract is ethanol extract.It is further preferred that the Chinese medicine extract is by following methods Prepare：
Take each Chinese herbal medicine 20g to crush respectively, mix, add 95% ethanol 100ml, soak 48h, 3 layers of gauze at room temperature Filtering, filtrate 4000r/min centrifugation 15min, takes supernatant, rotary evaporated to dryness at 40 DEG C, residue is dissolved in the second of 20ml 50% In alcohol, the Chinese medicine extract is obtained.
After the spore suspension of Chinese medicine extract of the present invention and the actinomyces of resisting rice blast bacteria is used cooperatively, its institute The inhibiting rate and preventive effect to Pyricularia oryzae shown, the spore for being all remarkably higher than the actinomyces that resisting rice blast bacteria is used alone hang The effect of supernatant liquid.Therefore, the invention provides the Chinese medicine extract in preventing and treating rice blast and/or to prepare preventing and treating rice blast Application in biological prevention and control agent.
Meanwhile the present invention provides a kind of botanical pesticide preparation of prevention and control rice blast according to application, by resisting rice blast bacteria Actinomyces spore suspension and the Chinese medicine extract composition.Wherein, the actinomyces are rice Antagonistic Actinomycetes CZ1367, deposit number are CCTCC No：M2016277, it is preserved on May 23rd, 2016 in China typical culture collection The heart.Preferably, the concentration of actinomyces is 10 in the spore suspension8Individual/mL.
Preferably, the spore suspension of the actinomyces is prepared by following methods：
Step 1, actinomyces are inoculated into test tube culture medium slant, culture medium prescription is：Potato 200g, grape Sugared 20g, agar 20g, water 1000mL, pH value 7.2, cultivated 7 days at 28 DEG C, obtain test tube kind；
Step 2, first prepare liquid seed culture medium, liquid seed culture medium formula is yeast extract 4g, glucose 4g, malt Cream 10g, agar 20g, water 1000mL, pH value 7.2；Load 100mL fluid nutrient mediums, 120 DEG C of sterilizings 30 in each triangular flask Minute, access 1cm after cooling2The test tube kind, it is 200r/min concussion and cultivates 7 days in rotating speed at 28 DEG C, obtains seed liquor；
Step 3, first prepare liquid fermentation medium, liquid fermentation medium formula is soy meal 20g, sucrose 10g, solvable Property starch 5g, peptone 2g, NaCl 2g, CaCO31g, MgSO4·7H2O 0.5g, KH2PO40.5g, water 1000mL, pH value 7.0；Load 100mL liquid fermentation mediums in each 500ml triangular flasks, 120 DEG C are sterilized 30 minutes, and 10mL is accessed after cooling The seed liquor obtained in step 2, in cultivate 6 days to obtain zymotic fluid on the shaking table that rotating speed is 220rpm at 28 DEG C, add in zymotic fluid Enter isometric ethanol, after shaking table vibrates 8 hours, the centrifuge 10 minutes for being 3000rpm with rotating speed, take supernatant and 50 DEG C boil off ethanol on Rotary Evaporators, are diluted with water to original fermentation liquor volume, obtain the spore suspension.
Botanical pesticide preparation of the present invention has to the multiple biological strains of Pyricularia oryzae, such as 62, RB3,131, rice farmer Higher inhibiting rate, while there is higher preventive effect and rate of growth in pot experiment and field experiment.Based on this, the present invention also carries The botanical pesticide preparation has been supplied in preventing and treating rice blast and has prepared the application in preventing and treating rice blast pesticidal preparations.
In addition, the invention provides the preparation method of the botanical pesticide preparation, including：Step 1, prepare actinomyces Spore suspension, and prepare the coptis, the wind-weed, moutan bark, river cassia bark, the root of purple-flowered peucedanum, radix ranunculi ternati, the mixed traditional Chinese medicine extraction of wilsonii Liquid；
Spore suspension unwrapping wire bacteria concentration prepared by step 2, set-up procedure 1, and Chinese medicine extract mixing, obtain the plant Material resource pesticidal preparations.
Specifically, step 1 can be carried out as follows：
Step 1.1, actinomyces are inoculated into test tube culture medium slant, culture medium prescription is：Potato 200g, Portugal Grape sugar 20g, agar 20g, water 1000mL, pH value 7.2, are cultivated 7 days at 28 DEG C, obtain test tube kind；
Step 1.2, first prepare liquid seed culture medium, liquid seed culture medium formula is yeast extract 4g, glucose 4g, wheat Bud cream 10g, agar 20g, water 1000mL, pH value 7.2；Load 100mL fluid nutrient mediums, 120 DEG C of sterilizings in each triangular flask 30 minutes, 1cm is accessed after cooling2The test tube kind, it is 200r/min concussion and cultivates 7 days in rotating speed at 28 DEG C, obtains seed liquor；
Step 1.3, first prepare liquid fermentation medium, liquid fermentation medium formula is soy meal 20g, sucrose 10g, can Soluble starch 5g, peptone 2g, NaCl 2g, CaCO31g, MgSO4·7H2O 0.5g, KH2PO40.5g, water 1000mL, pH value 7.0；Load 100mL liquid fermentation mediums in each 500ml triangular flasks, 120 DEG C are sterilized 30 minutes, and 10mL is accessed after cooling The seed liquor obtained in step 2, in cultivate 6 days to obtain zymotic fluid on the shaking table that rotating speed is 220rpm at 28 DEG C, add in zymotic fluid Enter isometric ethanol, after shaking table vibrates 8 hours, the centrifuge 10 minutes for being 3000rpm with rotating speed, take supernatant and 50 DEG C boil off ethanol on Rotary Evaporators, are diluted with water to original fermentation liquor volume, obtain the spore suspension, standby；
Step 1.4, take each Chinese herbal medicine 20g to crush respectively, mix, add 95% ethanol 100ml, soak 48h at room temperature, 3 layers of filtered through gauze, filtrate 4000r/min centrifugation 15min, take supernatant, rotary evaporated to dryness at 40 DEG C, residue is dissolved in 20ml In 50% ethanol, the Chinese medicine extract is obtained.Wherein, step 1.4 belongs to coordination with step 1.1-1.3.
From above technical scheme, the invention provides a kind of Chinese medicine for the actinomyces preventive effect for strengthening resisting rice blast bacteria Extract solution, the actinomyces of itself and resisting rice blast bacteria are engaged collaboration prevention and control Pyricularia oryzae, not only compensate for Antagonistic Actinomycetes preventive effect On deficiency, and avoid using chemical pesticide, still ensure that environmentally friendly, have no toxic side effect, compared to actinomyces more It is not easy pathogen is developed immunity to drugs.
Biomaterial preservation information explanation
CZ1367, Classification And Nomenclature：Actinomyces CZ1367, Actinomyces sp.CZ1367, protected on May 23rd, 2016 China typical culture collection center is hidden in, address is Wuhan, China, and Wuhan University, deposit number is CCTCC No： M2016277。
Embodiment
The embodiment of the invention discloses a kind of botanical pesticide preparation of prevention and control rice blast and preparation method thereof, ability Field technique personnel can use for reference present disclosure, be suitably modified technological parameter realization.In particular, it is all similar to replace Change and change apparent to those skilled in the art, they are considered as being included in the present invention.Institute of the present invention State product and preparation method thereof to be described by preferred embodiment, related personnel can be not substantially being departed from the present invention Hold, product as described herein, methods and applications are modified or suitably changed with combining in spirit and scope, to realize and answer Use the technology of the present invention.
Just a kind of botanical pesticide preparation of prevention and control rice blast provided by the present invention and preparation method thereof is done below Further illustrate.
Embodiment 1：Prepare Chinese medicine extract of the present invention
Take each 20g of the coptis, the wind-weed, moutan bark, river cassia bark, the root of purple-flowered peucedanum, radix ranunculi ternati, wilsonii to crush respectively, mix, add 95% ethanol 100ml, soaks 48h at room temperature, 3 layers of filtered through gauze, filtrate 4000r/min centrifugation 15min, takes supernatant, and 40 Rotary evaporated to dryness at DEG C, residue are dissolved in the ethanol of 20ml 50%, obtain the Chinese medicine extract.
Embodiment 2：Prepare botanical pesticide preparation of the present invention
1st, the spore suspension of the actinomyces of resisting rice blast bacteria is prepared
Step 1, actinomyces are inoculated into test tube culture medium slant, culture medium prescription is：Potato 200g, grape Sugared 20g, agar 20g, water 1000mL, pH value 7.2, cultivated 7 days at 28 DEG C, obtain test tube kind；
Step 2, first prepare liquid seed culture medium, liquid seed culture medium formula is yeast extract 4g, glucose 4g, malt Cream 10g, agar 20g, water 1000mL, pH value 7.2；Load 100mL fluid nutrient mediums, 120 DEG C of sterilizings 30 in each triangular flask Minute, access 1cm after cooling2The test tube kind, it is 200r/min concussion and cultivates 7 days in rotating speed at 28 DEG C, obtains seed liquor；
Step 3, first prepare liquid fermentation medium, liquid fermentation medium formula is soy meal 20g, sucrose 10g, solvable Property starch 5g, peptone 2g, NaCl 2g, CaCO31g, MgSO4·7H2O 0.5g, KH2PO40.5g, water 1000mL, pH value 7.0；Load 100mL liquid fermentation mediums in each 500ml triangular flasks, 120 DEG C are sterilized 30 minutes, and 10mL is accessed after cooling The seed liquor obtained in step 2, in cultivate 6 days to obtain zymotic fluid on the shaking table that rotating speed is 220rpm at 28 DEG C, add in zymotic fluid Enter isometric ethanol, after shaking table vibrates 8 hours, the centrifuge 10 minutes for being 3000rpm with rotating speed, take supernatant and 50 DEG C boil off ethanol on Rotary Evaporators, are diluted with water to original fermentation liquor volume, obtain the spore suspension.
2nd, the preparation of botanical pesticide preparation
The concentration of actinomyces is 10 in spore suspension in adjustment 18Individual/mL, the Chinese medicine extract obtained with embodiment 1 Mixing, botanical pesticide preparation as of the present invention.
Embodiment 3：Flat board antagonism bacteriostatic test
Processing 1：Using flat board face-off method, on a diameter of 90mm PDA culture medium flat board, apart from culture dish center 30mm Position on be inoculated with the actinomyces that activates, then in culture dish center inoculation Pyricularia oryzae.Sealed membrane seals culture dish, Culture more than 7 days is inverted in mold incubator under the conditions of 28 DEG C, each processing is repeated 3 times, and is compareed not connect actinomyces as CK, 7d Observation experiment result afterwards.Measurement fungus colony diameter simultaneously calculates Fungal hyphal growth inhibiting rate.
Processing 2：Chinese medicine extract is configured to 50mg/ml band medicine culture medium, in addition to culture medium is different, remaining step It is identical with processing 1.
Inhibiting rate calculates according to the following formula：
Mycelial growth inhibition rate %=(CK bacterium colonies average diameter-processing bacterium colony average diameter)/CK bacterium colonies average diameter × 100%
It the results are shown in Table 1.
Bacteriostasis rate of the different disposal of table 1 to rice blast difference biological strain
As can be seen from Table 1, after rice Antagonistic Actinomycetes CZ1367 is used cooperatively with Chinese medicine extract, to each rice blast diease occurrence Reason microspecies bacteriostasis rate is obviously improved.
Embodiment 4：Pot experiment
1. seed treatment：
Rice paddy seed carries out surface sterilization processing：With sterilized water washing after 75% ethanol postincubation 15s three times, 5% time Sterilized water is washed three times after sodium chlorate solution handles 15min.The rice paddy seed of surface sterilization is soaked in concentration as >=108Individual/ml After CZ1367 spore liquids (and the botanical pesticide preparation of embodiment 2) 30min, seedling raising is sowed after moisturizing vernalization.
2. the inoculation of pathogen and the administration of CZ1367 microbial inoculums
With the small barreled rice field soil of plastics, (soil first sterilizes, vermiculite of adding some points, vermiculite：Soil=1: 3, dredging soil Earth), every barrel of 3 caves of transplanting, per 5, cave rice shoot.Pathogen is inoculated with during rice plant length to tillering stage.With physiological races of rice blast fungus RB3 is pathogen), Seedling Inoculation：After germ is induced into production spore, rice blast spore suspension is sprayed on the rice seedlings of 2-3 leaf phases Liquid 106Individual/ml inoculations, overwork water droplet with blade, do not drip as standard, 25 DEG C, more than 95% humidity is handled one day, then 28 DEG C Culture.
3. experimental design
Processing 1:Clear water compares.
Processing 2:CZ1367, after being inoculated with magnaporthe grisea spore, sprayed for the first time when starting morbidity to clear water control group CZ1367 spore suspensions 108Individual/ml, water droplet is overworked with blade, does not drip as standard, hereafter sprayed every 7-10 days once, Spray 2-3 times.
Processing 3：CZ1367+ Chinese medicine extracts, in addition to spraying the difference of the botanical pesticide preparation in embodiment 2, remaining With processing 2, spraying concentration 108Individual/ml.
Processing 4:75% tricyclazole wettable powder (production of Jiangsu bumper harvest agrochemical Co., Ltd), except spraying 75% 3 ring Outside azoles wettable powder, remaining is the same as processing 2, spraying concentration 1g/L.
3. statistical result
Leaf pest disease index=∑ (the sick level number of sheets × represents numerical value)/total number of sheets/morbidity most heavy duty represents numerical value
Leaf pest preventive effect=(check plot disease index-treatment region disease index)/check plot disease index
4. result of the test
Rice ear sprouting period, preventive effect is investigated, the results are shown in Table 2.
Table 2 prevents and treats the results from pot experiment test of rice blast
As shown in Table 2, handle 1 CZ1367 in greenhouse test to rice blast preventive effect up to 80.49%, with compareing three The preventive effect of ring azoles is suitable, and the botanical pesticide preparation preventive effect of processing 2 will be significantly higher than the CZ1367 of processing 1.
Embodiment 5：Field district test
In rice blast heavier Liuyang City of Hunan Province great Wei Shan and Guidong County of Hunan Province water occur year after year for field test selection In rice field.4 processing are set altogether：A：CZ1367 108Individual/ml spore liquid 500ml/ mus, it is watered 30L sprayings；B：The plant of embodiment 2 Source pesticide preparation 500ml/ mus, it is watered 30L sprayings；C：70% tricyclazole (WP) 30g/ mus, it is watered 30L sprayings；D：Clear water compares. 4 repetitions are often handled, per plot area 30m2, respectively handle random district's groups arrangement.With protection row isolation between cell, experimental plot is pressed It is conventional to carry out rich water quality management.Seed soaking before sowing, preventing and treating leaf pest are carried out at the initial stage of a disease, 7 days after first time dispenser, then dispenser 1 time, sprayed using knapsack sprayer.
Disease investigation investigates hair with reference to pesticide field efficacy medicine test criterion (one) (GB/T17980.19-2000) before dispenser Sick radix, 8~10d progress after the 2nd dispenser is investigated to the preventive effect of leaf pest, the preventive effect of panicle blast is investigated then in panicle blast Investigated 1 time after feelings development stability.Per the 5 points of samplings of cell diagonal, every investigation 5 clumps, totally 25 clumps, the hair of investigation blade (fringe) Course of disease degree, calculate disease index and preventive effect.It is identical with standard used in the national rice blast test of pesticide effectiveness to investigate grade scale.During harvest Survey production.It the results are shown in Table 3 and table 4.
Grade scale：
Leaf pest (in units of blade)
0 grade：It is disease-free；
1 grade：Leaf spot lesion is less than 5, length<1cm sides
3 grades：Leaf spot lesion 6-10, part scab length>1cm；
5 grades：Leaf spot lesion 11-25, part scab is linked to be piece, accounts for leaf area 10%-25%；
7 grades：Leaf spot lesion more than 26, scab is linked to be piece, accounts for leaf area 26%-50%；
9 grades：Scab is linked to be piece, accounts for leaf area more than 50% or full leaf is withered.
Fringe pest (in units of fringe)
0 grade：It is disease-free；
1 grade：Loss less than 5% (indivedual branch stalk morbidities) per fringe；
3 grades：Loss 6%-20% (morbidity of 1/3 or so branch stalk) per fringe；
5 grades：Loss 21%-50% per fringe (fringe neck or main shaft morbidity, grain half are flat)；
7 grades：Loss 51%-70% per fringe (fringe neck is fallen ill, big portion's shrivelled kernel)；
9 grades：Loss 71%-100% per fringe (fringe neck is fallen ill, and causes dead ears).
Drug effect computational methods：
CK0The disease of not dispenser check plot refers to before-spray；
CK1The disease of not dispenser check plot refers to after-spray；
pt0The disease in-spray pre-treatment area refers to；
pt1The disease of-spray post processing zone refers to.
Table 3 prevents and treats the field district test result (Liuyang City) of rice blast
Table 4 prevents and treats the field district test result (Guidong County) of rice blast
Note：Numeral is all four repetition average values in table 3 and table 4.
There is preferable effect to rice blast by table 3 and the results showed that CZ1367 of table 4, in Liuyang City of Hunan Province The leaf pest preventive effect of great Wei Shan and Guidong County of Hunan Province is respectively 81.65% and 71.04%, fringe pest preventive effect is respectively 76.99%, 85.37%, rate of growth is respectively 7.36%, 8.37%, suitable with the preventive effect for compareing 75% tricyclazole wettable powder；
And leaf pest preventive effect of the botanical pesticide preparation in Liuyang City of Hunan Province great Wei Shan and Guidong County of Hunan Province is respectively 89.95% and 82.32%, fringe pest preventive effect is respectively 87.73%, 92.15%, and rate of growth is respectively 13.23%, 12.60%, effect Fruit is considerably better than CZ1367.
The method and its core concept described above for being only intended to understand the present invention, it is noted that for the art Those of ordinary skill for, under the premise without departing from the principles of the invention, some improvement and modification can be carried out to the present invention, These are improved and modification also falls into the protection domain of right of the present invention.

Claims (7)

1. a kind of botanical pesticide preparation of prevention and control rice blast, it is characterised in that by Chinese medicine extract and resisting rice blast bacteria Actinomyces spore suspension composition, the Chinese medicine extract be the coptis, the wind-weed, moutan bark, river cassia bark, the root of purple-flowered peucedanum, radix ranunculi ternati, The mixed extract of wilsonii, the actinomyces are rice Antagonistic Actinomycetes CZ1367, and deposit number is CCTCC No：M 2016277。
2. botanical pesticide preparation according to claim 1, it is characterised in that the coptis, the wind-weed, moutan bark, river cassia bark, The root of purple-flowered peucedanum, radix ranunculi ternati, the weight ratio of wilsonii are 1:1:1:1:1:1:1.
3. botanical pesticide preparation according to claim 1, it is characterised in that the Chinese medicine extract is prepared by following methods Obtain：
Take each Chinese herbal medicine 20g to crush respectively, mixing, add 95% ethanol 100ml, soak 48h at room temperature, 3 layers of filtered through gauze, Filtrate 4000r/min centrifuges 15min, takes supernatant, and rotary evaporated to dryness at 40 DEG C, residue is dissolved in the ethanol of 20ml 50%, Obtain the Chinese medicine extract.
4. botanical pesticide preparation described in claim 1-3 any one prevents and treats rice blast in preventing and treating rice blast and/or preparing Biological prevention and control agent in application.
5. botanical pesticide preparation according to claim 1, it is characterised in that the spore suspension of the actinomyces is by following Method prepares：
Step 1, actinomyces are inoculated into test tube culture medium slant, culture medium prescription is：Potato 200g, glucose 20g, agar 20g, water 1000mL, pH value 7.2, cultivated 7 days at 28 DEG C, obtain test tube kind；
Step 2, first prepare liquid seed culture medium, liquid seed culture medium formula is yeast extract 4g, glucose 4g, malt extract 10g, agar 20g, water 1000mL, pH value 7.2；Load 100mL fluid nutrient mediums in each triangular flask, 120 DEG C sterilize 30 points Clock, 1cm is accessed after cooling2The test tube kind, it is 200r/min concussion and cultivates 7 days in rotating speed at 28 DEG C, obtains seed liquor；
Step 3, liquid fermentation medium is first prepared, liquid fermentation medium formula is soy meal 20g, sucrose 10g, and solubility is formed sediment Powder 5g, peptone 2g, NaCl 2g, CaCO31g, MgSO4·7H2O 0.5g, KH2PO40.5g, water 1000mL, pH value 7.0； Load 100mL liquid fermentation mediums in each 500ml triangular flasks, 120 DEG C are sterilized 30 minutes, and 10mL steps 2 are accessed after cooling In obtained seed liquor, in cultivate 6 days to obtain zymotic fluid on the shaking table that rotating speed is 220rpm at 28 DEG C, the body such as addition in zymotic fluid Product ethanol, after shaking table vibrates 8 hours, the centrifuge 10 minutes for being 3000rpm with rotating speed, take supernatant and steamed in rotation 50 DEG C boil off ethanol on hair instrument, are diluted with water to original fermentation liquor volume, obtain the spore suspension.
6. the preparation method of botanical pesticide preparation described in claim 1, it is characterised in that including：
Step 1, prepare actinomyces spore suspension, and prepare the coptis, the wind-weed, moutan bark, river cassia bark, the root of purple-flowered peucedanum, radix ranunculi ternati, The mixed Chinese medicine extract of wilsonii；
Spore suspension unwrapping wire bacteria concentration prepared by step 2, set-up procedure 1, and Chinese medicine extract mixing, obtain the plant source Pesticidal preparations.
7. preparation method according to claim 6, it is characterised in that step 1 is：
Step 1.1, actinomyces are inoculated into test tube culture medium slant, culture medium prescription is：Potato 200g, glucose 20g, agar 20g, water 1000mL, pH value 7.2, cultivated 7 days at 28 DEG C, obtain test tube kind；
Step 1.2, first prepare liquid seed culture medium, liquid seed culture medium formula is yeast extract 4g, glucose 4g, malt extract 10g, agar 20g, water 1000mL, pH value 7.2；Load 100mL fluid nutrient mediums in each triangular flask, 120 DEG C sterilize 30 points Clock, 1cm is accessed after cooling2The test tube kind, it is 200r/min concussion and cultivates 7 days in rotating speed at 28 DEG C, obtains seed liquor；
Step 1.3, first prepare liquid fermentation medium, liquid fermentation medium formula is soy meal 20g, sucrose 10g, soluble Starch 5g, peptone 2g, NaCl 2g, CaCO31g, MgSO4·7H2O 0.5g, KH2PO40.5g, water 1000mL, pH value 7.0； Load 100mL liquid fermentation mediums in each 500ml triangular flasks, 120 DEG C are sterilized 30 minutes, and 10mL steps are accessed after cooling The seed liquor obtained in 2, in cultivate 6 days to obtain zymotic fluid on the shaking table that rotating speed is 220rpm at 28 DEG C, addition etc. in zymotic fluid Volume ethanol, after shaking table vibrates 8 hours, the centrifuge 10 minutes for being 3000rpm with rotating speed, take supernatant and rotating 50 DEG C boil off ethanol on evaporimeter, are diluted with water to original fermentation liquor volume, obtain the spore suspension, standby；
Step 1.4, take each Chinese herbal medicine 20g to crush respectively, mixing, add 95% ethanol 100ml, soak 48h at room temperature, 3 layers Filtered through gauze, filtrate 4000r/min centrifugation 15min, takes supernatant, rotary evaporated to dryness at 40 DEG C, residue is dissolved in 20ml In 50% ethanol, the Chinese medicine extract is obtained.
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Families Citing this family (5)

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CN107624795A (en) * 2017-09-25 2018-01-26 安徽省中日农业环保科技有限公司 A kind of prevention and controls of rice sheath blight disease
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CN110283852A (en) * 2019-07-04 2019-09-27 漳州市农业科学研究所 A kind of extract of soil actinomycetes fermenting liquid and preparation method thereof
CN110558337A (en) * 2019-07-19 2019-12-13 湖南科技学院 biocontrol preparation for preventing and treating rice blast and preparation method thereof
CN111670769B (en) * 2020-04-23 2022-05-06 湖南大学 Method for improving stress resistance of rice

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104126619A (en) * 2013-05-02 2014-11-05 天津蓟县青甸洼学雨水稻种植专业合作社 Traditional Chinese medicine formula for treating rice blast and preparation method thereof
CN104195064A (en) * 2014-06-20 2014-12-10 湖南大学 Paddy rice endophytic actinomycete realizing in-vitro efficient antagonism on rice blast pathogen
CN104230533A (en) * 2014-08-26 2014-12-24 中国水稻研究所 Special rice fertilizer containing efficient silicon and preparation method of special rice fertilizer
CN104982471A (en) * 2015-07-15 2015-10-21 广西宏华生物实业股份有限公司 A method for controlling rice blast of organic rice

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105567596B (en) * 2016-01-14 2019-01-18 中国水稻研究所 A kind of actinomyces and its application

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104126619A (en) * 2013-05-02 2014-11-05 天津蓟县青甸洼学雨水稻种植专业合作社 Traditional Chinese medicine formula for treating rice blast and preparation method thereof
CN104195064A (en) * 2014-06-20 2014-12-10 湖南大学 Paddy rice endophytic actinomycete realizing in-vitro efficient antagonism on rice blast pathogen
CN104230533A (en) * 2014-08-26 2014-12-24 中国水稻研究所 Special rice fertilizer containing efficient silicon and preparation method of special rice fertilizer
CN104982471A (en) * 2015-07-15 2015-10-21 广西宏华生物实业股份有限公司 A method for controlling rice blast of organic rice

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
对稻瘟菌具有抑制作用的植物源提取物的筛选研究;许春梅;《华中农业大学2006届硕士毕业论文》;20080715;第25-28页表3-1，第31页第4段 *

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