CN106359473A - Fermented bacteria liquid capable of preventing and controlling pawpaw mosaic diseases and preparation method thereof - Google Patents
Fermented bacteria liquid capable of preventing and controlling pawpaw mosaic diseases and preparation method thereof Download PDFInfo
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- CN106359473A CN106359473A CN201610745985.XA CN201610745985A CN106359473A CN 106359473 A CN106359473 A CN 106359473A CN 201610745985 A CN201610745985 A CN 201610745985A CN 106359473 A CN106359473 A CN 106359473A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/10—Animals; Substances produced thereby or obtained therefrom
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/40—Liliopsida [monocotyledons]
Abstract
The invention provides a fermented bacteria liquid capable of preventing and controlling pawpaw mosaic diseases and a preparation method thereof. The fermented bacteria liquid capable of preventing and controlling pawpaw mosaic diseases is prepared through mixed culture fermentation of Chinese herbal medicines including radix astragali, radix scutellariae, Coptis chinensis Franch, Lonicera japonica Thunb., Radix Paeoniae Rubra, licorice root, Heartleaf Houttuynia Herb, Chinese Pulsatilla Root, Rehmannia glutinosa Libosch, Pinellia Ternata, Radix Burpleuri, Eriobotrya japonica Thunb., Forsythia suspensa, folium isatidis, radix isatidis, Flos Lonicerae, Semen Cassiae, Flos Carthami, Radix Salviae Miltiorrhizae, Herba Leonuri and the like by using eurasian siskin streptomycete, dark yellow and purple streptomycete, bacillus siamensis, paenibacillus macquariensis, trichoderma fertile and other strains. The fermented bacteria liquid has the significant functions of preventing and controlling pawpaw mosaic diseases, and has the advantages of being easy to use, little in Chinese herbal medicines consumption and high in living bacteria content and the like.
Description
Technical field
The present invention relates to agriculture technical field of microbe application, in particular to a kind of Fructus Chaenomeliss mosaic disease of preventing and treating
Zymocyte liquid and preparation method thereof.
Background technology
Causal Organisms of Papaya Ringspot Mosaic Disease is Fructus Caricae ring class virus, and main host plants have the crops such as Cucurbitaceae, Fructus Caricae.Should
Disease causes harm seriously in provinces and regions such as China Guangxi, Hainan, Guangdong, Fujian and Taiwan.In Hainan, October to next year April temperature is suitable
Virus Fast-propagation in plant cell, is onset peak period.After plant morbidity, mortality rate is up to 90%.Disease plant result is few,
Fruit quality is poor, and diseased plant survives the winter difficulty.Due to there is no effective medical treatment at present, peasant household can only autumn sowing spring planting, then receive fruit
Surgery plant afterwards, make Fructus Caricae be changed into annual from perennial, economic benefit is badly damaged.
Field infected plant is main primary source of infection.Propagated by black peach aphid and cotton aphid, 7-28 days incubation period, warm dry
The morbidity of dry time is more serious, this growth with aphid and migrate relevant, and high temperature and unfavorable aphid activity cold season, fall ill relatively
Few.More closely fall ill apart from old Fructus Chaenomeliss garden heavier, on the contrary then light.
And in medical domain, Chinese medicine can have been confirmed by clinical and experiment with antibacterium and virus, can antibacterium and virus
Chinese herbal medicine more belong to diaphoretic medicine and antipyretic.Also some Chinese herbal medicine have Vermins-expelling vermins-proof effect；But directly utilize Chinese herbal medicine to soak
Bubble, dosage is big, and effect is not also notable；Accordingly, it is considered to directly utilize beneficial flora fermentation of Chinese herbal medicine, the thin of Chinese herbal medicine can be destroyed
Cell wall, greatly improves the utilization rate of the active ingredient of Chinese herbal medicine, and produces multiple somatomedins during the fermentation with prebiotic
Bacterium, the zymocyte liquid of the present invention has functions that significantly to prevent and treat Fructus Chaenomeliss mosaic disease, and has easy to use, and medium-height grass survival dose is few,
The advantages of viable bacteria content is high.
Content of the invention
It is an object of the invention to provide a kind of zymocyte liquid that can prevent and treat Fructus Chaenomeliss mosaic disease.The present invention is used for preventing and treating Fructus Chaenomeliss
The zymocyte liquid of leaf disease is on the basis of a large amount of investigation, and the design function according to product requires more than 250 kinds of Chinese herbal medicine are carried out
Screening, according to the monarch of traditional Chinese medicine, seven emotions compatibility, eighteen incompatible medicamentss, nineteen medicaments of mutual restraint scheduling theory, screening to filtering out one by one
Effective component of Chinese herbal medicine, antibacterial, anthelmintic action carry out extracting, verify, finally formula really.Again from the micro- life of 350 multiple beneficial
Filter out in thing 5 kinds can symbiotic co-existence, and the utilization rate of the active ingredient of Chinese herbal medicine can be greatly improved, in sweat
Middle generation multiple somatomedins probiotic bacteria.The zymocyte liquid of the present invention has functions that significantly to prevent and treat Fructus Chaenomeliss mosaic disease, and
Have easy to use, medium-height grass survival dose is few, the advantages of viable bacteria content is high.
To achieve these goals, the present invention employs the following technical solutions:
A kind of zymocyte liquid that can prevent and treat Fructus Chaenomeliss mosaic disease is it is characterised in that be made up of the raw material of following weight portion: Radix Astragali 2-4
Part, Radix Scutellariae 2-4 part, Rhizoma Coptidis 2-4 part, Flos Lonicerae 2-4 part, Radix Paeoniae Rubra 2-4 part, Radix Glycyrrhizae 2-4 part, Herba Houttuyniae 2-4 part, Radix Pulsatillae 2-6 part,
Radix Rehmanniae 2-6 part, Rhizoma Pinelliae 2-6 part, Radix Bupleuri 2-6 part, Folium Eriobotryae 2-6 part, Fructus Forsythiae 3-8 part, Folium Isatidiss 3-8 part, Radix Isatidis 3-8 part,
Flos Lonicerae 3-8 part, Semen Cassiae 3-8 part, Flos Carthami 3-8 part, Radix Salviae Miltiorrhizae 3-8 part, Herba Leonuri 1-3 part, soy molasses 40-60 part, bean cake
Powder 20-40 part, 850 parts of water, yellowbird streptomycete seed liquor 5-15 part, dark yellow purple streptomycete seed liquor 5-15 part, Lyceum spore bar
Fermented liquid 5-15 part, series bacillus fermentation liquid 5-15 part in Ma Kuo, push up spore Trichoderma spp. seed liquor 5-15 part；By said herbal medicine
Size-reduced, immersion, heating and thermal insulation, fermentation and the zymocyte liquid of preventing and treating Fructus Chaenomeliss mosaic disease is obtained.
It is a further object of the present invention to provide a kind of preparation method of the zymocyte liquid that can prevent and treat Fructus Chaenomeliss mosaic disease.
In order to achieve the above object, a kind of preparation method of the zymocyte liquid that can prevent and treat Fructus Chaenomeliss mosaic disease, walks including following
Rapid:
(1), various Chinese herbal medicine commercially are individually ground into 400 mesh fine powders；
(2), sequentially add in fermentation tank: Radix Astragali 2-4 part, Radix Scutellariae 2-4 part, Rhizoma Coptidis 2-4 part, Flos Lonicerae 2-4 part, Radix Paeoniae Rubra 2-4 part,
Radix Glycyrrhizae 2-4 part, Herba Houttuyniae 2-4 part, Radix Pulsatillae 2-6 part, Radix Rehmanniae 2-6 part, Rhizoma Pinelliae 2-6 part, Radix Bupleuri 2-6 part, Folium Eriobotryae 2-6 part,
Fructus Forsythiae 3-8 part, Folium Isatidiss 3-8 part, Radix Isatidis 3-8 part, Flos Lonicerae 3-8 part, Semen Cassiae 3-8 part, Flos Carthami 3-8 part, Radix Salviae Miltiorrhizae 3-8
Part, Herba Leonuri 1-3 part, soy molasses 40-60 part, bean cake powder 20-40 part, 850 parts of water, after soaking 30 minutes, it is heated to 95 DEG C,
Insulation two hours, is cooled to 30 DEG C；
(3), the coolant (2) produced in step adds top spore Trichoderma spp. seed liquor 5-15 part；30 DEG C -33 DEG C, open stirring 120-160r/
Min, logical filtrated air amount per minute and fermentating liquid volume are than for 0.8:1-1:1；After fermentation 36 hours, then add in fermentation tank
Yellowbird streptomycete seed liquor 5-15 part, dark yellow purple streptomycete seed liquor 5-15 part, 28-30 DEG C of aerobic culture 36-48 hour, stirring
200-160r/min, than for 1:1-1.2:1, logical filtrated air amount per minute detects that its bacterial content reaches 100g/ with fermentating liquid volume
L, you can add Lyceum fermentation of bacillus liquid 5-15 part, series bacillus fermentation liquid 5-15 part in Ma Kuo, terminates after mixing to send out
Ferment；
(4), release bacterium solution, packed products.
Wherein, preventing and treating Fructus Chaenomeliss mosaic disease zymocyte liquid using method be foliage-spray 1-5l/ mu, foliage-spray dilute
The multiple released is 50-100 times；Use once within every 7-10 days, continuous administration 5-10 time.
Wherein, yellowbird streptomycete seed liquor, dark yellow purple streptomycete seed liquor, Lyceum fermentation of bacillus liquid, class in Ma Kuo
Fermentation of bacillus liquid, the production of top spore Trichoderma spp. seed liquor can adopt following methods:
, the preparation of yellowbird streptomycete seed liquor
Take out yellowbird streptomyces species preservation pipe, draw flat board with Gause I solid medium and recovered, cultivate 7 days for 30 DEG C.
Under flat board, the streak inoculation of picking single bacterium colony is equipped with the Fructus Solani melongenae bottle of 150 milliliters of Gause I solid mediums, in incubator
In 30 DEG C cultivate 6-8 days, treat that lawn covers with Fructus Solani melongenae bottle, produce a large amount of spore powders and can use 500 milliliters of physiological saline solution to wash
De-, regulation spore concentration is 0.1 hundred million cfu/ml, as yellowbird streptomycete seed liquor.
, dark yellow purple streptomycete seed liquor preparation
Take out dark yellow purple streptomyces species preservation pipe, draw flat board with Gause I solid medium and recovered, 30 DEG C of cultures 7
My god.Under flat board, the streak inoculation of picking single bacterium colony, equipped with the Fructus Solani melongenae bottle of 150 milliliters of Gause I solid mediums, is being cultivated
Cultivate 6-8 days for 30 DEG C in case, treat that lawn covers with Fructus Solani melongenae bottle, produce the physiological saline solution that a large amount of spore powders can use 500 milliliters
Eluting, regulation spore concentration is 0.2 hundred million cfu/ml, as dark yellow purple streptomycete seed liquor.
3., the preparation of Lyceum fermentation of bacillus liquid
Take out Lyceum bacillus cereuss preservation pipe, draw flat board respectively with lb solid medium and recovered, cultivate 48 hours for 30 DEG C.?
Under flat board, picking single bacterium colony is seeded to equipped with lb solid medium, culture 48 hours in 30 DEG C of incubators, with 3000ml no
Bacterium water is seeded to the lawn eluting in three Fructus Solani melongenae bottles in the 500l fermentation tank equipped with 300l Lyceum bacillus culture medium,
Open stirring 120r/min, ventilation is 200l/min within first 12 hours, after 12 hours, ventilation is 320l/min, 30 DEG C of culture 26-
36 hours, treat that total spore content is not less than 6,000,000,000 cfu/ml, you can as Lyceum fermentation of bacillus liquid；
Wherein, described lb solid medium: yeast extract 5.0 g, peptone 10.0, nacl10.0 g, agar 20 g, water
1000ml, ph7.2；
Wherein, described Lyceum bacillus culture medium: glucose 5g/l, Semen Maydis powder 28 g/l, bean cake powder 45 g/l, magnesium sulfate
0.2 g/l, manganese sulfate 0.5 g/l, potassium dihydrogen phosphate 0.5 g/l, ph7.0.The condition of each medium sterilization is: 0.10-
0.15mpa, 121 DEG C sterilize 30 minutes.
4., in Ma Kuo series bacillus fermentation liquid preparation
Take out series bacillus preservation pipe in Ma Kuo, draw flat board respectively with na solid medium and recovered, 30 DEG C of cultures 48 are little
When.Under flat board, picking single bacterium colony is seeded to equipped with na solid medium, cultivates 48 hours in 30 DEG C of incubators, uses
3000ml sterilized water is seeded to the lawn eluting in three Fructus Solani melongenae bottles equipped with series bacillus culture medium in 300l Ma Kuo
In 500l fermentation tank, open stirring 120r/min, ventilation is 200l/min within first 12 hours, after 12 hours, ventilation is 320l/min
, 30 DEG C of culture 26-36 hours, treat that total spore content is not less than 6,000,000,000 cfu/ml, you can ferment as series bacillus in Ma Kuo
Liquid；
Wherein, na solid medium: (g/l) beef extract 3.0, peptone 5.0, glucose 2.5, agar 18, ph value 7.0
±0.1；
Wherein, series bacillus culture medium in described Ma Kuo: glucose 5g/l, Semen Maydis powder 20 g/l, rapeseed cake powder 35 g/l, sulfur
Sour magnesium 0.4 g/l, manganese sulfate 0.5 g/l, potassium dihydrogen phosphate 0.6 g/l, ph7.0.The condition of each medium sterilization is: 0.10-
0.15mpa, 121 DEG C sterilize 30 minutes.
5., push up the preparation of spore Trichoderma spp. seed liquor
Take out culture presevation pipe, draw flat board with pda solid medium and recovered, cultivate 48 hours for 30 DEG C.Picking under flat board
Single bacterium colony is rule to the Fructus Solani melongenae bottle equipped with pda solid medium, cultivates 3-5 days for 30 DEG C in incubator, treats that strain produces
After substantial amounts of spore, scraped with physiological saline solution, adjustment top spore reesei spores concentration is 0.5 hundred million cfu/ml, as top spore wood
Mould seed liquor；
Wherein, described pda solid medium: Rhizoma Solani tuber osi 200g, sucrose 20g, agar 20 g, water 1000ml.
Contain yellowbird streptomycete, dark yellow purple streptomycete in the zymocyte liquid of the present invention, push up spore Trichoderma spp., these three bacterium are in fermentation
Abundant cellulase, xylanase, the non-starch polysaccharide enzyme such as glucanase and amylase, egg can be produced during Chinese herbal medicine
White enzyme, destroys the cell wall of Chinese herbal medicine, the content of the active ingredient of Chinese herbal medicine can be improved several times, and produce during the fermentation
Raw canarius, the antibacterium of α-rubromycin, β-rubromycin, anthelmintic and antiviral antibiotic composition, greatly strengthen
Its antiviral and the activity of insect protected, organically combine effect of probiotic bacteria and Chinese herbal medicine, can greatly suppress papaw mosaic virus
With cut off its virus spread number of pest, thus can strengthen its preventing and treating papaw mosaic virus effect.
Series bacillus in Lyceum bacillus cereuss and Ma Kuo are contained, both spore are certainly in zymocyte liquid in the present invention
So boundary is widely distributed, can from soil separation and Culture, it can produce abundant amylase and protease, is that one kind has wide spectrum suppression
The ability that the antibacterial of bacterium activity produces secondary metabolite is stronger, the antibiotic of its secretion, antibacterial protein or polypeptides matter etc.
Play preferable Biological control effect.Both bacillus cereuss controlling plant diseases can come from it and can produce multiple antibacterials
Matter, including multiple compounds such as lipopeptid class, peptides, phospholipid, many alkenes, amino acidses and nucleic acids, these antibacterial mass-energy
The normal growth of suppression funguses, antibacterial, virus and bacterium substance etc..The lipopeptid that they produce in growth naturally and fermentation culture later stage
Class antibiotic is its most important antibiotic substance.Lipopeptide antibiotic includes iturin (iturins), general leather element
And Surfactin (surfactin), (fengycins) wherein iturin and general leather element have very strong antifungal and live
Property, and Surfactin has very high inhibitory activity to virus, tumor, mycoplasma；Two kinds of fermentation of bacillus Chinese herbal medicine
During, with Chinese herbal medicine as culture medium, some new Chinese herbal medicine functional group active factorses and somatomedin can be synthesized, through inspection
Survey, its Antagonistic protein and lipopeptide antibiotic, polypeptides matter substantially improves more than 20% than general culture medium, strengthens it
Cure the disease, the effect of diseases prevention.
The excellent effect of the present invention is: 1., technical scheme is by the research of Tiny ecosystem for many years and plantation warp
Test repetition test, meet the theory of Chinese medicine；It is to be sieved from more than 250 kinds of Chinese herbal medicine from invention Chinese herbal medicine formula
Choosing, according to the monarch of traditional Chinese medicine, seven emotions compatibility, eighteen incompatible medicamentss, nineteen medicaments of mutual restraint scheduling theory, screening to filtering out one by one
Effective component of Chinese herbal medicine, bacteriostasis carry out extracting, verify, finally determine formula, and being proved by probiotics fermention can available energy
Preventing and treating Fructus Chaenomeliss mosaic disease；
2., the Chinese herbal medicine in the preventing and treating Fructus Chaenomeliss mosaic disease zymocyte liquid of the present invention is in yellowbird streptomycete, dark yellow purple streptomycete, Lyceum
Bacillus cereuss, series bacillus in Ma Kuo, under the Fermentation of top spore Trichoderma spp., destroy the cell wall of Chinese herbal medicine, can be by medium-height grass
The content of the active ingredient of medicine improves 2-4 times, and produces canarius, α-rubromycin, β-rubromycin during the fermentation
Antibacterium, anthelmintic and antiviral antibiotic composition, greatly strengthen the activity of its antiviral and insect protected, organically combine
Probiotic bacteria and effect of Chinese herbal medicine, can greatly suppress papaw mosaic virus and the number of pest reducing its virus spread；
3., the various probiotic bacterias in the present invention are to filter out 5 kinds from 350 multiple beneficial microorganisms to greatly improve Chinese herbal medicine
Active ingredient utilization rate, and during the fermentation can symbiotic co-existence, promote mutually.This 5 kinds of bacterium be all can from soil,
On plant roots mark, separation obtains, and all has the effect promoting plant growing.
Specific embodiment
Embodiment 1
A kind of zymocyte liquid that can prevent and treat Fructus Chaenomeliss mosaic disease is it is characterised in that be made up of the raw material of following weight portion: 3 parts of the Radix Astragali,
3 parts of Radix Scutellariae, 3 parts of Rhizoma Coptidis, 3 parts of Flos Lonicerae, 3 parts of Radix Paeoniae Rubra, 3 parts of Radix Glycyrrhizae, 3 parts of Herba Houttuyniae, 5 parts of the Radix Pulsatillae, 5 parts of the Radix Rehmanniae, 5 parts of the Rhizoma Pinelliae,
4 parts of Radix Bupleuri, 5 parts of Folium Eriobotryae, 6 parts of Fructus Forsythiae, 6 parts of Folium Isatidiss, 6 parts of Radix Isatidis, 6 parts of Flos Lonicerae, 6 parts of Semen Cassiae, 6 parts of Flos Carthami, red
6 parts of ginseng, 2 parts of Herba Leonuri, 50 parts of soy molasses, 30 parts of bean cake powder, 850 parts of water, 10 parts of yellowbird streptomycete seed liquor, dark yellow purple chain
10 parts of mycete seed liquor, 10 parts of Lyceum fermentation of bacillus liquid, 10 parts of series bacillus fermentation liquid in Ma Kuo, push up spore Trichoderma spp. seed
10 parts of liquid；By size-reduced for said herbal medicine, immersion, heating and thermal insulation, fermentation and be obtained preventing and treating Fructus Chaenomeliss mosaic disease zymocyte liquid.
In order to achieve the above object, a kind of preparation method of the zymocyte liquid that can prevent and treat Fructus Chaenomeliss mosaic disease, walks including following
Rapid:
(1), various Chinese herbal medicine commercially are individually ground into 400 mesh fine powders；
(2), sequentially add in fermentation tank: 3 parts of the Radix Astragali, 3 parts of Radix Scutellariae, 3 parts of Rhizoma Coptidis, 3 parts of Flos Lonicerae, 3 parts of Radix Paeoniae Rubra, 3 parts of Radix Glycyrrhizae, fish
3 parts of raw meat grass, 5 parts of the Radix Pulsatillae, 5 parts of the Radix Rehmanniae, 5 parts of the Rhizoma Pinelliae, 4 parts of Radix Bupleuri, 5 parts of Folium Eriobotryae, 6 parts of Fructus Forsythiae, 6 parts of Folium Isatidiss, Radix Isatidis 6
Part, 6 parts of Flos Lonicerae, 6 parts of Semen Cassiae, 6 parts of Flos Carthami, 6 parts of Radix Salviae Miltiorrhizae, 2 parts of Herba Leonuri, 50 parts of soy molasses, 30 parts of bean cake powder, water
850 parts, after soaking 30 minutes, it is heated to 95 DEG C, be incubated two hours, be cooled to 30 DEG C；
(3), the coolant (2) produced in step adds 10 parts of the spore Trichoderma spp. seed liquor in top；30 DEG C -33 DEG C, open stirring 120-160r/
Min, logical filtrated air amount per minute and fermentating liquid volume are than for 0.8:1-1:1；After fermentation 36 hours, then add in fermentation tank
10 parts of yellowbird streptomycete seed liquor, dark yellow 10 parts of streptomycete seed liquor of purple, 28-30 DEG C of aerobic culture 36-48 hour, stirs 200-
160r/min, than for 1:1-1.2:1, logical filtrated air amount per minute detects that its bacterial content reaches 100g/l with fermentating liquid volume, that is,
10 parts of Lyceum fermentation of bacillus liquid can be added, 10 parts of series bacillus fermentation liquid in Ma Kuo, terminate fermentation after mixing；
(4), release bacterium solution, packed products.
Wherein, yellowbird streptomycete seed liquor, dark yellow purple streptomycete seed liquor, Lyceum fermentation of bacillus liquid, class in Ma Kuo
Fermentation of bacillus liquid, the production of top spore Trichoderma spp. seed liquor is using following production technology:
, the preparation of yellowbird streptomycete seed liquor
Take out yellowbird streptomyces species preservation pipe, draw flat board with Gause I solid medium and recovered, cultivate 7 days for 30 DEG C.
Under flat board, the streak inoculation of picking single bacterium colony is equipped with the Fructus Solani melongenae bottle of 150 milliliters of Gause I solid mediums, in incubator
In 30 DEG C cultivate 6-8 days, treat that lawn covers with Fructus Solani melongenae bottle, produce a large amount of spore powders and can use 500 milliliters of physiological saline solution to wash
De-, regulation spore concentration is 0.1 hundred million cfu/ml, as yellowbird streptomycete seed liquor.
, dark yellow purple streptomycete seed liquor preparation
Take out dark yellow purple streptomyces species preservation pipe, draw flat board with Gause I solid medium and recovered, 30 DEG C of cultures 7
My god.Under flat board, the streak inoculation of picking single bacterium colony, equipped with the Fructus Solani melongenae bottle of 150 milliliters of Gause I solid mediums, is being cultivated
Cultivate 6-8 days for 30 DEG C in case, treat that lawn covers with Fructus Solani melongenae bottle, produce the physiological saline solution that a large amount of spore powders can use 500 milliliters
Eluting, regulation spore concentration is 0.2 hundred million cfu/ml, as dark yellow purple streptomycete seed liquor.
3., the preparation of Lyceum fermentation of bacillus liquid
Take out Lyceum bacillus cereuss preservation pipe, draw flat board respectively with lb solid medium and recovered, cultivate 48 hours for 30 DEG C.?
Under flat board, picking single bacterium colony is seeded to equipped with lb solid medium, culture 48 hours in 30 DEG C of incubators, with 3000ml no
Bacterium water is seeded to the lawn eluting in three Fructus Solani melongenae bottles in the 500l fermentation tank equipped with 300l Lyceum bacillus culture medium,
Open stirring 120r/min, ventilation is 200l/min within first 12 hours, after 12 hours, ventilation is 320l/min, 30 DEG C of cultures 30
Hour, the total spore content of detection is 6,500,000,000 cfu/ml, that is, as Lyceum fermentation of bacillus liquid；
Wherein, described lb solid medium: yeast extract 5.0 g, peptone 10.0, nacl10.0 g, agar 20 g, water
1000ml, ph7.2；
Wherein, described Lyceum bacillus culture medium: glucose 5g/l, Semen Maydis powder 28 g/l, bean cake powder 45 g/l, magnesium sulfate
0.2 g/l, manganese sulfate 0.5 g/l, potassium dihydrogen phosphate 0.5 g/l, ph7.0.The condition of each medium sterilization is: 0.10-
0.15mpa, 121 DEG C sterilize 30 minutes.
4., in Ma Kuo series bacillus fermentation liquid preparation
Take out series bacillus preservation pipe in Ma Kuo, draw flat board respectively with na solid medium and recovered, 30 DEG C of cultures 48 are little
When.Under flat board, picking single bacterium colony is seeded to equipped with na solid medium, cultivates 48 hours in 30 DEG C of incubators, uses
3000ml sterilized water is seeded to the lawn eluting in three Fructus Solani melongenae bottles equipped with series bacillus culture medium in 300l Ma Kuo
In 500l fermentation tank, open stirring 120r/min, ventilation is 200l/min within first 12 hours, after 12 hours, ventilation is 320l/min
, cultivate 32 hours for 30 DEG C, the total spore content of detection is 6,200,000,000 cfu/ml, that is, as series bacillus fermentation liquid in Ma Kuo；
Wherein, na solid medium: (g/l) beef extract 3.0, peptone 5.0, glucose 2.5, agar 18, ph value 7.0
±0.1；
Wherein, series bacillus culture medium in described Ma Kuo: glucose 5g/l, Semen Maydis powder 20 g/l, rapeseed cake powder 35 g/l, sulfur
Sour magnesium 0.4 g/l, manganese sulfate 0.5 g/l, potassium dihydrogen phosphate 0.6 g/l, ph7.0.The condition of each medium sterilization is: 0.10-
0.15mpa, 121 DEG C sterilize 30 minutes.
5., push up the preparation of spore Trichoderma spp. seed liquor
Take out culture presevation pipe, draw flat board with pda solid medium and recovered, cultivate 48 hours for 30 DEG C.Picking under flat board
Single bacterium colony is rule to the Fructus Solani melongenae bottle equipped with pda solid medium, cultivates 3-5 days for 30 DEG C in incubator, treats that strain produces
After substantial amounts of spore, scraped with physiological saline solution, adjustment top spore reesei spores concentration is 0.5 hundred million cfu/ml, as top spore wood
Mould seed liquor；
Wherein, described pda culture medium: Rhizoma Solani tuber osi 200g, sucrose 20g, agar 20 g, water 1000ml.
Embodiment 2
Zymocyte liquid of the present invention detects to the prevention effect of Fructus Chaenomeliss mosaic disease, is tested on October 9th, 2015
2015 to December 9 was carried out in Zhanjiang, Guangdong Province Xuwen County Fructus Chaenomeliss base, and pilot region Fructus Chaenomeliss mosaic disease is fallen ill relatively
Weight, sickness rate has substantially achieved 25%；Carried out by the way of foliar spray.It is divided into two groups of process, test group: 10 mu, blade face
Spray 4l zymocyte liquid/mu, the multiple of the dilution of foliage-spray is 50 times；Use once within every 7 days, continuous administration 8 times.；Comparison
Group: 10 mu, sickness rate has reached 15%, directly by morbidity cutting down, and with 99% mineral oil (green grain husk) 150-200 times+48% poison with poison
1500 times of Tickss, every month is using twice；
Result of the test: prevention effect is shown in Table 1, and the Fructus Chaenomeliss mosaic disease sickness rate of test group is significantly lower than conventional treatment, and by original
Sickness rate is reduced to 5.2% for 25%, and does not cut Fructus Chaenomeliss, and therefore, its yield increased group is much higher, and yield increased group is high
57.07%, control the diffusion of mosaic disease completely, and most of morbidity Fructus Chaenomeliss have obtained recovery it was demonstrated that the zymocyte of the present invention
Liquid has significant effect of increasing production to preventing and treating Fructus Chaenomeliss mosaic disease with volume increase.
Table 1 zymocyte liquid is to the preventing and treating of Fructus Chaenomeliss mosaic disease and effect of increasing production
| Group | Initial sickness rate (%) | Sickness rate (%) | Preventive effect (%) | Yield (kg) | Rate of growth (%) |
| Test group | 25 | 5.2 | 79.2 | 3110 | 57.07 |
| Matched group | 15 | 12.4 | 1980 |
The above is only the preferred embodiment of the present invention it is noted that coming for those skilled in the art
Say, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should be regarded as
Protection scope of the present invention.
Claims (3)
1. a kind of zymocyte liquid that can prevent and treat Fructus Chaenomeliss mosaic disease is it is characterised in that be made up of the raw material of following weight portion: Radix Astragali 2-
4 parts, Radix Scutellariae 2-4 part, Rhizoma Coptidis 2-4 part, Flos Lonicerae 2-4 part, Radix Paeoniae Rubra 2-4 part, Radix Glycyrrhizae 2-4 part, Herba Houttuyniae 2-4 part, Radix Pulsatillae 2-6
Part, Radix Rehmanniae 2-6 part, Rhizoma Pinelliae 2-6 part, Radix Bupleuri 2-6 part, Folium Eriobotryae 2-6 part, Fructus Forsythiae 3-8 part, Folium Isatidiss 3-8 part, Radix Isatidis 3-8
Part, Flos Lonicerae 3-8 part, Semen Cassiae 3-8 part, Flos Carthami 3-8 part, Radix Salviae Miltiorrhizae 3-8 part, Herba Leonuri 1-3 part, soy molasses 40-60 part, bean
Dregs of rice powder 20-40 part, 850 parts of water, yellowbird streptomycete seed liquor 5-15 part, dark yellow purple streptomycete seed liquor 5-15 part, Lyceum spore
Bacillus fermentation liquid 5-15 part, series bacillus fermentation liquid 5-15 part in Ma Kuo, push up spore Trichoderma spp. seed liquor 5-15 part；By above-mentioned medium-height grass
Medicine is size-reduced, immersion, heating and thermal insulation, fermentation and be obtained preventing and treating Fructus Chaenomeliss mosaic disease zymocyte liquid.
2. the preparation method of the zymocyte liquid that can prevent and treat Fructus Chaenomeliss mosaic disease according to claim 1 is it is characterised in that include
Following steps:
(1), various Chinese herbal medicine commercially are individually ground into 400 mesh fine powders；
(2), sequentially add in fermentation tank: Radix Astragali 2-4 part, Radix Scutellariae 2-4 part, Rhizoma Coptidis 2-4 part, Flos Lonicerae 2-4 part, Radix Paeoniae Rubra 2-4 part,
Radix Glycyrrhizae 2-4 part, Herba Houttuyniae 2-4 part, Radix Pulsatillae 2-6 part, Radix Rehmanniae 2-6 part, Rhizoma Pinelliae 2-6 part, Radix Bupleuri 2-6 part, Folium Eriobotryae 2-6 part,
Fructus Forsythiae 3-8 part, Folium Isatidiss 3-8 part, Radix Isatidis 3-8 part, Flos Lonicerae 3-8 part, Semen Cassiae 3-8 part, Flos Carthami 3-8 part, Radix Salviae Miltiorrhizae 3-8
Part, Herba Leonuri 1-3 part, soy molasses 40-60 part, bean cake powder 20-40 part, 850 parts of water, after soaking 30 minutes, it is heated to 95 DEG C,
Insulation two hours, is cooled to 30 DEG C；
(3), the coolant (2) produced in step adds top spore Trichoderma spp. seed liquor 5-15 part；30 DEG C -33 DEG C, open stirring 120-160r/
Min, logical filtrated air amount per minute and fermentating liquid volume are than for 0.8:1-1:1；After fermentation 36 hours, then add in fermentation tank
Yellowbird streptomycete seed liquor 5-15 part, dark yellow purple streptomycete seed liquor 5-15 part, 28-30 DEG C of aerobic culture 36-48 hour, stirring
200-160r/min, logical filtrated air amount per minute and fermentating liquid volume, than for 1:1-1.2:1, detect that its bacterial content reaches
100g/l, you can add Lyceum fermentation of bacillus liquid 5-15 part, series bacillus fermentation liquid 5-15 part in Ma Kuo, tie after mixing
Bundle fermentation；
(4), release bacterium solution, packed products；
Wherein, yellowbird streptomycete seed liquor, dark yellow purple streptomycete seed liquor, Lyceum fermentation of bacillus liquid, class spore in Ma Kuo
Bacillus fermentation liquid, the production of top spore Trichoderma spp. seed liquor adopts following methods:
, the preparation of yellowbird streptomycete seed liquor
Take out yellowbird streptomyces species preservation pipe, draw flat board with Gause I solid medium and recovered, cultivate 7 days for 30 DEG C,
Under flat board, the streak inoculation of picking single bacterium colony is equipped with the Fructus Solani melongenae bottle of 150 milliliters of Gause I solid mediums, in incubator
In 30 DEG C cultivate 6-8 days, treat that lawn covers with Fructus Solani melongenae bottle, produce a large amount of spore powders and can use 500 milliliters of physiological saline solution to wash
De-, regulation spore concentration is 0.1 hundred million cfu/ml, as yellowbird streptomycete seed liquor；
, dark yellow purple streptomycete seed liquor preparation
Take out dark yellow purple streptomyces species preservation pipe, draw flat board with Gause I solid medium and recovered, 30 DEG C of cultures 7
My god, picking single bacterium colony streak inoculation under flat board, equipped with the Fructus Solani melongenae bottle of 150 milliliters of Gause I solid mediums, is being cultivated
Cultivate 6-8 days for 30 DEG C in case, treat that lawn covers with Fructus Solani melongenae bottle, produce the physiological saline solution that a large amount of spore powders can use 500 milliliters
Eluting, regulation spore concentration is 0.2 hundred million cfu/ml, as dark yellow purple streptomycete seed liquor；
3., the preparation of Lyceum fermentation of bacillus liquid
Take out Lyceum bacillus cereuss preservation pipe, draw flat board respectively with lb solid medium and recovered, cultivate 48 hours for 30 DEG C,
Under flat board, picking single bacterium colony is seeded to equipped with lb solid medium, culture 48 hours in 30 DEG C of incubators, with 3000ml no
Bacterium water is seeded to the lawn eluting in three Fructus Solani melongenae bottles in the 500l fermentation tank equipped with 300l Lyceum bacillus culture medium,
Open stirring 120r/min, ventilation is 200l/min within first 12 hours, after 12 hours, ventilation is 320l/min, 30 DEG C of culture 26-
36 hours, treat that total spore content is not less than 6,000,000,000 cfu/ml, you can as Lyceum fermentation of bacillus liquid；
Wherein, described lb solid medium: yeast extract 5.0 g, peptone 10.0, nacl10.0 g, agar 20 g, water
1000ml, ph7.2；
Wherein, described Lyceum bacillus culture medium: glucose 5g/l, Semen Maydis powder 28 g/l, bean cake powder 45 g/l, magnesium sulfate
0.2 g/l, manganese sulfate 0.5 g/l, potassium dihydrogen phosphate 0.5 g/l, ph7.0；The condition of each medium sterilization is: 0.10-
0.15mpa, 121 DEG C sterilize 30 minutes；
4., in Ma Kuo series bacillus fermentation liquid preparation
Take out series bacillus preservation pipe in Ma Kuo, draw flat board respectively with na solid medium and recovered, 30 DEG C of cultures 48 are little
When, under flat board, picking single bacterium colony is seeded to equipped with na solid medium, cultivates 48 hours in 30 DEG C of incubators, uses
3000ml sterilized water is seeded to the lawn eluting in three Fructus Solani melongenae bottles equipped with series bacillus culture medium in 300l Ma Kuo
In 500l fermentation tank, open stirring 120r/min, ventilation is 200l/min within first 12 hours, after 12 hours, ventilation is 320l/min
, 30 DEG C of culture 26-36 hours, treat that total spore content is not less than 6,000,000,000 cfu/ml, you can ferment as series bacillus in Ma Kuo
Liquid；
Wherein, na solid medium: (g/l) beef extract 3.0, peptone 5.0, glucose 2.5, agar 18, ph value 7.0
±0.1；
Wherein, series bacillus culture medium in described Ma Kuo: glucose 5g/l, Semen Maydis powder 20 g/l, rapeseed cake powder 35 g/l, sulfur
Sour magnesium 0.4 g/l, manganese sulfate 0.5 g/l, potassium dihydrogen phosphate 0.6 g/l, ph7.0, the condition of each medium sterilization is: 0.10-
0.15mpa, 121 DEG C sterilize 30 minutes；
5., push up the preparation of spore Trichoderma spp. seed liquor
Take out culture presevation pipe, draw flat board with pda solid medium and recovered, cultivate 48 hours for 30 DEG C, picking under flat board
Single bacterium colony is rule to the Fructus Solani melongenae bottle equipped with pda solid medium, cultivates 3-5 days for 30 DEG C in incubator, treats that strain produces
After substantial amounts of spore, scraped with physiological saline solution, adjustment top spore reesei spores concentration is 0.5 hundred million cfu/ml, as top spore wood
Mould seed liquor；
Wherein, described pda solid medium: Rhizoma Solani tuber osi 200g, sucrose 20g, agar 20 g, water 1000ml.
3. a kind of zymocyte liquid that can prevent and treat Fructus Chaenomeliss mosaic disease according to claim 1 is it is characterised in that prevent and treat Fructus Chaenomeliss
The using method of the zymocyte liquid of leaf disease is foliage-spray 1-5l/ mu, and the multiple of the dilution of foliage-spray is 50-100 times；Every 7-
Use once within 10 days, continuous administration 5-10 time.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108250002A (en) * | 2018-04-14 | 2018-07-06 | 潍坊奥丰作物植保服务有限公司 | A kind of complex micro organism fungicide containing Chinese herbal medicine and preparation method thereof |
| CN117343860A (en) * | 2023-07-31 | 2024-01-05 | 沈阳农业大学 | Streptomyces SN0246 and method for preparing polyketide caner tetracyclic ketone by using same |
-
2016
- 2016-08-29 CN CN201610745985.XA patent/CN106359473A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108250002A (en) * | 2018-04-14 | 2018-07-06 | 潍坊奥丰作物植保服务有限公司 | A kind of complex micro organism fungicide containing Chinese herbal medicine and preparation method thereof |
| CN117343860A (en) * | 2023-07-31 | 2024-01-05 | 沈阳农业大学 | Streptomyces SN0246 and method for preparing polyketide caner tetracyclic ketone by using same |
| CN117343860B (en) * | 2023-07-31 | 2024-04-16 | 沈阳农业大学 | Streptomyces SN0246 and method for preparing polyketide caner tetracyclic ketone by using same |
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Application publication date: 20170201 |