CN104745510B - A kind of lactobacillus fermenti and its application in fungal diseases of plants preventing and treating - Google Patents
A kind of lactobacillus fermenti and its application in fungal diseases of plants preventing and treating Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
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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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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
Abstract
A kind of application the invention discloses lactobacillus fermenti and its in fungal diseases of plants preventing and treating, belongs to biological prevention and control agent field.The bacteria strain is isolated in the soil planted under tomato, and its Classification And Nomenclature is lactobacillus fermenti, and the Classification system of strain is：Lactobacillus fermentum, join the microorganism of evidence：NJHYHWGY20130877, preservation date are on 01 17th, 2014, and collection numbering of registering on the books is CGMCC No.8735.The fermented culture of lactobacillus fermenti obtains zymotic fluid, zymotic fluid concentration then is prepared into lactein concentrate, then diluted and be sprayed on corps leaf surface；The bacterial strain works well in terms of farm crop fungus disease is suppressed, while has the function of promoting crop growth, meets ecological agriculture requirement.
Description
Technical field
The invention belongs to biological prevention and control agent field, and in particular to a kind of lactic acid bacteria and its answering in fungal diseases of plants preventing and treating
With.
Technical background
Today's society, agricultural sustainable development have turned into world's most countries and have dominated trend.Agricultural problem is as economical
Social development matter of utmost importance, various countries' agenda is put on.China is used as a large agricultural country, and secure agricultural production is concerning national economy
It is stable with social harmony.Largely although agricultural product can be made significantly to increase production using agricultural chemicals such as chemical fertilizer, agricultural chemicals, also band
Many adverse effects are carried out.For this problem, there is the friendly biological pesticide of green ecological in succession in China, is used with Traditional Agricultural
Chemicals is compared, and is had the advantages that low toxicity, selectively strong, efficient, low-residual, is easily decomposed.Mainly include Natural Enemies of Insects, plant
Source pesticide, microbial pesticide, genetically modified organism, biochemical pesticides and antibiotic agricultural chemicals.
Lactobacillus is also known as Bacillus acidi lactici, is a category for lactic acid bacteria.Cell is generally in regular elongated rod shape, 0.5~1.2 μ m
1.0~10.0 μm, sometimes to be spherical, into short chain.Gram-positive, spore is not given birth to, cell is rare to be moved with peritrichous.It is facultative to detest
Oxygen, sometimes micro- aerobic, bacterium colony projection, full edge, colourless, 2~5mm of diameter on nutrient agar.Chemoheterotrophic bacteria is, it is necessary to nutritious
Culture medium.Glycometabolism is decomposed in fermentation, and more than 50% is lactic acid in end-product.Azymic lactate, nitrate is not reduced, not liquid
Change gelatin, catalase and oxidizing ferment are all negative, rare to cause a disease.30~40 DEG C of optimum temperature, acid resistance is strong, and optimal pH 5.5~
Remain to survive under the conditions of 6.0, pH3.0~4.5.
Lactic acid bacteria is distributed widely in environment, particularly in animal, vegetables and food, generally inhabits bird and vertebrate
Alimentary canal, and in the urethra of mammal.Plant surface, soil, dairy products, meat products, beer, grape wine, fruit juice,
In brewer's wort, sewage and human and animal excreta, separate.Because lactic acid bacteria can produce a large amount of bateriostatics in fermentating metabolism
Matter, including organic acid, enzyme, lactein, hydrogen peroxide, carbon dioxide, biacetyl etc., thus can effectively suppress spoilage organisms and
The growth of pathogenic bacteria, and there is growth promoting function.In addition, lactic acid bacteria has abundant species diversity, be not only research classification,
The ideal material of biochemical, heredity, molecular biology and genetic engineering, and have in fields such as farming and animal husbandry, industry, food and medicines
There is high application value.
The content of the invention
The purpose of the present invention is to propose to one plant of lactobacillus fermenti, it is a further object of the present invention to provide above-mentioned fermentation side
Method, further object of the present invention are to provide application of the lactobacillus fermenti in terms of fungal diseases of plants suppression.
Technical scheme is such as：One strains of lactic acid bacteria, its Classification And Nomenclature are lactobacillus fermenti, the Classification system of strain
It is：Lactobacillus fermentum, join the microorganism of evidence：NJHYHWGY20130877, preservation date are 01 month 2014
17, collection numbering of registering on the books was CGMCC No.8735.
Present invention also offers the fermentation process of above-mentioned lactobacillus fermenti, and it is comprised the following steps that：By lactobacillus fermenti
CGMCC No.8735 are inoculated in solid medium, 30~40 DEG C of 12~24h of culture；A bacterium is chosen in solid medium
Fall, picking thalline, in conical flask of the access equipped with seed liquid culture medium, it is 100~180r/min to control rotating speed, 30~40 DEG C of bars
10~22h is cultivated under part and obtains lactobacillus fermenti seed liquor；Take in conical flask of the seed liquor access equipped with fermentation medium, control turns
Speed is 120~160r/min, and cultivating 24~36h under the conditions of 30~40 DEG C obtains zymotic fluid.
It is preferred that the component of above-mentioned solid medium is：9~11g/L of peptone, 9~11g/L of beef extract, yeast extract 4.5~
5.5g/L, 1.5~2.5g/L of DisodiumHydrogen Citrate, 18~22g/L of glucose, 4.5~5.5g/L of sodium acetate, dipotassium hydrogen phosphate
1.5~2.5g/L, 0.5~0.6g/L of magnesium sulfate, 0.2~0.3g/L of manganese sulfate, 15~25g/L of agar.
It is preferred that the component of above-mentioned seed liquid culture medium is：9~12g/L of peptone, 8~12g/L of beef extract, yeast extract 4~
6g/L, 16~24g/L of glucose, 1.5~2.5g/L of DisodiumHydrogen Citrate, 4~6g/L of sodium acetate, dipotassium hydrogen phosphate 1.5~
2.5g/L, 0.5~0.6g/L of magnesium sulfate, 0.2~0.3g/L of manganese sulfate.
It is preferred that the component of above-mentioned fermentation medium is：20~30g/L of sucrose, yeast extract 15~25g/L, KCl0.6~
1.0g/L, 1.5~2.5g/L of DisodiumHydrogen Citrate, 4~6g/L of sodium acetate, dipotassium hydrogen phosphate 1.5~2.5g/L, pH 5.5~
6.5。
It is preferred that the liquid amount of above-mentioned seed liquid culture medium, which is seed liquor culture volume, accounts for the 16~40% of conical flask capacity；
The liquid amount of fermentation medium is that fermentation medium volume accounts for the 16~40% of conical flask capacity, seed liquor and fermentation medium
Volume ratio is 1：(10~100).
Present invention also offers application of the above-mentioned lactobacillus fermenti in fungal diseases of plants preventing and treating, wherein plant mycosis
Evil bacterial strain preferred cotton verticillium wilt, graw mold of tomato or rice sheath blight disease；Its application process, will to prepare lactein concentrate
The lactein concentrate dilutes 1000~1500 times, is sprayed on corps leaf surface；
Wherein, the lactein concentration liquid and preparation method thereof concretely comprises the following steps, by zymotic fluid obtained by above-mentioned fermentation 8000
Under the conditions of~12000r/min, 20~40min is centrifuged, lower floor's bacterium mud is discarded, it is standby to obtain fermented supernatant fluid；Fermented supernatant fluid is existed
Concentrate is concentrated to give under vacuum condition, wherein volume concentration multiple is 10~20 times；Ammonium sulfate is added into concentrate,
Whirlpool shakes 1~2h, and ammonium sulfate quality saturation degree is 40~50% wherein in solution；Under the conditions of 18000~20000r/min from
25~40min of the heart, supernatant is removed, take lower sediment to be dissolved in the citrate buffer solution that pH is 4.0~5.0, obtain lactic acid bacteria
The concentrate of element, wherein precipitation and the ratio of citrate buffer solution are 1：(100~1000).
The antibacterial bacteriostatic diameter for being verified as, lactein concentrate suppression pathogen being surveyed using Odontothrips loti, is calculated
Bacteriostasis rate；With oese picking pathogen mycelium in centrifuge tube, whirlpool concussion crushes to obtain pathogen hypha fluid, pipettes bacterium
Filament liquid, is spread evenly across in PDA culture medium, and its middle plateform specification is diameter 90mm, high 16mm；In the flat board containing pathogen
On be equidistantly placed Oxford cup 4, after standing 5~10min, the concentration of 100~200 μ L lacteins is added into each Oxford cup
Liquid, wherein Oxford cup specification are 6.0 ± 0.1mm of internal diameter, external diameter 8.0 ± 0.1mm, high 10.0 ± 0.1mm；After 28 DEG C of culture 24h
Survey antibacterial circle diameter.Pathogen inhibiting rate is calculated, wherein bacteriostasis rate calculation formula is：
Growth of pathogenic bacteria inhibiting rate (%)=(antibacterial circle diameter-cups and dishes diameter)/(control colony diameter-cups and dishes diameter)
× 100%
The microorganism of above-mentioned lactobacillus fermenti Lactobacillus fermentum ginsengs evidence：NJHYHWGY20130877, it is
By experimental group from plantation tomato soil in it is isolated.
CGMCC No.8735 bacterial strains have following properties：
1st, form and cultural characteristic
Cultivated on MRS culture mediums, bacterium colony is flat, circular or irregularly arrive coarse, usually transparent, non-pigment；Gram
Positive bacteria, it is shaft-like, gemma is not produced, size is 0.4~1.0 × 2.5~3.0 μm, paired or chaining, is not moved；Heterofermentation, from
Glucose produces sour aerogenesis；Lactobacillus fermenti amphimicrobian, its thalli growth and metabolite generation will appropraite conditions：Growth temperature
Spend for 15~45 DEG C, optimum temperature is 34~40 DEG C, likes slant acidity, pH5.5~7.0, optimal pH 6.0~6.8.
2nd, physiological and biochemical property：
The major physiological biochemical character of lactobacillus fermenti CGMCC No.8735 bacterial strains is shown in Table 1：
The physiological and biochemical property of the bacterial strain of table 1
Note：+：Positive or growth；—：Feminine gender does not grow
Beneficial effect：
The invention discloses one plant of lactobacillus fermenti CGMCC No.8735 and its in crop plants fungal disease preventing and treating side
The application in face, wherein have obvious inhibiting effect to cotton verticillium wilt, it is inhibited to graw mold of tomato or rice sheath blight disease.
Preservation information
The microorganism of above-mentioned lactobacillus fermenti Lactobacillus fermentum ginsengs evidence：NJHYHWGY20130877 is
By this laboratory seed selection and it is preserved in common micro-organisms center (court of Beijing of China General Microbiological culture presevation administration committee
The positive institute 3 of area's North Star West Road 1, Institute of Microorganism, Academia Sinica), it is referred to as CGMCC, and the numbering registered on the books is
CGMCC No.8735, preservation date are：On 01 17th, 2014.
Embodiment
The present invention provides a kind of lactobacillus fermenti Lactobacillus fermentum CGMCC No.8735 and its true
Application in terms of fungus diseases suppression, example is set forth below and is further described.
Example one：
(1) strain culturing
Lactobacillus fermenti CGMCC No.8735 are inoculated in solid medium, 30 DEG C of culture 12h.From solid medium
One bacterium colony of upper selection, picking thalline, access are equipped with the 250mL conical flasks of seed liquid culture medium, wherein seed liquid culture medium
Volume is 16% (in 250mL conical flask load 40mL seeds liquid culture medium) of conical flask capacity, rotating speed 100r/min, 30 DEG C of bars
10h is cultivated under part and obtains seed liquor；Take in 250mL conical flasks of the seed liquor access equipped with fermentation medium, wherein fermentation medium
Volume is 16% (in 250mL conical flask load 40mL seeds liquid culture medium) of conical flask capacity, seed liquor and fermentation medium
Volume ratio be 1：100, rotating speed 120r/min, culture 28h obtains zymotic fluid under the conditions of 30 DEG C.
Above-mentioned solid medium component is (g/L)：Peptone 9, beef extract 9, yeast extract 4.5, glucose 18, hydrogen citrate
Disodium 1.5, sodium acetate 4.5, dipotassium hydrogen phosphate 1.5, magnesium sulfate 0.5, manganese sulfate 0.2, agar 15.
Seed culture medium component is (g/L)：Peptone 9, beef extract 8, yeast extract 4, glucose 16, DisodiumHydrogen Citrate
1.5, sodium acetate 4, dipotassium hydrogen phosphate 1.5, magnesium sulfate 0.5, manganese sulfate 0.2.
Fermentation medium component is (g/L)：Yeast extract 15, sucrose 20, DisodiumHydrogen Citrate 1.5, sodium acetate 4, phosphoric acid hydrogen
Dipotassium 1.5, KCl 0.6, pH5.5.
(2) prepared by fermented supernatant fluid
By zymotic fluid under the conditions of 8000r/min, 20min is centrifuged, lower floor's bacterium mud is discarded, takes fermented supernatant fluid standby.
(3) prepared by lactein concentrate
Fermented supernatant fluid is concentrated to give concentrate under vacuum, wherein volume concentration multiple is 10 times；To concentration
Ammonium sulfate is added in liquid, whirlpool concussion 1h, ammonium sulfate saturation degree is 40% wherein in solution；Under the conditions of 18000r/min from
Heart 25min, supernatant is removed, take lower sediment to be dissolved in the citrate buffer solution that pH is 4.0, obtain the concentration of lactein
Liquid, wherein precipitation and the ratio of citrate buffer solution are 1：100.
(4) bacteriostasis rate calculates
The bacteriostatic diameter of lactein concentrate suppression pathogen is surveyed using Odontothrips loti, calculates bacteriostasis rate.Use oese
Picking pathogen mycelium the broken pathogen hypha fluid of whirlpool concussion, pipettes hypha fluid, in 1.5mL centrifuge tubes
It is even to be coated in PDA culture medium；4 Oxford cups are equidistantly placed on the flat board containing pathogen, after standing 5min, to each ox
100 μ L lactein concentrates are added in the cup of Tianjin；Antibacterial circle diameter is surveyed after 28 DEG C of culture 24h.Calculate pathogen inhibiting rate：
Above-mentioned growth of pathogenic bacteria inhibiting rate (%)=(antibacterial circle diameter-cups and dishes diameter)/(control colony diameter-cups and dishes
Diameter) × 100%
Flat board specification is (mm)：Diameter 90 is high by 16.
The amount that PDA culture medium is poured into flat board is (mL)：25.
Oxford cup specification is (mm)：Internal diameter 6.0 ± 0.1, external diameter 8.0 ± 0.1 are high by 10.0 ± 0.1.
Concrete outcome is shown in Table 1：
Bacteriostasis rate of the table 1 to different fungal diseases
Note：Carbendazim and procymidone concentration are 10 μ g/mL
Lactobacillus fermenti CGMCC No.8735 fermented liquids can effectively suppress cotton verticillium wilt, tomato ash as shown in Table 1
Mildew or rice banded sclerotial blight pathogen.It is wherein best to the inhibition of cotton verticillium wilt, inhibiting rate 59.14%.
Example two：
(1) strain culturing
Lactobacillus fermenti CGMCC No.8735 are inoculated in solid medium, 36 DEG C of culture 18h.From solid medium
One bacterium colony of upper selection, picking thalline, access are equipped with the 250mL conical flasks of seed liquid culture medium, wherein seed liquid culture medium
Volume is the 32% of conical flask capacity, rotating speed 140r/min, and culture 16h obtains seed liquor under the conditions of 36 DEG C；Take seed liquor access dress
Have in the 250mL conical flasks of fermentation medium, wherein fermentation medium volume is the 32% of conical flask capacity, seed liquor and fermentation
The volume ratio of culture medium is 1：20, rotating speed 140r/min, culture 28h obtains zymotic fluid under the conditions of 36 DEG C.
Above-mentioned solid medium component is (g/L)：Peptone 10, beef extract 10, yeast extract 5, glucose 20, hydrogen citrate
Disodium 2, sodium acetate 5, dipotassium hydrogen phosphate 2, magnesium sulfate 0.55, manganese sulfate 0.25, agar 18.
Seed culture medium component is (g/L)：Peptone 10, beef extract 10, yeast extract 5, glucose 20, DisodiumHydrogen Citrate
2, sodium acetate 5, dipotassium hydrogen phosphate 2, magnesium sulfate 0.55, manganese sulfate 0.25.
Fermentation medium component is (g/L)：Yeast extract 18, sucrose 23, DisodiumHydrogen Citrate 2, sodium acetate 5, phosphoric acid hydrogen two
Potassium 2, KCl 0.85, pH6.0.
(2) prepared by fermented supernatant fluid
By zymotic fluid under the conditions of 10000r/min, 30min is centrifuged, lower floor's bacterium mud is discarded, takes fermented supernatant fluid standby.
(3) prepared by lactein concentrate
Fermented supernatant fluid is concentrated to give concentrate under vacuum, wherein volume concentration multiple is 15 times；To concentration
Ammonium sulfate is added in liquid, whirlpool concussion 1.5h, ammonium sulfate saturation degree is 45% wherein in solution；Under the conditions of 19000r/min
32min is centrifuged, removes supernatant, takes lower sediment to be dissolved in the citrate buffer solution that pH is 4.5, obtains the concentration of lactein
Liquid, wherein precipitation and the ratio of citrate buffer solution are 1：1000.
(4) bacteriostasis rate calculates
The bacteriostatic diameter of lactein concentrate suppression pathogen is surveyed using Odontothrips loti, calculates bacteriostasis rate.Use oese
Picking pathogen mycelium the broken pathogen hypha fluid of whirlpool concussion, pipettes hypha fluid, in 1.5mL centrifuge tubes
It is even to be coated in PDA culture medium；4 Oxford cups are equidistantly placed on the flat board containing pathogen, after standing 5min, to each ox
150 μ L lactein concentrates are added in the cup of Tianjin；Antibacterial circle diameter is surveyed after 28 DEG C of culture 24h.Calculate pathogen inhibiting rate：
Above-mentioned growth of pathogenic bacteria inhibiting rate (%)=(antibacterial circle diameter-cups and dishes diameter)/(control colony diameter-cups and dishes
Diameter) × 100%
Flat board specification is (mm)：Diameter 90 is high by 16.
The amount that PDA culture medium is poured into flat board is (mL)：25.
Oxford cup specification is (mm)：Internal diameter 6.0 ± 0.1, external diameter 8.0 ± 0.1 are high by 10.0 ± 0.1.
Concrete outcome is shown in Table 2：
Bacteriostasis rate of the table 2 to different fungal diseases
Note：Carbendazim and procymidone concentration are 10 μ g/mL
Lactobacillus fermenti CGMCC No.8735 fermented liquids can effectively suppress cotton verticillium wilt, tomato ash as shown in Table 2
Mildew or rice banded sclerotial blight pathogen.It is wherein best to the inhibition of cotton verticillium wilt, inhibiting rate 71.41%.
Example three：
(1) strain culturing
Lactobacillus fermenti CGMCC No.8735 are inoculated in solid medium, 40 DEG C of culture 24h.From solid medium
One bacterium colony of upper selection, picking thalline, access are equipped with the 250mL conical flasks of seed liquid culture medium, wherein seed liquid culture medium
It is the 40% of conical flask capacity, rotating speed 180r/min, culture 22h obtains seed liquor under the conditions of 40 DEG C；Seed liquor access is taken equipped with hair
In the 250mL conical flasks of ferment culture medium, wherein fermentation medium volume is the 40% of conical flask capacity, seed liquor and fermented and cultured
The volume ratio of base is 1：10, rotating speed 160r/min, culture 36h obtains zymotic fluid under the conditions of 40 DEG C.
Above-mentioned solid medium component is (g/L)：Peptone 11, beef extract 11, yeast extract 5.5, glucose 22, citric acid
Disodium hydrogen 2.5, sodium acetate 5.5, dipotassium hydrogen phosphate 2.5, magnesium sulfate 0.6, manganese sulfate 0.3, agar 20.
Seed culture medium component is (g/L)：Peptone 12, beef extract 12, yeast extract 6, glucose 24, DisodiumHydrogen Citrate
2.5, sodium acetate 6, dipotassium hydrogen phosphate 2.5, magnesium sulfate 0.6, manganese sulfate 0.3.
Fermentation medium component is (g/L)：Yeast extract 25, sucrose 30, DisodiumHydrogen Citrate 2.5, sodium acetate 6, phosphoric acid hydrogen
Dipotassium 2.5, KCl 1.0, pH6.5.
(2) prepared by fermented supernatant fluid
By zymotic fluid under the conditions of 12000r/min, 40min is centrifuged, lower floor's bacterium mud is discarded, takes fermented supernatant fluid standby.
(3) prepared by lactein concentrate
Fermented supernatant fluid is concentrated to give concentrate under vacuum, wherein volume concentration multiple is 20 times；To concentration
Ammonium sulfate is added in liquid, whirlpool concussion 2h, ammonium sulfate saturation degree is 50% wherein in solution；Under the conditions of 20000r/min from
Heart 40min, supernatant is removed, take lower sediment to be dissolved in the citrate buffer solution that pH is 5.0, obtain the concentration of lactein
Liquid, wherein precipitation and the ratio of citrate buffer solution are 1：500.
(4) bacteriostasis rate calculates
The bacteriostatic diameter of lactein concentrate suppression pathogen is surveyed using Odontothrips loti, calculates bacteriostasis rate.Use oese
Picking pathogen mycelium the broken pathogen hypha fluid of whirlpool concussion, pipettes hypha fluid, in 1.5mL centrifuge tubes
It is even to be coated in PDA culture medium；4 Oxford cups are equidistantly placed on the flat board containing pathogen, after standing 10min, to each
200 μ L lactein concentrates are added in Oxford cup；Antibacterial circle diameter is surveyed after 28 DEG C of culture 24h.Calculate pathogen inhibiting rate：
Above-mentioned growth of pathogenic bacteria inhibiting rate (%)=(antibacterial circle diameter-cups and dishes diameter)/(control colony diameter-cups and dishes
Diameter) × 100%
Flat board specification is (mm)：Diameter 90 is high by 16.
The amount that PDA culture medium is poured into flat board is (mL)：25.
Oxford cup specification is (mm)：Internal diameter 6.0 ± 0.1, external diameter 8.0 ± 0.1 are high by 10.0 ± 0.1.
Concrete outcome is shown in Table 3：
Bacteriostasis rate of the table 3 to different fungal diseases
Note：Carbendazim and procymidone concentration are 10 μ g/mL
Lactobacillus fermenti CGMCC No.8735 fermented liquids can effectively suppress cotton verticillium wilt, tomato ash as shown in Table 3
Mildew or rice banded sclerotial blight pathogen.It is wherein best to the inhibition of cotton verticillium wilt, inhibiting rate 61.32%.
The present invention is used for green bio and prevented and treated, and is had broad application prospects in agriculture field.By to lactobacillus fermenti
The process exploitation of lactobacillus fermenti Lactobacillus fermentum CGMCC No.8735 fermentation lactic acid producing rhzomorphs, is reduced
Chemical pesticide usage amount, agricultural product security is improved, be advantageous to environmental protection and agricultural sustainable development.
Claims (4)
1. one plant of lactobacillus fermenti, the Classification system of strain are：Lactobacillus fermentum, join the microorganism of evidence：
NJHYHWGY20130877, preservation date are on 01 17th, 2014, and collection numbering of registering on the books is CGMCC
No.8735。
2. a kind of fermentation process using lactobacillus fermenti as claimed in claim 1, it is comprised the following steps that：By lactobacillus fermenti
CGMCC No.8735 are inoculated in solid medium, 30~40 DEG C of 12~24h of culture；A bacterium is chosen in solid medium
Fall, picking thalline, in conical flask of the access equipped with seed liquid culture medium, it is 100~180r/min to control rotating speed, 30~40 DEG C of bars
10~22h is cultivated under part and obtains lactobacillus fermenti seed liquor；Take in conical flask of the seed liquor access equipped with fermentation medium, control turns
Speed is 120~160r/min, and cultivating 24~36h under the conditions of 30~40 DEG C obtains zymotic fluid；The group of wherein described solid medium
It is divided into：9~11g/L of peptone, 9~11g/L of beef extract, 4.5~5.5g/L of yeast extract, 1.5~2.5g/L of DisodiumHydrogen Citrate,
18~22g/L of glucose, 4.5~5.5g/L of sodium acetate, 1.5~2.5g/L of dipotassium hydrogen phosphate, 0.5~0.6g/L of magnesium sulfate, sulphur
Sour 0.2~0.3g/L of manganese, 15~25g/L of agar；The component of described seed liquid culture medium is：9~12g/L of peptone, beef
8~12g/L of cream, 4~6g/L of yeast extract, 16~24g/L of glucose, 1.5~2.5g/L of DisodiumHydrogen Citrate, 4~6g/ of sodium acetate
L, 1.5~2.5g/L of dipotassium hydrogen phosphate, 0.5~0.6g/L of magnesium sulfate, 0.2~0.3g/L of manganese sulfate；Described fermentation medium
Component be：20~30g/L of sucrose, yeast extract 15~25g/L, KCl 0.6~1.0g/L, 1.5~2.5g/ of DisodiumHydrogen Citrate
L, 4~6g/L of sodium acetate, dipotassium hydrogen phosphate 1.5~2.5g/L, pH 5.5~6.5.
3. fermentation process as claimed in claim 2, it is characterised in that seed liquor culture volume for conical flask capacity 16~
40%；Fermentation medium volume is the 16~40% of conical flask capacity, and the volume ratio of seed liquor and fermentation medium is 1：(10~
100)。
4. a kind of lactobacillus fermenti as claimed in claim 1 is prevented in cotton verticillium wilt, graw mold of tomato or rice banded sclerotial blight disease
Control the application of aspect.
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| CN109452284A (en) * | 2018-11-19 | 2019-03-12 | 南京工业大学 | A kind of preparation process of amino-oligosaccharide and bacillus compound preparation |
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| CN101525633A (en) * | 2009-04-13 | 2009-09-09 | 天津科技大学 | Method for preparing plant pathomycete zymotic fluid produced by lactobacillus |
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