CN110511891A - A kind of and anti-tobacco root rot and the microbial bacterial agent of balck shank and its preparation method and application - Google Patents
A kind of and anti-tobacco root rot and the microbial bacterial agent of balck shank and its preparation method and application Download PDFInfo
- Publication number
- CN110511891A CN110511891A CN201910789604.1A CN201910789604A CN110511891A CN 110511891 A CN110511891 A CN 110511891A CN 201910789604 A CN201910789604 A CN 201910789604A CN 110511891 A CN110511891 A CN 110511891A
- Authority
- CN
- China
- Prior art keywords
- bacillus
- bacillus subtilis
- gram
- bacterial agent
- microbial bacterial
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- 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
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Virology (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Environmental Sciences (AREA)
- Agronomy & Crop Science (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Pest Control & Pesticides (AREA)
- Biomedical Technology (AREA)
- Dentistry (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Plant Pathology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The present invention provides a kind of and anti-tobacco root rot and the microbial bacterial agent of balck shank and its preparation method and application, belong to technical field of biological fertilizer.And the microbial bacterial agent of anti-tobacco root rot and balck shank, it is characterised in that it includes gram lining institute series bacillus Paenibacillus kribbensis that bacillus subtilis Bacillus subtilis and deposit number that deposit number is CGMCC No.10248 are CGMCC No.17248；Concentration >=1 × 10 of the bacillus subtilis9CFU/mL；Concentration >=0.5 × 10 of described gram of lining institute series bacillus9CFU/mL.Apply the disease incidence that tobacco root rot and balck shank can not only be effectively reduced in microbial bacterial agent provided by the invention, moreover it is possible to be obviously promoted the growth of cigarette strain and the raising of yield.
Description
Technical field
The invention belongs to technical field of biological fertilizer, and in particular to a kind of and anti-tobacco root rot and balck shank microorganism
Microbial inoculum and its preparation method and application.
Background technique
Tobacco diseases are one of the principal elements for restricting tobacco high quality and stable yield.As countries in the world are food-safe and environment
Safety is increasingly paid close attention to, so that microbial bacterial agent obtains better opportunity to develop.According to international biological and ecological methods to prevent plant disease, pests, and erosion Manufacturers Association (IBMA)
Estimation, whole world biological control goods producer is more than 350, more than 1000 kinds of biological and ecological methods to prevent plant disease, pests, and erosion products at present.Biological pesticide is due to its resource
Be easy to get, be safe and environment-friendly, is long-acting, noresidue the advantages that, be substitution chemical pesticide, solve environmental pollution problem, realize it is nuisanceless and
The important leverage of green agriculture production.
Microorganism is very extensive in application agriculturally, such as being capable of improvement using microorganism fertilizer made of microorganism
The problems such as soil hardening caused by fertilizer, fertility decline and environmental pollution, and then improve the yield and quality of crops；Mushroom pesticide
Using the metabolite of thallus and they, desinsection, sterilizing, in terms of have remarkable result；Also there is mushroom
Soil conditioner, can reconcile the physicochemical property of soil, improve its heat preservation, water conservation, fertilizer conservation performance, and can significantly change
The structure and environment of kind soil.
Dominant mechanism using microbial control disease is that it is colonized in crop root, and one is pass through a large amount of artificial applications
Beneficial bacterium captures the ecological site of the easily infected germ of root system in advance, establishes protective barrier, plant is protected not infected by germ；
One is the invasions by screening the bacterial strain for having the antagonisms such as competition, inhibition, parasitism with germ, to disease resistance；Both
' antibiotic ' can be secreted and deteriorate germ living environment, change microbial bacteria group structure in soil, improve plant disease resistance.
The development and application of microorganism has become the inexorable trend of tobacco business sustainable development.Biological control gradually replaces
Chemical prevention can effectively solve tobacco pest and disease damage and Pesticide Residue, reduce the injury to human body, be a kind of efficient, peace
Complete and permanent control method.Tobacco micro-ecological environment plays a significant role the formation of its quality, at present China's tobacco-growing soil
Since the increase year by year of chemical fertilizer usage amount causes tobacco planting environment gradually to deteriorate, strong by development growth-promoting, resistance
Microbial bacterial agent is the effective means for improving Tobacco ecological environment, keeping ecological balance.Increase matter in degrading nicotine and flavouring
Aspect, the development and utilization of microorganism also show that its unique superiority, and degrading nicotine bacterium not only contributes to improve quality of tobacco,
Improve the utilization rate of tobacco leaf, moreover it is possible to pollution on the environment in tobacco leaf process be effectively reduced.
Currently, there are many kinds of the types of microbial bacterial agent, for example, the patent of publication number CN102888356A discloses one kind
Microbial bacterial agent containing bacillus subtilis enhances Muskmelon Plants by promoting the growth and development of Muskmelon Plants and root system
Disease resistance, and achieve the purpose that the disease incidence for reducing Muskmelon Fusarium wilt；The patent of publication number CN101676242A discloses one kind
Microbial bacterial agent containing bacillus subtilis LJ2 can improve soil, reduce crops while increasing soil fertility
The generation that can eliminate anthracnose is used for a long time in anthracnose disease incidence；The patent of publication number CN109913394A discloses waxy
The microbial-bacterial fertilizer of bacillus Lzh-X7 preparation has very strong antagonism to pathogen fusarium moniliforme, prevents and treats root-rot
The generation of disease.However the microbial bacterial agent reported at present is relatively simple in terms of disease-resistant type, can not achieve and applies a kind of micro- life
Object microbial inoculum achievees the purpose that two or more disease.
Summary of the invention
In view of this, the purpose of the present invention is to provide the microbial bacterial agent of a kind of and anti-tobacco root rot and balck shank and
Preparation method and application.
The present invention provides a kind of and anti-tobacco root rot and balck shank microbial bacterial agents, including bacillus subtilis
Bacillussubtilis and gram lining institute series bacillus Paenibacillus kribbensis；The bacillus subtilis
Concentration >=1 × 109CFU/mL；Concentration >=0.5 × 10 of described gram of lining institute series bacillus9CFU/mL；
The deposit number of the bacillus subtilis is CGMCC No.10248；The guarantor of described gram of lining institute series bacillus
Hiding number is CGMCC No.17248.
Preferably, the concentration of the bacillus subtilis is 1.5~3 × 109CFU/mL；Described gram of lining institute class gemma bar
The concentration of bacterium is 0.8~2 × 109CFU/mL。
The present invention provides the preparation methods of the microbial bacterial agent, comprising the following steps:
1) bacillus subtilis or gram lining institute series bacillus streak inoculation are into first cell culture medium, in 28~30 DEG C of work
Change 24~28h of culture, the bacillus subtilis activated or gram lining institute series bacillus；
The first cell culture medium is that every 1000mL water includes peptone 10g, beef extract 3g, NaCl 5g and agar 18g；Institute
The pH value for stating first cell culture medium is 7.0~7.4；
2) bacillus subtilis of the activation or a gram lining institute series bacillus are inoculated into secondary medium, in 28
~30 DEG C of 24~48h of shake flask fermentation obtain bacillus subtilis or gram lining institute series bacillus seed fermentation liquid；The second level
Culture medium is in every 1000mL water including starch 30g, sucrose 5g, bean powder 15g, dipotassium hydrogen phosphate 2.5g, manganese sulfate 0.2g and ferment
Female cream 1g；The pH value of the secondary medium is 7.0~7.4；
3) bacillus subtilis or gram lining institute series bacillus seed fermentation liquid are seeded in secondary medium
30~35 DEG C fermented and cultured 2~4 days, ventilate during fermented and cultured, into culture for 24 hours in, ventilatory capacity be 10~20m3/
H, after culture for 24 hours, ventilatory capacity is 15~24m3/ h obtains bacillus subtilis fermentation liquor or gram lining institute's series bacillus fermentation
Liquid；
4) bacillus subtilis fermentation liquor and gram lining institute series bacillus fermentation liquid are mixed, obtains simultaneous anti-tobacco
The microbial bacterial agent of root rot and balck shank.
Preferably, the inoculum concentration of the bacillus subtilis or gram lining institute series bacillus seed fermentation liquid be 3%~
10%.
Preferably, the method for the ventilation is stirring；The frequency of the stirring is 1~2 time/h, and the time stirred every time is
10~20min.
Preferably, the body when mixing of the bacillus subtilis fermentation liquor and gram lining institute series bacillus fermentation liquid
Product is than being 1:1.
The microbial bacterial agent provided by the invention or the microbial bacterial agent of the method preparation are in anti-tobacco diseases
Using.
Preferably, the tobacco diseases include tobacco root rot and balck shank.
Provided by the invention a kind of and anti-tobacco root rot and balck shank microbial bacterial agent, including deposit number is
The bacillus subtilis Bacillus subtilis and deposit number of CGMCC No.10248 be CGMCC No.17248 gram in
Cloth institute series bacillus Paenibacillus kribbensis；Wherein, bacillus subtilis can simultaneous anti-sickle-like bacteria and tobacco epidemic disease
Mould, a gram lining institute series bacillus can inhibit sickle-like bacteria.It can be effective it is demonstrated experimentally that applying microbial bacterial agent provided by the invention
The disease incidence of tobacco root rot and balck shank is reduced, relative control effect is respectively 68.5%~80.6% and 48.4%~49.1%.
Using microbial bacterial agent provided by the invention, stem girth higher than the cigarette strain of control group, the number of blade be increased simultaneously, respectively
4.0cm, 0.2cm and 0.6, under, in, top leaf area increase obvious, illustrate that cigarette can be obviously promoted by spraying microbial bacterial agent
The growth of strain and the raising of yield.
Biomaterial preservation information
Bacillus subtilis (Bacillus subtilis), it is general to be deposited in China Committee for Culture Collection of Microorganisms
Logical microorganism center, preservation time are on December 29th, 2014, and preservation address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3,
Institute of Microorganism, Academia Sinica, deposit number are CGMCC No.10248, and strain number is three squares -07.
Gram lining institute series bacillus (Paenibacillus kribbensis), is deposited in Chinese microorganism strain preservation
Administration committee's common micro-organisms center, preservation time are on February 20th, 2019, and preservation address is the Chaoyang District, Beijing City North Star
The institute 3 of West Road 1, Institute of Microorganism, Academia Sinica, deposit number are CGMCC No.17248, and strain number is three squares-
021。
Detailed description of the invention
Fig. 1 is bacillus subtilis and gram lining institute's series bacillus to the bacteriostatic experiment result of sickle-like bacteria；
Fig. 2 is bacillus subtilis and gram lining institute's series bacillus to the bacteriostatic experiment result of Phytophthora nicotianae.
Specific embodiment
The present invention provides a kind of and anti-tobacco root rot and balck shank microbial bacterial agents, including bacillus subtilis
Bacillus subtilis and gram lining institute series bacillus Paenibacillus kribbensis；The bacillus subtilis
Concentration >=1 × 10 of bacterium9CFU/mL；Concentration >=0.5 × 10 of described gram of lining institute series bacillus9CFU/mL；The withered grass bud
The deposit number of spore bacillus is CGMCC No.10248；The deposit number of described gram of lining institute series bacillus is CGMCC
No.17248。
In the present invention, the dosage form of the microbial bacterial agent is preferably aqua.In the microbial bacterial agent, the withered grass
The final concentration of bacillus is preferably 1.5~3 × 109CFU/mL；The concentration of described gram of lining institute series bacillus is preferably 0.8
~2 × 109CFU/mL.Bacillus subtilis energy and anti-sickle-like bacteria and Phytophthora nicotianae, a gram lining institute series bacillus can inhibit sickle
Knife bacterium, wherein sickle-like bacteria is the pathogen of tobacco root rot, and Phytophthora nicotianae is the pathogenic bacteria of balck shank.The microbial bacterial agent
It further include the acceptable auxiliary material in microbial bacterial agent field.
The present invention provides the preparation method of the microbial bacterial agent of described and anti-tobacco root rot and balck shank, including it is following
Step:
1) bacillus subtilis or gram lining institute series bacillus streak inoculation are into first cell culture medium, in 28~30 DEG C of work
Change 24~28h of culture, the bacillus subtilis activated or gram lining institute series bacillus；
The first cell culture medium is that every 1000mL water includes peptone 10g, beef extract 3g, NaCl 5g and agar 18g；Institute
The pH value for stating first cell culture medium is 7.0~7.4；
2) bacillus subtilis of the activation or a gram lining institute series bacillus are inoculated into secondary medium, in 28
~30 DEG C of 24~48h of shake flask fermentation obtain bacillus subtilis or gram lining institute series bacillus seed fermentation liquid；The second level
Culture medium is in every 1000mL water including starch 30g, sucrose 5g, bean powder 15g, dipotassium hydrogen phosphate 2.5g, manganese sulfate 0.2g and ferment
Female cream 1g；The pH value of the secondary medium is 7.0~7.4；
3) bacillus subtilis or gram lining institute series bacillus seed fermentation liquid are seeded in secondary medium
30~35 DEG C fermented and cultured 2~4 days, ventilate during fermented and cultured, into culture for 24 hours in, ventilatory capacity be 10~20m3/
H, after culture for 24 hours, ventilatory capacity is 15~24m3/ h obtains bacillus subtilis fermentation liquor or gram lining institute's series bacillus fermentation
Liquid；
4) bacillus subtilis fermentation liquor and gram lining institute series bacillus fermentation liquid are mixed, obtains simultaneous anti-tobacco
The microbial bacterial agent of root rot and balck shank.
The present invention by bacillus subtilis or gram lining institute series bacillus streak inoculation into first cell culture medium, in 28~
30 DEG C of 24~28h of activation culture, the bacillus subtilis activated or gram lining institute series bacillus.The present invention is to described stroke
The method of line inoculation is not particularly limited, using streak inoculation method known in the art.The temperature of the activation culture
Preferably 29 DEG C of degree, the time of the activation culture is preferably 26h.The activation culture is conducive to provide growth vigor.
First cell culture medium is all made of when bacillus subtilis and gram lining institute series bacillus activation culture.The present invention trains the level-one
The preparation method for supporting base is not particularly limited, using culture medium preparation method known in the art.
After the bacillus subtilis activated or gram lining institute series bacillus, the present invention is by the withered grass bud of the activation
Spore bacillus or a gram lining institute series bacillus are inoculated into secondary medium, in 28~30 DEG C of 24~48h of shake flask fermentation, are obtained withered
Careless bacillus or gram lining institute series bacillus seed fermentation liquid.
In the present invention, the inoculum concentration of the inoculation is 2~15%, more preferably 10%.The temperature of the shake flask fermentation
Preferably 29 DEG C, the time of the shake flask fermentation is 36h.The shake flask fermentation promotes bacillus subtilis or gram lining institute class bud
Spore bacillus increases thalline quantity, increases strain quantity.
Obtain bacillus subtilis or gram lining institute series bacillus seed fermentation liquid, the present invention is by the bacillus subtilis
Bacterium or gram lining institute series bacillus seed fermentation liquid be seeded in secondary medium 30~35 DEG C fermented and cultured 2~4 days, hair
It ventilates during ferment culture, into culture for 24 hours, ventilatory capacity is 10~20m3/ h, culture for 24 hours after, ventilatory capacity be 15~
24m3/ h obtains bacillus subtilis fermentation liquor or gram lining institute series bacillus fermentation liquid.
The culture medium that the fermented and cultured uses is above-mentioned secondary medium.The temperature of the fermented and cultured is preferably 32
DEG C, the time of the fermented and cultured is 3d.Into in culture for 24 hours, ventilatory capacity is preferably 15m3/ h, after culture for 24 hours, ventilatory capacity is excellent
It is selected as 18~22m3/ h, more preferably 20m3/h.The method of the ventilation preferably stirs；The frequency of the stirring is preferably 1~
2 times/h, the time stirred every time is preferably 10~20min.The bacillus subtilis or gram lining institute series bacillus seed
The inoculum concentration of fermentation liquid is preferably 3%~10%, and more preferably 5~8%.
After fermented and cultured, cell concentration in fermentation liquid is detected.The concentration of bacillus subtilis is 2 × 109CFU/
ML or more, while gram lining institute series bacillus is 1 × 109Enter subsequent preparation when CFU/mL or more.It is tied when fermentation liquid detects
Fruit does not meet above-mentioned cell concentration requirement, then returning charge continues fermented and cultured, until reaching the requirement of above-mentioned cell concentration.
After obtaining bacillus subtilis fermentation liquor and gram lining institute series bacillus fermentation liquid, the present invention is by the withered grass bud
Spore bacillus fermentation liquid and gram lining institute series bacillus fermentation liquid mixing, sterile filling obtain simultaneous anti-tobacco root rot and black shin
The microbial bacterial agent of disease.
In the present invention, the mixing of the bacillus subtilis fermentation liquor and gram lining institute series bacillus fermentation liquid
When volume ratio be preferably 1:1.
The microbial bacterial agent provided by the invention or the microbial bacterial agent of the method preparation are in anti-tobacco diseases
Using.The tobacco diseases preferably include tobacco root rot and balck shank.
In the present invention, in anti-tobacco diseases, preferably the microbial bacterial agent is administered simultaneously with fertilizer.When application,
After 500 times of liquid of the microbial bacteria dilution agent, spray on tobacco.The amount of spraying of microbial bacterial agent after dilution is preferably 30~
50L/ mus.The method of administration of the fertilizer is not particularly limited, using tobacco fertilizer application method known in the art.
Below with reference to embodiment to the microbial bacterial agent of provided by the invention a kind of and anti-tobacco root rot and balck shank and
Preparation method and application are described in detail, but they cannot be interpreted as limiting the scope of the present invention.
Embodiment 1
The screening of tobacco black shank and root rot Antagonistic Fungi
Pedotheque is carried out to be serially diluted coating culture acquisition bacterium single colonie.By tobacco black shank pathogen sickle-like bacteria
It is activated with Pathogens Causing Root Rot Disease Phytophthora nicotianae Breda, bacterium piece is made in colony edge punching with punch (diameter 5mm), respectively
Switching for 24 hours, using face-off method, pipettes 1mL in advance at 28 DEG C of beef-protein medium in PDA plate center, 28 DEG C of cultures
The bacterium solution of culture for 24 hours, is added in Oxford cup, four Oxford cups of each plating, two of them bacillus subtilis, in addition
Two inoculation gram lining institute series bacillus, inoculation are placed in incubator and cultivate 3~5 days for 28 DEG C, observe antibacterial situation.
Antibacterial the result is shown in Figure 1 and Fig. 2.Fig. 1 is bacillus subtilis and gram suppression of the lining institute's series bacillus to sickle-like bacteria
Bacterium experimental result；Fig. 2 is bacillus subtilis and gram lining institute's series bacillus to the bacteriostatic experiment result of Phytophthora nicotianae.By scheming
1 it is found that bacillus subtilis and gram lining institute's series bacillus all have inhibiting effect to sickle-like bacteria, as shown in Figure 2, withered grass bud
Spore bacillus is inhibited to the growth of Phytophthora nicotianae, and gram lining institute's series bacillus imitates the growth inhibition of Phytophthora nicotianae
Fruit is poor.
Embodiment 2
Pot experiment
For studying object tobacco K326.
The soil of tobacco root rot is taken for examination soil
Experimental design test sets 4 processing, 3 repetitions, and every basin fills soil 3kg, microbial bacterial agent is taken to be diluted with water to 500
Again, in being manured into soil by the way of pouring root, after a week, the health tobacco cultivated in advance in healthy soil is transplanted to potting
In, then spray a microbial bacterial agent.Disease incidence is observed after three months, is during which normally watered.
Handle A bacillus subtilis microbial agent
Handle B grams of lining institute series bacillus microbial inoculum
Handle C bacillus subtilis+gram lining institute series bacillus mix bacterium agent
Handle D blank control
Test results and analysis
Obtained by pot experiment: the root rot average attack rate for handling A is 16.1%, and processing B is 19.5%, and processing C is
8.4%, processing D is 47.4%, it is seen that two kinds of bacterium all have root rot control efficiency, but two bacterium combined effects are more preferably.
Embodiment 3
It is dark green No. 1 for examination crop test tobacco bred.
For examination soil testing soil is red earth, soil organism 19.6g/kg, full nitrogen 1.02g/kg, alkali-hydrolyzable nitrogen
89.6mg/kg, available phosphorus 31.2mg/kg, available potassium 79.8mg/kg, pH value 5.3.
Experimental design test sets 4 processing, 3 repetitions, experimental plot area 30m2, arranged using random district's groups, surrounding
If protection row.
Handle A microbial bacterial agent+conventional fertilizer application；
Handle microbial bacterial agent+conventional fertilizer application of B sterilizing；
Handle C conventional fertilizer application；
It handles D blank control (not applying fertilizer).
Fertilising is recorded with management
After handling the microbial bacterial agent of A, B, 500 times of microbial bacteria dilution agent of sterilizing, sprayed after transplanting, the amount of spraying is
30070mL/ mus.
Tobacco conventional fertilizer application: tobacco was transplanted on 2 5th, 2019,5 days 2 months basal dressing 666.7m2It applies decomposed organic
Fertile 1000kg, sheep dung 200kg, Special compound fertilizer for tobacco (15:10:20) 15kg, March 15 group phase every 666.7m2Use tobacco
Special complex fertilizer (15:10:20) 12.5kg.Tobacco terminates in harvesting on June 3rd, 2019.
Test results and analysis
As it can be seen from table 1 processing A ratio processing B, C, D flue-cured tobacco plant height, stem girth, the number of blade increased, under, in,
Top leaf area obviously increases；Processing A ratio processing B plant height, stem girth, the number of blade increased, respectively 4.0cm, 0.2cm and
0.6, under, in, top leaf area increase obvious, illustrate to spray growth and yield that microbial bacterial agent can be obviously promoted cigarette strain
Raising.
Influence of 1 different disposal of table to flue-cured tobacco economical character
From table 2 it can be seen that tobacco root rot and balck shank can be effectively reduced in application microbial bacterial agent of the invention
Disease incidence, relative control effect are respectively 80.6% and 49.1%.
Influence of 2 different disposal of table to tobacco root rot and balck shank disease incidence
Embodiment 4
It is K326 for examination crop test tobacco bred.
For examination soil testing soil is plaster field, soil organism 27.9g/kg, full nitrogen 1.48g/kg, alkali-hydrolyzable nitrogen
108mg/kg, available phosphorus 35.4mg/kg, available potassium 109mg/kg, pH value 5.5.
Experimental design is the same as embodiment 2
Fertilising is recorded with management
After handling the microbial bacterial agent of A, B, 500 times of microbial bacteria dilution agent of sterilizing, sprayed after transplanting, the amount of spraying is
30070mL/ mus.
Tobacco conventional fertilizer application: tobacco was transplanted on 2 5th, 2019,5 days 2 months basal dressing 666.7m2It applies decomposed organic
Fertile 1000kg, sheep dung 200kg, Special compound fertilizer for tobacco (15:10:20) 15kg, March 15 group phase every 666.7m2Use tobacco
Special complex fertilizer (15:10:20) 12.5kg.Tobacco terminates in harvesting on June 3rd, 2019.
Test results and analysis
(1) influence of the microbial bacterial agent to tobacco K326 disease
From table 3 it can be seen that the root rot of tobacco K326 and black can be effectively reduced in application microbial bacterial agent of the invention
Shin disease disease incidence, relative control effect are respectively 68.5% and 48.4%.
Protection effect of 3 different disposal of table to tobacco K326 root rot and balck shank
(2) influence of the microbial bacterial agent to tobacco K326 yield
(table 4) is calculated according to effective yield, handles A (microbial bacterial agent+conventional fertilizer application) yield highest, average 666.7m2
Yield is 160.23kg, 666.7m more every than processing B (aseptic substrate+conventional fertilizer application)2Yield 146.01kg increases 14.22kg, increases
Produce 9.74%；It is 666.7m more every than processing C (conventional fertilizer application)2Yield 145.12kg increases 15.11kg, volume increase 10.41%；Than processing
D (blank control) every 666.7m2Yield 98.45kg increases 61.78kg, volume increase 62.75%.
Protection effect of 4 different disposal of table to tobacco K326 root rot and balck shank
By above-mentioned experiment it is found that using microbial bacterial agent provided by the invention, compared with only conventional fertilizer application, effectively reduce
The disease incidence of tobacco black shank and root rot, the relative control effect in tobacco root rot reaches 50% or more, and mentions significantly
The high yield of tobacco.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (8)
1. a kind of and anti-tobacco root rot and balck shank microbial bacterial agent, which is characterized in that including bacillus subtilis
Bacillussubtilis and gram lining institute series bacillus Paenibacillus kribbensis；The bacillus subtilis
Concentration >=1 × 109CFU/mL；Concentration >=0.5 × 10 of described gram of lining institute series bacillus9CFU/mL；
The deposit number of the bacillus subtilis is CGMCC No.10248；The preservation of described gram of lining institute series bacillus is compiled
Number be CGMCCNo.17248.
2. microbial bacterial agent according to claim 1, which is characterized in that the concentration of the bacillus subtilis is 1.5~3
×109CFU/mL；The concentration of described gram of lining institute series bacillus is 0.8~2 × 109CFU/mL。
3. the preparation method of microbial bacterial agent as claimed in claim 1 or 2, which comprises the following steps:
1) bacillus subtilis or gram lining institute series bacillus streak inoculation are trained into first cell culture medium in 28~30 DEG C of activation
Support 24~28h, the bacillus subtilis activated or gram lining institute series bacillus；
The first cell culture medium is that every 1000mL water includes peptone 10g, beef extract 3g, NaCl 5g and agar 18g；Described one
The pH value of grade culture medium is 7.0~7.4；
2) bacillus subtilis of the activation or a gram lining institute series bacillus are inoculated into secondary medium, in 28~30
DEG C 24~48h of shake flask fermentation, obtains bacillus subtilis or gram lining institute series bacillus seed fermentation liquid；The second level culture
Base is in every 1000mL water including starch 30g, sucrose 5g, bean powder 15g, dipotassium hydrogen phosphate 2.5g, manganese sulfate 0.2g and yeast extract
1g；The pH value of the secondary medium is 7.0~7.4；
3) bacillus subtilis or gram lining institute series bacillus seed fermentation liquid are seeded in secondary medium 30
~35 DEG C fermented and cultured 2~4 days, ventilate during fermented and cultured, into culture for 24 hours in, ventilatory capacity be 10~20m3/ h, training
After supporting for 24 hours, ventilatory capacity is 15~24m3/ h obtains bacillus subtilis fermentation liquor or gram lining institute series bacillus fermentation liquid；
4) bacillus subtilis fermentation liquor and gram lining institute series bacillus fermentation liquid are mixed, obtains simultaneous anti-tobacco root-rot
The microbial bacterial agent of disease and balck shank.
4. preparation method according to claim 3, which is characterized in that the bacillus subtilis or gram lining institute class gemma
The inoculum concentration of bacillus seed fermentation liquid is 3%~10%.
5. preparation method according to claim 3, which is characterized in that the method for the ventilation is stirring；The stirring
Frequency is 1~2 time/h, and the time stirred every time is 10~20min.
6. preparation method according to claim 3, which is characterized in that the bacillus subtilis fermentation liquor and Ke Li
Volume ratio is 1:1 when the mixing of cloth institute series bacillus fermentation liquid.
7. microbial bacterial agent as claimed in claim 1 or 2 or the microbial bacterial agent of claim 3~6 any one method preparation exist
Application in anti-tobacco diseases.
8. application according to claim 7, which is characterized in that the tobacco diseases include tobacco root rot and balck shank.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910789604.1A CN110511891A (en) | 2019-08-26 | 2019-08-26 | A kind of and anti-tobacco root rot and the microbial bacterial agent of balck shank and its preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910789604.1A CN110511891A (en) | 2019-08-26 | 2019-08-26 | A kind of and anti-tobacco root rot and the microbial bacterial agent of balck shank and its preparation method and application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110511891A true CN110511891A (en) | 2019-11-29 |
Family
ID=68627889
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910789604.1A Withdrawn CN110511891A (en) | 2019-08-26 | 2019-08-26 | A kind of and anti-tobacco root rot and the microbial bacterial agent of balck shank and its preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110511891A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110747153A (en) * | 2019-12-10 | 2020-02-04 | 吉安职业技术学院 | Paenibacillus kribbensis viable bacteria-free fermentation broth and application thereof in rice planting |
| CN111793498A (en) * | 2020-07-31 | 2020-10-20 | 福建三炬生物科技股份有限公司 | Microbial agent and preparation method and application thereof |
| CN111944716A (en) * | 2020-08-03 | 2020-11-17 | 北京恩格兰环境技术有限责任公司 | Special compound microbial agent for tobacco seedling culture and preparation method and application thereof |
| CN112225595A (en) * | 2020-10-21 | 2021-01-15 | 福建三炬生物科技股份有限公司 | Water-soluble compound microbial fertilizer and preparation method thereof |
| CN113151085A (en) * | 2021-04-23 | 2021-07-23 | 贵州省烟草公司遵义市公司 | Compound microbial agent for preventing and treating soil-borne diseases of tobacco and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20040092067A (en) * | 2003-04-24 | 2004-11-03 | 고도화학(주) | Novel strain Paenibacillus kribbensis AP-2 producing flocculant polysaccharide material |
| CN104651277A (en) * | 2015-02-09 | 2015-05-27 | 福建三炬生物科技股份有限公司 | Straw composting bacterium agent as well as preparation method and application thereof |
| CN105052471A (en) * | 2015-08-03 | 2015-11-18 | 河南省农业科学院 | A method for cultivating tobacco disease-resistant strong seedlings by utilizing beneficial biological bacteria |
| CN110066756A (en) * | 2019-05-29 | 2019-07-30 | 漳州三炬生物技术有限公司 | One plant of gram of lining institute's series bacillus and its preparation and application |
-
2019
- 2019-08-26 CN CN201910789604.1A patent/CN110511891A/en not_active Withdrawn
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20040092067A (en) * | 2003-04-24 | 2004-11-03 | 고도화학(주) | Novel strain Paenibacillus kribbensis AP-2 producing flocculant polysaccharide material |
| CN104651277A (en) * | 2015-02-09 | 2015-05-27 | 福建三炬生物科技股份有限公司 | Straw composting bacterium agent as well as preparation method and application thereof |
| CN105052471A (en) * | 2015-08-03 | 2015-11-18 | 河南省农业科学院 | A method for cultivating tobacco disease-resistant strong seedlings by utilizing beneficial biological bacteria |
| CN110066756A (en) * | 2019-05-29 | 2019-07-30 | 漳州三炬生物技术有限公司 | One plant of gram of lining institute's series bacillus and its preparation and application |
Non-Patent Citations (2)
| Title |
|---|
| 张中义: "《植物病原真菌学》", 31 March 1988, 四川科学技术出版社 * |
| 李桂兰: "《山西地道药材黄芪质量控制研究》", 31 December 2014, 山西科学技术出版社 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110747153A (en) * | 2019-12-10 | 2020-02-04 | 吉安职业技术学院 | Paenibacillus kribbensis viable bacteria-free fermentation broth and application thereof in rice planting |
| CN111793498A (en) * | 2020-07-31 | 2020-10-20 | 福建三炬生物科技股份有限公司 | Microbial agent and preparation method and application thereof |
| CN111944716A (en) * | 2020-08-03 | 2020-11-17 | 北京恩格兰环境技术有限责任公司 | Special compound microbial agent for tobacco seedling culture and preparation method and application thereof |
| CN112225595A (en) * | 2020-10-21 | 2021-01-15 | 福建三炬生物科技股份有限公司 | Water-soluble compound microbial fertilizer and preparation method thereof |
| CN113151085A (en) * | 2021-04-23 | 2021-07-23 | 贵州省烟草公司遵义市公司 | Compound microbial agent for preventing and treating soil-borne diseases of tobacco and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110511891A (en) | A kind of and anti-tobacco root rot and the microbial bacterial agent of balck shank and its preparation method and application | |
| CN102964178B (en) | Composite microbial bactericide, and preparation method and application thereof | |
| CN106591193B (en) | One plant of bacillus amyloliquefaciens with the degeneration-resistant effect of wide spectrum growth-promoting | |
| CN102515927B (en) | Drug effect type microbial fertilizer, preparation method thereof and application thereof | |
| CN102001869B (en) | Method for preparing biologic herbicide-containing fertilizer from biogas slurry | |
| CN102726454B (en) | Biological control agent for fusarium wilt and preparation and use method of biological control agent | |
| CN102010825B (en) | Microorganism compound bactericide for controlling cucumber fusarium wilt and preparation method thereof | |
| WO2011050547A1 (en) | Biocontrol strain for cucumber and watermelon continuous cropping wilt diseases and microbial organic fertilizer thereof | |
| CN106119155B (en) | For preventing and treating the bacterial strain WXX-2 and microbial inoculum of peanut root rot | |
| CN102584359B (en) | Composite biological agent and preparation method thereof | |
| CN101575574A (en) | Trichoderma harzianum composite bacteria culture and application of trichoderma harzianum composite bacteria culture in aspect of plant protection | |
| CN103493677A (en) | Method of cutting propagation of peony immature stem | |
| CN103396954B (en) | Biological prevention and control bacterial strain for preventing and controlling rice sheath blight, biological organic fertilizer, and preparation method of biological organic fertilizer | |
| CN103820348A (en) | Plant growth-promoting bacteria and preparation method and application of fungicide | |
| CN107502572A (en) | Application of the bacillus licheniformis in straw degradative, the microbial bacterial agent comprising the bacterium and its application | |
| CN100424164C (en) | Bacteria for improving agricultural production environment and the quality of products and yield, and its preparation method | |
| CN107056436A (en) | It is a kind of to utilize the method that waste material is raw material production bio-bacterial manure | |
| CN104480046A (en) | Brevibacillus laterosporus strain and application thereof | |
| CN103304305A (en) | Compound-microorganism functional enteromorpha foliar fertilizer and preparation method thereof | |
| CN106119143A (en) | A kind of disease resistance growth promotion microbial-bacterial fertilizer and preparation method thereof | |
| CN106465734A (en) | A kind of preventing and treating tobacco bacterial wilt fermentation liquid and preparation method thereof | |
| CN102515977A (en) | Water-holding root-promoting halophilic alkaliphilic microorganism root dipping liquid and preparation and application thereof | |
| CN102533603B (en) | Pseudomonas sp. 841P-3 for preventing cotton verticillium wilt and use thereof | |
| CN103255064B (en) | Fungal agent for prevention and control of watermelon fusarium wilt and preparation method thereof | |
| CN103141519A (en) | Preparation method and application of anti-biological inoculants for controlling crop soil-borne diseases |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20191129 |
|
| WW01 | Invention patent application withdrawn after publication |