CN102787084B - The Methylotrophic Bacillus strain 4-L-16 of a kind of preventing and controlling banana fusarium wilt and application thereof - Google Patents
The Methylotrophic Bacillus strain 4-L-16 of a kind of preventing and controlling banana fusarium wilt and application thereof Download PDFInfo
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Abstract
The invention discloses Methylotrophic Bacillus strain 4 L 16 of a kind of preventing and controlling banana fusarium wilt, preservation entitled Methylotrophic bacillus 4 L 16 of this bacterial strain, Classification And Nomenclature is Bacillus methylotrophicus4 L 16, depositary institution is China typical culture collection center, preservation date is on June 26th, 2012, and preserving number is CCTCC NO:M2012248.This bacterial strain is respectively provided with stronger antagonism to No. 1, No. 4 biological strains of banana blight and other banana common disease kinds.Also disclosing this bacterial strain and having the purposes in the medicine of preventing and controlling banana fusarium wilt effect and the microorganism formulation containing this bacterial strain in preparation, this microorganism formulation not only has more preferable antagonism to multiple banana pathogens, also has facilitation to plant strain growth.
Description
Technical field
The present invention relates to a kind of bacterial strain preventing and treating banana blight and application thereof, be specifically related to a kind of prevention and control banana withered
The Methylotrophic Bacillus strain 4-L-16 of disease of withering and application thereof.
Background technology
Banana blight is by Jian Bao sickle-like bacteria Cuba specialized form (Fusarium oxysporium f.sp.
Cubense, FOC) the crushing soil-borne disease that causes of disease fungus, from 1874 first in Australia's discovery
Since, once caused large area to endanger in 1896 in Panama, gradually reached all over the world afterwards, be so far
An insurmountable world technology difficult problem.At present, at bananas such as China Guangdong, Hainan, Guangxi, Yunnan and Fujian
Main producing region, banana blight is to be caused by FOC No. 4 biological strains, and dwarf banana droop is by FOC
No. 1 biological strain causes, the morbidity any of several broadleaf plants garden incidence of disease typically at 30-50%, grave illness district up to more than 90%,
Cause the banana underproduction, even have no harvest, be a main limiting factor in banana production.
So far preferable disease-resistant variety and chemical agent be there is no to this banana blight, therefore, in conjunction with cultivation step,
Administration has the microbial strains of antagonistic effect to this disease, thus suppresses soil banana blight quantity, changes soil
The microbial ecological structure of earth, promotes Banana Growth, improves banana broad spectrum resistance, becomes a kind of preferably raw
Thing prophylactico-therapeutic measures.
Current Antagonistic Fungi source is to be obtained with screening by the separation of soil, the microorganism of plant own, this method
Obtain antagonistic microbe and be usually present the shortcomings such as colonization ability is weak, breeding coefficient is low, Antagonistic Fungi antimicrobial spectrum is narrower.
Particularly in soil, the Antagonistic Fungi of isolated tends not to a large amount of field planting in plant body, and control effect does not shows
Write.Therefore using other approach to obtain and being easy to breeding, the wide Antagonistic Fungi of antibacterial range is particularly important.
Content of the invention
First technical problem to be solved by this invention is that the methyl providing a kind of preventing and controlling banana fusarium wilt is sought
Supporting type Bacillus strain 4-L-16, this strain isolation is in the organic zymotic fluid with brown sugar water, it is easy to field planting
Breeding, antibacterial range is extensive, to No. 1, No. 4 biological strains of banana blight and other banana common disease kinds such as perfume
The elongated leaf spot fungi of any of several broadleaf plants, Glorosprium musarum Cookeet Mass, the big Cercospora Sojina Hara of banana, the mould leaf spot fungi of mango rod method, litchi
Branch multiple pathogen such as anthrax-bacilus, mango anthrax-bacilus is respectively provided with stronger antagonism.
Second technical problem to be solved by this invention is that the methyl providing above-mentioned preventing and controlling banana fusarium wilt is sought
Support type Bacillus strain 4-L-16, in preparation, there is the purposes in the medicine of preventing and controlling banana fusarium wilt effect.
3rd technical problem to be solved by this invention is to provide the preparation of a kind of preventing and controlling banana fusarium wilt, should
Preparation contains the Methylotrophic Bacillus strain 4-L-16 of above-mentioned preventing and controlling banana fusarium wilt, and said preparation is to banana
No. 1, No. 4 biological strains of droop and other elongated leaf spot fungi of banana common disease kind such as banana, banana anthracnose
Bacterium, the big Cercospora Sojina Hara of banana, the mould leaf spot fungi of mango rod method, lichee anthrax-bacilus, mango anthrax-bacilus etc. are multiple
Pathogen has more preferable antagonism, also has growth-promoting functions to plant strain growth simultaneously.
The present invention finally to solve the technical problem that the preparation being to be to provide above-mentioned preventing and controlling banana fusarium wilt preparation
Method.
First technical problem to be solved of the present invention is achieved by the following technical solution: Yi Zhongfang
The Methylotrophic Bacillus strain 4-L-16 of control banana blight, the entitled methylotrophy of preservation of this bacterial strain
Type bacillus 4-L-16, Classification And Nomenclature is Bacillus methylotrophicus4-L-16, during depositary institution is
State's Type Tissue Collection, preservation date is on June 26th, 2012, and preserving number is CCTCC NO:M
2012248。
The Methylotrophic Bacillus strain 4-L-16 of the preventing and controlling banana fusarium wilt in the present invention is to use flat board
Method of dilution butteron on plate is isolatable from the mixed fermentation liquid of culture matrix cake fertilizer, brown sugar and water, and through flat board opposite culture method
Screening obtains, and wherein cake fertilizer is preferably one or more in peanut cake, rape cake and soya-bean cake, concrete bolter
Select the process can be refering to the description in embodiment in detail below.
Second technical problem of the present invention is achieved by the following technical solution: above-mentioned prevention and control banana is withered
Sick Methylotrophic Bacillus strain 4-L-16 has in the medicine of preventing and controlling banana fusarium wilt effect in preparation
Purposes.
This medicine not only flat board antagonism to No. 1, No. 4 biological strain of banana blight bacteria；Long to banana
The big Cercospora Sojina Hara of shape leaf spot fungi, Glorosprium musarum Cookeet Mass, banana, the mould leaf spot fungi of mango rod method, lichee charcoal
Multiple pathogens such as subcutaneous ulcer bacterium, mango anthrax-bacilus also there is antagonism.
3rd technical problem of the present invention is achieved by the following technical solution: a kind of prevention and control banana is withered
Sick preparation, the Methylotrophic Bacillus strain 4-L-16 containing above-mentioned preventing and controlling banana fusarium wilt.
As a modification of the present invention: the preparation of preventing and controlling banana fusarium wilt of the present invention, possibly together with cultivation
Matrix cake fertilizer, brown sugar and water, wherein the gross weight of cake fertilizer and brown sugar and the gross weight of water are than for 1:4, culture medium
The C/N ratio of matter is 15 ~ 25:1.
Described cake fertilizer preferably is selected from one or more in peanut cake, soya-bean cake and rape cake.
Last technical problem of the present invention is achieved by the following technical solution: above-mentioned prevention and control banana is withered
The preparation method of disease of withering preparation, containing following steps:
(1) by brown sugar water soluble in water for brown sugar, in cake fertilizer is placed in breathable's package body and be placed on can be saturating
Cake fertilizer in gas package body is placed in brown sugar water or is directly dissolved in brown sugar water by cake fertilizer, and regulation pH value is
6 ~ 8, it is placed at shady and cool ventilation, seal and preserve, the brown sugar water solution of cake fertilizer must be contained；
(2) methyl of the preventing and controlling banana fusarium wilt described in the brown sugar water solution containing cake fertilizer and claim 1 is sought
Support type Bacillus strain 4-L-16 to ferment 7 ~ 15 days under the conditions of 25 ~ 30 DEG C, and stirring keeps ventilation frequently
To completing sweat, obtain red tan solution, filter the banana vascular wilt resistant fermentation liquor i.e. obtaining containing multiple-microorganism.
The brown sugar water solution containing cake fertilizer described in step of the present invention (2) and the methylotrophy of preventing and controlling banana fusarium wilt
The volume ratio of type Bacillus strain 4-L-16 is preferably 8 ~ 12:1.
Compared with prior art, present invention have the advantage that
(1) present invention utilizes the raw materials such as culture matrix cake fertilizer, brown sugar and water according to the direct liquid of optimization of C/C composites first
Fermentation, by separating the microorganism producing in zymotic fluid, screening obtains a strain to banana blight 1
Number, No. 4 biological strains and other multiple pathogens be respectively provided with the Methylotrophic bacillus of preferable antagonistic effect
4-L-16；
(2) owing to bacterium source produces in biofermentation, it is easy to field planting is bred, banana can be effectively suppressed to wither
No. 1, No. 4 biological strains of germ and the growth of other banana common disease kinds, environmental safety is good, withered to banana
Disease of withering prevention and control are significant；
(3) the Methylotrophic bacillus 4-L-16 in the present invention is made micro-life together with other organic matters
No. 1, No. 4 biological strains of banana blight and other multiple pathogens are had more preferable antagonism effect by thing preparation
Really, also growth-promoting functions is had to plant strain growth simultaneously；
(4) the bacterial strain 4-L-16 strain culturing condition of the preventing and controlling banana fusarium wilt in the present invention is simple simultaneously, breeding
Speed is fast, it is easy to preserves, it is easy to industrialized production, has good development prospect.
Brief description
Fig. 1 is the antagonistic effect to No. 1 biological strain of Fusarium oxysporum for the 4-L-16 bacterial strain in embodiment 2；
Fig. 2 is the antagonistic effect to No. 4 biological strains of Fusarium oxysporum for the 4-L-16 bacterial strain in embodiment 2；
Fig. 3 be in embodiment 2 4-L-16 bacterial strain to banana elongated leaf spot fungi, the mould leaf spot of mango rod method
The fungistatic effect figure of bacterium, banana big gray leaf spot bacterium, lichee anthrax-bacilus, mango anthrax-bacilus and musae,
Wherein A represents the elongated leaf spot fungi of banana；B represents the mould leaf spot fungi of mango rod method；It is big that C represents banana
Gray leaf spot bacterium；D represents lichee anthrax-bacilus；E represents mango anthrax-bacilus；F represents musae；
Fig. 4 is that in embodiment 3, the 4-L-16 of bacillus containing Methylotrophic microbial bacterial agent is withered for seedling stage to dwarf banana
The control effect figure of disease of withering；
Fig. 5 be in embodiment 4 4-L-16 of bacillus containing Methylotrophic microbial bacterial agent to Brazil any of several broadleaf plants seedling stage
The control effect figure of droop.
Detailed description of the invention
Embodiment 1
The source of bacterial strain, separation and qualification
This bacterial strain is by isolated in rape cake, brown sugar and water by fermentation liquid, and the preparation method of concrete zymotic fluid is as follows:
Brown sugar 155g is dissolved in 1000mL water, is dissolved in rape cake 95g in brown sugar water, the pH of regulation brown sugar water
Value is 7.0, is placed in shady and cool ventilative place, seals and preserves, must contain the brown sugar water solution of cake fertilizer；Red by containing cake fertilizer
Sugar aqueous solution is placed in 5L volumetric flask, and at 180rpm, under the conditions of 30 DEG C, concussion is cultivated 7 days, filters red
The zymotic fluid that brown solution agent completes for fermentation.
1. the separation of Antagonistic Fungi in zymotic fluid, primary dcreening operation
Use dilution plating procedure, dilution factor (10-1～10-3), use PDA culture medium to enter fungi
Row separates；Carrying out to bacterium separates LB culture medium；Gause I culture medium separates to actinomycetic,
Weight/mass percentage composition is added to be 3% potassium bichromate suppression bacterium, the growth of fungi before separating.With liquid-transfering gun respectively
Draw above-mentioned difference dilution solution 100 μ L, drip on culture medium flat plate, with the coating of L-type glass rod uniformly,
Slightly dry, be inverted culture dish, cultivate 3 days in 25～28 DEG C of incubators, according to the bacterium colony growing on flat board
Color, form, size etc., choose different fungies, bacterium and actinomycetic single bacterium colony to new culture medium
Upper purifying is cultivated, bacterium, actinomyces LB medium culture, fungi PDA medium culture.Use
Flat board opposite culture method primary dcreening operation antagonistic strain, primary dcreening operation obtains 28 strain Antagonistic Fungis altogether, by lyophilized for bacterial classification-20 DEG C of preservations,
For screening further.
2. the multiple sieve of Antagonistic Fungi in zymotic fluid
Use flat board opposite culture method, detect the Antagonistic Fungi after primary dcreening operation respectively to No. 1 biological strain of Fusarium oxysporum
With the bacteriostatic activity of No. 4 biological strains, make with the antibacterial bandwidth (cm) between antagonistic strain and pathogen
For the strong and weak index of antagonism.
After antagonistic strain to be measured is cultivated 72 hours in 28 DEG C of incubators, put down at potato dextrose medium
Plate central authorities access the FOC1/FOC4 mycelia block of 5mm, in distance FOC1/FOC4 mycelia block surrounding 2.5cm
Place, inoculates Antagonistic Fungi 5 μ L to be measured, is placed in 28 DEG C of incubators and cultivates, and only to inoculate pathogen for comparison, treats
When comparison just covers with culture dish.Antibacterial bandwidth (cm) between measurement antagonistic strain and pathogen is as short of money
The strong and weak index of anti-effect.
Final screening obtains the bacterial strain 4-L-16 all having preferable antagonistic effect to FOC1 and FOC4 pathogen.
3. the qualification of Antagonistic Fungi
The Main Biological of Methylotrophic bacillus 4-L-16 is: this bacterium is gram-positive bacteria, training
Foster condition is LB culture medium, 37 DEG C, pH7.0.Detection 16sRNA sequence, and sequence is submitted to online
Carry out BLAST comparison on GENBANK, find it in Bacillus amyloliquefaciens
The similarity bacterium of two kinds of bacterium of subsp.Plantarum and Bacillus methylotrophicus reaches 99.859%, then
Identifying final determination through Physiology and biochemistry, this bacterium is Bacillus methylotrophicus, and its 16sRNA sequence can
With refering to shown in sequence table SEQ ID NO:1.
Table 1 bacterial strain 4-L-16 phenotype and physio-biochemical characteristics-enzyme are lived, carbon assimilation
Note :+: positive reaction；-: negative reaction；W: weak positive reaction
Table 2 bacterial strain 4-L-16 physio-biochemical characteristics-carbon assimilation
Note :+: positive reaction；-: negative reaction；W: weak positive reaction
This bacterial strain is sent to after China typical culture collection center carries out preservation on June 26th, 2012, warp
Separating the preservation entitled Methylotrophic bacillus 4-L-16 identifying this bacterial strain 4-L-16, Classification And Nomenclature is
Bacillus methylotrophicus4-L-16, depositary institution is China typical culture collection center, preservation day
Phase is on June 26th, 2012, and preserving number is CCTCC NO:M2012248.
The prevention and control effect to banana blight bacteria for the embodiment 2 Methylotrophic bacillus 4-L-16
The flat board antagonism to No. 1 microspecies FOC1 of banana blight bacteria for the 2.1 Methylotrophic bacillus 4-L-16
Test
Methylotrophic bacillus 4-L-16 is activated on LB culture medium, is placed in 37 DEG C of incubators training
After supporting 72 hours, access the FOC1 mycelia block of 5mm in potato glucose PDA culture medium flat plate central authorities,
At distance FOC1 mycelia block 2.5cm, inoculate Methylotrophic bacillus 4-L-16 nutrient solution 5 μ L,
It is placed in 28 DEG C of incubators cultivation 5 ~ 7 days with the flat board not meeting Methylotrophic bacillus 4-L-16 for comparison,
Measuring antibacterial bandwidth is 9mm, has obvious inhibiting effect to No. 1 microspecies of banana blight bacteria, is shown in Table 3
And Fig. 1.
The flat board antagonism to No. 4 microspecies FOC4 of banana blight bacteria for the 2.2 Methylotrophic bacillus 4-L-16
Test
Methylotrophic bacillus 4-L-16 is activated on LB culture medium, is placed in 37 DEG C of incubators training
After supporting 72 hours, access the FOC4 mycelia block of 5mm in potato glucose PDA culture medium flat plate central authorities,
At distance FOC4 mycelia block 2.5cm, inoculate Methylotrophic bacillus 4-L-16 nutrient solution 5 μ L,
It with the flat board that do not meets Methylotrophic bacillus 4-L-16 for comparison, is placed in 28 DEG C of incubators cultivation 5 ~ 7
My god, measuring antibacterial bandwidth is 12mm, has obvious inhibiting effect to No. 4 microspecies of banana blight bacteria, sees
Table 3 and Fig. 2.
The inhibitory action to No. 1, No. 4 biological strain of banana blight for the table 3 Methylotrophic bacillus 4-L-16
Note :-: unrestraint effect；+: antibacterial band diameter 1～4mm；++: antibacterial band diameter 4～8mm；
+++: antibacterial band diameter ＞ 8mm.
The antagonism to multiple pathogens for the 2.3 Methylotrophic bacillus 4-L-16
Use flat board face-off method, elongated leaf spot fungi, the mould leaf spot fungi of mango rod method, banana anthrax to banana
The antagonistic effect of 6 kinds of pathogens such as germ, the big Cercospora Sojina Hara of banana, lichee anthrax-bacilus, mango anthrax-bacilus, should
Six kinds of pathogens are by Chinese Academy of Tropical Agricultural Sciences's resource environment and Plant Protection Institute Xie Yixian researcher
There is provided.Methylotrophic bacillus 4-L-16 is activated on LB culture medium, is placed in 37 DEG C of incubators
After cultivating 72 hours, access the target bacterium mycelia of 5mm in potato glucose PDA culture medium flat plate central authorities
Block, at distance target bacterium mycelia block 2.5cm, inoculates Methylotrophic bacillus 4-L-16 nutrient solution 5 μ L,
It with the flat board that do not meets Methylotrophic bacillus 4-L-16 for comparison, is placed in 28 DEG C of incubators cultivation 5 ~ 7
My god, the antibacterial bandwidth of detection measurement.As shown in table 4 and Fig. 3.
The inhibitory action to multiple pathogens for the table 4 Methylotrophic bacillus 4-L-16
Note :-: unrestraint effect；+: antibacterial band diameter 1～4mm；++: antibacterial band diameter 4～8mm；
+++: antibacterial band diameter ＞ 8mm.
The biologic product of embodiment 3 4-L-16 of bacillus containing Methylotrophic
First by the Methylotrophic bacillus 4-L-16 bacterial strain for preparing in embodiment 1 at LB culture medium
On activate, after being placed in 37 DEG C of incubators cultivation 72 hours, drawn on LB culture medium by liquid-transfering gun
Thalline 5 ~ 10 μ L, is placed in the brown sugar water solution that 100mL weight/mass percentage composition is 10%, shaken cultivation,
Cultivation temperature is 30 DEG C, and shaking speed is 180rpm, and incubation time is 3 ~ 5 days, i.e. obtains containing methylotrophy
The seed liquor of type bacillus 4-L-16 bacterial strain, then by this seed liquor with the ratio of 1:10, joins dish
In the mixed liquor of seedcake and brown sugar, the preparation ratio of this mixed liquor is: the C/N according to rape cake and nitrogen content,
And the phosphorus content of brown sugar, the C/N using material in conventional method regulation mixed liquor is 15/1, and adds suitable
The water of amount.
By fermentation 1000mL microbial inoculum and as a example by making the gross weight of rape cake and brown sugar gross weight/water be 1/4, need
Adding seed liquor 100mL, rape cake 141g, brown sugar 109g, add water 900mL.The zymotic fluid preparing is placed in
Temperature is 30 DEG C, shaking speed is shaken cultivation 7 ~ 15 days under the conditions of 180rpm；Under natural conditions, if needing
Want bulk fermentation also permissible, use the good tun of ventilation type according to above ratio, be placed at shady and cool ventilation,
Direct fermentation, but to ensure each stirring 5 ~ 10min sooner or later, it is ensured that throughput, ferments 15 days；Use
Large-scale fermentation fills when fermenting, and temperatures above and mixing speed are fermented 7 days, and fermentation obtains after completing
Red tan solution filter i.e. obtain containing a large amount of Methylotrophic bacillus 4-L-16 bacterial strains and other multiple beneficial micro-
Biological microbial bacterial agent.
The biologic product of embodiment 4 4-L-16 of bacillus containing Methylotrophic
First by the Methylotrophic bacillus 4-L-16 bacterial strain for preparing in embodiment 1 at LB culture medium
On activate, after being placed in 37 DEG C of incubators cultivation 72 hours, drawn on LB culture medium by liquid-transfering gun
Thalline 5 ~ 10 μ L, is placed in the brown sugar water solution that 100mL weight/mass percentage composition is 10%, shaken cultivation,
Cultivation temperature is 30 DEG C, and shaking speed is 180rpm, and incubation time is 3 ~ 5 days, i.e. obtains containing methylotrophy
The seed liquor of type bacillus 4-L-16 bacterial strain, then by this seed liquor with the ratio of 1:10, joins flower
In the mixed liquor of raw cake and brown sugar, the preparation ratio of this mixed liquor is: the C/N according to peanut cake and nitrogen content,
And the phosphorus content of brown sugar, the C/N using material in conventional method regulation mixed liquor is 20/1, and adds suitable
The water of amount.
By fermentation 1000mL microbial inoculum and as a example by making the gross weight of peanut cake and brown sugar gross weight/water be 1/4, need
Adding seed liquor 80mL, peanut cake 88g, brown sugar 162g, add water 900mL.The zymotic fluid preparing is placed in
Temperature is 30 DEG C, shaking speed is shaken cultivation 7 days under the conditions of 180rpm；Under natural conditions, if desired greatly
Amount fermentation is also permissible, uses the good tun of ventilation type according to above ratio, is placed at shady and cool ventilation, directly
Fermentation, but to ensure each stirring 5 ~ 10min sooner or later, it is ensured that throughput, ferments 15 days；When having ready conditions
Large-scale fermentation is used to fill, temperatures above and mixing speed, to ferment 7 days, fermentation obtains bronzing after completing
Solution filters i.e. to obtain and obtains containing a large amount of Methylotrophic bacillus 4-L-16 bacterial strains and other multiple beneficial microorganisms
Microbial bacterial agent.
The biologic product of embodiment 5 4-L-16 of bacillus containing Methylotrophic
First by the Methylotrophic bacillus 4-L-16 bacterial strain for preparing in embodiment 1 at LB culture medium
On activate, after being placed in 37 DEG C of incubators cultivation 72 hours, drawn on LB culture medium by liquid-transfering gun
Thalline 5 ~ 10 μ L, is placed in the brown sugar water solution that 100mL weight/mass percentage composition is 10%, shaken cultivation,
Cultivation temperature is 30 DEG C, and shaking speed is 180rpm, and incubation time is 3 ~ 5 days, i.e. obtains containing methylotrophy
The seed liquor of type bacillus 4-L-16 bacterial strain, then by this seed liquor with the ratio of 1:10, joins flower
In the mixed liquor of raw cake and brown sugar, the preparation ratio of this mixed liquor is: the C/N according to soya-bean cake and nitrogen content,
And the phosphorus content of brown sugar, the C/N using material in conventional method regulation mixed liquor is 25/1, and adds suitable
The water of amount.
By fermentation 1000mL microbial inoculum and as a example by making the gross weight of soya-bean cake and brown sugar gross weight/water be 1/4, need to add
Entering seed liquor 120mL, soya-bean cake 62g, brown sugar 188g, add water 900mL.The zymotic fluid preparing is placed in temperature
Degree is 30 DEG C, shaking speed is shaken cultivation 7 days under the conditions of 180rpm；Under natural conditions, if desired in a large number
Ferment also permissible, use the good tun of ventilation type according to above ratio, be placed at shady and cool ventilation, directly send out
Ferment, but to ensure each stirring 5 ~ 10min sooner or later, it is ensured that throughput, ferments 15 days；Adopt when having ready conditions
Filling with large-scale fermentation, temperatures above and mixing speed, fermenting 7 days, it is molten that fermentation obtains bronzing after completing
Liquid filters i.e. to obtain and obtains micro-containing a large amount of Methylotrophic bacillus 4-L-16 bacterial strains and other multiple beneficial microorganisms
Bacteria agent.
Dwarf banana droop in seedling stage is prevented by the embodiment 6 4-L-16 microbial bacterial agent of bacillus containing Methylotrophic
Control effect
Test site: China tropic Agriculture Academy Sciences tropic Biotechnology Research Institute's warmhouse booth.
Test kind: No. 1 dwarf banana of bronze, is purchased from Zu Pei factory of Guangxi academy of agricultural sciences, height of seedling about 20cm, growth
Healthy and strong, five leaves wholeheartedly, without pest and disease damage.
Pathogen: No. 1 biological strain Fusarium oxysopoyum of Fusarium oxysporum Cuba specialized form
F.sp.cubence race 1(FOC1), by Chinese Academy of Tropical Agricultural Sciences's resource environment and Plant Protection Institute
Xie Yixian researcher provides.FOC1 is seeded on PDA culture medium, incubated l0d at 25 DEG C,
Spore washes and is made into concentration in sterilized water is 1 × 105Individual/mL spore suspension, for inoculation.
For examination soil: laterite, pick up from Haikou City Longhua District.
Microorganism formulation: the 4-L-16 microbial bacteria of bacillus containing Methylotrophic that embodiment 3 fermentation prepares
Agent
Test process: (1) CK: do not execute pathogen, do not execute microbial inoculum；(2) pathogen comparison: execute pathogen,
Do not execute microbial inoculum；(3) microbial inoculum process: execute pathogen, executes microbial inoculum.Each processes 20 strain any of several broadleaf plants seedlings.
Test method: choose the consistent Banana Seedlings of size growing way, seedling is hindered root, being then immersed in concentration is
5×10630min in individual/mL spore suspension, CK are immersed in equivalent clear water.After immersion, seedling replanting is extremely
High 10cm, diameter 12cm plastic tub in, apply microbial inoculum (diluting 50 times) by different disposal, every basin 100mL,
CK and pathogen comparison apply equivalent clear water, repeat zymotic fluid and the clear water applying equivalent every 7 days, examination
Testing period, the potted plant control measures of each process are all consistent, and every day observes the incidence of each process, treats pathogen pair
According to the incidence of disease reach more than 80% when, investigate each process disease index, measurement plant height and calculating relatively anti-
Effect.
Result of the test: observe through following the tracks of, any of several broadleaf plants seedling starts morbidity, when transplanting 22 days, pathogen after transplanting 10 days
Any of several broadleaf plants seedling incidence of disease of control treatment reaches 85%, investigates the disease index of each process, and the pathogen comparison state of an illness refers to
Number is 26.42, and the disease index that microbial inoculum is processed is 12.85, and relative control effect is 51.36%.Microbial inoculum process simultaneously
Rear average plant height is 29.27, exceeds 8.05cm than pathogen comparison, shows that this microbial inoculum has rush to plant strain growth
Raw effect.Such as Fig. 4, shown in table 5.
The control effect to dwarf banana droop in seedling stage for the table 5 4-L-16 microbial bacterial agent of bacillus containing Methylotrophic
The prevention and control to Brazil's any of several broadleaf plants droop in seedling stage for the embodiment 7 4-L-16 microbial bacterial agent of bacillus containing Methylotrophic
Effect
Test site: China tropic Agriculture Academy Sciences tropic Biotechnology Research Institute's warmhouse booth.
Test kind: Brazil any of several broadleaf plants, is purchased from Tropical China academy of sciences Zu Pei center, height of seedling about 15cm, growth
Healthy and strong, 3 leaves wholeheartedly, without pest and disease damage.
Pathogen: No. 4 biological strain Fusarium oxysopoyum of Fusarium oxysporum Cuba specialized form
F.sp.cubence race4(FOC4), provided by Agricultural University Of South China professor Jiang Zide.FOC4 is seeded in
On PDA culture medium, incubated l0d at 25 DEG C, spore being washed and is made into concentration in sterilized water is 5 × 106Individual
/ mL spore suspension, for inoculation.
For examination soil: laterite, pick up from Haikou City Longhua District.
Microorganism formulation: the 4-L-16 microorganism of bacillus containing Methylotrophic that in embodiment 3, fermentation prepares
Microbial inoculum
Test process: (1) CK: do not execute pathogen, do not execute microbial inoculum；(2) pathogen comparison: execute pathogen,
Do not execute microbial inoculum；(3) microbial inoculum process: execute pathogen, executes microbial inoculum.Each processes 20 strain any of several broadleaf plants seedlings.
Test method: choose the consistent Banana Seedlings of size growing way, seedling is hindered root, being then immersed in concentration is
5×10630min in individual/mL spore suspension, CK are immersed in equivalent clear water.After immersion, seedling replanting is extremely
High 10cm, diameter 12cm plastic tub in, apply microbial inoculum (diluting 50 times) by different disposal, every basin 100mL,
CK and pathogen comparison apply equivalent clear water, repeat zymotic fluid and the clear water applying equivalent every 7 days, examination
Testing period, the potted plant control measures of each process are all consistent, and every day observes the incidence of each process, treats pathogen pair
According to the incidence of disease reach 80% when, investigate the disease index of each process, measurement plant height simultaneously calculates relative control effect.
Result of the test: observe through following the tracks of, any of several broadleaf plants seedling starts morbidity after transplanting 14 days, when transplanting 30 days, sick
Any of several broadleaf plants seedling incidence of disease of former bacterium control treatment reaches 90%, investigates the disease index of each process, pathogen comparison disease
Feelings index is 34.28, and the disease index that microbial inoculum is processed is 11.43, and relative control effect is 66.56%.Simultaneously through adjusting
The average plant height looking into plant after each process plant height microbial inoculum is processed is 20.67cm, and the average plant height of pathogen comparison
For 15.46cm, show that this microbial inoculum has certain growth-promoting functions to Banana Growth；As shown in Fig. 5 and Biao 6.
The prevention and control to banana droop in seedling stage for the table 6 4-L-16 microbial bacterial agent of bacillus containing Methylotrophic are imitated
Really
Above-described embodiment is the present invention preferably embodiment, but embodiments of the present invention are not by above-mentioned enforcement
The restriction of example, the change made, modifies, replaces under other any Spirit Essence without departing from the present invention and principle
Generation, combination, simplification, all should be the substitute mode of equivalence, be included in protection scope of the present invention.
Claims (2)
1. a Methylotrophic Bacillus strain 4-L-16 for preventing and controlling banana fusarium wilt, is characterized in that: should
The preservation entitled Methylotrophic bacillus 4-L-16 of bacterial strain, Classification And Nomenclature is Bacillus
Methylotrophicus 4-L-16, depositary institution is China typical culture collection center, and preservation date is 2012
In on June 26, in, preserving number is CCTCC NO:M 2012248.
2. the Methylotrophic Bacillus strain 4-L-16 of the preventing and controlling banana fusarium wilt described in claim 1 exists
Preparation has the purposes in the medicine of preventing and controlling banana fusarium wilt effect.
Priority Applications (1)
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| CN103045515B (en) * | 2012-12-27 | 2015-12-09 | 陕西恒田化工有限公司 | Bacteria agent of a kind of Methylotrophic genus bacillus and its preparation method and application |
| CN103981131B (en) * | 2014-05-14 | 2016-08-31 | 中国农业科学院农产品加工研究所 | One strain Methylotrophic bacillus cereus and the application in degrading zearalenone thereof |
| ES2561908B2 (en) * | 2014-07-31 | 2016-12-01 | Universidad De Granada | Use of Bacillus methylotrophicus as a plant growth stimulant and biological control medium, and strains isolated from that species |
| CN105039204B (en) * | 2015-06-16 | 2018-04-20 | 安徽农业大学 | The strain of Methylotrophic bacillus and its application |
| CN105925504A (en) * | 2016-05-20 | 2016-09-07 | 南京林业大学 | Bacillus methylotrophicus ZY3A and application thereof to promotion of leaf lettuce growth |
| CN111557306B (en) * | 2020-05-21 | 2021-05-18 | 中国热带农业科学院南亚热带作物研究所 | Nano-drug preparation and application thereof in prevention and control of banana vascular wilt |
| CN116716190B (en) * | 2023-06-02 | 2024-04-05 | 云南省农业科学院农业环境资源研究所 | Chaetomium spirochaeta 201 and application thereof |
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