CN106172506A - A kind of prevention or the method for the treatment of banana Panama disease - Google Patents
A kind of prevention or the method for the treatment of banana Panama disease Download PDFInfo
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- CN106172506A CN106172506A CN201610544299.6A CN201610544299A CN106172506A CN 106172506 A CN106172506 A CN 106172506A CN 201610544299 A CN201610544299 A CN 201610544299A CN 106172506 A CN106172506 A CN 106172506A
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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
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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
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/12—Powders or granules
- A01N25/14—Powders or granules wettable
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Abstract
A kind of prevention or the method for the treatment of banana Panama disease, use bacillus subtilis HMB19198 preparation for Fructus Musae.Good prevention effect can be reached, a kind of microbial bacterial agent, the most more ensure that the preventing and treating of banana Panama disease is high-efficiency environment friendly, free of contamination；Manufacture method is the simplest simultaneously, and cost is the lowest, uses technology highly developed, is the one more preferably means of preventing and treating banana Panama disease.
Description
Technical field
The present invention relates to microorganism field, particularly relate to bacillus subtilis HMB19198 preparation and take at preventing and treating Fructus Musae bar
The method of horse disease.
Background technology
Fructus Musae (formal name used at school: Musa nana Lour.) Musaceae banana.Torrid areas is extensively cultivated edible.Fructus Musae
Taste is fragrant, rich in nutrition, can gather in the crops all the year round, be paid attention to the most very much in Temperate Region in China.Plant is large-scale draft, sends from root stock, by
The false bar of high 3～6 meters (10～20 chi) is formed at sheath bottom；Leaf Long Circle to oval, have up to 3～3.5 meter (10
～11.5 chis), wide 65 centimeters (26 cun), 10～20 pieces of bunches of raw stem tops.Spike is big, false masthead end extract out, spend majority, light
Yellow；Infructescence is curved hangs down, 10～20 string as a result, about 50～150.After plant result withered, root stock the sucker that grows continues
Breeding, each stock can be lived for many years.Originate in the southeast, Asia: all there is cultivation in the areas such as TaiWan, China, Hainan, Guangdong, Guangxi.
Banana Panama disease (Panama disease), has another name called: banana blight, is a kind of by soilborne Fructus Musae
Infectious disease, Fructus Musae progressively Panama of catching an illness is dead, and because soil is left over, Fructus Musae can not be planted in more than 30 years in Infected regions, is
" incurable disease " of Fructus Musae, is also the maximum killer threatening whole world banana planting industry.
Banana Panama disease has the destructive disease of generation in south China number province Fructus Musae producing region, and it shows Adult plant
Diseased plant first presents special yellow in lower blade and outer sheath, and the initial stage occurs at blade edge, then by portion to middle rib
Extension, contrasts notable with the dark green part of blade.Also having whole leaf to turn to be yellow, infected leaves is the most wilting, by xanthochromia brown and
Withered, its last a piece of top is extracted out the latest and maybe can not be extracted out, and last diseased plant is withered.Although having the most withered, but
Fruit development is bad, low quality.Maternal plant falls ill, and after portion (i.e. cauloid) is withered on the ground, its underground part (i.e. bulb) is the most immediately
Withered, remain to grow sprouting, continued growth, the middle and late growth stage to be arrived just manifests symptom.
It has been investigated that, banana Panama disease is that one infects the fascicular fungus-caused soil-borne disease of banana plant.It is
A kind of destructive disease, is International Plant quarantine object, Fusarium oxysporum Cuba specialized form infects and cause.In South China number formulary
There is generation in province Fructus Musae producing region.Mainly having No. 1 and No. 4 microspecies of cloth, No. 1 microspecies mainly cause harm dwarf banana, and No. 4 microspecies cause harm dwarf banana
And Fructus Musae.The general underproduction reaches more than 20%, and serious field even has no harvest.
The pathogen of banana Panama disease inside soil can remaining more than 30 years, do not have any pesticide can remove handle
It eradicates, so while there are some simple Oryza sativa L. crop rotations or organic with some, in the improvement of soil effectively, but
That is all locality, it is desired nonetheless to with disease-resistant, or the kind of resistance to disease is main.
In current banana Panama disease preventing and treating, those skilled in the art of the present technique the most do not find " specific drug ".At this
In field, technical staff's the most continuous test is groped middle searching and can more preferably be prevented and treated the preparation of banana Panama disease, Qi Zhongyi
It is so to be main by integrated control: 1, strict implement quarantine system strictly limits any of several broadleaf plants Seedling and abaca Seedling and incidental soil thereof
Inputted by lesion；2, choosing plants disease-free seedlings and disease-resistant variety for guaranteeing that Jiao Yuan, without Panama disease, should select the anosis any of several broadleaf plants Seedling of cultivation, the most just
Method is that the area of never banana Panama disease is introduced a fine variety.Additionally, select the kind of anti-Panama disease as far as possible；3, block lesion,
Prevent disease from spreading.Jiao Yuan finds fragmentary diseased plant, uproop immediately and diseased plant chopped, loads in plastic bag, adds stone
Ash also seals bag mouth, removes and allows it rot away from Jiao Yuan；4, sick soil processes and infects source to remove, and stops to propagate chance,
It is a critically important measure that sick soil processes；5, strengthen cultivation management and enrich fertilizer, dig trenches to drain the water away, strengthen plant premunition.Flat
Ground severe disease any of several broadleaf plants garden with good conditionsi can waterflooding leisure half a year or with Oryza sativa L. crop rotation.6, fiting chemical medicament is prevented and treated accordingly.But by
In chemical agent there is the drawback of essence in it, as the property of medicine is single, easily cause high poison, high residual, cause banana quality to reduce, simultaneously
It is easily generated Drug resistance.Because long-term in areal, use a kind of pesticide continuously, or use the pesticide that the mechanism of action is identical, evil
Worm, pathogenic bacteria or weeds to pesticide resistance will raising, and then produce certain Drug resistance.
In recent years, biological control method increasingly comes into one's own due to advantages such as efficient, environmental protection, pollution-free, noresidues.Cause
How this, search out that can to prevent and treat banana Panama disease bacteria agent be that those skilled in the art need the technology of solution at present badly and ask
Topic.
Summary of the invention
It is an object of the invention to provide a kind of prevention or the method for the treatment of banana Panama disease.
For reaching above-mentioned purpose, the technical scheme is that and be achieved in that:
A kind of prevention or the method for the treatment of banana Panama disease, including: use bacillus subtilis HMB19198 for Fructus Musae
Preparation.
Bacillus subtilis HMB19198 preparation is used for Fructus Musae.
Preferably, the living spores content of described bacillus subtilis HMB19198 preparation is: 1,000,000,000 living spores/gram hay bud
Spore bacillus wettable powder.
Preferably, the consumption of described bacillus subtilis HMB19198 is 50g/ strain-200g/ strain.
Preferably, use described in as root-pouring method.
The bacillus subtilis formulation of the present invention is a kind of solution utilizing Biological control banana Panama disease, this bacterium
Agent has preferable prevention effect to banana Panama disease, a kind of microbial bacterial agent, the most more ensure that Fructus Musae bar
Preventing and treating by horse disease is high-efficiency environment friendly, free of contamination；Manufacture method is the simplest simultaneously, and cost is the lowest, uses technology
Highly developed, it is the one more preferably means of preventing and treating banana Panama disease.
Detailed description of the invention
The core of the present invention is to provide a kind of prevention or the method for the treatment of banana Panama disease, it is possible to effectively preventing Fructus Musae
Panama disease.
Below in conjunction with the embodiment of the present invention, the technical scheme in the embodiment of the present invention is clearly and completely described,
Obviously, described embodiment is only a part of embodiment of the present invention rather than whole embodiments.Based in the present invention
Embodiment, the every other embodiment that those of ordinary skill in the art are obtained under not making creative work premise, all
Belong to the scope of protection of the invention.
Embodiment one: the bacillus subtilis wettable powder security test to banana seedlings.
The purpose of the present embodiment is to measure the safety of bacillus subtilis wettable powder preventing and treating Fructus Musae, for its preventing and treating perfume
Any of several broadleaf plants Panama disease provides foundation.
The present embodiment for study thing be Fructus Musae, wherein crop varieties is respectively high foot and pauses Rhizoma Anemones flaccidae, belongs to high dry type perfume (or spice) tooth
Any of several broadleaf plants, for one of Gaozhou City of Guangdong Province improved seeds；Embanked field plantain, belongs to plantain type, for one of the local varieties of Xinhui of Guangdong Province；In
The big dwarf banana in mountain, belongs to dwarf banana type, and for one of the local varieties of Guangdong Zhong Shan, above banana seedlings is all purchased for from academy of agricultural sciences of Guangdong Province
Examination.
The reagent agent of the present embodiment is the microbial bacterial agent utilizing bacillus subtilis HMB19198 to produce, this microorganism
The active component of microbial inoculum is bacillus subtilis HMB19198 spore and its Extracellular metabolism.The present embodiment use section Lv Feng
1,000,000,000 living spores/gram bacillus subtilis that biochemical technology company limited produces with bacillus subtilis HMB19198 spore is wettable
Property powder, its consumption is typically up to 50g/ strain, by its gradient design test dose of 1,2,3 times, i.e. 50,100,200g/
Strain.Comparison medicament is 10,000,000,000 spores/gram bacillus subtilis, according to the design concentration of test medicine, according to effective ingredient equivalent
Principle, is defined as 5g/ strain.To process as blank without medicament clear water.
The condition of culture of the present embodiment is to carry out in indoor, plants the tissue cultured seedling of above-mentioned 3 banana varieties in nutritive cube
In, add appropriate decomposed manure as culture matrix not planting the fertile field soil of Fructus Musae, Routine Management, cultivate to the Fructus Musae 5 leaf phase
Standby.
The application method of the present embodiment is root-pouring method, is uniformly poured onto potted plant banana root soil layer, waters after being diluted by medicinal liquid
Filling amount have substantially pouring moistening with soil layer but without hydrops till.
The test method of the present embodiment is as follows: selecting the number of blade 5 leaf, the banana seedlings that cauloid height is basically identical, along Fructus Musae
Shoot root periphery digs deep about 10cm ditch, carries out process of spreading manuer in holes.With clear water comparison, this confession examination pesticide 1,000,000,000 spore/gram bacillus subtilis
Bacterium WP preparation 50,100,200g/ strain, compare medicine 10,000,000,000 spore/gram bacillus subtilis WP preparation 5g/ strain, dispenser bonnet thin soil
Layer.Often process 4 repetitions, often repeat 5 strain banana seedlings, the most often process totally 20 strain banana seedlings.Disposed within, conventional management of the use of water, examination
The most separately apply fertilizer during testing material.
The investigation method that the present embodiment uses is as follows: in this test, after dispenser 5 days, observation Leaf of banana, each medicament of comparison
Processing and whether blade is had poisoning, if any poisoning, time, symptom and the order of severity that record poisoning produces, according to NY/T
In 1965.1-2010, poisoning grade scale carries out classification, cultivates to dispenser 21 days, observes with or without situations such as deformity, excessive growth, as
There is a poisoning, continue to observe its development degree, if can recover and recovery situation etc., calculate its number of blade respectively, measure its vacation
Stem height (cm), the diversity between comparison process and CK, it is calculated as follows safety coefficient I, evaluates the shadow to banana seedlings growth
Ring.
On the maximum of crop, without affecting, test dose/medicament field is the highest recommends dosage to safety coefficient I=medicament
The present embodiment processes range estimation in latter 5 days in test, for examination 1,000,000,000 spores/gram bacillus subtilis WP preparation 50,100,
200g/ strain, and comparison medicament 10,000,000,000 spore/gram bacillus subtilis WP preparation 5g/ strain spread manuer in holes process after, 3 kinds of perfume (or spice) that are put to the test
Any of several broadleaf plants is all safely that blade all shows as strong green without poisoning, without the variable colors such as yellow, albefaction, chlorisis, rust staining, withered speckle, necrosis now
As, leaf margin is normal, compares CK no significant difference with clear water.Dispenser processes latter 21 days and observes, and each process Fructus Musae is different without excessive growth, blade
The abnormal phenomenon such as often curling, cauloid is without hanging contracting, and the number of blade, cauloid height are estimated and CK zero difference, illustrates for examination typically up to
Consumption and 2 times, under the conditions of 4 times all to target crop banana seedlings safety without poisoning.
Test result see table:
Table 1, confession examination 1,000,000,000 spores/gram bacillus subtilis WP pouring root safety evaluatio to banana seedlings
Note: same breed, with the identical person of lower case after column data, represents through the not notable (p-of DMRT legally constituted authority meter comparing difference
0.05)
Knowable to the data of the present embodiment result, test medicine 1,000,000,000 spore/gram bacillus subtilis WP preparation 50,100,
200g/ strain is spreaded manuer in holes, to after " high foot pause Rhizoma Anemones flaccidae ", " furrow head plantain " and " the big dwarf banana in middle mountain " banana seedlings root irrigation 21 days, and blade
Number be respectively 6.5-7.1,5.6-6.1,5.0-5.6 leaf/strains, cauloid height aspect be respectively 67.6-40.7,32.67-33.8,
31.8-33.4cm all with compare medicament 10,000,000,000 spore/gram bacillus subtilis WP preparation 5g/ strain spread manuer in holes and clear water comparison CK
The number of blade in district is slightly better than CK without significance difference XOR, but all with compare medicament, CK difference within " ± 10% ", and without other
Symptom, according to NY/T1965.1-2010 required standard, illustrates under this test dose and method, 1,000,000,000 spores/gram bacillus subtilis
Bacterium WP supplies examination banana variety safety to 3 kinds.
Initial data is as follows:
Table 2 10 hundred million spore/gram bacillus subtilis WP on the impact of the Fructus Musae second number of blade (perforated method, after medicine 21 days, leaf/
Strain)
Table 3 10 hundred million spore/gram bacillus subtilis WP on the impact of banana seedlings cauloid height (perforated method, after medicine 21 days,
Unit: cm)
Embodiment two: bacillus wettable powder preventing and treating banana Panama disease indoor biological determination of activity.
The purpose of the present embodiment is to measure 20,000,000,000 spores/gram bacillus subtilis wettable powder to banana Panama disease
Indoor biological activity, determines the optimal using dosage of medicament and rational use technology.
The present embodiment for examination target be banana Panama pathogenic bacteria, be Fusarium oxysporum Cuba specialized form Fusarium o ×
Ysporum f.sp.cubense4 biological strain B2 bacterial strain, is ground by Plant Pathology system of agricultural college of Agricultural University Of South China fungus
Studying carefully room to provide, activation is for examination.
The reagent agent of the present embodiment is the microbial bacterial agent utilizing bacillus subtilis HMB19198 to produce, this microorganism
The active component of microbial inoculum is bacillus subtilis HMB19198 spore and its Extracellular metabolism.The present embodiment use section Lv Feng
Biochemical technology company limited with bacillus subtilis HMB19198 spore produce 20,000,000,000 living spores/a gram bacillus subtilis can
WP；The 200000000000 spores/gram hay provided by Wuhan City, Hubei Province Tian Hui biological engineering company limited selected by comparison medicament
Bacillus cereus；And select emulsifying agent 0203B to use as other solvents.
The medicament of the present embodiment is formulated as follows:
According to pilot study, the experimental concentration of design experiment medicament and comparison medicament bacillus subtilis is 1 × 104、1×
105、1×106、1×107、1×108Cfu/mL (bacteriostasis rate about 20%-90%, containing concentration in PDA culture medium, lower same).
Take test medicine 20,000,000,000 spore/gram bacillus subtilis (R31) former powder and comparison medicament 200,000,000,000 spore/gram withered
Grass bacillus cereus mother's medicine, prepares bacillus subtilis with the clear water containing 0.05%0203B emulsifying agent respectively and becomes 2 × 109Cfu/mL,
The medicinal liquid of other concentration, is diluted by upper level concentration liquid.
The chemicals treatment of the present embodiment is as follows: under aseptic technique, processes the sterilizing that will melt in advance according to test
PDA culture medium is quantitatively adding in aseptic conical flask, in medicinal liquid with culture volume than 1: 19 ratio, depends on from low concentration to high concentration
Secondary quantitative absorption medicinal liquid, is separately added in above-mentioned conical flask, makes the pastille PDA culture medium of above-mentioned concentration, is subsequently poured into diameter
For the culture dish of 9cm, every ware culture medium 20mL, make the pastille flat board of respective concentration.Every concentration sets 4 repetitions, often repeats 1
Culture dish.
The present embodiment inoculation is with cultivation: by the banana Panama cause of disease bacterium of activation culture 7d, aseptically use diameter
The sterilizing card punch of 5mm, from colony edge and take bacterium cake, with inoculator by pure culture biscuits involvng inoculation in pastille flat board central authorities, mycelia faces
On, covering ware lid, be placed in temperature 25 ± 1 DEG C, humidity 80%-90% incubator is dark cultivates 7d " Invest, Then Investigate " continuously.
Fact-finding process is as follows: after cultivating 7d, and in comparison culture dish, pathogenic bacteria has covered culture dish more than 90%, with ruler with ten
Word interior extrapolation method vertical survey colony diameter is each once, and unit is cm.
The statistics of the present embodiment is as follows with the method for analysis: often processes 4 repetitions, takes its meansigma methods D, is calculated as follows each
Processing for examination banana Panama pathogenic bacteria mycelial growth inhibition rate, unit is percentage ratio (%)
D=D1-D2(wherein D: bacterium colony increases diameter；D1: colony diameter；D2: bacterium cake diameter)
I=(D0-Dt)/D0×100
I: mycelial growth inhibition rate；D0=blank bacterium colony increases diameter, Dt=chemicals treatment bacterium colony increases diameter
Preventive effect probability value according to each drug concentration logarithm value and correspondence makees linear regression analysis, seeks virulence regression equation,
Calculate concentration EC in the suppression of each single dose and each proportioning thereof50、EC90And 95% confidence limit (mg/L).
The present embodiment interpretation of result:
Each colony diameter data that process are shown in Table 1, EC50Values etc. are done index result of calculation and are shown in Table 2,.Result shows, after cultivation
7d, CK colony diameter increases 7.81cm, and reagent agent 20,000,000,000 spore/gram bacillus subtilis former powder suppression is taken for examination Fructus Musae bar
The EC50 value of horse disease bacterium mycelial growth and EC90 value are respectively 1.64 × 105CFU/mL and 156.78 × 105CFU/mL, compare medicine
Agent 200,000,000,000 spore/gram bacillus subtilis starter medicine is respectively 1.81 to EC50 and the EC90 value of banana Panama pathogenic bacteria of the same race ×
105CFU/mL and 128.40 × 105CFU/mL, EC50 value with for examination the former powder of bacillus subtilis basically identical (± 10% with
In), meet the regular activated feature of this series products.
Table 4 bacillus subtilis indoor biological activity assay data (cm, Plating) to banana Panama pathogenic bacteria
Note: colony diameter value added=colony diameter value (0.5cm) bacteriostasis rate (%) (CK value added-process value added)/
CK value added × 100%
Table 5 bacillus subtilis virulence (Plating) to banana Panama pathogenic bacteria
Note: EC50/EC90 machine 95% confidence limit concentration unit is × 105CFU/mL
The present embodiment conclusion be the former powder of bacillus subtilis be EC50 value and EC90 to banana Panama pathogenic bacteria inhibitory activity
Value is respectively 1.64CFU/mL and 1.56.78CFU/mL, inconspicuous with the biological activity difference of female medicine of the like product used,
I.e. reach it and prevent and treat level.
Although, the present invention is described in detail the most with a general description of the specific embodiments, but
On the basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to the scope of protection of present invention.
Claims (4)
1. a prevention or the method for the treatment of banana Panama disease, it is characterised in that including: use bacillus subtilis for Fructus Musae
HMB19198 preparation.
Prevention the most according to claim 1 or the method for the treatment of banana Panama disease, it is characterised in that described hay spore
The living spores content of bacillus HMB19198 preparation is: 1,000,000,000 living spores/gram bacillus subtilis wettable powder.
Prevention the most according to claim 2 or the method for the treatment of banana Panama disease, it is characterised in that described hay spore
The consumption of bacillus HMB19198 is 50g/ strain-200g/ strain.
Prevention the most according to claim 1 or treatment banana Panama disease method, it is characterised in that described in use into fill
Root method.
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Cited By (4)
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| CN106973943A (en) * | 2017-04-18 | 2017-07-25 | 柏玉兰 | The antimicrobial plant vaccine and preparation method of Panama disease can be prevented and treated |
| CN106986378A (en) * | 2017-04-15 | 2017-07-28 | 柏玉兰 | Panama disease microbe soil restoration agent can be prevented and treated |
| CN112457125A (en) * | 2020-12-22 | 2021-03-09 | 广西壮族自治区农业科学院 | Banana wilt prevention bio-organic fertilizer and preparation method and application thereof |
| CN113265480A (en) * | 2021-06-08 | 2021-08-17 | 河北省农林科学院植物保护研究所 | Specific primer of bacillus subtilis HMB19198 strain, Taqman probe and application |
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| CN112457125A (en) * | 2020-12-22 | 2021-03-09 | 广西壮族自治区农业科学院 | Banana wilt prevention bio-organic fertilizer and preparation method and application thereof |
| CN113265480A (en) * | 2021-06-08 | 2021-08-17 | 河北省农林科学院植物保护研究所 | Specific primer of bacillus subtilis HMB19198 strain, Taqman probe and application |
| CN113265480B (en) * | 2021-06-08 | 2022-03-18 | 河北省农林科学院植物保护研究所 | Specific primer of bacillus subtilis HMB19198 strain, Taqman probe and application |
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