CN105385643A - Compound microbial agent for preventing and treating banana wilt disease and prevention and treatment method - Google Patents
Compound microbial agent for preventing and treating banana wilt disease and prevention and treatment method Download PDFInfo
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- CN105385643A CN105385643A CN201511009699.9A CN201511009699A CN105385643A CN 105385643 A CN105385643 A CN 105385643A CN 201511009699 A CN201511009699 A CN 201511009699A CN 105385643 A CN105385643 A CN 105385643A
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- banana blight
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protecting plants
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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
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
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- 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
- C12N1/14—Fungi; Culture media therefor
Abstract
The invention relates to the field of banana cultivation, in particular to a compound microbial agent for preventing and treating a banana wilt disease and a prevention and treatment method. The compound microbial agent comprises paenibacillus polymyxa, bacillus subtilis, bacillus thuringiensis, pseudomonas stutzeri, bacillus amyloliquefaciens, cold-resistant brevibacteriaceae, trichoderma and metarrhizium anisopliae. By means of a reasonable proportion, all strains are mutually influenced and promoted, and the prevention and treatment effect on the banana wilt disease is greatly improved. The prevention and treatment method of the banana wilt disease can effectively adjust the composition of bacterial colonies in soil, promote growth and propagation of probiotics and antagonistic bacteria in the soil, rapidly restore and stabilize the balance of the bacterial colonies in the soil, prevent and treat occurrence and spread of the banana wilt disease, effectively decrease the occurrence rate of the banana wilt disease and maintain the stable and high yield of bananas.
Description
Technical field
The present invention relates to banana cultivation field, refer to a kind of composite fungus agent and the prevention and controls of preventing and treating banana blight especially.
Background technology
Banana is famous Perenniporia martius fruit, and all there is plantation countries and regions, about 120, the whole world, and its annual production is more than 100,000,000 tons.China is banana production and consumption big country, and there are Guangdong, Guangxi, Fujian, Hainan and Yunnan in main banana plantation, defines large-scale industrial pattern.Banana blight is by Fusarium oxysporum Cuba specialized form (Fusariumoxysporiumf.sp.cubense, FOC) destructive disease in the banana production caused, is distributed widely in the banana plantation in Asia, Africa, Australia and tropical America.Generally, namely there is blight diseased plant in 5 months in banana planting, to seedling damage greatly.Since there is the disease of banana blight No. 4 microspecies from 1996 in China, disease with geometricprogression in the main banana producing region expansion of China, in Guangdong, Hainan, the big area outburst such as Guangxi and popular, make traditional banana growing areas slump of disastrous proportions.Traditional chemical pesticide is not only not good enough to the prevention effect of banana blight, and medicament also can be stranded in soil, water body, cause environmental pollution, cause serious threat to human health, so it is extremely necessary to obtain a kind of method of preventing and treating banana blight safely and efficiently.
Summary of the invention
The present invention proposes a kind of composite fungus agent and the prevention and controls of preventing and treating banana blight, effectively can prevent and treat banana blight, reduce the sickness rate of continuous cropping banana garden blight.
Technical scheme of the present invention is achieved in that
A kind of composite bacteria, according to cell count gauge, described composite bacteria comprises following bacterial strain: Paenibacillus polymyxa 1-10 part, subtilis 1-15 part, bacillus thuringiensis 1-15 part, Pseudomonas stutzeri 5-10 part, bacillus amyloliquefaciens 5-10 part, brevibacterium frigoritolerans 5-10 part, wooden mould 5-10 part, green muscardine fungus 1-10 part.
Further, according to cell count gauge, described composite bacteria comprises following bacterial strain: Paenibacillus polymyxa 3-7 part, subtilis 8-12 part, bacillus thuringiensis 8-12 part, Pseudomonas stutzeri 6-8 part, bacillus amyloliquefaciens 6-8 part, brevibacterium frigoritolerans 6-8 part, wooden mould 6-8 part, green muscardine fungus 3-7 part.
The application of above-mentioned composite bacteria in control banana blight.
Prevent and treat a composite fungus agent for banana blight, containing above-described composite bacteria.
Further, in the composite fungus agent of described control banana blight, total bacterial strain concentration is>=10
8cfu/mL.
A prevention and controls for banana blight, comprises the following steps:
After seedling plantation, get above-described composite fungus agent and be administered to Jiao Yuanzhong.
Further, in the prevention and controls of above-mentioned banana blight, when described composite fungus agent uses, being diluted to total bacterial strain concentration is 10
4-10
6cfu/mL.
Further, the application process of described composite fungus agent is that filling root or ground, the end spray; Consumption is 1-2.5kg/ strain, and every 15-30 days uses once.
Further, the prevention and controls of above-mentioned banana blight, also comprises front step: before Banana Seedlings is transplanted to land for growing field crops, first sprays mycocide large Tanaka, shines, then Banana Seedlings is transplanted to large Tanaka through spacious.
Further, described mycocide is dioxide peroxide, potassium permanganate or dazomet; Mass concentration used is 50-500mg/L, and consumption is every square metre of 100-300mg.
Beneficial effect of the present invention:
Paenibacillus polymyxa all has certain restraining effect to plurality of plant diseases, experiment proves, Paenibacillus polymyxa has two large functions: (1) effectively can prevent and treat vegetative bacteria and fungoid soil-borne disease by filling with root, the bacterium in leaf portion and fungal disease can be made obviously to reduce simultaneously; (2) to plant, there is obvious growth promotion, production-increasing function.
Subtilis is extensively distributed in the organism of soil and corruption, can improve soil aggregate, improvement soil, alleviate continuous cropping obstacle, suppress the breeding of pathogenic bacteria and the invasion and attack to plant root in soil, reduce plant soil-borne diseases, prevent the outburst of multiple disease and pest.
Bacillus thuringiensis is a kind of widely used environmental type microbial pesticide, all has toxic action to 200 various pests, and to environment and water source pollution-free, to person poultry harmless, but its less stable, the longevity of residure is short, desinsection speed is slow.
Bacillus amyloliquefaciens is made scientific researches and is produced a series of meta-bolites in himself process of growth, as microbiotic, antimicrobial protein polypeptide, phytase, Surfactin, plant growth hormones etc., not only there is the ability suppressing Plant diseases, in plant surface, Colonization inside plants and conduction, effect of significant growth promoting effects can also be had to plant.
Wood is mould is a kind of important Antagonistic Fungi in biological control, can be used to the main mechanism of controlling disease or suppression cause of disease, its antagonistic action can be classified into five large classes usually, namely produces microbiotic, nutrient competition, micro-parasitism, cell wall breakdown ferment and inducing plant and produces resistance.Except the effect in biological control, wood is mould to be had various plants and impels effect significantly, can improve plant seed vigor, increase seedlings root active, promote plants grow taller, improve leaf chlorophyll and soluble protein content.
Green muscardine fungus can parasitize a class disinsection fungal of various pests, is a kind of insect pathogenic bacteria of wide spectrum, to person poultry harmless, also has free from environmental pollution, noresidue, the advantage that insect can not develop immunity to drugs simultaneously.
Pseudomonas stutzeri and brevibacterium frigoritolerans all have certain bacteriostatic action.
Composite fungus agent of the present invention, by multiple bacterial strain is carried out rational proportion, has again the effect influencing each other, mutually promote, greatly increases the prevention effect of banana blight between each bacterial strain.Meanwhile, after being administered in soil, the metabolism of each bacterial strain and breeding activity, the bacterium colony composition in soil and Bacterial community can be affected again, maintain the colony balance in soil, suppress the generation of banana blight and spread, to banana blight, there is good prophylactic effect.The prevention and controls of banana blight of the present invention, before transplanting, first carrying out disinfection to field soil with killing agent, effectively can reduce the pathogenic bacteria content in soil; After Banana Seedlings is transplanted in field soil, regularly use composite fungus agent pathogenic bacteria being had to inhibition, the bacterium colony in soil can be effectively regulated to form, promote the growth and breeding of probiotic bacterium and Antagonistic Fungi in soil, Quick-return colony balance in stable soil, prevent the generation of banana blight and spread, promoting the growth of Banana Seedlings, effectively reducing the incidence of banana blight.
Accompanying drawing explanation
In order to be illustrated more clearly in the embodiment of the present invention or technical scheme of the prior art, be briefly described to the accompanying drawing used required in embodiment or description of the prior art below, apparently, accompanying drawing in the following describes is only some embodiments of the present invention, for those of ordinary skill in the art, under the prerequisite not paying creative work, other accompanying drawing can also be obtained according to these accompanying drawings.
Fig. 1 prevents and treats the composite fungus agent of banana blight to banana blight germ fungistatic effect for a kind of described in the embodiment of the present invention 1;
Fig. 2 prevents and treats the composite fungus agent of banana blight to banana blight germ fungistatic effect for a kind of described in the embodiment of the present invention 2;
Fig. 3 prevents and treats the composite fungus agent of banana blight to banana blight germ fungistatic effect for a kind of described in the embodiment of the present invention 3;
Fig. 4 prevents and treats the composite fungus agent of banana blight to banana blight germ fungistatic effect for a kind of described in the embodiment of the present invention 4;
Fig. 5 prevents and treats the composite fungus agent of banana blight to banana blight germ fungistatic effect for a kind of described in the embodiment of the present invention 5;
Fig. 6 prevents and treats the composite fungus agent of banana blight to banana blight germ fungistatic effect for a kind of described in the embodiment of the present invention 6;
Fig. 7 is that different treatment affects Banana Seedlings blade MDA;
Fig. 8 is the impact of different treatment on Banana Seedlings blade POD activity;
Fig. 9 is the impact of different treatment on Banana Seedlings sod activity;
Figure 10 is the impact of different treatment on Banana Seedlings proline content.
In Fig. 1-Fig. 6, a is banana blight germ, and b is the composite fungus agent described in the embodiment of the present invention 1; C is the composite fungus agent described in the embodiment of the present invention 2; D is the composite fungus agent described in the embodiment of the present invention 3; E is the composite fungus agent described in the embodiment of the present invention 4; F is the composite fungus agent described in the embodiment of the present invention 5; G is the composite fungus agent described in the embodiment of the present invention 6.
Embodiment
Below in conjunction with the embodiment of the present invention, be clearly and completely described the technical scheme in the present invention, obviously, described embodiment is only the present invention's part embodiment, instead of whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art, not making the every other embodiment obtained under creative work prerequisite, belong to the scope of protection of the invention.
Embodiment 1
A kind of composite bacteria, according to cell count gauge, described composite bacteria comprises following bacterial strain: Paenibacillus polymyxa 1 part, subtilis 1 part, bacillus thuringiensis 1 part, Pseudomonas stutzeri 5 parts, bacillus amyloliquefaciens 5 parts, brevibacterium frigoritolerans 5 parts, mould 5 parts of wood, green muscardine fungus 1 part.
Above-described composite bacteria can be used for the control of banana blight.
Prevent and treat a composite fungus agent for banana blight, containing above-described composite bacteria; Total bacterial strain concentration is 10
8cfu/mL.
A prevention and controls for banana blight, comprises the following steps:
Before Banana Seedlings is transplanted to land for growing field crops, first spray mycocide dioxide peroxide, potassium permanganate or dazomet large Tanaka, mass concentration used is 50mg/L, and consumption is every square metre of 100mg; Shine through spacious, then Banana Seedlings is transplanted to large Tanaka; After seedling plantation, getting above-described composite fungus agent, to be diluted to total bacterial strain concentration be 10
4cfu/mL, is administered to Jiao Yuanzhong, and application process is that filling root or ground, the end spray; Consumption is 1kg/ strain, within every 15 days, uses once.
Embodiment 2
A kind of composite bacteria, according to cell count gauge, described composite bacteria comprises following bacterial strain: Paenibacillus polymyxa 10 parts, subtilis 15 parts, bacillus thuringiensis 15 parts, Pseudomonas stutzeri 10 parts, bacillus amyloliquefaciens 10 parts, brevibacterium frigoritolerans 10 parts, mould 10 parts of wood, green muscardine fungus 10 parts.
Further, according to cell count gauge, described composite bacteria comprises following bacterial strain: Paenibacillus polymyxa 3-7 part, subtilis 8-12 part, bacillus thuringiensis 8-12 part, Pseudomonas stutzeri 6-8 part, bacillus amyloliquefaciens 6-8 part, brevibacterium frigoritolerans 6-8 part, wooden mould 6-8 part, green muscardine fungus 3-7 part.
The application of above-described composite bacteria in control banana blight.
Prevent and treat a composite fungus agent for banana blight, containing above-described composite bacteria; Total bacterial strain concentration is 10
9cfu/mL.
A prevention and controls for banana blight, comprises the following steps:
Before Banana Seedlings is transplanted to land for growing field crops, first spray mycocide dioxide peroxide, potassium permanganate or dazomet large Tanaka, mass concentration used is 500mg/L, and consumption is every square metre of 300mg; Shine through spacious, then Banana Seedlings is transplanted to large Tanaka; After seedling plantation, getting above-described composite fungus agent, to be diluted to total bacterial strain concentration be 10
6cfu/mL, is administered to Jiao Yuanzhong, and application process is that filling root or ground, the end spray; Consumption is 2.5kg/ strain, within every 30 days, uses once.
Embodiment 3
A kind of composite bacteria, according to cell count gauge, described composite bacteria comprises following bacterial strain: Paenibacillus polymyxa 5 parts, subtilis 10 parts, bacillus thuringiensis 10 parts, Pseudomonas stutzeri 7 parts, bacillus amyloliquefaciens 7 parts, brevibacterium frigoritolerans 7 parts, mould 7 parts of wood, green muscardine fungus 5 parts.
Above-described composite bacteria can be used for the control in banana blight.
Prevent and treat a composite fungus agent for banana blight, containing above-described composite bacteria; Total bacterial strain concentration is 10
8cfu/mL.
A prevention and controls for banana blight, comprises the following steps:
Before Banana Seedlings is transplanted to land for growing field crops, first spray mycocide dioxide peroxide, potassium permanganate or dazomet large Tanaka, mass concentration used is 250mg/L, and consumption is every square metre of 200mg; Shine through spacious, then Banana Seedlings is transplanted to large Tanaka; After seedling plantation, getting above-described composite fungus agent, to be diluted to total bacterial strain concentration be 10
5cfu/mL, is administered to Jiao Yuanzhong, and application process is that filling root or ground, the end spray; Consumption is 2kg/ strain, within every 20 days, uses once.
Embodiment 4
A kind of composite bacteria, according to cell count gauge, described composite bacteria comprises following bacterial strain: Paenibacillus polymyxa 3 parts, subtilis 8 parts, bacillus thuringiensis 8 parts, Pseudomonas stutzeri 6 parts, bacillus amyloliquefaciens 6 parts, brevibacterium frigoritolerans 6 parts, mould 6 parts of wood, green muscardine fungus 3 parts.
Above-described composite bacteria can be used for the control in banana blight.
Prevent and treat a composite fungus agent for banana blight, containing above-described composite bacteria; Total bacterial strain concentration is 10
9cfu/mL.
A prevention and controls for banana blight, comprises the following steps:
Before Banana Seedlings is transplanted to land for growing field crops, first spray mycocide dioxide peroxide, potassium permanganate or dazomet large Tanaka, mass concentration used is 150mg/L, and consumption is every square metre of 200mg; Shine through spacious, then Banana Seedlings is transplanted to large Tanaka; After seedling plantation, getting above-described composite fungus agent, to be diluted to total bacterial strain concentration be 10
4cfu/mL, is administered to Jiao Yuanzhong, and application process is that filling root or ground, the end spray; Consumption is 1.5kg/ strain, within every 25 days, uses once.
Embodiment 5
A kind of composite bacteria, according to cell count gauge, described composite bacteria comprises following bacterial strain: Paenibacillus polymyxa 7 parts, subtilis 12 parts, bacillus thuringiensis 12 parts, Pseudomonas stutzeri 8 parts, bacillus amyloliquefaciens 8 parts, brevibacterium frigoritolerans 8 parts, mould 8 parts of wood, green muscardine fungus 7 parts.
Above-described composite bacteria can be used for the control in banana blight.
Prevent and treat a composite fungus agent for banana blight, containing above-described composite bacteria; Total bacterial strain concentration is 10
8cfu/mL.
A prevention and controls for banana blight, comprises the following steps:
Before Banana Seedlings is transplanted to land for growing field crops, first spray mycocide dioxide peroxide, potassium permanganate or dazomet large Tanaka, mass concentration used is 300mg/L, and consumption is every square metre of 250mg; Shine through spacious, then Banana Seedlings is transplanted to large Tanaka; After seedling plantation, getting above-described composite fungus agent, to be diluted to total bacterial strain concentration be 10
5cfu/mL, is administered to Jiao Yuanzhong, and application process is that filling root or ground, the end spray; Consumption is 2.5kg/ strain, within every 30 days, uses once.
Embodiment 6
A kind of composite bacteria, according to cell count gauge, described composite bacteria comprises following bacterial strain: Paenibacillus polymyxa 1 part, subtilis 15 parts, bacillus thuringiensis 10 parts, Pseudomonas stutzeri 8 parts, bacillus amyloliquefaciens 10 parts, brevibacterium frigoritolerans 8 parts, mould 6 parts of wood, green muscardine fungus 5 parts.
Above-described composite bacteria can be used for preventing and treating banana blight.
Prevent and treat a composite fungus agent for banana blight, containing above-described composite bacteria; Total bacterial strain concentration is 10
8cfu/mL.
A prevention and controls for banana blight, comprises the following steps:
Before Banana Seedlings is transplanted to land for growing field crops, first spray mycocide dioxide peroxide, potassium permanganate or dazomet large Tanaka, mass concentration used is 250mg/L, and consumption is every square metre of 200mg; Shine through spacious, then Banana Seedlings is transplanted to large Tanaka; After seedling plantation, getting above-described composite fungus agent, to be diluted to total bacterial strain concentration be 10
6cfu/mL, is administered to Jiao Yuanzhong, and application process is that filling root or ground, the end spray; Consumption is 1kg/ strain, within every 200 days, uses once.
Composite fungus agent of the present invention is to the inhibition of banana blight germ
Different composite fungus agents is prepared respectively according to following scheme, banana blight bacteria (the Fusariumoxysporumf.sp.cubenseSnyder & Hansen that diameter is 5mm is inoculated in the PDA culture medium flat plate centre being 9cm at diameter, referred to as Foc) the mycelia block of No. 4 physiological strains, by obtained different mix bacterium agents to its bacteriostatic activity.Again the composite fungus agent after activation is seeded in the surrounding apart from target bacterial strain mycelia block 2 ~ 3cm place, every plating four composite fungus agents, not connect the banana blight bacteria of composite fungus agent as blank, each process arranges three repetitions, cultivate 7d at 28 DEG C after, measure the width of antibacterial band.Its result is as shown in table 1 and Fig. 1 to Fig. 6.
Scheme 1: obtain composite fungus agent according to the composite bacteria formula described in embodiment 1;
Scheme 2: obtain composite fungus agent according to the composite bacteria formula described in embodiment 2;
Scheme 3: obtain composite fungus agent according to the composite bacteria formula described in embodiment 3;
Scheme 4: obtain composite fungus agent according to the composite bacteria formula described in embodiment 4;
Scheme 5: obtain composite fungus agent according to the composite bacteria formula described in embodiment 5;
Scheme 6: obtain composite fungus agent according to the composite bacteria formula described in embodiment 6;
Scheme 7: remove Paenibacillus polymyxa in the composite bacteria formula described in embodiment 2;
Scheme 8: remove subtilis in the composite bacteria formula described in embodiment 2;
Scheme 9: remove bacillus thuringiensis in the composite bacteria formula described in embodiment 2;
Scheme 10: remove Pseudomonas stutzeri in the composite bacteria formula described in embodiment 2;
Scheme 11: remove bacillus amyloliquefaciens in the composite bacteria formula described in embodiment 2;
Scheme 12: remove brevibacterium frigoritolerans in the composite bacteria formula described in embodiment 2;
Scheme 13: remove wood in the composite bacteria formula described in embodiment 2 mould;
Scheme 14: remove green muscardine fungus in the composite bacteria formula described in embodiment 2.
Table 1 different composite bacterium is to the antibacterial bandwidth of banana blight germ
As can be seen from table 1 and Fig. 1-6, the composite fungus agent of the present invention's 6 embodiment gained all has good inhibition to banana blight germ.Removing wherein any one component in composite fungus agent, obviously decline to the width of its antibacterial band, there is significant difference in its fungistatic effect and 6 embodiments.Illustrate that composite fungus agent of the present invention is by combination matching, can have good restraining effect to banana blight germ, remove wherein any one component, its inhibition all has obvious reduction.
Composite fungus agent of the present invention is to the prevention effect of potted plant Banana Seedlings
Getting the plant height of planting in black nutritive alms bowl is the banana seedlings of 25cm, is divided into 15 process at random.After FOC4 being inoculated in Banana Seedlings root 7d, the composite fungus agent of gained in difference application program 1 to 14, process is compared with clear water, often process a community (every community 5 basin, every basin 1 young plant), repeat for 3 times, each process is at 0d, 1d, 2d, 3d, 5d clip Banana Seedlings second leaf (from top to bottom) respectively, repeat second period after 5d, repeat 3 cycles altogether; In time there is withered or false stem cracking in clear water contrast any of several broadleaf plants seedling, in combination upper leaf wilting and dissect disease index and prevention effect that false cauline bundle browning level calculates each process.
Disease index grade scale is with reference to " planting disease organon " (third edition) of Fang Zhongda
1 grade: blade is asymptomatic, dissect bulb, false stem tissue white, have no browning, Gen Duobai, minority brown stain.
2 grades: the yellow patch of small area appears in peripheral bottom 1 one 2 blade, and account for less than 1/2 of leaf area, or organize browning area to account for less than 1/4 of bulb area, false stem tissue has no browning, the browning that root has.
3 grades: the yellow patch of big area appears in peripheral blade, accounts for more than 1/2 of leaf area, withering appears in blade or bulb organizes browning area to account for 1/4 ~ 1/2 of bulb area, and false stem tissue has no browning, the browning that root has.
4 grades: yellowing leaf is wilted, death, and bulb organizes browning area to account for about 1/2 of bulb area, and false stem organizes top to have no browning, and light brown mottled or brown linear pathology, root blackening brown appear in bottom, there is slight wilting in plant.
5 grades: plant occur wilt, withered, bulb organize more than l/2 to entire area browning or rot, there is the base portion frangibility atrophy of brown strip pathology or rotten root chocolate or occur rotten in false stem top and the bottom.
Table 2 different treatment method is to the prevention effect of potted plant banana seedlings
As can be seen from Table 2, the composite fungus agent of the present invention's 6 embodiment gained all has significant prevention effect to potted plant banana seedlings banana blight, and prevention effect can reach more than 94%.Remove wherein any one component in composite fungus agent, its prevention effect obviously declines, and all below 50%, all there is significant difference with 6 embodiments.Illustrate that composite fungus agent of the present invention is by combination matching, has significant preventive and therapeutic effect to the banana blight of potted plant Banana Seedlings, remove wherein any one component, its preventive and therapeutic effect all has obvious reduction.
Composite fungus agent of the present invention is on the impact of potted plant Banana Seedlings physical signs
Getting the plant height of planting in black nutritive alms bowl is the banana seedlings of 25cm, is divided into 15 process at random.After FOC4 being inoculated in Banana Seedlings root 7d, the composite fungus agent of gained in process 1-14 difference application program 1 to 14, process 15 does blank process with clear water, often process a community (every community 5 basin, every basin 1 young plant), repeat for 3 times, each process is at 0d, 1d, 2d, 3d, 5d clip Banana Seedlings second leaf (from top to bottom) respectively, repeat second period after 5d, repeat 3 cycles altogether; Arteries and veins and leaf margin part in removal, take 0.5g Leaf of banana, shred fast and be placed in mortar, add 8ml50mmolL-1pH7.8 phosphoric acid buffer (PBS, containing 1%PVPP, cross-linking polyethylene pyrrolidone), in the rear 4 DEG C of centrifugal 20min of 12000rpm of ice bath grinding, supernatant liquor is enzyme liquid, and 4 DEG C save backup.For measuring the activity of the content of mda (MDA) and proline(Pro) (Pro) and peroxidase POD, superoxide-dismutase SOD.
(1) mensuration of mda (MDA) content
By thiobarbituricacidα-(TBA) method.Get enzyme liquid 2ml (contrast is 2ml distilled water), add 2ml0.6%TBA (with 10% trichoroacetic acid(TCA) preparation) solution, cool rapidly after boiling water bath reacting 15min, centrifugal 10min under 9000rpm.Get supernatant liquor, measure 450,532, light absorption value under 600nm wavelength.Calculate MDA concentration according to the following formula, and then calculate the content in its unit fresh weight tissue.Its result is as shown in table 3 and Fig. 7.
MDA concentration (μm oL/L)=6.45 (OD
532-OD
600)-0.56OD
450
Table 3 different treatment is on the impact of MDA content
The trend first declining and rise afterwards is presented at the MDA content of process each process in 5 days.First day starts overall MDA value and starts to decline, at the 2nd day, the MDA content for the treatment of group 1-14 all reaches Schwellenwert, and the MDA value change of each process has higher consistence, and the MAD for the treatment of group 1-6 reduces degree is not only significantly higher than control group 15, and apparently higher than treatment group 7-14, reach conspicuous level.Illustrate that the composite fungus agent described in the present invention's 6 embodiments can reduce the degree of injury of cytolemma, its successful is better than other treatment group.
(2) peroxidase POD determination of activity
Employing guaiacol method measures.Methyl catechol solution: 0.05molL
-1the PBS100ml of PH7.8, adds methyl catechol 250 μ L, and heated and stirred, to dissolving, is stored in refrigerator stand-by.Every 3ml reaction system is containing methyl catechol solution 250 μ L, 30%H
2o
2after 5 μ L, enzyme liquid 50 μ L mixes.In wavelength 470nm place colorimetric, replace the zeroing of enzyme liquid with equivalent PBS, represent enzymic activity size with per minute absorbancy changing value, unit is △ OD
470mg
-1fwmin.Its result is as shown in table 4 and Fig. 8.
Table 4 different treatment is on the impact of Banana Seedlings blade POD
Banana Seedlings POD content after the process of different bacterium liquid is totally downtrending after first rising.Each process compares with control group, whole POD value all reaches maximum on the 2nd day in process, and compared with control group 15, the POD level for the treatment of group 1-14 all has rising in various degree, and the degree that treatment group 1-6 raises is apparently higher than treatment group 7-14, significant difference.Illustrate that the mixed bacteria liquid of the present invention's 6 embodiment gained can the histocyte of available protecting seedling, alleviate the injury of active oxygen to plant tissue, effect is better than other treatment group.
(3) mensuration of superoxide-dismutase SOD activity
Adopt chlorination nitrate blue tetrazolium (nitrobluetetrazolium, NBT) method.Every 3ml reaction system contains 0.05molL
-1pBS1.5ml, 130mmolL
-1methionine(Met) (Met) solution 0.3ml, 750 μm of olL
-1nBT solution 0.3ml, 100 μm of olL
-1eDTA-Na
2solution 0.3ml, 50 μ L enzyme liquid (2 control tube replace enzyme liquid with PBS) and 0.25ml distilled water, finally add 20 μm of olL
-1riboflavin solution 0.3ml, mixes immediately, and 1 control tube is put dark place, and other each pipe reacts 10min under 4000lx daylight, the light absorption value of assaying reaction liquid 560nm.1 unit of enzyme activity is defined as to suppress the enzyme amount needed for NBT photochemical reduction 50%.Its result is as shown in table 5 and Fig. 9.
Table 5 different treatment affects Banana Seedlings foliar SOD
From Fig. 3-4, after using different bacterium liquid, SOD content is in rear downtrending of first rising, from first day, treatment group 1-14 compares compared with control group 15, SOD content raise significantly improve, and process after 2 days SOD level reach peak value, the 5th day time, SOD value drops to Schwellenwert again.At the 2nd day, treatment group 1-6 rose significantly than the SOD content for the treatment of group 7-14, and difference reaches conspicuous level.Result shows, and treatment group 1-14 Banana Seedlings resistance of oxidation is improved than control group, and treatment group 1-6 Banana Seedlings resistance of oxidation is apparently higher than treatment group 7-14.Illustrate that the composite fungus agent of embodiment of the present invention 1-6 gained can significantly improve the resistance of oxidation of Banana Seedlings, reduce seedling damage.
(4) mensuration of proline(Pro) (Pro) content
The mensuration of Pro content adopts acid ninhydrine colorimetry.Take 0.5g blade, add 5ml3% sulfosalisylic acid solution, sealing, extract 10min in boiling water bath, (will often shake in leaching process), cooled and filtered is in clean test tube.Draw 2ml filtrate in another clean band glass plug test tube, add 2ml Glacial acetic acid and 2ml acid ninhydrine reagent, in boiling water bath, heat 30min.Add 4ml toluene after cooling, eddy mixer shakes 30s, standing a moment makes solution layering, gets upper strata red solution in cuvette, is blank with toluene, and on spectrophotometer, 520nm measures light absorption value, with standard P ro drawing standard curve.Its result is as shown in table 6 and Figure 10.
Table 6 different treatment affects Banana Seedlings blade Pro
Compared with blank group, dissociate Pro content is in rear downtrending of first rising.Process the 2nd day, between each process, dissociate Pro value was all significantly higher than contrast, and treatment group 1-6 dissociate Pro content elevated-levels is apparently higher than treatment group 7-14.Result shows: treatment group 1-14 shows satisfactory stability in the accumulation of promotion permeate substance, and the successful for the treatment of group 1-6 is better than treatment group 7-14.Illustrate that the composite fungus agent of the present invention's 6 embodiment gained can significantly improve dissociate Pro content, its action effect is not only better than blank group 15, and is significantly better than other treatment group.
Banana blight prevention and controls of the present invention is to the prevention effect of banana blight
Choose 15 banana growing areas that area is identical, working condition is close respectively, experimental group establishes 6 groups, carry out the control of banana blight respectively according to the method provided in the present invention's 6 embodiments in cultivating and growing process, control group establishes 10 groups, control group 1-8 composite fungus agent is prepared according to scheme 7-15, other prevention and controls are identical with embodiment 2, control group 9 groups only carries out transplanting pre-treatment, do not use composite fungus agent, control group 10 only uses composite fungus agent, do not carry out transplanting pre-treatment, using the implantation methods of routine as space management.Continuous plantation 5 years, add up each sickness rate for the treatment of group banana blight and the output of banana, its result is as shown in table 3.
The different prevention and controls of table 3 is on the impact of banana blight and output
As can be seen from Table 3, continuously plantation 5 years, the sickness rate of 6 experimental group banana blights all remains on lower level, and after each control group and space management group plant 5 years continuously, banana blight sickness rate obviously raises.The sickness rate of 6 experimental group banana blights is not only starkly lower than space management group, is also starkly lower than 10 control groups, illustrates that prevention and controls of the present invention effectively can reduce the sickness rate of banana blight.
Only adopt and transplant pre-treatment than the control group using separately composite fungus agent, its banana blight sickness rate obviously raises, illustrate that composite fungus agent of the present invention has certain prevention effect to banana blight, its preventive effect is better than only carrying out transplanting pre-treatment with sterilant, but still lower than the treatment process of 6 experimental group.The present invention is obviously better than being used alone wherein a kind of by transplanting the prevention and controls that pre-treatment and composite fungus agent combine, illustrate two kinds of methods with the use of, there is the effect of synergy, the prevention effect of banana blight significantly improved.
The prevention and controls provided in control group 1-8 is provided, although the sickness rate of banana blight comparatively space management group have obvious reduction, still far above 6 experimental group.Illustrate that the prevention effect of any one bacterial strain to banana blight removed in composite fungus agent also obviously reduces.Illustrate that each bacterial strain in composite fungus agent of the present invention influences each other, mutually promote, jointly play a role, lack wherein any one bacterial strain, its prevention effect reduces all greatly.
In addition, by the plantation of continuous 5 years, adopt prevention and controls of the present invention, banana annual production is all stabilized in higher level, and other treatment group all have reduction in various degree, and along with the increase of Planting Years, output reduces more and more obvious.Illustrate that prevention and controls of the present invention not only can not affect the normal growth of banana, and can maintain the stable and high yields of banana, action effect is obviously better than other treatment group, has economic benefit significantly.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (10)
1. a composite bacteria, it is characterized in that, according to cell count gauge, described composite bacteria comprises following bacterial strain: Paenibacillus polymyxa 1-10 part, subtilis 1-15 part, bacillus thuringiensis 1-15 part, Pseudomonas stutzeri 5-10 part, bacillus amyloliquefaciens 5-10 part, brevibacterium frigoritolerans 5-10 part, the mould 5-10 part of wood, green muscardine fungus 1-10 part.
2. composite bacteria as claimed in claim 1, it is characterized in that, according to cell count gauge, described composite bacteria comprises following bacterial strain: Paenibacillus polymyxa 3-7 part, subtilis 8-12 part, bacillus thuringiensis 8-12 part, Pseudomonas stutzeri 6-8 part, bacillus amyloliquefaciens 6-8 part, brevibacterium frigoritolerans 6-8 part, the mould 6-8 part of wood, green muscardine fungus 3-7 part.
3. the application of composite bacteria as claimed in claim 1 or 2 in control banana blight.
4. prevent and treat a composite fungus agent for banana blight, it is characterized in that, containing the composite bacteria in claim 1-2 described in any one.
5. the composite fungus agent of control banana blight as claimed in claim 4, it is characterized in that, in described composite fungus agent, total bacterial strain concentration is>=10
8cfu/mL.
6. a prevention and controls for banana blight, is characterized in that, comprises the following steps:
After seedling plantation, the composite fungus agent got in claim 4 or 5 described in any one is administered to Jiao Yuanzhong.
7. the prevention and controls of banana blight according to claim 6, is characterized in that, when described composite fungus agent uses, being diluted to total bacterial strain concentration is 10
4-10
6cfu/mL.
8. the prevention and controls of the banana blight according to claim 6 or 7, is characterized in that, the application process of described composite fungus agent is that filling root or ground, the end spray; Consumption is 1-2.5kg/ strain, and every 15-30 days uses once.
9. the prevention and controls of banana blight according to claim 6, is characterized in that, also comprises front step: before Banana Seedlings is transplanted to land for growing field crops, first sprays mycocide large Tanaka, shines, then Banana Seedlings is transplanted to large Tanaka through spacious.
10. the prevention and controls of banana blight according to claim 9, is characterized in that: described mycocide is dioxide peroxide, potassium permanganate or dazomet; The mass concentration of mycocide used is 50-500mg/L, and consumption is every square metre of 100-300mg.
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