CN117286046A - Pseudomonas BWT8-8 and application thereof in plant disease and pest control - Google Patents
Pseudomonas BWT8-8 and application thereof in plant disease and pest control Download PDFInfo
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- 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
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/27—Pseudomonas
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P1/00—Disinfectants; Antimicrobial compounds or mixtures thereof
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P21/00—Plant growth regulators
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P5/00—Nematocides
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/38—Pseudomonas
Abstract
The invention belongs to the technical field of biological control of plant diseases and insect pests, and particularly relates to a pseudomonas BWT8-8 defending strain and application thereof in plant disease and insect pest control. The defending pseudomonas BWT8-8 provided by the invention is separated from the root knot nematode body and is preserved in the Guangdong province microorganism strain collection center, and the preservation number is GDMCCNO.62299. The fermentation liquor for defending the pseudomonas has obvious lethal effect on plant root knot nematodes, is diluted by 10 times for use, and has the corrected mortality rate of the nematodes as high as 90.73%. In the greenhouse potting test, the relative control effect of the fermentation liquor can reach 38.27 percent. In addition, the strain BWT8-8 fermentation broth also has active substances for resisting plant pathogenic bacteria, and has remarkable inhibition effect on xanthomonas citri, acidovorax avenae subspecies watermelon and Lawsonia solanacearum. Therefore, the method can be used as a potential biocontrol bacterial resource to be applied to plant disease and pest control.
Description
Technical Field
The invention belongs to the technical field of biological control of plant diseases and insect pests, and particularly relates to a pseudomonas (Pseudomonas protegens) BWT8-8 defending strain and application thereof in plant disease and insect pest control.
Background
The lag of high-density single planting and water and fertilizer management technology causes the outbreak of root-knot nematode diseases and soil-borne diseases of crops to become serious. According to statistics, the economic loss caused by the root-knot nematodes is up to 12% each year, and the serious land parcels are up to 30% -40% and even more than 70%. At present, the prevention and treatment of plant root-knot nematodes still takes chemical nematicides or soil fumigants as main materials, but most of the plant root-knot nematodes have the problems of drug resistance and ecological environment pollution, and some highly toxic nematicides threaten the health of people and livestock. With the importance of people on food safety and the continuous improvement of green development concepts, chemical nematicides are continuously limited in use. Therefore, there is an urgent need to find new drug substitutes.
Compared with chemical nematicides, the biological nematicide technology utilizes the relationship of natural enemies in biospheres to prevent and treat plant root-knot nematodes by screening and modifying strains which have predatory, parasitic, nutritional and space occupation effects on the root-knot nematodes and can generate nematicidal active substances. Therefore, the method has the advantages of long duration of control effect, almost no great harm to agricultural ecology, and accordance with the novel concept of environmental protection, health and sustainable development of novel agriculture. However, the current biocontrol resources for preventing and controlling plant root-knot nematodes are very limited, and new biocontrol resources are to be further excavated.
Disclosure of Invention
In order to better find a new chemical nematicide substitute, the invention aims to provide a plant for defending pseudomonas (Pseudomonas protegens) BWT8-8, and the plant can be used in the control process of plant root-knot nematodes, and the growth and development capacity of the cabbages can be obviously improved.
In order to achieve the above purpose, the invention adopts the following technical scheme:
a strain of the Pseudomonas BWT8-8Pseudomonas protegens is deposited with the Guangdong province microorganism strain collection center with the deposit number of GDMCC No.62299 and the deposit date of 2022, 3 and 18 days.
The defenses against pseudomonas BWT8-8 are isolated from the root knot nematode body.
The invention also provides a biocontrol microbial inoculum which comprises one or more of a culture of the defensive pseudomonas BWT8-8, a bacterial strain metabolite thereof, a supernatant obtained by filtering and separating bacterial liquid thereof and a crude enzyme liquid extracted by culturing the same.
Preferably, the culture of the defenses pseudomonas BWT8-8 is obtained by activating the defenses pseudomonas BWT8-8 and then inoculating the defenses pseudomonas BWT8-8 into an NB culture medium for fermentation culture.
The invention also provides application of the biocontrol microbial inoculum in plant root-knot nematode control.
The invention also provides application of the biocontrol microbial inoculum in preventing and controlling plant pathogenic bacteria.
Preferably, the plant comprises citrus, cantaloupe, tomato.
Preferably, the pathogenic bacteria include xanthomonas citri Xanthomonas citri subsp.citri, xcc (citrus canker pathogenic bacteria), avians subsp. Acidovorax avenae subsp.citrulli (cantaloupe angular leaf spot pathogenic bacteria) and ralstonia solanacearum Ralstonia solanacearum (tomato bacterial wilt pathogenic bacteria).
The invention also provides application of the biocontrol microbial inoculum in promoting growth and development of cabbages.
Compared with the prior art, the invention has the following beneficial effects: according to the invention, the defensive pseudomonas is separated from the root-knot nematode for the first time, the fermentation culture can prevent and treat the root-knot nematode, inhibit the growth of multiple bacteria such as xanthomonas citri, acidovorax avenae subspecies citrulli, and ralstonia solanacearum, and can promote the growth and development process of the cabbages.
Drawings
FIG. 1 colony growth of dead root-knot nematodes on NA plates;
FIG. 2 colony morphology on NA plates against Pseudomonas BWT 8-8;
FIG. 3 effects of the fermentation broth of defenses against Pseudomonas BWT8-8 on the growth and development of tomato plants;
FIG. 4 is a graph showing the effect of the fermentation broth of Pseudomonas BWT8-8 in inhibiting xanthomonas citri (Xanthomonas citri subsp. Citri, xcc);
FIG. 5 is a graph showing the effect of the fermentation broth of Pseudomonas defenses BWT8-8 on inhibition of the sub-species Citrulli (Acidovorax avenae subsp. Citrulli) of Acidovorax Avicularis;
FIG. 6 is a graph showing the effect of the fermentation broth of Pseudomonas BWT8-8 on inhibiting L.solanacearum (Ralstonia solanacearum);
FIG. 7 shows the effect of protecting against the growth and development of P.bWT 8-8 broth on cabbage.
Detailed Description
The present invention will be further explained with reference to specific examples, but it should be noted that the following examples are only for explaining the present invention, and are not intended to limit the present invention, and all technical solutions identical or similar to the present invention are within the scope of the present invention. The specific techniques or conditions are not noted in this example and are practiced according to methods and apparatus conventional in the art; the reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
The invention relates to a culture medium and a preparation method thereof, wherein the culture medium comprises the following components:
NA culture medium and preparation method thereof: 10g/L peptone, 3g/L beef extract powder, 5g/L sodium chloride and 15g/L agar powder; the preparation method comprises the following steps: taking 18g of the culture medium, adding deionized water to 1000mL, stirring and heating to dissolve completely, adjusting the pH of the solution to 7.2, subpackaging in triangular flasks, sterilizing at 121 ℃ under high pressure for 15min, and pouring into a disposable culture dish for standby.
The NB medium is NA medium with agar powder removed.
Test example 1 isolation, identification and preservation of protection against Pseudomonas BWT8-8
Isolation of BWT8-8 Strain: the defensive pseudomonas BWT8-8 provided by the invention is separated from the root-knot nematode body. The specific separation steps are as follows: the root system of the tomato plant with serious disease is taken and washed clean by tap water, three-stage water is used for washing for 3 times, water is wiped off, the root system with root nodules is sheared into small sections with the length of about 1.5cm by scissors, the small sections are put into a triangular flask, 0.5% sodium hypochlorite solution which submerges the root nodules is added, the triangular flask is sealed by a preservative film, the triangular flask is shaken forcefully for 3min, after the completion, the suspension is poured into a cover screen (20 meshes, 60 meshes, 100 meshes, 300 meshes and 500 meshes in sequence from top to bottom), tap water is used for repeatedly washing, and finally, the insect eggs and the larvae on the 500 meshes screen are collected by sterile water.
Then 1mL of worm eggs are put into a culture dish (phi=9 cm), 13mL of sterile water is added, the culture dish is placed into a 27 ℃ incubator for dark culture, the survival condition of the root-knot nematodes in the culture dish is observed every 24 hours after 7 days, dead and stiff root-knot nematodes are put into 75% alcohol for disinfection by forceps, then the root-knot nematodes are put into sterile water for washing for 4 times, inoculated into NA culture medium and dark cultured for about 4 days at 28 ℃, and after colonies grow out (figure 1), colonies with different forms and colors are picked up for scribing (specific process is that the colonies with different forms and colors are picked up and scribed on an NA plate by bamboo sticks, single colony is repeatedly picked up for scribing until pure strains are obtained), and the pure strains are obtained for 4 times.
Identification of BWT8-8 Strain: the apparent identification of the strain is carried out according to colony morphology, thallus color, transparency, glossiness and the like. As shown in FIG. 2, the colony of the defensive pseudomonas BWT8-8 on the NA culture medium is light yellow, round, neat in edge and opaque, and the surface of the thallus is smooth, moist and glossy.
16S rDNA sequencing analysis: the DNA of the Pseudomonas BWT8-8 strain of the present invention was extracted according to the specification of the OMEGA kit, and the above DNA was used as a template,PCR amplification was performed using 16S rDNA universal primer 27F/1492R. The reaction system was 25. Mu.L, green Tap Mix 12.5. Mu.L, primers 27F/1492R each 0.5. Mu.L, template DNA 1. Mu. L, ddH 2 O10.5. Mu.L. Reaction conditions: pre-denaturation at 95℃for 3min; denaturation at 95℃for 30s; annealing at 55 ℃ for 30s; extending at 72 ℃ for 60s, and 30 cycles in total; extending at 72℃for 5min. The PCR products were sequenced. The sequence similarity of the defensive pseudomonas BWT8-8 and Pseudomonas protegens is 99.78% by 16S rDNA sequencing analysis (the result is shown as SEQ ID NO. 1). The strain BWT8-8 was identified as a defensive Pseudomonas strain in combination with the morphological characteristics of the strain (Pseudomonas protegens).
Wherein, the primer sequence of 27F is: 5'-AGAGTTTGATCCTGGCTCAG-3';
the primer sequence of 1492R is: 5'-TACGGCTACCTTGTTACGACTT-3';
CGAGCGGCAGCACGGGTACTTGTACCTGGTGGCGAGCGGCGGACGGGCGTGCGGCAGCACGGGTACTTGTACCTGGTGGCGAGCGGCGGACGGGTGAGTAATGCCTAGGAATCTGCCTAGTAGTGGGGGATAACGTCCGGAAACGGGCGCTAATACCGCATACGTCCTACGGGAGAAAGTGGGGGATCTTCGGACCTCACGCTATTAGATGAGCCTAGGTCGGATTAGCTAGTTGGTGAGGTAATGGCTCACCAAGGCGACGATCCGTAACTGGTCTGAGAGGATGATCAGTCACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGCGAAAGCCTGATCCAGCCATGCCGCGTGTGTGAAGAAGGTCTTCGGATTGTAAAGCACTTTAAGTTGGGAGGAAGGGCAGTTACCTAATACGTGATTGTTTTGACGTTACCGACAGAATAAGCACCGGCTAACTCTGTGCCAGCAGCCGCGGTAATACAGAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGCGCGTAGGTGGTTTGTTAAGTTGGATGTGAAAGCCCCGGGCTCAACCTGGGAACTGCATCCAAAACTGGCAAGCTAGAGTATGGTAGAGGGTGGTGGAATTTCCTGTGTAGCGGTGAAATGCGTAGATATAGGAAGGAACACCAGTGGCGAAGGCGACCACCTGGACTGATACTGACACTGAGGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGTCAACTAGCCGTTGGGAGCCTTGAGCTCTTAGTGGCGCAGCTAACGCATTAAGTTGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGCCTTGACATCCAATGAACTTTCTAGAGATAGATTGGTGCCTTCGGGAACATTGAGACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGTAACGAGCGCAACCCTTGTCCTTAGTTACCAGCACGTTATGGTGGGCACTCTAAGGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCATCATGGCCCTTACGGCCTGGGCTACACACGTGCTACAATGGTCGGTACAAAGGGTTGCCAAGCCGCGAGGTGGAGCTAATCCCATAAAACCGATCGTAGTCCGGATCGCAGTCTGCAACTCGACTGCGTGAAGTCGGAATCGCTAGTAATCGCGAATCAGAATGTCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTGCAGCAGACGTAGCTAG(SEQ ID NO.1)；
preservation of BWT8-8 Strain: the identified single colonies were made into bacterial suspensions with 50% glycerol in distilled water and stored at-80℃in ultra-low temperature.
Test example 2 lethal Effect of Pseudomonas BWT8-8 fermentation broth against Meloidogyne incognita
1. The test process comprises the following steps: and (3) picking a single colony of the pseudomonas BWT8-8, inoculating the single colony into an NB liquid culture medium, and culturing for 36h at 30 ℃ and 180 r/min. Centrifuging at 10000r/min for 10min after completion, collecting supernatant, and sterilizing with 0.22 μm microporous filter membrane for 3 times. In a sterile 24-well cell culture plate, 1mL of the filtered sterilized fermentation stock solution and 10-fold dilution of the fermentation solution are added to each well of an experimental group, an equal amount of the filtered sterilized NB liquid medium is added to a control group, 100 μl (about 50 nematodes) of the two-instar larva suspension is added to each well, the mixture is placed in a constant temperature incubator at 27 ℃ after sealing for 24 hours and 48 hours, the death condition of the nematodes is observed, and the corrected mortality rate of the nematodes is calculated. Each treatment was repeated 4 times and the results averaged.
Corrected mortality (%) = (control survival rate-treatment survival rate) ×100/control survival rate
2. Test results: the results are shown in Table 1.
TABLE 1 poisoning of the meloidogyne incognita by the Pseudomonas BWT8-8 fermentation broth
| Treatment of | Mortality (%) | Mortality (%) |
| Stock solution | 100 | 100 |
| 10-fold dilution | 88.25 | 90.73 |
As can be seen from Table 1, the defenses against Pseudomonas have a remarkable lethal effect on meloidogyne incognita. The corrected mortality of the stock solution reaches 100% after 24 hours, the corrected mortality of the 10-time diluent reaches 88.25%, and the corrected mortality of the 10-time diluent is improved to 90.73% after 48 hours.
Test example 3 greenhouse pot control of plant root knot nematode by protection against Pseudomonas BWT8-8 fermentation broth
1. The test process comprises the following steps: the three-leaf one-heart tomato seedlings are transplanted into a flowerpot containing 200 g/pot sterilized soil (soil: sand volume ratio=2:1), watered until the soil water content reaches about 80%, cultured for 3d, 5mL of a second-age larva suspension with the density of about 400 pieces/mL is evenly injected around the rhizosphere of the tomato seedlings by a pipette, about 2000 second-age larvas are inoculated into each pot, the culture is continued for 3d, 20 mL/plant of a pseudomonas defensis BWT8-8 fermentation liquor is taken by a syringe (the fermentation liquor preparation method is the same as that of test example 2) around the root system of the tomato seedlings to be used as an experimental group, and the control group is connected with an equal amount of NB sterilized culture medium or an equal amount of sterile water. Each group was tested 3 replicates of 10 tomato seedlings each time. After 15d of cultivation, the cells were inoculated again by the same inoculation method and the same inoculation amount. The whole test period is inoculated with 2 times of fermentation liquid of the pseudomonas BWT8-8, and other management is carried out according to the high-quality cultivation management measures of tomatoes. FIG. 3 shows the growth of tomato plants after inoculation with the first P.defenses BWT8-8 broth.
Root knot index and relative control efficiency statistics: after 35d inoculation of nematodes, statistical analysis of tomato root knot number, root knot index and relative control effect was performed using Liu Zhongliang agricultural grading standard (Liu Zhongliang, high jun Jie, etc. bacterial dreg composting treatment and control effect on tomato root knot nematode disease [ J ]. North gardening, 2020 (06): 49-54.). Here, root knot index=Σ (number of individual stage disease plants×grade)/total number of investigation×100, relative control effect (%) = (control root knot index-treated root knot index)/control root knot index×100%.
2. Test results: the specific test results are shown in Table 2 below.
TABLE 2 disease conditions of tomato plants after 35d inoculation of the protective Pseudomonas BWT8-8 fermentation broth
| Treatment of | Root knot count (number) of each plant | Root knot index | Relative control effect (%) |
| NB sterilization medium | 46.00±1.00 | 81.00±1.00 | — |
| BWT8-8 fermentation broth | 33.00±3.00* | 50.00±1.00 | 38.27 |
Note that: table x represents: * P is more than 0.01 and less than or equal to 0.05.
As can be seen from Table 2, the treatment of the defensive Pseudomonas BWT8-8 fermentation broth significantly reduced the root knot number of tomato plants compared with the NB sterilization medium control, and the tomato root knot index was reduced from 81.00% to 50.00% 35d after tomato transplantation, with a relative control effect of 38.27%.
Test example 4 antagonism effect of the fermentation broth against pseudomonas BWT8-8 on phytopathogenic bacteria 1. Test procedure: activating and fermenting and culturing the strain: the A.citri, A.oat acidophilus subspecies watermelon species, B.solani and P.defenses BWT8-8 were activated with NA plates and then cultivated with NB fermentation at 30℃for 36h.
Preparing bacterial suspension and fermentation liquor: diluting pathogenic bacteria liquid (i.e. bacteria liquid before sterilizing microporous filter membrane) to mother liquid with OD=1.0, and diluting to 10 -3 Standby; the preparation method of the fermentation broth for defending against Pseudomonas BWT8-8 is the same as that of test example 2.
Antagonism experiment: 100 mu L of pathogenic bacteria diluted bacterial liquid is uniformly coated on an NA solid culture medium, a blue gun head is used for punching on a flat plate, then 50 mu L of fermentation liquid for defending pseudomonas BWT8-8 is taken and cultured in the holes for 2d at 30 ℃, the size of a bacteriostasis ring is observed, and a photographing record result is carried out.
Test results: the bacteriostatic effect of the defenses against pseudomonas BWT8-8 against phytopathogenic bacteria is shown in figures 4, 5 and 6. As can be seen from fig. 4, 5 and 6, the defensive pseudomonas BWT8-8 shows good inhibition effect on xanthomonas citri (pathogenic bacteria of citrus canker), acidovorax avenae subspecies (pathogenic bacteria of cantaloupe angular leaf spot) and ralstonia solanacearum (pathogenic bacteria of tomato bacterial wilt), and forms obvious inhibition zone on each inhibition flat plate. The method shows that the BWT8-8 and the fermentation liquor thereof can be used for preventing and controlling plant bacterial diseases such as citrus canker, cantaloupe angular leaf spot, tomato bacterial wilt and the like.
Test example 5 growth promoting effect of protective pseudomonas BWT8-8 inoculant on chinese cabbage greenhouse potting test 1. Preparation of inoculant: activating the defensive pseudomonas BWT8-8 by an NA flat plate, inoculating the defensive pseudomonas BWT8-8 into an NB liquid culture medium, placing the NB liquid culture medium into a shaking table at 30 ℃ and 200r/min for fermentation culture for 48 hours, and obtaining bacterial liquid of the defensive pseudomonas BWT8-8 after the completion.
2. The test process comprises the following steps: washing root system matrix of three-leaf one-core young Chinese cabbage, transplanting into a flowerpot containing 200 g/pot sterilized soil (soil: sand volume ratio=2:1), watering until the soil water content reaches about 80%, placing in a greenhouse for culturing for 3d, taking 30 mL/plant of protective pseudomonas BWT8-8 bacterial liquid by a syringe, watering around the root system of the young Chinese cabbage, continuously placing in the greenhouse for culturing, treating the control group by using sterile water, namely, after culturing for 3d, taking 30 mL/plant of sterile water by the syringe, watering around the root system of the young Chinese cabbage, and placing in the greenhouse for culturing. In the experimental group and the control group, 1 plant is transplanted in each pot, each treatment is repeated for 4 times, after 7 days, the growth condition of the cabbages is recorded by photographing, the agronomic character indexes of the cabbages are measured, and the growth and development pictures of the cabbages are shown in figure 7.
3 test results: the specific test results are shown in Table 3.
TABLE 3 protection against Pseudomonas BWT8-8 microbial agent growth promoting conditions of Brassica Capitata
Note that: table x represents: * P 0.01< 0.05, p 0.001< 0.01.
As shown in Table 3, compared with the sterile water control, the defensive pseudomonas BWT8-8 can obviously promote the growth and development of the cabbages, and after the microbial inoculum is applied for 7 days, the leaf length, the leaf width, the leaf area and the individual plant weight of the cabbages are obviously higher than those of the sterile water control, and particularly, the leaf width, the leaf area and the individual plant weight are most obvious, and the difference reaches extremely obvious level.
Finally, it should be noted that the above-mentioned embodiments are merely illustrative of the principles, performances and effects of the present invention, and are not meant to limit the invention. Modifications and variations may be made to the above-described embodiments by those skilled in the art without departing from the spirit and scope of the invention. Accordingly, it is intended that all equivalent modifications and variations of the invention be covered by the claims, which are within the ordinary skill of the art, be within the spirit and scope of the present disclosure.
Claims (9)
1. A strain of defensive pseudomonas BWT8-8, deposited with the cantonese province microorganism strain collection under accession number GDMCC No.62299.
2. The pseudomonas BWT8-8 defenses of claim 1 isolated from a root knot nematode.
3. A biocontrol microbial agent comprising one or more of the culture of the defensive pseudomonas BWT8-8, a strain metabolite thereof, a supernatant of a bacterial liquid filtration separation thereof, and a crude enzyme liquid extracted from the culture thereof according to claim 1.
4. The biocontrol agent according to claim 3, wherein said culture of pseudomonas defenses BWT8-8 is obtained by activating said pseudomonas defenses BWT8-8 and then inoculating it to NB medium for fermentation culture.
5. Use of the biocontrol microbial agents as claimed in claim 3 for controlling plant root knot nematodes.
6. Use of a biocontrol agent as claimed in claim 3 for controlling phytopathogenic bacteria.
7. The use according to claim 6, wherein the plant comprises citrus, cantaloupe, tomato.
8. The use according to claim 6, wherein the pathogenic bacteria comprise xanthomonas citri, acidovorax avenae subspecies citrulli, lausum solanacearum.
9. Use of the biocontrol microbial agent of claim 3 for promoting growth and development of pakchoi.
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