CN114891679B - Pseudomonas fluorescens and application thereof in preventing and controlling cherry branch diseases - Google Patents
Pseudomonas fluorescens and application thereof in preventing and controlling cherry branch diseases Download PDFInfo
- Publication number
- CN114891679B CN114891679B CN202210545263.5A CN202210545263A CN114891679B CN 114891679 B CN114891679 B CN 114891679B CN 202210545263 A CN202210545263 A CN 202210545263A CN 114891679 B CN114891679 B CN 114891679B
- Authority
- CN
- China
- Prior art keywords
- cherry
- pseudomonas fluorescens
- cherry branch
- diseases
- strain
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/27—Pseudomonas
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Virology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Agronomy & Crop Science (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Environmental Sciences (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses pseudomonas fluorescens and application thereof in preventing and controlling cherry branch diseases. The invention screens fluorescent pseudomonas (Pseudomonas fluorescens) GH2-1 with stronger inhibiting effect on cherry branch diseases from healthy plant rhizosphere soil in a cherry producing area, and the strain is preserved in China general microbiological culture collection center (CGMCC) with a preservation number of 14743 in the year 09/26 of 2017. The strain and the fermentation supernatant thereof have antagonistic effect on cherry branch disease bacteria, and the bacteriostasis rates are 90.98 percent and 87.45 percent respectively; meanwhile, the control effect of the 300-time diluent of the strain liquid preparation on the cherry branch diseases is 64.3%, the occurrence of the cherry branch diseases can be effectively controlled, microbial pesticides aiming at the cherry branch diseases can be developed, and the green and efficient control of the cherry branch diseases is realized.
Description
Technical Field
The invention relates to pseudomonas fluorescens and application thereof in preventing and controlling cherry branch diseases, belonging to the technical field of microorganisms.
Background
Cherry branch disease is a common disease in cherry growth period, and especially serious disease occurs after seedling transplanting and in the first three years of the growth period. The disease is caused by infection of a cranberry base shell (Diaporthe vaccinii), and once the cranberry is infected by the disease, the yield of the cherry is often reduced, and the planting benefit is affected; if the disease is not found timely, the cherry trees almost all die with the continuous accumulation of pathogenic bacteria, and the harvest is disabled.
Aiming at the disease, physical measures such as pruning and garden cleaning are mainly adopted at present, but the method can not effectively remove pathogenic matters, especially the pathogenic matters are often infected in healthy plants near the pruning, and the pathogenic matters can be continuously infected although the pruning is pruned, so that cherry fruiting in the coming year is seriously influenced. Although the chemical pesticide sprayed can relieve the disease condition of the dry branch disease to a certain extent, the index of the pesticide with poor permeability is often not permanent. The use of chemical pesticides in large quantities also presents a series of problems, such as: causes drug resistance of the cranberry base shells, increases the control difficulty, causes environmental pollution, and seriously threatens human health due to cherry pesticide residues. Biological control is increasingly favored because of the advantages of high control effect, strong specificity, good drug effect durability, environmental friendliness, difficult generation of drug resistance and the like. However, few reports on biological control of cherry branch diseases are reported at present.
Pseudomonas fluorescens (Pseudomonas fluorescens) is a bacterium belonging to the genus Pseudomonas of the order Pseudomonas, the family Pseudomonas, belonging to the genus Proteus, and is widely used in biological control of plant diseases. At present, the method is reported to be used for preventing and treating coptis southern blight (CN 202110929652.3), tobacco angular leaf spot (CN 200910088952.2), tobacco wildfire (CN 200810224386.9), various bacterial soft rot (CN 201810449282.1) and the like, but no report of preventing and treating cherry branch diseases by using pseudomonas fluorescens is found.
Disclosure of Invention
Aiming at the problems, the invention provides pseudomonas fluorescens (Pseudomonas fluorescens) GH2-1, which has antagonistic effect on cherry branch disease bacteria, can effectively control the occurrence of cherry branch diseases, can develop microbial pesticides aiming at the cherry branch diseases, and realizes green and efficient control of the cherry branch diseases.
The invention firstly separates from healthy plant rhizosphere soil in a disease area of a cherry producing area, and through a flat plate counter culture method experiment, screens out pseudomonas fluorescens (Pseudomonas fluorescens) GH2-1 with stronger antagonism effect on pathogenic bacteria of cherry branches, namely cowberry fruit, from a base shell (Diaporthe vaccinii), and then the strain is preserved in China general microbiological culture collection center (CGMCC No. 14743) at the 09 th month 26 of 2017. Experiments show that: the bacterial strain GH2-1 and the metabolite thereof can effectively inhibit the growth of cherry branch disease bacteria, and have wide application prospects in the prevention and treatment of cherry branch diseases.
The invention also provides a biological agent of the pseudomonas fluorescens (Pseudomonas fluorescens) GH 2-1.
The preparation method of the biological agent comprises the following steps: after Pseudomonas fluorescens GH2-1 is used for preparing seed liquid, the seed liquid is inoculated into a fermentation culture medium for fermentation culture for 30-36 hours at the temperature of 25-30 ℃ to obtain fermentation liquor, and the fermentation liquor can be directly used (liquid preparation) or added with other auxiliary agents, and is prepared into solid preparations such as wettable powder and the like by spray drying.
Seed liquid culture medium formula (mass ratio): peptone 0.8-1.2%, beef powder 0.2-0.4%, sodium chloride 0.4-0.6%, and water for the rest, and adjusting pH 7.3+ -0.1. Preferred seed liquid culture medium formula (mass ratio): 1% of peptone, 0.3% of beef powder, 0.5% of sodium chloride and the balance of water, and regulating the pH value to be 7.3+/-0.1.
Fermentation medium formula (mass ratio): 1.5 to 2.5 percent of sucrose, 1.5 to 2.5 percent of soluble starch, 1.0 to 2.0 percent of yeast extract powder, 0.3 to 0.5 percent of fish peptone, 0.03 to 0.08 percent of magnesium sulfate, 0.03 to 0.08 percent of potassium dihydrogen phosphate, 0.5 to 0.8 percent of calcium carbonate and the balance of bean product wastewater, and the pH value is adjusted to 7.0 to 7.2. Preferably, the fermentation medium formula (mass ratio): 2% of sucrose, 2% of soluble starch, 1.5% of yeast extract powder, 0.4% of fish peptone, 0.05% of magnesium sulfate, 0.05% of monopotassium phosphate, 0.6% of calcium carbonate and the balance of bean product wastewater, and regulating the pH to 7.0-7.2.
The invention adopts the bean product waste water as the fermentation medium instead of water, and the bean product waste water contains abundant proteins, saccharides, minerals, vitamins and the like, so that nutrient substances can be provided, and the protein content in the bean product waste water is controlled to be 16-18%.
The invention also provides application of the pseudomonas fluorescens GH2-1 strain, the metabolite and the biological agent in antagonizing the cranberry interstice crust (Diaporthe vaccinii) of pathogenic bacteria of cherry branch diseases and preventing and controlling the cherry branch diseases.
The invention has the technical effects that:
1. good effect of preventing and controlling cherry branch diseases
The plate counter experiment shows that: the pseudomonas fluorescens GH2-1 strain and the fermentation supernatant thereof have strong antagonism effect on cherry branch pathogenic bacteria cowberry fruit inter-seat shells (Diaporthe vaccinii), and the bacteriostasis rates are 90.98% and 87.45% respectively; meanwhile, the control effect of the 300-time diluent field of the strain liquid preparation on cherry branch diseases is 64.3%, and the occurrence of cherry branch diseases can be effectively controlled. Has wide application prospect in preventing and controlling cherry branch diseases.
2. The strain is easy to ferment and produce, has a simple use mode, and is suitable for large-area popularization and use.
Drawings
FIG. 1 shows the plate inhibition effect of different strains on cherry branch pathogenic bacteria cranberry mesothelium (Diaporthe vaccinii);
FIG. 2 is a comparison of the bacteriostasis rates of different strains on the cherry branch pathogenic bacteria cranberry interstice housing (Diaporthe vaccinii);
FIG. 3 shows the plate inhibition effect of fermentation supernatants of different strains on cherry branch pathogenic bacteria cranberry interstice shells (Diaporthe vaccinii);
FIG. 4 is a comparison of the bacteriostasis rate of fermentation supernatants of different strains against cherry branch pathogenic bacteria cranberry interstice shells (Diaporthe vaccinii);
FIG. 5 shows the morphology of Pseudomonas fluorescens GH2-1 single colony.
Detailed Description
The effects thereof are described below with reference to examples.
Example 1: isolation and purification of Pseudomonas fluorescens (Pseudomonas fluorescens) Strain GH2-1
The Pseudomonas fluorescens (Pseudomonas fluorescens) strain GH2-1 is obtained by separating healthy plant rhizosphere soil of a disease area of a cherry producing area of a county smelting origin from a Weifang of Shandong province Lin/264 by adopting a dilution coating method, and the specific separation method comprises the following steps of:
(1) Soil sample collection
Collecting healthy cherry root systems in cherry branch disease attack areas by adopting a five-point sampling method, and placing the healthy cherry root systems in a freshness protection package to be brought back to a laboratory; and lightly brushing the root system of the cherry tree by using a hairbrush, and collecting soil, namely rhizosphere soil.
(2) Preparation of soil suspension
Accurately weighing 1g of rhizosphere soil in a triangular flask containing 99mL of sterile water, placing the triangular flask containing glass beads in a shaking table at 28 ℃ and uniformly shaking at 120-140 rpm for 20-30 min, and then placing the triangular flask in a water bath at 55-60 ℃ for incubation for 25min to obtain soil suspension;
(3) Dilution coating
Sequentially diluting the bacterial sample into 10 by 10-fold dilution method -1 ～10 -6 A diluted bacterial sample; respectively from 10 -4 、10 -5 And 10 -6 100 μl of the diluted bacterial sample was pipetted onto a plate in NA medium, three replicates per gradient;
(4) Culture purification
Culturing at 28 ℃ for 24-30 hours, then picking out microbial colonies with different forms on the NA culture medium, marking on a NA culture medium flat plate, and observing the growth condition of the bacterial colonies at regular time; and then adopting a plate streaking method to select single bacteria for purification, and respectively numbering and storing.
By the method, 14 strains are initially separated according to different colony morphologies, and the numbers of the 14 strains are GH2-1 to GH2-14 respectively.
Example 2: screening of cherry branch disease high-efficiency antagonistic strain
(1) Primary screen
Screening antagonistic strains for efficiently antagonizing pathogenic cowberry fruit space shells (Diaporthe vaccinii) of cherry branch diseases by using a plate counter culture method.
Culturing 14 strains to be detected, which are separated and purified in the embodiment 1, on NA culture medium in a streaking way, and culturing at the constant temperature of 28 ℃ for 24 hours; and (3) picking a loop of fungus moss by using an inoculating loop, inoculating the loop of fungus moss into a 50mL triangular flask containing 10mL of NB culture medium, and uniformly shaking for 36h at 120rpm/min in a shaking table at 28 ℃ to obtain fungus liquid of the strain to be selected.
Selecting a cherry branch disease fungus strain, performing activation culture on a PDA culture medium, and performing constant-temperature culture at 28 ℃ for 60 hours; a puncher is used for punching a bacterial cake with the diameter of 5mm at the edge of the activated cherry branch disease bacteria, and the bacterial cake is inoculated in the center of a PDA flat plate; inoculating 10 mu L of strain liquid of the selected strain on two sides 2.0cm away from the center by using inoculating loops; the control group is inoculated with 10 mu L of NB culture medium at the same distance on the other two sides of the cherry branch disease fungus cake inoculated with 5 mm; each treatment was repeated 3 times; after the treatment is finished, culturing at the constant temperature of 28 ℃, observing the inhibition effect of the selected strain on pathogenic bacteria day by day, measuring the size of the disease spots of the experimental group after the pathogenic bacteria of the control group grow on the flat plate, and calculating the bacteriostasis rate.
Antibacterial ratio = [ (control colony diameter-cake diameter) - (treated colony diameter-cake diameter) ]/(control colony diameter-cake diameter) ×100%.
The results showed that 6 strains among the 14 strains isolated in example 1 had inhibitory effects on the growth of cherry branch disease bacteria, GH2-1, GH2-2, GH2-4, GH2-5, GH2-6 and GH2-9 (FIG. 1, table 1), respectively; the calculation and statistics of the bacteriostasis rate of different strains show that the inhibition effect of the strain GH2-1 on cherry branch disease bacteria is as high as 90.98 percent and is obviously higher than that of other strains (table 1 and figure 2).
TABLE 1 statistics of the antibacterial Rate of different strains
(2) Double screen
The inhibition effect of the fermentation supernatants of GH2-1, GH2-2, GH2-4, GH2-5, GH2-6 and GH2-9 strains on the cranberry interstice shell (Diaporthe vaccinii) of cherry branch pathogenic bacteria is compared by using a toxic medium method. The specific method comprises the following steps:
carrying out streak culture on the 6 strains to be tested screened by the primary screening on an NA culture medium, and carrying out constant-temperature culture at 28 ℃ for 24 hours; transferring a loop by using a fungus inoculating loop, inoculating the loop into a triangular flask (50 mL) filled with 10mL of NB culture medium, and carrying out constant-temperature shaking culture at 28 ℃ and 120rpm/min for 24 hours to obtain seed liquid; inoculating the seed solution into a triangular flask (250 mL) filled with 100mL NA liquid culture medium according to 1% inoculum size, and performing constant-temperature shaking culture at 28 ℃ and 120r/min for 36h to obtain fermentation liquor; the fermentation broth was centrifuged and filtered through a 0.22 μm bacterial filter to give a fermentation supernatant.
Uniformly mixing fermentation supernatant and PDA culture medium at 45-50 ℃ according to the volume ratio of 1:19, pouring into a culture dish, cooling, and placing pathogenic bacteria cakes with the diameter of 5mm in the center of a flat plate; sterile water is added into the control group according to the same proportion to replace fermentation supernatant; each treatment was repeated 3 times; after the treatment is finished, culturing at the constant temperature of 28 ℃, observing the inhibition effect of 6 strain fermentation supernatants on pathogenic bacteria day by day, measuring the size of the disease spots of the experimental group after the pathogenic bacteria of the control group grow on a flat plate, and calculating the bacteriostasis rate.
The results show that the fermentation supernatants of GH2-1, GH2-2, GH2-4, GH2-5, GH2-6 and GH2-9 have inhibition effect on the growth of cherry branch disease bacteria; wherein, the inhibition rates of the GH2-1 and GH2-5 fermentation supernatants on cherry branch disease bacteria are 87.45% and 83.92%, respectively, which are significantly higher than the antibacterial activity of other strain fermentation broths, but have no significant difference (see Table 2 and FIG. 3-4). Combining the inhibiting result of the strain GH2-1 on cherry branch disease bacteria, we select GH2-1 as our functional strain for subsequent research and development.
TABLE 2 statistics of bacteriostasis rates of fermentation supernatants of different strains
The results show that the bacterial strain GH2-1 and the metabolic products thereof can effectively inhibit the growth of the cherry branch pathogenic cranberry interstice crust (Diaporthe vaccinii), and have wide application prospects in prevention and treatment of cherry branch diseases.
Example 3: identification of Strain GH2-1
(1) Microbiological characteristics: after the strain grows on NA culture medium, the colony is irregular, the surface is moist, smooth, sticky, semitransparent and has yellow-green pigment (figure 5).
(2) Molecular biological Properties
The 16S rDNA gene sequence of this strain was determined as follows (SEQ-1): TAAGATCGCTGGGGGCAGGCCTAACACATGCAAGTCGAGCGGTAGAGAGAAGCTTGCTTCTCTTGAGAGCGGCGGACGGGTGAGTAAAGCCTAGGAATCTGCCTGGTAGTGGGGGATAACGTTCGGAAACGGACGCTAATACCGCATACGTCCTACGGGAGAAAGCAGGGGACCTTCGGGCCTTGCGCTCCCAGATGAGCCTAGGTCGGATTAGCTAGTTGGTGAGGTAATGGCTCACCAAGGCGACGATCCGTAACTGGTCTGAGAGGATGATCAGTCACACTGGAAGTGAGACTCGGTCCAACCCTACGGGGCGCAGTGGGGAATATTGGACAATGGGCGAAAGCCTGATCCAGCCATGCCGCGTGTGTGAAGAAGGTCTTCGGATTGTAAAGCACTTTAMGTTGGGAGGAAGGGCATTAACCTAATACGTTAGTGTTTCGACGTTACCGACAGAATAAGCACCGGCTAACTCTGTGCCAGCAGCCGCGGTAATACAGAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGCGCGTAGGTGGTTTGTTAAGTTGGATGTGAAGGCCCCGGGCTCAACCTGGGAACTGCATCCAAAACTGACTGACTAGAGTATGGTAGAGGGTGGTGGAATTTCCTGTGTAGYGGTGAAATGCGTTGATATAGGAAGGAACACCAGTGGTGAAGGCGACCACCTGGACTAATACTGACACTGAGGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGTCAACTAGCCGTTGGGTGCCTTGAGCTCTTAGTGGCGCAGCTAACGCATTAAGTTGACCGCCTGGAGAGTACGGTCGCAAGGTTAAAACTCAAATGAATTGACGGGGGCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGCCTTGACATCCAATGAACTTTCTAGAGATAGATTGGTGCCTTCGGGAACATTGAGACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGTAACGAGCGCAACCCTTGTCGATAGTTACCAGCACGTAATGGTGGGCACTCTAAGGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCATCATGGCCCTTACGGCCTGGCCTACACACGTGCTACAATGGTCGGTACAGAGGGTTGCCAAGCCGCGAGGTGGAGCTAATCCCACAAAACCGATCGTAGTCCGGATCGCAGTCTGCAACTCGACTGCGTGAAGTCGGAATCGCTAGTAATCGCGAATCAGAATGTCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTGCACCAGAAGTAGCTAGTCTAACCTTCGGGAGGACGGTTACCACGGTGTGATTCATGACTGGAGTGAAGTCGTAACAAGGTAGCCGTAGGGGAACCTGCGGCTGGAACAGGTCCGG
The bacterial strain GH2-1 is identified as pseudomonas fluorescens (Pseudomonas fluorescens) by microbiological characteristics and molecular biological characteristics, and the bacterial strain is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of 14743 in the year 09 in 2017 and 26.
Example 4: preparation process of pseudomonas fluorescens GH2-1 liquid preparation
Seed liquid culture medium formula (mass ratio): 1% of peptone, 0.3% of beef powder, 0.5% of sodium chloride and the balance of water, and regulating the pH value to be 7.3+/-0.1.
Fermentation medium formula (mass ratio): 2% of sucrose, 2% of soluble starch, 1.5% of yeast extract powder, 0.4% of fish peptone, 0.05% of magnesium sulfate, 0.05% of monopotassium phosphate, 0.6% of calcium carbonate and the balance of bean product wastewater (protein content 17%), and regulating the pH to 7.0-7.2.
The preparation process of the pseudomonas fluorescens GH2-1 liquid preparation comprises the following steps:
(1) Seed liquid obtaining: the Pseudomonas fluorescens GH2-1 is selected from the inclined surface of the test tube, and a small amount of bacteria are moved into a seed liquid culture medium for constant-temperature shaking culture at 28 ℃ and 120r/min for 24 hours, and the seed liquid is the seed liquid;
(2) And (3) carrying out mass fermentation culture: sterilizing the fermentation medium material in a fermentation tank (the temperature is 121-124 ℃ for 30 min); under the aseptic condition, the cultured seed liquid (5 percent according to the inoculation volume ratio) is poured into a fermentation tank, and is fermented and cultured for 30-36 hours at the temperature of 28 ℃, the bacterial count can reach about 650 hundred million/ml, the transformation rate can reach about 80 percent, and the fermentation liquid is GH2-1 liquid preparation.
Example 5: pseudomonas fluorescens GH2-1 liquid preparation for preventing and controlling cherry branch diseases
The embodiment provides a related experiment of pseudomonas fluorescens GH2-1 liquid preparation aiming at cherry branch diseases.
(1) Test agent
GH2-1 liquid formulation (prepared in example 4).
(2) Test material and control object
The test material is cherry seedling; the control object is cherry branch disease and pathogenic cranberry base shells (Diaporthe vaccinii).
(3) Test ground conditions
The experimental area is set in Weifang city Lin/264,264 and county cherry producing area. The diseases of cherry branches and trunks are serious in the place in the past year, and particularly the survival rate of the cherry branches and trunks infected after the cherry seedlings are transplanted is low; the test for preventing and controlling cherry branch diseases in forests is carried out in 3-4 months in 2020. All the test cells have consistent cultivation conditions and management measures, and cherry branch diseases are in the initial stage of disease occurrence during application.
(4) Test design and arrangement
The test designs that the Pseudomonas fluorescens GH2-1 liquid preparation is 300 times diluted and does not apply the medicine to carry out the contrast to carry out 2 treatments, and each cell is 200m 2 Repeating for 3 times, 6 cells, each randomly arranged. The root soaking treatment is carried out for the first time in 3 months and 15 days in 2020 after the drug is applied twice; and after 15d of transplanting and seedling-recovering, the second application is carried out at the end of 3 months in 2020, and the current mode is root irrigation, and the application amount of each cell is 100 kg. And (5) investigation of the disease condition is carried out at the end of 4 months.
(5) Test investigation and calculation method
And counting the disease state after the last medicine application interval of 30d, and calculating the prevention and treatment effect.
The disease incidence stage number of cherry branch is shown in Table 3, and is expressed in terms of single plant.
TABLE 3 disease incidence grading Standard for cherry branches
The efficacy is calculated according to the following disease index and prevention and treatment effect:
disease index DI (%) = [ Σ (disease grade) x number of plants at the grade) ]/[ (highest value of disease grade x total number of investigation) ]x100%;
control effect (%) = (control disease index-treatment disease index)/control disease index x 100%.
(6) Results
The disease index and the control effect of each treated cherry branch disease are shown in table 4.
TABLE 4 disease index and control Effect of cherry branch diseases after treatment
The results in Table 4 show that the liquid preparation of the Pseudomonas fluorescens (P.fluoroscens) strain GH2-1 has better control effect on cherry branch diseases, and the control effect is 64.3 percent after 30d of the last application. The further shows that the strain GH2-1 has wide application prospect in cherry branch disease control.
SEQUENCE LISTING
<110> Shandong province forestry science institute
<120> Pseudomonas fluorescens and application thereof in preventing and controlling cherry branch diseases
<130> 0
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1503
<212> DNA
<213> 16S rDNA Gene sequencing of Pseudomonas fluorescens (Pseudomonas fluorescens) GH2-1
<400> 1
taagatcgct gggggcaggc ctaacacatg caagtcgagc ggtagagaga agcttgcttc 60
tcttgagagc ggcggacggg tgagtaaagc ctaggaatct gcctggtagt gggggataac 120
gttcggaaac ggacgctaat accgcatacg tcctacggga gaaagcaggg gaccttcggg 180
ccttgcgctc ccagatgagc ctaggtcgga ttagctagtt ggtgaggtaa tggctcacca 240
aggcgacgat ccgtaactgg tctgagagga tgatcagtca cactggaagt gagactcggt 300
ccaaccctac ggggcgcagt ggggaatatt ggacaatggg cgaaagcctg atccagccat 360
gccgcgtgtg tgaagaaggt cttcggattg taaagcactt tamgttggga ggaagggcat 420
taacctaata cgttagtgtt tcgacgttac cgacagaata agcaccggct aactctgtgc 480
cagcagccgc ggtaatacag agggtgcaag cgttaatcgg aattactggg cgtaaagcgc 540
gcgtaggtgg tttgttaagt tggatgtgaa ggccccgggc tcaacctggg aactgcatcc 600
aaaactgact gactagagta tggtagaggg tggtggaatt tcctgtgtag yggtgaaatg 660
cgttgatata ggaaggaaca ccagtggtga aggcgaccac ctggactaat actgacactg 720
aggtgcgaaa gcgtggggag caaacaggat tagataccct ggtagtccac gccgtaaacg 780
atgtcaacta gccgttgggt gccttgagct cttagtggcg cagctaacgc attaagttga 840
ccgcctggag agtacggtcg caaggttaaa actcaaatga attgacgggg gccgcacaag 900
cggtggagca tgtggtttaa ttcgaagcaa cgcgaagaac cttaccaggc cttgacatcc 960
aatgaacttt ctagagatag attggtgcct tcgggaacat tgagacaggt gctgcatggc 1020
tgtcgtcagc tcgtgtcgtg agatgttggg ttaagtcccg taacgagcgc aacccttgtc 1080
gatagttacc agcacgtaat ggtgggcact ctaaggagac tgccggtgac aaaccggagg 1140
aaggtgggga tgacgtcaag tcatcatggc ccttacggcc tggcctacac acgtgctaca 1200
atggtcggta cagagggttg ccaagccgcg aggtggagct aatcccacaa aaccgatcgt 1260
agtccggatc gcagtctgca actcgactgc gtgaagtcgg aatcgctagt aatcgcgaat 1320
cagaatgtcg cggtgaatac gttcccgggc cttgtacaca ccgcccgtca caccatggga 1380
gtgggttgca ccagaagtag ctagtctaac cttcgggagg acggttacca cggtgtgatt 1440
catgactgga gtgaagtcgt aacaaggtag ccgtagggga acctgcggct ggaacaggtc 1500
cgg 1503
Claims (9)
1. Pseudomonas fluorescens @Pseudomonas fluorescens) GH2-1, the preservation number of the strain is CGMCC NO.14743.
2. The Pseudomonas fluorescens GH2-1 strain and its metabolite in antagonizing pathogenic bacteria of cherry branch diseaseDiaporthe vaccinii) Is used in the field of applications.
3. A Pseudomonas fluorescens strain according to claim 1Pseudomonas fluorescens) Biological control preparation with GH2-1 as main effective component.
4. A method of preparing a biocontrol formulation as claimed in claim 3, wherein said method of preparation comprises: and (3) preparing seed liquid by using pseudomonas fluorescens GH2-1, inoculating the seed liquid into a fermentation medium, and fermenting and culturing for 30-36 hours at the temperature of 25-30 ℃ to obtain fermentation liquid.
5. The method of claim 4, wherein the seed liquid culture medium comprises the following components in percentage by mass: peptone 0.8-1.2%, beef powder 0.2-0.4%, sodium chloride 0.4-0.6%, and water for the rest, and adjusting pH 7.3+ -0.1.
6. The method according to claim 4, wherein the fermentation medium comprises the following components in percentage by mass: 1.5-2.5% of sucrose, 1.5-2.5% of soluble starch, 1.0-2.0% of yeast extract powder, 0.3-0.5% of fish peptone, 0.03-0.08% of magnesium sulfate, 0.03-0.08% of potassium dihydrogen phosphate, 0.5-0.8% of calcium carbonate and the balance of bean product wastewater, wherein the pH is adjusted to 7.0-7.2.
7. The method of claim 6, wherein the protein content of the bean product wastewater is controlled to be 16-18%.
8. Use of the biocontrol formulation of claim 3 for controlling cherry branch diseases.
9. The use according to claim 8, wherein the cherry branch disease is a disease caused by cranberry seed shellsDiaporthe vaccinii) Cherry branch diseases caused by infection.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210545263.5A CN114891679B (en) | 2022-05-19 | 2022-05-19 | Pseudomonas fluorescens and application thereof in preventing and controlling cherry branch diseases |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210545263.5A CN114891679B (en) | 2022-05-19 | 2022-05-19 | Pseudomonas fluorescens and application thereof in preventing and controlling cherry branch diseases |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114891679A CN114891679A (en) | 2022-08-12 |
| CN114891679B true CN114891679B (en) | 2023-06-20 |
Family
ID=82723952
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210545263.5A Active CN114891679B (en) | 2022-05-19 | 2022-05-19 | Pseudomonas fluorescens and application thereof in preventing and controlling cherry branch diseases |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114891679B (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012108598A1 (en) * | 2011-02-10 | 2012-08-16 | 경북대학교 산학협력단 | Novel photorhabdus temperata j5 and method for pest control using same |
| CN107502584A (en) * | 2017-10-16 | 2017-12-22 | 山东省林业科学研究院 | One fluorescent pseudomonads and its application in walnut black spot is prevented and treated |
| CN112359000A (en) * | 2020-11-27 | 2021-02-12 | 江苏省农业科学院 | Efficient biocontrol pseudomonas and application thereof in prevention and control of rice diseases |
| CN114085788A (en) * | 2021-10-27 | 2022-02-25 | 天津市农业科学院 | Pseudomonas japonicae strain and application thereof |
| CN114381391A (en) * | 2021-12-21 | 2022-04-22 | 天津农学院 | Pseudomonas aeruginosa and application thereof in inhibition of cucumber soil-borne diseases |
| CN114456986A (en) * | 2022-03-09 | 2022-05-10 | 山东国伟林业防治有限公司 | Broad-spectrum antibacterial Bacillus aryabhattai and application thereof |
-
2022
- 2022-05-19 CN CN202210545263.5A patent/CN114891679B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012108598A1 (en) * | 2011-02-10 | 2012-08-16 | 경북대학교 산학협력단 | Novel photorhabdus temperata j5 and method for pest control using same |
| CN107502584A (en) * | 2017-10-16 | 2017-12-22 | 山东省林业科学研究院 | One fluorescent pseudomonads and its application in walnut black spot is prevented and treated |
| CN112359000A (en) * | 2020-11-27 | 2021-02-12 | 江苏省农业科学院 | Efficient biocontrol pseudomonas and application thereof in prevention and control of rice diseases |
| CN114085788A (en) * | 2021-10-27 | 2022-02-25 | 天津市农业科学院 | Pseudomonas japonicae strain and application thereof |
| CN114381391A (en) * | 2021-12-21 | 2022-04-22 | 天津农学院 | Pseudomonas aeruginosa and application thereof in inhibition of cucumber soil-borne diseases |
| CN114456986A (en) * | 2022-03-09 | 2022-05-10 | 山东国伟林业防治有限公司 | Broad-spectrum antibacterial Bacillus aryabhattai and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 樱桃枝干病害病原鉴定及生物防治;张淑静等;山东林业科技(第4期);第9-14页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114891679A (en) | 2022-08-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN114456986B (en) | Broad-spectrum antibacterial bacillus aryabhattai and application thereof | |
| CN111690578B (en) | Salt and alkali resistant Siamese bacillus and production method and application of viable bacteria preparation thereof | |
| CN105062920A (en) | Streptomyces polychromogene bacterial strain, and applications thereof | |
| CN115369062A (en) | Tomato bacterial wilt antagonistic bacterium WJB0802 and application thereof | |
| CN112342173A (en) | Bacillus belgii and application thereof | |
| CN108913625B (en) | Salt-tolerant streptomycete, microbial inoculum thereof and application of microbial inoculum thereof in promoting plant growth | |
| CN113755389A (en) | Bacillus belgii and application thereof | |
| CN110791459B (en) | Bacillus subtilis for preventing and controlling continuous cropping lily soil-borne blight and application thereof | |
| CN115851553B (en) | Streptomyces virginiae capable of preventing and treating clubroot and application thereof | |
| CN116694505A (en) | Streptomyces crimson capable of preventing and treating clubroot and application thereof | |
| CN114032182B (en) | Fungus with functions of antagonizing pathogenic bacteria of garlic root rot and promoting growth | |
| CN114933980B (en) | Streptomyces shallowus HJB-XTBG45 for preventing and treating rhizoma polygonati root rot and application thereof | |
| CN114456973B (en) | Streptomyces rochei in tobacco and application thereof in prevention and control of tobacco diseases | |
| CN114891679B (en) | Pseudomonas fluorescens and application thereof in preventing and controlling cherry branch diseases | |
| CN116240126A (en) | Multifunctional bacillus belgium SB10 and application thereof | |
| CN107629970B (en) | Microorganism for preventing and treating ginseng erythroderma and application | |
| CN112831430A (en) | Comamonas aquosus strain, biocontrol microbial inoculum and preparation method and application thereof | |
| CN114747595B (en) | Application of Burkholderia cepacia JT79 in controlling meloidogyne incognita | |
| CN115851534B (en) | Bacillus belicus strain for tobacco rhizosphere and application of bacillus belicus strain in prevention and control of tobacco diseases | |
| CN116042461B (en) | Streptomyces rohder and application thereof in disease prevention and control of medlar | |
| CN112877222B (en) | Strain for antagonizing sclerotinia rot of asarum and application thereof | |
| CN114774288B (en) | Trichoderma harzianum and application thereof, and microbial agent for resisting anthracnose and promoting growth and development of strawberries | |
| CN116179640B (en) | Method for efficiently screening antagonistic bacteria of banana vascular wilt | |
| CN116769607B (en) | Application of endophytic fungus A21-1-1 in preventing and controlling rice blast | |
| CN117431166B (en) | Novel trichoderma brevicompactum strain TB2 and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |