CN114467975B - Application of staphylococcus equi in prevention and treatment of fruit and vegetable diseases - Google Patents
Application of staphylococcus equi in prevention and treatment of fruit and vegetable diseases Download PDFInfo
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- 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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Abstract
The invention discloses an application of Staphylococcus equi in preventing and treating fruit and vegetable diseases, the strain is named as Staphylococcus equi (Staphylococcus equi) F1, and the preservation number is CGMCC No.21658; the fruit and vegetable diseases are apple tree canker, pear tree canker, peach brown rot, apple anthracnose leaf blight or apple ring rot. The invention also provides an apple tree rot biocontrol microbial inoculum prepared by the supernatant fluid fermented by the strain F1 or the bacterial suspension of the strain F1 and a method for inducing the disease resistance of fruit tree branches. The staphylococcus equisimilis F1 has a remarkable control effect on apple tree canker, can effectively inhibit hypha growth and spore germination of apple tree canker, and can also be used for preventing diseases such as fruit tree canker and the like in advance, so that the morbidity of fruit trees and the treatment cost are greatly reduced.
Description
Technical Field
The invention relates to the technical field of microorganisms, and particularly relates to application of staphylococcus equinus in prevention and treatment of fruit and vegetable diseases.
Background
Apple tree rot caused by Humicola melanosporum (Vslsa mali) is a devastating disease with the greatest threat to the apple industry, and can damage main branches and trunks of apple trees, cause cortex rot, cause dead branches and dead trees, and even damage gardens. In 2008, nationwide investigation shows that the overall incidence rate of the rot disease of the apple main producing area in China is 52%, and the incidence rate of partial areas is more than 85%; in 2011, apple tree canker occurs rampant again in the tobacco plateau apple producing district in China, which causes serious economic loss. Since the apple tree canker is discovered for the first time in China in 1961, the apple tree canker has caused 4 pandemics, and related researchers indicate that the apple industry in China faces the threat of the 5 th pandemics of cankers.
At present, no disease-resistant planting resource for rot disease is available in production, and the disease cannot be effectively controlled by surgical operation treatment. The rot pathogen is a typical filamentous nutritional type bacterium, is mainly infected or long-term hidden through shearing mouths, various micro wounds, dead tissues of epidermis and the like, and is difficult to control by a chemical agent. Recently, researches find that the rot pathogen can rapidly grow in xylem of apple trees and longitudinally expand, but does not present obvious external symptoms, and the rot pathogen can not cause the tree body to suffer from diseases until the rot pathogen reaches cortex tissues; since germs exist in xylem, only the germs in cortex can be removed by surgical operation such as scraping, but hypha in xylem cannot be removed, which is also the main reason of causing serious scar of rot disease.
Due to the lack of control strategies and technologies with strong pertinence and good effect, the rot disease is always a great control problem in apple production in China. The agent can effectively prevent the apple tree from rotting, but the lasting period of the agent cannot maintain the whole growth period of the fruit tree, the measure is time-consuming and labor-consuming, the application in many apple production areas is limited, and the use of a large amount of chemical pesticides does not meet the development strategy of weight reduction and drug reduction of modern agriculture in China.
The biological control agent for preventing and treating the apple tree canker is environment-friendly, and can avoid a series of problems caused by chemical prevention and treatment. However, no report of using staphylococcus equi to prevent and treat apple tree canker exists in the prior art at present. How to solve the problems is a technical problem to be solved in the field of microorganisms at present.
Disclosure of Invention
In order to solve the problems, the invention provides the application of Staphylococcus equinus (Staphylococcus equirum) F1 in fruit tree disease control, and the Staphylococcus equinus F1 has high-efficiency antibacterial effect on various fruit and vegetable diseases (such as apple tree rot, pear tree rot, peach brown rot, apple anthracnose leaf blight and apple ring rot), has a wide antibacterial spectrum, can be widely applied to control of fruit and vegetable diseases, and has the advantages of stable control effect and environmental friendliness.
The technical scheme of the invention is as follows:
in a first aspect, the invention provides an application of Staphylococcus equi in preventing and treating fruit and vegetable diseases, wherein the strain is named as Staphylococcus equi (Staphylococcus equi) F1 with the preservation number of CGMCC No.21658; the fruit and vegetable diseases are apple tree canker, pear tree canker, peach brown rot, apple anthracnose leaf blight or apple ring rot.
Staphylococcus equine ventriculi F1 was deposited in China general microbiological culture Collection center (CGMCC) at 18.1.2021, with a collection number of CGMCC No.21658 and a collection address of: xilu No. 1 Hospital No. 3, beijing, chaoyang, north.
In a second aspect, the invention also provides an application of the biocontrol microbial inoculum prepared by the staphylococcus equinus F1 in fruit and vegetable disease control, wherein the active ingredient of the biocontrol microbial inoculum is strain F1 fermentation supernatant or strain F1 bacterial suspension. The fruit and vegetable diseases comprise apple tree rot, pear tree rot, peach brown rot, apple anthracnose leaf blight and apple ring spot.
The invention also provides a biological control microbial inoculum for the apple tree canker, and the active ingredient is fermentation supernatant or bacterial suspension of Staphylococcus equi (Staphylococcus equi) F1.
Preferably, in the apple tree canker biocontrol microbial inoculum, the concentration of Staphylococcus equinus (Staphylococcus equiorum) strain F1 is 10 5 -10 9 cfu·mL -1 。
Further preferably, the apple tree rot biocontrol agent is in a concentration of 10 9 cfu·mL -1 The Staphylococcus equine gastritis (Staphylococcus equirum) strain F1 suspension.
The preparation method of the apple tree rot biocontrol microbial inoculum provided by the invention comprises the following steps:
(1) Inoculating the strain F1 in a PDA culture medium in a streak manner, and performing activation culture at a constant temperature of 28 ℃ for 24h to obtain an activated single strain F1 colony a;
(2) Inoculating the activated strain F1 single colony a into PDB culture medium with bottling amount of 80mL/250mL for 180r min -1 Performing shake culture at constant temperature of 28 ℃ for 48 hours to obtain a strain F1 fermentation seed solution b;
(3) Inoculating the fermentation seed liquid b into a PDB culture medium according to the inoculation amount of 1 9 cfu·mL -1 Cell culture fluid c of the F1 strain;
(4) Will 10 9 cfu·mL -1 Cell culture solution c of F1 strain at 4 deg.C and 12000 r.min -1 Centrifuging for 15min, and filtering the collected supernatant with 0.22 μm microporous membrane to obtain strain F1 fermentation supernatant as biocontrol microbial inoculum; collecting F1 thallus precipitate, resuspending with sterile water to obtain 10 9 cfu·mL -1 The F1 bacterial suspension is the biological control agent.
In a third aspect, the invention also provides a method for inducing disease resistance of fruit tree branches, which comprises the following steps: and applying the bacterial liquid of the staphylococcus equine gastri to the surface of the fruit tree branch.
Preferably, in the method for inducing disease resistance of fruit tree branches, the specific application method is as follows: spraying or smearing the bacterial liquid of the staphylococcus equorum on the surfaces of the branches of the fruit trees.
Preferably, the concentration of the staphylococcus equine gastric bacteria liquid is not less than 10 9 cfu·mL -1 。
Has the advantages that:
1. the staphylococcus equi F1 provided by the invention has a remarkable control effect on apple tree canker, and the research of the embodiment shows that the bacterial strain F1 can effectively inhibit the hypha growth and the spore germination of apple tree canker, and the control effect on the apple tree canker can reach more than 65%; the strain F1 can induce the activity of a plurality of defense related genes in apple tissues to be increased, and the expression level of disease-resistant related genes of fruit trees is improved.
2. The apple tree rot biocontrol microbial inoculum provided by the invention has the characteristics of stable control effect, environmental friendliness and the like, is suitable for large-scale production, is simple in fermentation process, low in production cost and has good market prospect.
3. The method for inducing the disease resistance of the fruit tree branches is simple to operate and low in cost, and can be widely used for preventing diseases such as fruit tree rot and the like in advance, so that the disease incidence of the fruit trees and the treatment cost are greatly reduced.
Drawings
FIG. 1 is the colony morphology of strain F1 on LB medium;
FIG. 2 is a schematic diagram showing PCR amplification of 16S rDNA sequence of DNA extracted from strain F1;
wherein, M in the figure is nucleic acid Marker DL2000; FIG. 1 shows the result of amplification of the 16S rDNA sequence of the genomic DNA of the strain F1;
FIG. 3 is a graph showing the effect of strain F1 on hyphal growth (a) and spore germination (b) of rot pathogen of apple trees;
wherein, A in the figure is the inhibiting effect of the strain F1 on apple tree canker; in the figure, B is the influence of the thalli and the supernatant of the strain F1 on the germination of the rot pathogen spores;
FIG. 4 shows the control effect of the supernatant of the strain F1 on apple tree canker;
wherein, A in the figure is a control of inoculating putrefaction bacteria after inoculating a PDB culture medium on apple branches; in the figure, B is the control effect of the supernatant of the strain F1 on apple tree canker;
FIG. 5 is a graph of the effect of bacterial suspension treatment with strain F1 on defense-related enzyme activity in apple shoot tissue;
FIG. 6 shows the effect of bacterial suspension treatment of strain F1 on the expression of pathogenesis-related protein genes in apple shoot tissue.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention. In the present invention, the equipment and materials used are commercially available or commonly used in the art, if not specified. The methods in the following examples are conventional in the art unless otherwise specified. The reagents used are commercially available, unless otherwise specified.
Culture medium for experiment and formula thereof
1. LB medium: 10g of tryptone, 5g of yeast extract and 5g of NaCl, and adjusting the pH to 7.0.
2. PDA culture medium: peeling potatoes, weighing 200g, cutting into small pieces, boiling in water for 15-20 min, filtering with four layers of gauze, adding 20g of glucose and 15g of agar powder, fixing the volume to 1000mL, naturally adjusting the pH value, and sterilizing with high-pressure steam at 121 ℃ for 20min.
3. PDB culture medium: peeling potato, weighing 200g, cutting into small pieces, boiling in water for 15-20 min, filtering with four layers of gauze, adding glucose 20g, diluting to 1000mL, keeping pH natural, and sterilizing with high pressure steam at 121 deg.C for 20min.
Example 1 isolation screening and Strain identification of Staphylococcus equine F1
1. Isolation and selection of strains
Mature and healthy fruits are collected from Shandong commercial Fuji apple orchard, the fruit tissues are ground after the surfaces are disinfected, sterile water is added, a conventional gradient dilution coating separation method is adopted, PDA culture media are used for culture at 28 ℃, colonies with obvious colony morphology differences are picked up, the colonies are purified and stored on the PDA culture media, primary screening and repeated screening of antagonistic bacteria are carried out by taking apple tree rotting pathogenic bacteria as target pathogenic bacteria, and finally a bacterial strain with the bacteriostatic action and the remarkable prevention and control effect is obtained and named as F1.
2. Identification of Strain F1
(1) Morphological characteristics
As shown in the results of FIG. 1, the bacterial colony of the strain F1 on LB medium is small, pale, round (slightly irregular) in shape, opaque, and smooth with wet surface, and the bacterial body is identified as gram-positive bacteria.
(2) Physiological and biochemical characteristics
The strain F1 can utilize glucose, sucrose, mannitol and the like as carbon sources, but cannot utilize galactose, lactose and the like as carbon sources; the catalase, V-P, methyl red and urease have positive reaction, can liquefy the Mingzhi, hydrolyze starch and do not form a mycoderm, and the characteristics are consistent with those of staphylococcus equi.
(3) And (3) gene sequence identification:
A. the experimental method comprises the following steps: extracting the genome DNA of the strain F1 by using the kit, taking the genome DNA as a DNA template, and performing amplification by using a universal Primer F:5'-AGAGTTTGATCCTGGCTCAG-3', primer R:5'-AAGGAGGTGATCCAGCCGCA-3' PCR amplification of DNA template was performed.
PCR amplification reaction (50. Mu.L): DNA template 2. Mu.L, upstream primer 2. Mu.L, downstream primer 2. Mu.L, taq enzyme (TaKaRa) 1. Mu.L, dNTP 1. Mu.L, 2 XTaq buffer 25. Mu.L, ddH 2 O17. Mu.L. PCR amplification procedure: pre-denaturation at 95 ℃ for 5min, denaturation at 95 ℃ for 30s, annealing at 58 ℃ for 30s, extension at 72 ℃ for 2min, 35 cycles, and extension at 72 ℃ for 10min. The amplification product (5. Mu.L) was detected by electrophoresis on a 1% agarose gel. As shown in FIG. 2, the PCR amplification product was recovered and then transferred to Shanghai bioengineering Co., ltd for sequence determination.
B. Results and analysis: the determination result shows that the length of the 16S rDNA sequence amplification fragment is 1431bp, and the sequence is shown as the sequence table SEQ ID NO: 1. The resulting sequences were aligned with the nucleic acid sequences in GenBank using BLAST software in the NCBI database (http:// www.ncbi.nlm.nih.gov) to show 99.9% homology to the 16 rDNA sequences of Staphylococcus equine (Staphylococcus equivus) C4044 strain and C2060 strain (Accession NO: KF439736 and KF 439734).
Based on the above morphological characteristics, physiological and biochemical characteristics and the results of sequence analysis, the strain F1 was identified as Staphylococcus equinus (Staphylococcus equivum).
Example 2 inhibition of various plant pathogenic bacteria by Staphylococcus equine Strain F1
A. The experimental method comprises the following steps: a suspension of Staphylococcus equisimilis (Staphylococcus equivum) F1 was prepared to a final concentration of 10 9 cfu·mL -1 So as not to add the strain F1PDA served as a control, and was inoculated with each pathogen in activated culture.
Bacteriostasis rate = (control colony diameter-treated colony diameter)/control colony diameter × 100%.
B. Results and analysis: as shown in the results in Table 1, the strain F1 has obvious inhibiting effect on main plant pathogenic bacteria such as botrytis cinerea, apple tree rot, apple ring rot, apple colletotrichum gloeosporioides and the like, shows good broad-spectrum antibacterial property, and has the bacteriostasis rate in 4d culture as shown in Table 1.
TABLE 1 inhibitory Effect of Staphylococcus equi (Staphylococcus equi) Strain F1 on major phytopathogens
Example 3 Effect of Staphylococcus equine F1 on mycelial growth and spore germination of Alternaria mali
(1) Effect of Strain F1 on the growth of hyphae of decay fungi
A. The experimental method comprises the following steps:
preparing a PDA culture medium, preparing a flat plate, streaking and inoculating the F1 strain at a position which is about 2cm away from the center of one side of the flat plate, culturing at 28 ℃ for 2d, inoculating activated apple tree canker at the opposite position of the other side, performing dark culture at a constant temperature of 28 ℃ for 3d by taking the flat plate which is not inoculated with the F1 strain and only inoculated with the canker as a control, and measuring the diameter of a bacterial colony.
B. Results and analysis:
the result of FIG. 3A shows that the strain F1 can obviously inhibit the growth of hyphae of the rot pathogen, after 3 days of culture, the average diameter of a contrast colony is 8.8cm, the diameter of the colony in a counter petri dish is 4.7cm, and the bacteriostasis rate is 46.6%.
(2) Effect of Strain F1 on spore germination of Cordycephe
A. The experimental method comprises the following steps:
preparing spore suspension of rot pathogen, culturing pathogenic bacteria on PDA plate, inducing under black light (365 nm) when hypha grows over the plate, and observing the generation of black conidiophore after about 2 weeksReleasing yellow conidium angle, preparing conidium suspension, and adjusting concentration to 10 6 Each is mL -1 。
Inoculating seed fermentation liquid of the strain F1 into a PDB culture medium, performing shake culture at a constant temperature of 28 ℃ for 48h to obtain a cell culture liquid of the strain F1, and adjusting the concentration to 10 9 cfu·mL -1 Centrifuging and filtering to obtain supernatant of F1 strain, resuspending the strain in sterile water, and making into suspension 10 with different concentrations 9 cfu·mL -1 ，10 7 cfu·mL -1 And 10 5 cfu·mL -1 . Respectively mixing the bacterial suspension and supernatant of the F1 strain with the conidiospore suspension 10 of the rot fungi with equal volume 6 Each is mL -1 Mixing, placing in dark condition at 25 deg.C, and observing germination rate of rot pathogen spore after 12 hr.
B. Results and analysis:
as shown in the results of FIG. 3B, the control group (CK) exhibited a conidium germination rate of 80.25% and the F1 strain exhibited a significant inhibitory effect on conidium germination at a concentration of 10 9 cfu·mL -1 The inhibition effect of the F1 thalli is most obvious, the spore germination rate is only 3.25%, and the inhibition rate is as high as 95.95%; the germination rate of the spores treated by the F1 supernatant is 12.13%, and the inhibition rate of the F1 supernatant on the germination of the spores of the rot pathogen is 84.89%.
Example 4 prevention and treatment effects of fermentation supernatant of Staphylococcus equine F1 on apple Tree rot
1. The experimental method comprises the following steps:
inoculating seed fermentation liquid of the strain F1 into a PDB culture medium, performing shake culture at a constant temperature of 28 ℃ for 48h to obtain a cell culture liquid of the strain F1, and adjusting the concentration to 10 9 cfu·mL -1 And centrifuging and filtering to obtain the supernatant of the F1 strain. The putrefaction bacteria are activated and cultured on PDA for 3d, and a fungus cake with the diameter of 5mm is taken at the edge of the colony for standby.
Selecting 1-2 year-old healthy Fuji branches with consistent growth vigor, scalding with an electric iron to make the wounds about 3mm long and 1mm deep, inoculating 20 mu L of strain F1 supernatant to each wound, airing at room temperature, placing the branches in an incubator with the relative humidity of 90% at 25 ℃, inoculating the activated rotten germ cakes after 48h interval, and inoculating PDB culture medium and then inoculating rotten germs to serve as a control. Lesion length was observed and measured 4d after inoculation.
Control effect = (control group lesion diameter-treatment group lesion diameter)/control group lesion diameter × 100%
2. Results and analysis of the experiments
As shown in figure 4, after the branches of the control group are inoculated with the rot pathogen, the scab rapidly expands, the average length of the scab is 4.38cm at 4d, while in the treatment group inoculated with the supernatant of the strain F1, the scab slowly expands, the length of the scab is only 1.52cm, and the prevention and treatment effect is 65.3%.
Example 5 preparation of biocontrol microbial inoculum for apple tree rot
The embodiment provides an apple tree rot biocontrol microbial inoculum prepared by adopting a strain F1, and the preparation method specifically comprises the following steps:
(1) Inoculating the strain F1 in a PDA culture medium in a streak manner, and performing activation culture at a constant temperature of 28 ℃ for 24h to obtain an activated single strain F1 colony a;
(2) Inoculating the activated strain F1 single colony a into PDB culture medium with bottling amount of 80mL/250mL for 180 r.min -1 Performing shake culture at constant temperature of 28 ℃ for 12h to obtain a strain F1 fermentation seed solution b;
(3) Inoculating the fermentation seed liquid b into a PDB culture medium according to the inoculation amount of 1 9 cfu·mL -1 Cell culture fluid c of the F1 strain;
(4) Will 10 9 cfu·mL -1 Cell culture solution c of F1 strain at 4 deg.C and 12000 r.min -1 Centrifuging for 15min, and resuspending the thallus precipitate with sterile water to obtain 10 9 cfu·mL -1 The bacterial suspension is the biological control bacterial agent.
In addition, the above-mentioned reference numeral 10 9 cfu·mL -1 The bacterial suspension is generally diluted 0-10000 times when in use.
Example 5 prevention and treatment Effect of Staphylococcus equine F1 suspensions of different concentrations on apple Tree rot disease
1. The experimental method comprises the following steps:
preparation of suspensions of the Strain F1 at different concentrations, 10 7 cfu·mL -1 ，10 8 cfu·mL -1 And 10 9 cfu·mL -1 . Selecting 1-2 year-old healthy Fuji branches with consistent growth, scalding with an electric iron to make the wound length about 3mm and depth 1mm, inoculating 20 μ L of strain F1 bacterial suspension 10 to each wound 5 cfu·mL -1 ，10 7 cfu·mL -1 And 10 9 cfu·mL -1 The branches dried at room temperature are placed in an incubator with the relative humidity of 90% at 25 ℃, activated rotting pathogen cakes are inoculated after 48 hours of interval, and the treatment of inoculating PDB culture medium and then inoculating rotting pathogen is used as a contrast. And observing and measuring the length of the lesion spots 5 days after inoculation, counting the diameters of the lesion spots, and calculating the prevention and treatment effect.
Control effect = (control group lesion diameter-treatment group lesion diameter)/control group lesion diameter × 100%.
2. Results and analysis of the experiments
As shown in the results of Table 2, the control effect of the F1 bacterial suspensions with different concentrations is over 65 percent. Therein, 10 9 cfu·mL -1 The prevention and treatment effect of the F1 bacterial suspension is the highest and reaches over 75 percent.
TABLE 2 prevention and control effect of suspension of different concentrations of the F1 strain on apple tree canker
Example 6 Effect of Staphylococcus equine F1 bacterial suspension on defense-related enzyme Activity in apple tissue
1. The experimental method comprises the following steps:
selecting healthy Fuji apple branches according to the method of example 4, and uniformly spraying 10 9 cfu·mL -1 The F1 bacterial suspension of (1), and the branches treated with sterile water are used as a control. Taking branch phloem tissue at 0, 12, 24, 48, 72, 120h after treatment, respectively, and measuring appleTendency of defense-related enzyme activity in shoot tissue. Wherein, POD (peroxidase) and SOD (superoxide dismutase) activity are respectively measured by guaiacol method and xanthine oxidase method; PAL (phenylalanine ammonia lyase) and PPO (polyphenol oxidase) activity determination adopts phenylalanine method and catechol method, respectively.
2. Results and analysis of the experiments
The results are shown in FIG. 5, 10 9 cfu·mL -1 After the apple branches are treated by the F1 bacterial suspension, the activities of Peroxidase (POD), superoxide dismutase (SOD), phenylalanine ammonia acid lyase (PAL) and polyphenol oxidase (PPO) in the branch tissues are obviously improved to different degrees compared with a control, which shows that the staphylococcus equi F1 can induce the resistance of the apple branches to diseases by improving the activity of defense-related enzymes in the branches.
Example 7 Effect of Staphylococcus equine F1 bacterial suspension on Gene expression of Process-related proteins in apple tissue
1. The experimental method comprises the following steps:
selecting healthy Fuji apple branches according to the method of example 4, and uniformly spraying 10 9 cfu·mL -1 The F1 bacterial suspension of (1), and the branches treated with sterile water are used as a control. Taking branch phloem tissue 0, 24 and 48 hours after treatment, extracting total RNA of a sample, carrying out reverse transcription, and determining the expression condition of disease course related protein genes in the apple branch tissue by using an RT-qPCR technology.
2. Results and analysis of the experiments
The results are shown in FIG. 6, with a concentration of 10 9 cfu·mL -1 After the apple branches are treated by the F1 bacterial suspension, the expression quantity of the disease course related protein genes MdPR1 and MdPR5 in branch tissues is obviously increased compared with that of a control, the expression quantity of the 48h MdPR1 and MdPR5 after treatment is the highest, and the expression quantity is respectively increased by 8.33 times and 16.32 times compared with that of the control, which shows that staphylococcus equi F1 can induce the disease course related protein genes in the apple branches to be up-regulated and expressed, so that the resistance of the apple branches to diseases is improved.
The foregoing is directed to preferred embodiments of the present invention, other and further embodiments of the invention may be devised without departing from the basic scope thereof, and the scope thereof is determined by the claims that follow. However, any simple modification, equivalent change and modification of the above embodiments according to the technical essence of the present invention are within the protection scope of the technical solution of the present invention.
Sequence listing
<110> Qingdao agricultural university
<120> application of staphylococcus equi strain in prevention and treatment of fruit and vegetable diseases
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1431
<212> DNA
<213> 16S rDNA of Staphylococcus equine F1 (1696 rDNA of Staphylococcus equivum F1)
<400> 1
tatacatgca agtcgagcga acggataagg agcttgctcc tttgaagtta gcggcggacg 60
ggtgagtaac acgtgggtaa cctacctata agactggaat aacttcggga aaccggagct 120
aatgccggat aacatttgga accgcatggt tctaaagtaa aagatggttt tgctatcact 180
tatagatgga cccgcgccgt attagctagt tggtaaggta acggcttacc aaggcaacga 240
tacgtagccg acctgagagg gtgatcggcc acactggaac tgagacacgg tccagactcc 300
tacgggaggc agcagtaggg aatcttccgc aatggacgaa agtctgacgg agcaacgccg 360
cgtgagtgat gaaggttttc ggatcgtaaa actctgttat tagggaagaa caaatgtgta 420
agtaactgtg cacatcttga cggtacctaa tcagaaagcc acggctaact acgtgccagc 480
agccgcggta atacgtaggt ggcaagcgtt atccggaatt attgggcgta aagcgcgcgt 540
aggcggtttc ttaagtctga tgtgaaagcc cacggctcaa ccgtggaggg tcattggaaa 600
ctgggaaact tgagtacaga agaggaaagt ggaattccat gtgtagcggt gaaatgcgca 660
gagatatgga ggaacaccag tggcgaaggc gactttctgg tctgtaactg acgctgatgt 720
gcgaaagcgt ggggatcaaa caggattaga taccctggta gtccacgccg taaacgatga 780
gtgctaagtg ttagggggtt tccgcccctt agtgctgcag ctaacgcatt aagcactccg 840
cctggggagt acgaccgcaa ggttgaaact caaaggaatt gacggggacc cgcacaagcg 900
gtggagcatg tggtttaatt cgaagcaacg cgaagaacct taccaaatct tgacatcctt 960
tgaaaactct agagatagag ccttcccctt cgggggacaa agtgacaggt ggtgcatggt 1020
tgtcgtcagc tcgtgtcgtg agatgttggg ttaagtcccg caacgagcgc aacccttaaa 1080
cttagttgcc agcatttagt tgggcactct aggttgactg ccggtgacaa accggaggaa 1140
ggtggggatg acgtcaaatc atcatgcccc ttatgatttg ggctacacac gtgctacaat 1200
ggacaataca aagggcagct aaaccgcgag gtcatgcaaa tcccataaag ttgttctcag 1260
ttcggattgt agtctgcaac tcgactacat gaagctggaa tcgctagtaa tcgtagatca 1320
gcatgctacg gtgaatacgt tcccgggtct tgtacacacc gcccgtcaca ccacgagagt 1380
ttgtaacacc cgaagccggt ggagtaacca tttatggagc tagccgtcga a 1431
Claims (4)
1. The application of the staphylococcus equine stomach in preventing and treating the fruit and vegetable diseases is characterized in that the strain is named as staphylococcus equine stomach (A)Staphylococcus equorum) F1 with the preservation number of CGMCC No.21658; the fruit and vegetable diseases are apple tree canker (A)Valsa mali) Pear rot pathogen: (Valsa pyri) Monilinia fructicola (C.fructicola)Monilinia fructicola) Anthracnose of apple leaf blight bacteria (A)Glomerella cingulata) Or apples Physalospora piricola (A)Botryosphaeria dothidea) The concentration of the staphylococcus equine gastric bacteria liquid is not less than 10 9 cfu·mL -1 。
2. Biological control bacterium for apple tree rot diseaseThe agent is characterized in that the active ingredient of the apple tree rot disease biocontrol microbial inoculum is staphylococcus equi (A), (B), (C) and (C)Staphylococcus equorum) The fermentation supernatant or bacterial suspension of F1, the preservation number of which is CGMCC No.21658; the concentration of Staphylococcus equine strain F1 was 10 5 -10 9 cfu·mL -1 The disease of apple tree canker is apple tree canker: (Valsa mali）。
3. A method for inducing disease resistance of fruit tree branches is characterized by comprising the following steps: applying the bacterial liquid of staphylococcus equi according to claim 1 to the surface of branches of fruit trees, wherein the concentration of the bacterial liquid of staphylococcus equi is not less than 10 9 cfu·mL -1 Inducing fruit tree branches to produce apple tree canker (Valsa mali) The disease resistance of (1).
4. The method according to claim 3, wherein the application method is spraying to the surface of the fruit tree branches or smearing the bacterial liquid of staphylococcus equine gastri according to claim 1.
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