CN114467975A

CN114467975A - Application of staphylococcus equi in prevention and treatment of fruit and vegetable diseases

Info

Publication number: CN114467975A
Application number: CN202210337146.XA
Authority: CN
Inventors: 王彩霞; 郝柏慧; 许宇昕; 李世昱; 孙子豪; 李保华
Original assignee: Qingdao Agricultural University
Current assignee: Qingdao Agricultural University
Priority date: 2022-04-01
Filing date: 2022-04-01
Publication date: 2022-05-13
Anticipated expiration: 2042-04-01
Also published as: CN114467975B

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 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. The invention also provides an apple tree rot biocontrol microbial inoculum prepared by fermenting the supernatant or the suspension of the strain F1 by the strain F1 and a method for inducing the disease resistance of fruit tree branches. The staphylococcus equine gastritis F1 has a remarkable prevention and treatment 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

Application of staphylococcus equisimilis in preventing and treating fruit and vegetable diseases

Technical Field

The invention relates to the technical field of microorganisms, in particular 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 first discovery of apple tree canker in 1961, the apple tree canker has caused 4 pandemics in China, and related researchers point out that the apple industry in China is facing the threat of 5 th pandemics of canker.

At present, no rot disease-resistant planting resource is available in production, and the disease cannot be effectively controlled by surgical 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 equi (Staphylococcus equi) F1 in the prevention and treatment of fruit tree diseases, and the Staphylococcus equi 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 the prevention and treatment of fruit and vegetable diseases, and has the advantages of stable prevention and treatment 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 gastric F1 was deposited in China general microbiological culture Collection center (CGMCC) at 18.1.2021, with the collection number of CGMCC No.21658 and the deposition 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 preventing and treating fruit and vegetable diseases, wherein the active ingredient of the biocontrol microbial inoculum is a supernatant obtained by fermenting the strain F1 or a bacterial suspension of the strain F1. The fruit and vegetable diseases comprise apple tree rot, pear tree rot, peach brown rot, apple anthracnose leaf blight and apple ring rot.

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, the concentration of Staphylococcus equinus (Staphylococcus equiorum) strain F1 in the apple tree rot biocontrol microbial inoculum is 10⁵-10⁹cfu·mL^-1。

Further preferably, the apple tree rot biocontrol agent is in a concentration of 10⁹cfu·mL^-1The Staphylococcus equine gastric (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 the constant temperature of 28 ℃ for 24h to obtain an activated strain F1 single 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^-1Carrying out shake culture at the constant temperature of 28 ℃ for 48h to obtain a strain F1 fermentation seed liquid b;

(3) inoculating the fermentation seed liquid b into a PDB culture medium according to the inoculation amount of 1:100 for amplification culture, and performing constant-temperature shaking culture at 28 ℃ for 48h to obtain 10⁹cfu·mL^-1Cell culture solution c of F1 strain;

(4) will 10⁹cfu·mL^-1Cell culture solution c of F1 strain at 4 ℃ at 12000 r.min^-1Centrifuging for 15min, and filtering the collected supernatant with 0.22 μm microporous membrane to obtain strain F1 fermented supernatant as biocontrol microbial inoculum; collecting F1 thallus precipitate, and resuspending with sterile water to obtain 10⁹cfu·mL^-1The 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 equine gastri on the surface of the fruit tree branch.

Preferably, the concentration of the staphylococcus equine gastric bacteria liquid is not less than 10⁹cfu·mL^-1。

Has the advantages that:

1. the staphylococcus equi F1 provided by the invention has a remarkable prevention and treatment effect on apple tree canker, and the research of the embodiment shows that the strain F1 can effectively inhibit the hypha growth and spore germination of apple tree canker, and the prevention effect on the apple tree canker can reach more than 65%; the strain F1 can induce the increase of the activity of a plurality of defense related genes in apple tissues and improve the expression level of disease-resistant related genes of fruit trees.

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 provided by the invention 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 and treatment cost of the fruit trees are greatly reduced.

Drawings

FIG. 1 is the colony morphology of strain F1 on LB medium;

FIG. 2 is a schematic diagram of PCR amplification of 16S rDNA sequence of DNA extracted from strain F1;

wherein, M in the figure is nucleic acid Marker DL 2000; FIG. 1 shows the result of amplification of the 16S rDNA sequence of the genomic DNA of strain F1;

FIG. 3 is a graph showing the effect of strain F1 on hyphal growth (a) and spore germination (b) of apple rot;

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 putrescence bacteria spores;

FIG. 4 shows the control effect of the supernatant of 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 disease process-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 culture medium: 10g of tryptone, 5g of yeast extract and 5g of NaCl, and the pH value is adjusted 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, keeping the pH value natural, and sterilizing with high-pressure steam at 121 ℃ for 20 min.

3. PDB 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, diluting to 1000mL, keeping the pH value natural, and sterilizing with high-pressure steam at 121 ℃ for 20 min.

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 strain F1 was small in size, pale white, round (slightly irregular) in shape, opaque, and smooth and moist in surface on LB medium, and was identified as a gram-positive bacterium.

(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: the genomic DNA of the strain F1 was extracted using the kit and used as a DNA template, followed by the addition of the universal Primer F: 5'-AGAGTTTGATCCTGGCTCAG-3', Primer R: 5'-AAGGAGGTGATCCAGCCGCA-3' the DNA template is PCR amplified.

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, ddH₂O17. 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 10 min. 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 16S rDNA sequences of Staphylococcus aureus (Staphylococcus equum) 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 equi (Staphylococcus equi) F1 was prepared to have a final concentration of 10⁹cfu·mL^-1The control was performed by inoculating each pathogen in activated culture, using PDA without the addition of the strain F1.

The inhibition ratio is (control colony diameter-treatment colony diameter)/control colony diameter × 100%.

B. Results and analysis: as shown in the results of Table 1, the strain F1 has significant inhibitory effect on main plant pathogenic bacteria such as fruit and vegetable botrytis cinerea, apple tree rot, apple ring rot, apple anthracnose leaf blight and the like, and shows good broad-spectrum antibacterial property, and the bacteriostatic rate of the strain in 4d culture is shown in Table 1.

TABLE 1 inhibitory Effect of Staphylococcus equi (Staphylococcus equi) strain F1 on the major phytopathogens

Example 3 Effect of Staphylococcus equi F1 on mycelial growth and spore germination of Micrococcus vallismortis

(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 an F1 strain at a position which is about 2cm away from the center of one side of the flat plate, culturing at 28 ℃ for 2 days, inoculating activated apple tree canker at the opposite position of the other side, using the flat plate which is not inoculated with the F1 strain and is only inoculated with the canker as a control, culturing at 28 ℃ for 3 days in a dark and constant temperature mode, and measuring the colony diameter.

B. Results and analysis:

the result of FIG. 3A shows that the strain F1 can obviously inhibit the growth of hyphae of rot pathogen, after 3 days of culture, the average diameter of the contrast colony is 8.8cm, the diameter of the colony in the opposite culture dish is 4.7cm, and the bacteriostasis rate is 46.6%.

(2) Effect of Strain F1 on spore germination of Pythium putrescens

A. The experimental method comprises the following steps:

preparing spore suspension of putrefaction pathogen, culturing pathogenic bacteria on PDA plate, inducing under black light (365nm) when hypha grows over the plate, allowing generation of black conidium apparatus and release of yellow conidium angle after about 2 weeks, configuring conidium suspension, and adjusting concentration to 10⁶each.mL^-1。

Inoculating seed fermentation liquid of the strain F1 in PDB culture medium, shake culturing at 28 deg.C for 48h to obtain cell culture liquid of F1 strain, and adjusting the concentration to 10⁹cfu·mL^-1Centrifuging and filtering to obtain supernatant of F1 strain, resuspending the strain in sterile water, and making into suspension 10 with different concentrations⁹cfu·mL^-1，10⁷cfu·mL^-1And 10⁵cfu·mL^-1. Respectively mixing bacterial suspension and supernatant of F1 strain with equal volume of conidium suspension 10 of rot pathogen⁶each.mL^-1Mixing, 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 10⁹cfu·mL^-1The 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 disease

1. The experimental method comprises the following steps:

inoculating seed fermentation liquid of the strain F1 in PDB culture medium, shake culturing at 28 deg.C for 48h to obtain cell culture liquid of F1 strain, and adjusting the concentration to 10⁹cfu·mL^-1And 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, wherein the length of a wound is about 3mm, the depth of the wound is 1mm, 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 a PDB culture medium and then inoculating rotten germs to serve as a control. Lesion length was observed and measured 4d after inoculation.

The control effect is (control group lesion diameter-treatment group lesion diameter)/control group lesion diameter multiplied by 100%

2. Results and analysis of the experiments

As shown in figure 4, the disease spots of the branches of the control group rapidly spread after being inoculated with the rot pathogen, the length of the disease spots is averagely 4.38cm at 4d, while the disease spots spread slowly in the treated group inoculated with the supernatant of the strain F1, the length of the disease spots is only 1.52cm, and the control 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) streaking and inoculating the strain F1 in a PDA culture medium, and performing activation culture at a constant temperature of 28 ℃ for 24h to obtain an activated strain F1 single colony a;

(2) inoculating the activated strain F1 single colony a into PDB culture medium with bottling amount of 80mL/250mL for 180r min^-1Performing shake culture at constant temperature of 28 ℃ for 12h to obtain a strain F1 fermentation seed liquid b;

(3) inoculating the fermentation seed liquid b into a PDB culture medium according to the inoculation amount of 1:100 for amplification culture, and performing constant-temperature shaking culture at 28 ℃ for 48 hours to obtain 10⁹cfu·mL^-1Cell culture solution c of F1 strain;

(4) will 10⁹cfu·mL^-1Cell culture solution c of F1 strain at 4 ℃ at 12000 r.min^-1Centrifuging for 15min, and resuspending the thallus precipitate with sterile water to obtain 10⁹cfu·mL^-1The bacterial suspension is the biological control agent.

In addition, the above-mentioned reference numeral 10⁹cfu·mL^-1The bacterial suspension is generally diluted 0-10000 times.

Example 5 prevention and treatment of apple Tree rot disease by Staphylococcus equi F1 bacterial suspensions of different concentrations

1. The experimental method comprises the following steps:

preparation of suspensions of Strain F1 at various concentrations, 10⁷cfu·mL^-1，10⁸cfu·mL^-1And 10⁹cfu·mL^-1. Selecting 1-2 year-old healthy Fuji branches with consistent growth, scalding with an electric iron to make the wound about 3mm long and 1mm deep, inoculating 20 μ L of strain F1 bacterial suspension 10 to each wound⁵cfu·mL^-1，10⁷cfu·mL^-1And 10⁹cfu·mL^-1The 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.

The control effect is (control group lesion diameter-treatment group lesion diameter)/control group lesion diameter multiplied by 100%.

2. Results and analysis of the experiments

As shown in the results of Table 2, the control effect of the F1 bacterial suspension with different concentrations is over 65%. Therein, 10⁹cfu·mL^-1The prevention and treatment effect of the F1 bacterial suspension is the highest and reaches more than 75%.

TABLE 2 prevention and control of apple tree canker by suspension of strain F1 in different concentrations

Example 6 Effect of Staphylococcus equine F1 bacterial suspension on defense-related enzymatic 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 10⁹cfu·mL^-1The suspension of F1 was used as a control with shoots treated with sterile water. And (4) taking

branch phloem tissues

0, 12, 24, 48, 72 and 120 hours after treatment respectively, and determining the change trend of the defense related enzyme activity in the apple branch tissues. 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, 10⁹cfu·mL^-1After the apple branches are treated by the F1 bacterial suspension, the activities of Peroxidase (POD), superoxide dismutase (SOD), phenylalanine ammonia acid ammonia lyase (PAL) and polyphenol oxidase (PPO) in the branch tissues are all obviously improved to different degrees compared with a control, which shows that 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 10⁹cfu·mL^-1The suspension of F1 was used as a control with shoots treated with sterile water. 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 10⁹cfu·mL^-1After the apple branches are treated by the F1 bacterial suspension, the expression quantity of 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 disease-course-related protein genes MdPR1 and MdPR5 is highest 48h after treatment, and 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 up-regulated expression of the disease-course-related protein genes in the apple branches, 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

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<213> 16S rDNA of Staphylococcus equine F1 (16S rDNA of Staphylococcus equirum F1)

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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

1. The application of the Staphylococcus equi in preventing and treating the fruit and vegetable diseases is characterized in that 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.

2. The apple tree rot biocontrol microbial inoculum is characterized in that the active ingredient of the apple tree rot biocontrol microbial inoculum is fermentation supernatant or bacterial suspension of Staphylococcus equi (Staphylococcus equi) F1, and the preservation number of the apple tree rot biocontrol microbial inoculum is CGMCC No. 21658.

3. The apple tree rot biocontrol microbial inoculum of claim 2, wherein the concentration of staphylococcus equinus strain F1 is 10⁵-10⁹cfu·mL^-1。

4. A method for inducing disease resistance of fruit tree branches is characterized by comprising the following steps: applying the bacterial liquid of staphylococcus equi-gastric according to claim 1 to the surface of fruit tree branches.

5. The method according to claim 4, 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.

6. The method according to claim 5, wherein the concentration of the Staphylococcus equine gastral bacteria liquid used is not less than 10⁹cfu·mL^-1。