CN114574381B - Antagonistic strain JYC314 and application thereof - Google Patents

Abstract

The invention relates to an antagonistic strain JYC314 and application thereof, belonging to the technical field of biological control of plant diseases.

Description

Antagonistic strain JYC314 and application thereof

Technical Field

The invention belongs to the technical field of biological control of plant diseases, particularly relates to an antagonistic strain JYC314 for inhibiting the growth of streptomyces solani and phytophthora infestans, and belongs to the technical field of biological control of plant diseases.

Background

The potato scab is a typical soil-borne disease caused by streptomyces spp infection, and the streptomyces only infects the epidermis of potatoes and does not penetrate into potato flesh, scabs on the surfaces of tubers to form scab spots, so that the appearance quality of the potato blocks is reduced, and the development of the potato industry, particularly the potato industry, is seriously influenced. At present, scholars at home and abroad report a plurality of methods for preventing and treating potato scab, and mainly focus on aspects of breeding of disease-resistant varieties, regulating water, adjusting soil pH value, crop rotation and applying green manure, chemical prevention, biological prevention and the like. The species of potato scab pathogens identified at present are 26 species, such as S.scabies, S.acidiscabies, S.turigidicabes, etc., as can be seen from the literature. According to the reports of related data, antagonistic strains having a certain inhibiting effect on potato scab pathogenic bacteria mainly comprise: bacillus simplex, Bacillus belgii, Bacillus carlsbergii, Bacillus amyloliquefaciens, Paecilomyces gossypii, Trichoderma asperellum, Candida citricola and the like, but not antagonistic strains all have an inhibitory effect on all Streptomyces scabies. For potato scab biocontrol resources, the biocontrol resources are deficient, the main bacillus is taken as a main bacillus, the antibacterial spectrum is single, the antibacterial effect is not obvious, and the scab pathogens are various in species and complex in composition, so that the potato scab biocontrol resources provide challenges for prevention and treatment of potato scab. Therefore, it is necessary to screen more biocontrol bacteria having a good inhibitory effect on various streptomyces scabies.

The potato late blight is a destructive disease caused by phytophthora infestans (Mont.) debry) of oomycetes, and the late blight can infect the whole potato plant, and leaves, petioles, stems and tubers can be damaged. At present, chemical control still occupies an important position in the control measures of potato late blight at home and abroad. In the epidemic period of the late blight, chemical prevention and control are the main measures for controlling the spread of the potato late blight at present, and the prevention and control of the potato late blight are guided by spraying medicaments such as metalaxyl, dimethomorph, propamocarb fluopicolide and the like and combining the use of an early warning system, so that good effects of preventing diseases and increasing yield can be achieved. Although chemical control is an effective means for preventing potato diseases and is one of the most widely adopted measures, the long-term frequent use of chemical agents easily pollutes the environment, so that the pesticide residue of agricultural products exceeds the standard; meanwhile, pathogenic bacteria can also generate drug resistance and even drug resistance to chemical agents, so that the prevention and treatment effect of late blight is reduced and even the late blight completely loses efficacy.

The biological prevention and control of plant diseases is to control the harm of pathogenic microorganisms by utilizing the antagonism of beneficial microorganisms originally existing in nature to target pathogenic microorganisms, has smaller environmental risk, and is a prevention and control technology which is friendly to the environment. With the attention of people on food safety and ecological safety, biological control becomes one of important strategies for realizing the sustainable development of modern agriculture, and the beneficial microorganisms are utilized to control plant diseases, so that the method has a wide prospect.

Disclosure of Invention

The invention aims to provide an antagonistic strain which can obviously inhibit the growth of various streptomyces scabies and phytophthora infestans and can effectively prevent and treat potato scab and late blight.

In order to realize the purpose, the invention is realized by the following technical scheme:

the invention provides an antagonistic strain JYC314 (pseudomonas aeruginosa) which is currently deposited in China General Microbiological Culture Collection Center (CGMCC)) at the microbial institute of China academy of sciences No. 3 of North Cheng West Lu 1 of the sunward area in Beijing, wherein the deposit number is CGMCC NO.24047, and the preservation date is 12 months and 06 days 2021.

The invention provides application of the antagonistic strain JYC314 in plant disease control; the plant disease is late blight or scab of potato.

The invention provides application of the antagonistic strain JYC314 in preparation of plant disease or pathogen inhibitor; pathogenic bacteria are Streptomyces scabiosus or Phytophthora infestans; the plant disease is late blight or scab of potato.

The invention also provides a pathogenic bacteria inhibitor, which comprises the antagonistic strain JYC314 and/or a bacterial liquid of the antagonistic strain JYC 314.

The invention has the beneficial effects that:

the antagonistic strain is sampled and separated from the rhizosphere soil of healthy plants in a field with a serious potato scab disease, screened and verified by a flat plate antagonism method and a paper method, and identified mainly from the aspects of morphology, physiology, biochemistry and molecular biology, so that a new biocontrol resource is provided for the prevention and treatment of the potato scab disease and the late blight. The antagonistic strain JYC314 can obviously inhibit the growth of various streptomyces scab and phytophthora infestans, and can effectively prevent and treat potato scab and late blight.

Drawings

FIG. 1 shows the inhibition effect of strain JYC314 on Streptomyces solani-plate confrontation test;

FIG. 2 shows the inhibition effect of JYC314 strain on potato late blight pathogen-plate confrontation test;

FIG. 3 is a phylogenetic tree constructed by strain JYC314 based on the 16SrDNA sequence;

FIG. 4 shows the inhibition effect of a suspension of strain JYC314 on various Streptomyces solani Leptosphaera-paper sheet method;

FIG. 5 shows the inhibition effect of JYC314 on the growth of various Phytophthora infestans, by hypha weight method;

FIG. 6 shows morphological identification of strain JYC 314;

FIG. 7 is a physiological and biochemical assay, glycolysis assay, of strain JYC 314; a: comparison; b: antagonistic strain JYC 314;

FIG. 8 is a physiological and biochemical assay of strain JYC 314-hydrolysis assay of macromolecular substances; a: comparison; b: antagonistic strain JYC 314;

FIG. 9 is an IMViC assay, a physiological and biochemical assay of strain JYC 314; a: comparison; b: antagonistic strain JYC 314.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, preferred embodiments of the present invention will be described in detail below to facilitate understanding of the skilled person.

Example 1: separation and purification of potato scab antagonistic strain JYC314

1. Soil sample collection

Collecting rhizosphere soil of healthy plants from potato scab disease field in Yunnan, collecting depth of 10-20cm, sealing with sterile self-sealing bag, taking back to laboratory, and storing in refrigerator at 4 deg.C for separating antagonistic strains.

2. Target pathogenic microorganism for test

3 strains of streptomyces solani scab (S.scabies, S.acidiscabies and S.turigiiscabies) and 3 strains of potato late blight bacteria (P.infestans), wherein the strain numbers and the types of physiological race are SNK-Q9(1.2.3.4.5.6.7.8.9.10.11), DQ-1(1.2.3.4.5.10) and ML-S13-418(1.2.4.5.6.10), and the strains are provided by the research laboratory of potato diseases of plant protection institute of Yunnan agricultural university.

3. Test medium

The streptomyces scabies culture adopts a Gao's first culture medium or an oat agar culture medium, and the bacterial culture adopts a beef extract peptone culture medium. The phytophthora infestans is cultured by adopting a rye tomato culture medium. The specific culture medium formula is as follows:

oat agar medium (OMA): 20g of oatmeal, 20g of agar strips and 1000ml of water, and the pH value is 7.2-7.4;

beef extract peptone medium (NA): 3g of beef extract, 10g of peptone, 1000ml of water, 20g of agar strips and pH 7.4-7.6;

rye tomato medium: 70g of rye, 150m of tomato juice, 850ml of water, 1.2g of calcium carbonate and 17g of agar strips, and the pH value is natural;

4. isolation and purification of antagonistic Strain JYC314

Separating strains by heat treatment soil dilution separation method, selecting single colony strain, streaking, culturing, continuously purifying for 3 times, mixing in 70% glycerol at a ratio of 1:1, and storing in refrigerator at-80 deg.C for use.

Preparation of OMA Medium: weighing the substances according to the formula of the culture medium, soaking the agar strips and the oatmeal in 400ml of water for 30min, crushing the oatmeal by using a juicer, heating and boiling the oatmeal and the agar strips together, cooling the culture medium, adjusting the pH of the culture medium to 7.2-7.4 by using 1% sodium hydroxide solution, subpackaging the culture medium into 2 1000ml triangular bottles, wherein each bottle contains 500ml, sealing the bottles, placing the bottles in an autoclave for sterilization at 121 ℃ for 30min, sterilizing once again when the air pressure is reduced to 0Mpa, pouring the culture medium into a 90mm culture plate, cooling and drying the culture medium on an ultraclean workbench for later use.

6. Preparation of a streptomyces suspension: the streptomycete strain bacterial suspension stored in the laboratory is sucked on the surface of an OMA culture medium flat plate, a sterile metal spreader is uniformly smeared, a sealing film is sealed and then is inverted into a microbial incubator, the culture is performed for 7 to 10 days at 28 ℃ in a dark environment, 2ml of sterile water is added into a culture dish on a clean bench after the culture medium surface is full of streptomycete scabies, the streptomycete is scraped from the culture medium surface by using a sterile spatula, the liquid in the culture dish is sucked by a pipetting gun to wash the culture medium surface for 2 to 3 times, and the liquid in the dish is transferred into a 5ml sterile centrifuge tube for standby.

7. Screening of antagonistic strains

The antagonistic action of the strain JYC314 on the potato streptomyces scaber is preliminarily measured by adopting a plate confronting culture method. The specific operation method comprises the following steps: inoculating the purified strain JYC314 to an oat agar culture medium plate coated with a streptomyces scab bacterial suspension, sealing a sealing film, placing the sealed plate in an incubator at 28 ℃ for dark culture for 7 days, taking an OMA plate only inoculated with the streptomyces scab bacterial suspension as a control, and screening out a strain with a good bacteriostatic effect according to whether a bacteriostatic zone appears on the culture medium plate or not and the size of the bacteriostatic zone when the culture time is up; the results of the plate confronting culture method show that the 3 streptomyces scabies in the control culture medium can normally grow in the oat agar culture medium and can be distributed on the whole culture medium surface, and no bacteriostatic rings appear; the surfaces of the culture mediums inoculated with the JYC314 strain all have inhibition zones, which shows that the JYC314 strain can inhibit the growth of S.tuligiensis, S.acidiscabies and S.scabies of the streptomyces scabies, and has inhibition on 3 streptomyces scabies (figure 1).

The antagonistic action of the strain JYC314 on phytophthora infestans is measured by adopting a plate confronting culture method, a sterile puncher is used for respectively punching a rye tomato culture medium plate full of phytophthora infestans hyphae and an independent rye tomato culture medium plate, a fungus cake containing the phytophthora infestans hyphae is reversely buckled in a small hole in the center of the rye tomato culture medium plate, the thallus of the strain to be measured is picked to the rye tomato culture medium plate containing the phytophthora infestans fungus cake, the rye tomato culture medium plate is placed in a microorganism incubator at 20 ℃ for dark culture for 7-10 days, and the colony growth diameter is measured by adopting a cross measurement method after the culture is finished. The results of plate confrontation culture show that phytophthora infestans in a control culture medium can normally grow and can grow on the whole rye tomato culture medium plate, the colony growth diameter is 56.19mm, the colony growth diameters of the phytophthora infestans in a 3-point inoculation method and a 4-point inoculation method are 14.27mm and 0mm respectively, and the significant difference (P <0.05) is obtained compared with the control, which indicates that the strain JYC314 has a significant inhibition effect on the growth of phytophthora infestans (Table 1 and figure 2).

TABLE 1 inhibitory Effect of JYC314 Strain on potato late blight pathogen

Note: lower case letters in the table indicate significance of difference at the 0.05 level

In conclusion, according to the results of plate confrontation culture, a strain which has an inhibiting effect on the growth of the streptomyces solani scab and phytophthora infestans is obtained and named as JYC 314.

Example 2: molecular biological identification of antagonistic strain JYC314

1. Extraction of bacterial genomic DNA

The single colony of the strain JYC314 prepared and selected in the example 1 and purified and cultured is inoculated on a beef extract peptone culture plate, after dark culture for 120 hours at 28 ℃, the genome DNA of the strain JYC314 is extracted by adopting a Shanghai worker EZUP column type bacterial genome DNA extraction kit, and the specific steps are as follows:

(1) 0.5ml to 1ml of bacterial liquid cultured overnight is taken and added into a 1.5ml centrifuge tube, the room temperature is 8000rpm, the centrifugation is carried out for 1min, and the supernatant and the cell of the mobile phone are discarded. Adding 100 microliters of BufferDigestinon and 80 microliters of 20mg/ml lysozyme solution, carrying out water bath at 37 ℃ for 30min, adding 20 microliters of protease K solution after the water bath is finished, and uniformly mixing by shaking. And carrying out water bath at 56 ℃ for 30min until the cells are completely lysed. Adding 20 microliter 10mg/ml RNaseA solution after water bath, standing at room temperature for 2-5min,

(2) 200 microliters of BufferBD was added, mixed well by inversion, and then subjected to a water bath at 70 ℃ for 10 min.

(3) Add 200. mu.l of absolute ethanol and mix well by inversion.

(4) Putting the adsorption column into a collecting tube, adding the solution and the semitransparent fibrous suspended matters into the adsorption column by using a pipette, standing for 2min, centrifuging at 12000rpm for 1min at room temperature, and pouring out waste liquid in the collecting tube.

(5) Putting the adsorption column into a collecting pipe, adding 500 microliter PW Solution, centrifuging at 10000rpm for 30s, and pouring off the filtrate;

(6) putting the adsorption column into a collecting pipe, adding 500 microliter of Wash Solution, centrifuging at 10000rpm for 30s, and pouring off the filtrate;

(7) putting the adsorption column into a collecting pipe, adding 500 microliters of PW Solution, and centrifuging at 12000rpm for 2min to pour the filtrate;

(8) the adsorption column was taken out, and put into 1 new 1.5ml centrifuge tube, 50-100. mu.l of CE Buffer was added and left to stand for 3min, and centrifuged at 12000rpm for 2min, and the DNA solution was collected. The extracted DNA was stored at-20 ℃.

2.16 amplification of SrDNA sequences

16SrDNA of the strain JYC314 is amplified by using the genomic DNA of the strain JYC314 as a template and using universal primers 27F (5 '-AGTTTGATCMTGGCTCAG-3') and 1492R (5'-GGTTACCTTGTTACGACTT-3'). The PCR amplification conditions are pre-denaturation 94 ℃/4min, denaturation 94 ℃/45s, annealing 55 ℃/45s, extension 72 ℃/1min, circulation 30 times, and final extension 72 ℃/10 min. The PCR reaction system is as follows:

TABLE 2 PCR reaction System

The PCR amplification product is detected by agarose gel electrophoresis, the result shows that a bright band can be obtained, about 1500bp amplification product is obtained, the primer is used for sequencing the amplification gene segment after the electrophoresis band is recovered, the sequencing work is finished by Shanghai biological engineering Limited company, the nucleotide sequence length of the 16SrDNA of the strain is 1429bp, and the 16SrDNA gene sequencing result is as follows:

ATGGCTCAGATTGAACGCTGGCGGCAGGCCTAACACATGCAAGTCGAGCGGATGAAGGGAGCTTGCTCCTGGATTCAGCGGCGGACG GGTGAGTAATGCCTAGGAATCTGCCTGGTAGTGGGGGATAACGTCCGGAAACGGGCGCTAATACCGCATACGTCCTGAGGGAGAAAGTGGG GGATCTTCGGACCTCACGCTATCAGATGAGCCTAGGTCGGATTAGCTAGTTGGTGGGGTAAAGGCCTACCAAGGCGACGATCCGTAACTGG TCTGAGAGGATGATCAGTCACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGCGAAAG CCTGATCCAGCCATGCCGCGTGTGTGAAGAAGGTCTTCGGATTGTAAAGCACTTTAAGTTGGGAGGAAGGGCAGTAAGTTAATACCTTGCT GTTTTGACGTTACCAACAGAATAAGCACCGGCTAACTTCGTGCCAGCAGCCGCGGTAATACGAAGGGTGCAAGCGTTAATCGGAATTACTG GGCGTAAAGCGCGCGTAGGTGGTTCAGCAAGTTGGATGTGAAATCCCCGGGCTCAACCTGGGAACTGCATCCAAAACTACTGAGCTAGAGT ACGGTAGAGGGTGGTGGAATTTCCTGTGTAGCGGTGAAATGCGTAGATATAGGAAGGAACACCAGTGGCGAAGGCGACCACCTGGACTGAT ACTGACACTGAGGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGTCGACTAGCCGTTGGGAT CCTTGAGATCTTAGTGGCGCAGCTAACGCGATAAGTCGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACGGGGGC CCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCTGGCCTTGACATGCTGAGAACTTTCCAGAGATGGAT TGGTGCCTTCGGGAACTCAGACACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGTAACGAGCGCAAC CCTTGTCCTTAGTTACCAGCACCTCGGGTGGGCACTCTAAGGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCATC ATGGCCCTTACGGCCAGGGCTACACACGTGCTACAATGGTCGGTACAAAGGGTTGCCAAGCCGCGAGGTGGAGCTAATCCCATAAAACCGA TCGTAGTCCGGATCGCAGTCTGCAACTCGACTGCGTGAAGTCGGAATCGCTAGTAATCGTGAATCAGAATGTCACGGTGAATACGTTCCCG GGCCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTGCTCCAGAAGTAGCTAGTCTAACCGCAA

the sequencing results were entered into the National Center for Biotechnology Information (NCBI) database and compared with other 16SrDNA sequences in GenBank using BLAST software, which showed that the 16SrDNA sequence of strain JYC314 completely coincided with the 16SrDNA sequences of several strains of Pseudomonas aeruginosa. A similar sequence and a sequence of the strain JYC314 are selected, a phylogenetic evolutionary tree (figure 3) is constructed by using a Neighbor-Joining Method through a systematic differentiation analysis software MEGA7.0, the result shows that the strain JYC314 and the Pseudomonas aeruginosa strain S-04 MT626658.1 are in the same phylogenetic branch, the evolutionary relationship is recent, the homology reaches 99.31 percent, and the strain JYC314 is preliminarily judged to be the Pseudomonas aeruginosa (Pseudomonas aeruginosa).

Example 3: effect of antagonistic strain JYC314 bacterial suspension on growth of streptomyces scabies

1. Materials and methods

Test materials:

test strains: (ii) s.scabies, s.acidiscabies, s.turigiiscabies;

other materials: 18mm qualitative circular filter paper sheets, metal coating rods, 90mm culture dishes, sealing films, pipette guns, centrifuge tubes, sterile water, juice extractors, metal scrapers, metal tweezers and digital display vernier calipers;

the test method comprises the following steps:

OMA medium and Streptomyces suspensions were prepared as in example 1.

Preparing antagonistic strain suspension: and picking the purified antagonistic strain by using an aseptic gun head into a 5ml aseptic centrifuge tube containing 1ml of aseptic water, and performing vortex oscillation for 30 seconds by using an oscillation mixer for later use.

A streptomyces scabies inhibition test is carried out by adopting a paper sheet method: sucking 100 mul of streptomyces suspension into an OMA culture medium plate, uniformly smearing the streptomyces suspension by using a sterile metal coating rod, clamping a sterile filter paper sheet by a sterile forceps, placing the sterile filter paper sheet in the center of the culture plate, slightly pressing the filter paper sheet along the periphery to enable the filter paper sheet to be tightly attached to the surface of the culture medium, sucking 50 mul of bacterial suspension of a strain to be detected by a liquid transfer gun, slowly releasing the bacterial suspension of the strain to be detected on the filter paper sheet, after the surface of the filter paper sheet is dried in the air, sealing a sealing film, placing the filter paper sheet in a microorganism incubator, carrying out dark culture at 28 ℃ for 7-10 days, inoculating sterile water on the OMA culture medium plate coated with the streptomyces scab suspension, placing the filter paper sheet as a contrast, and measuring the diameter of an inhibition zone by adopting a cross measurement method after the culture time is cut off.

2. Results and analysis

The inhibition of 3 kinds of streptomyces solanacearum S.turigidicabes, S.acidiscabies and S.scabies by the strain JYC314 was tested again by the bacteriostatic circle method. The results show that: the determination result is that the control 1 and the control 2 can grow over the surface of the culture medium, and no inhibition zone appears. The surfaces of the culture media added with the JYC314 bacterial suspension to be detected have inhibition zones, the diameters of the inhibition zones are respectively 27.75mm, 27.50mm and 38.66mm, and the inhibition zones have significant differences (p <0.05) compared with a control (0mm), which shows that the JYC314 can significantly inhibit the growth of scab pathogens S.scab, S.turigis, S.acidiscabies, and has the most obvious inhibition effect on S.scab. (FIG. 4)

TABLE 3 Effect of the Strain JYC314 on the growth of the pathogenic bacteria of potato scab

Note: in the table, the lower case indicates the significance of the difference at the level of 0.05

Example 4: effect of antagonistic strain JYC314 on the growth of various phytophthora infestans

1. Materials and methods

Numbers of phytophthora infestans strains and physiological race types tested: SNK-Q9(1.2.3.4.5.6.7.8.9.10.11), DQ-1(1.2.3.4.5.10), ML-S13-418 (1.2.4.5.6.10).

Other materials: a rake type bamboo stick, a 5mm puncher, a 90mm culture dish, a sealing film and a ten thousandth balance.

The test method comprises the following steps:

the rye tomato medium was prepared as in example 1.

The antagonistic effect of the strain JYC314 on phytophthora infestans is measured by adopting a plate confronting culture method. The specific operation method comprises the following steps: respectively punching a rye tomato culture medium plate full of phytophthora infestans hyphae and an independent rye tomato culture medium plate by using an aseptic puncher, reversely buckling a fungus cake containing the phytophthora infestans hyphae in a small hole in the center of the rye tomato culture medium plate, picking thalli of a strain to be detected to the rye tomato culture medium plate containing the phytophthora infestans fungus cake, placing the rye tomato culture medium plate on a microbial incubator at 20 ℃ for dark culture for 7-10 days, stripping the phytophthora infestans hyphae in the culture dish from the surface of the rye tomato culture medium by using a rake type bamboo stick after the culture is finished, placing a ten-thousandth balance for weighing, and taking the weight of the hyphae as an index for measuring the growth of the hyphae.

2. Results and analysis

The weights of ML-S13-418, DQ-1, SNK-Q93 phytophthora infestans hyphae under the stress of the strain JYC314 were determined. The results show that 3 strains in the control culture can normally grow, the weights of hyphae are respectively 0.0169g, 0.0488g and 0.0618g, and the weights of ML-S13-418, DQ-1 and SNK-Q9 hyphae in the culture medium inoculated with the strain JYC314 are all obviously lower than the control (p is less than 0.05), which indicates that the hyphae growth is obviously inhibited under the stress of the strain JYC314, and the hyphae growth inhibition is increasingly obvious along with the increase of the inoculation amount of the JYC 314.

TABLE 4 influence of the Strain JYC314 on the amount of hyphae growth of the bacterial species of potato late blight

Note: in the table, the lower case indicates the significance of the difference at the level of 0.05

Example 5: morphological identification of antagonistic strain JYC314

And selecting single colonies of the separated and purified JYC314 strain, streaking the single colonies on a beef extract peptone culture medium, culturing for 48 hours, observing morphological characteristics of the colonies, and simultaneously determining the cell wall type of the strain by using a gram staining method. The result of morphological observation shows that the strain JYC314 grows well on a beef extract peptone culture medium, bacterial colonies are milky white or grey white, round, smooth and moist, have a raised center and irregular edges, the thalli are rod-shaped, have the size of 0.6-0.8 mu m multiplied by 2.0-5.0 mu m, are gram-positive, produce spores, and grow aerobically or facultative anaerobically. The colony periphery is light green, and the green color deepens with the prolonging of the culture time, and turns to purple or brown at the later stage (FIG. 6).

Example 6: physiological and biochemical determination results of antagonistic strain JYC314

See the handbook of identification of common bacteria systems for measuring the physiological and biochemical indexes of the thallus, such as saccharide fermentation, gelatin liquefaction test, hydrogen sulfide generation test, starch hydrolysis test, methyl red test, VP test, indole generation test, litmus milk test, citrate utilization test, urease generation test and the like. The results of physiological and biochemical tests show that the strain JYC314 can decompose glucose to produce acid, produce gelatinase to decompose gelatin, utilize citrate and produce H ₂ S gas, the test result of the milk litmus is positive; the strain JYC314 does not degrade lactose, does not hydrolyze starch, does not produce urease to decompose urea, and has negative results of methyl red test, indole production test and VP test (see table 5 and figures 7-9).

TABLE 5 physiological and biochemical characteristics of the antagonistic Strain JYC314

Note: in the table, "+" indicates a positive result; "-" indicates a negative result.

In conclusion, according to the sequence analysis, morphological characteristics and physiological and biochemical characteristics of the strain JYC 31416SrDNA, the strain JYC314 is judged to belong to the Pseudomonas aeruginosa (Pseudomonas aeruginosa) by combining the general bacteria system identification handbook, and the strain has obvious inhibiting effect on various streptomyces solani scab and phytophthora infestans, and can effectively prevent and treat the potato scab and late blight.

Finally, it is noted that the above-mentioned preferred embodiments illustrate rather than limit the invention, and that, although the invention has been described in detail with reference to the above-mentioned preferred embodiments, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the scope of the invention as defined by the appended claims.

SEQUENCE LISTING

<110> Yunnan university of agriculture

<120> antagonistic strain JYC314 and application thereof

<130> 2021

<160> 1

<170> PatentIn version 3.5

<210> 1

<211> 1429

<212> DNA

<213> Artificial sequence

<400> 1

atggctcaga ttgaacgctg gcggcaggcc taacacatgc aagtcgagcg gatgaaggga 60

gcttgctcct ggattcagcg gcggacgggt gagtaatgcc taggaatctg cctggtagtg 120

ggggataacg tccggaaacg ggcgctaata ccgcatacgt cctgagggag aaagtggggg 180

atcttcggac ctcacgctat cagatgagcc taggtcggat tagctagttg gtggggtaaa 240

ggcctaccaa ggcgacgatc cgtaactggt ctgagaggat gatcagtcac actggaactg 300

agacacggtc cagactccta cgggaggcag cagtggggaa tattggacaa tgggcgaaag 360

cctgatccag ccatgccgcg tgtgtgaaga aggtcttcgg attgtaaagc actttaagtt 420

gggaggaagg gcagtaagtt aataccttgc tgttttgacg ttaccaacag aataagcacc 480

ggctaacttc gtgccagcag ccgcggtaat acgaagggtg caagcgttaa tcggaattac 540

tgggcgtaaa gcgcgcgtag gtggttcagc aagttggatg tgaaatcccc gggctcaacc 600

tgggaactgc atccaaaact actgagctag agtacggtag agggtggtgg aatttcctgt 660

gtagcggtga aatgcgtaga tataggaagg aacaccagtg gcgaaggcga ccacctggac 720

tgatactgac actgaggtgc gaaagcgtgg ggagcaaaca ggattagata ccctggtagt 780

ccacgccgta aacgatgtcg actagccgtt gggatccttg agatcttagt ggcgcagcta 840

acgcgataag tcgaccgcct ggggagtacg gccgcaaggt taaaactcaa atgaattgac 900

gggggcccgc acaagcggtg gagcatgtgg tttaattcga agcaacgcga agaaccttac 960

ctggccttga catgctgaga actttccaga gatggattgg tgccttcggg aactcagaca 1020

caggtgctgc atggctgtcg tcagctcgtg tcgtgagatg ttgggttaag tcccgtaacg 1080

agcgcaaccc ttgtccttag ttaccagcac ctcgggtggg cactctaagg agactgccgg 1140

tgacaaaccg gaggaaggtg gggatgacgt caagtcatca tggcccttac ggccagggct 1200

acacacgtgc tacaatggtc ggtacaaagg gttgccaagc cgcgaggtgg agctaatccc 1260

ataaaaccga tcgtagtccg gatcgcagtc tgcaactcga ctgcgtgaag tcggaatcgc 1320

tagtaatcgt gaatcagaat gtcacggtga atacgttccc gggccttgta cacaccgccc 1380

gtcacaccat gggagtgggt tgctccagaa gtagctagtc taaccgcaa 1429

Claims (4)

1. Antagonistic strain Pseudomonas aeruginosa (Pseudomonas aeruginosa) JYC314, characterized by: the preservation number is CGMCC NO. 24047.

2. The antagonistic strain Pseudomonas aeruginosa (Pseudomonas aeruginosa) JYC314 as claimed in claim 1 for use in the prevention and treatment of potato late blight or potato scab.

3. The use of the antagonistic strain Pseudomonas aeruginosa (Pseudomonas aeruginosa) JYC314 as claimed in claim 1 for the preparation of a pathogen inhibitor, wherein: the pathogenic bacteria are Streptomyces scabies or Phytophthora infestans.

4. A pathogen inhibitor characterized by: a bacterial suspension comprising the antagonistic strain Pseudomonas aeruginosa (Pseudomonas aeruginosa) JYC314 or the antagonistic strain Pseudomonas aeruginosa (Pseudomonas aeruginosa) JYC314 of claim 1.

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