CN117551733A - Kiwi fruit pollen canker detection method and kit thereof - Google Patents
Kiwi fruit pollen canker detection method and kit thereof Download PDFInfo
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- CN117551733A CN117551733A CN202311475243.6A CN202311475243A CN117551733A CN 117551733 A CN117551733 A CN 117551733A CN 202311475243 A CN202311475243 A CN 202311475243A CN 117551733 A CN117551733 A CN 117551733A
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- 244000298697 Actinidia deliciosa Species 0.000 title claims abstract description 62
- 235000009436 Actinidia deliciosa Nutrition 0.000 title claims abstract description 62
- 238000001514 detection method Methods 0.000 title claims abstract description 36
- 208000025865 Ulcer Diseases 0.000 claims abstract description 9
- 239000001963 growth medium Substances 0.000 claims description 26
- 238000000034 method Methods 0.000 claims description 17
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 15
- 239000002609 medium Substances 0.000 claims description 15
- 238000000825 ultraviolet detection Methods 0.000 claims description 10
- 208000015181 infectious disease Diseases 0.000 claims description 8
- 241000894006 Bacteria Species 0.000 claims description 7
- 238000012258 culturing Methods 0.000 claims description 7
- 229920001817 Agar Polymers 0.000 claims description 6
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 claims description 6
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 6
- 239000001888 Peptone Substances 0.000 claims description 6
- 108010080698 Peptones Proteins 0.000 claims description 6
- 239000008272 agar Substances 0.000 claims description 6
- 239000012153 distilled water Substances 0.000 claims description 6
- 235000011187 glycerol Nutrition 0.000 claims description 6
- 235000019319 peptone Nutrition 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 238000003794 Gram staining Methods 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- 229930186147 Cephalosporin Natural products 0.000 claims description 3
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 claims description 3
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical group C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims description 3
- 239000005018 casein Substances 0.000 claims description 3
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 claims description 3
- 235000021240 caseins Nutrition 0.000 claims description 3
- 229940124587 cephalosporin Drugs 0.000 claims description 3
- 150000001780 cephalosporins Chemical class 0.000 claims description 3
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 3
- 229910001629 magnesium chloride Inorganic materials 0.000 claims description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 3
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 3
- OTYBMLCTZGSZBG-UHFFFAOYSA-L potassium sulfate Chemical compound [K+].[K+].[O-]S([O-])(=O)=O OTYBMLCTZGSZBG-UHFFFAOYSA-L 0.000 claims description 3
- 229910052939 potassium sulfate Inorganic materials 0.000 claims description 3
- 235000011151 potassium sulphates Nutrition 0.000 claims description 3
- 208000035285 Allergic Seasonal Rhinitis Diseases 0.000 claims 4
- 206010048908 Seasonal allergy Diseases 0.000 claims 4
- 230000001954 sterilising effect Effects 0.000 claims 1
- 238000004659 sterilization and disinfection Methods 0.000 claims 1
- 231100000397 ulcer Toxicity 0.000 abstract description 3
- 230000001580 bacterial effect Effects 0.000 description 7
- 239000000523 sample Substances 0.000 description 5
- 235000009434 Actinidia chinensis Nutrition 0.000 description 4
- 235000013399 edible fruits Nutrition 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 4
- 241000589615 Pseudomonas syringae Species 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 230000002146 bilateral effect Effects 0.000 description 2
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- 230000000694 effects Effects 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- 238000000265 homogenisation Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 239000012804 pollen sample Substances 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000197 pyrolysis Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- BDHFUVZGWQCTTF-UHFFFAOYSA-M sulfonate Chemical compound [O-]S(=O)=O BDHFUVZGWQCTTF-UHFFFAOYSA-M 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/38—Caps; Covers; Plugs; Pouring means
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/045—Culture media therefor
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/38—Pseudomonas
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention relates to the technical field of detection methods of kiwi fruit pollen ulcer, in particular to a detection method of kiwi fruit pollen ulcer and a kit thereof.
Description
Technical Field
The invention relates to a method for detecting kiwi fruit pollen canker, in particular to a method for detecting kiwi fruit pollen canker and a kit thereof.
Background
The kiwi canker is a destructive bacterial disease which seriously threatens the production of kiwi fruits and is listed as a national forest plant quarantine object. The disease has fierce appearance, the epidemic year causes the whole garden to be endangered, and serious economic loss is caused, the occurrence hazard of the disease not only reduces the yield, but also causes the fruit peel to become thicker, the fruit becomes sour, the fruit becomes smaller, the fruit shape is different, the quality is reduced, and the commodity value is reduced. Therefore, the method is very necessary for timely detection and treatment of the kiwi pollen ulcer.
After the first discovery of kiwi fruit canker in California and Jinggang county of Japan in 1980, expert scholars in the United states, new Zealand, japan and China developed the identification of kiwi fruit canker, and most of kiwi fruit canker bacteria were considered to belong to Pseudomonas syringae. Therefore, the detection operation of the kiwi fruit pollen canker can be realized based on the detection of pseudomonas syringae.
In the prior art, the detection operation of pseudomonas syringae in the kiwi fruit pollen canker is mostly adopted as a Chinese invention CN201911290716.9 method for rapidly detecting the canker of kiwi fruit pollen, and DNA and PCR specific amplification and other methods are obtained through tissue homogenization, bacterial culture and high-temperature pyrolysis, so that the rapid identification of the canker of kiwi fruit pollen samples is realized, and the development of a rapid detection method of pathogenic bacteria based on DNA is promoted. The detection mode is accurate, but needs to spend a great deal of time and detection cost, so that the cost and complexity of the detection of the kiwi fruit pollen canker are improved, and the timely discovery and treatment operation of the kiwi fruit pollen canker are not facilitated.
Based on the reasons, the invention provides a method for detecting the kiwi fruit pollen canker and a kit thereof, which solve the defects of the prior art.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a kiwi fruit pollen canker detection method and a kit thereof.
In order to solve the technical problems, the invention provides the following technical scheme:
the kiwi fruit pollen canker detection method comprises the following steps:
s1: collecting and processing samples of kiwi fruit tree infection parts;
s2: preparing a culture medium, wherein the culture medium comprises a cultivation medium and an identification medium;
s3: filling the extracted kiwi fruit tree infection part sample into a culture medium for culturing strains;
s4: observing the colonies cultured on the culture medium, and carrying out irradiation identification on the colonies on the culture medium through ultraviolet detection equipment;
s5: extracting the fluorescent bacteria after irradiation of the ultraviolet detection equipment, and filling the extracted bacteria on an identification culture medium for culture;
s6: performing a gram staining procedure on the identification medium and observing the results;
s7: if red is observed, the kiwi fruit is infected with kiwi fruit pollen canker, otherwise, the kiwi fruit is not infected with kiwi fruit pollen canker.
As a preferable technical scheme of the invention, in the step 1, after the sample of the kiwi fruit infection part is collected, the sample is crushed.
As a preferable technical scheme of the invention, the culture medium in the step 2 comprises 4 g of peptone, 0.3 g of dipotassium hydrogen phosphate, 0.3 g of magnesium sulfate, 3 g of agar, 2 ml of glycerol and 200 ml of distilled water, and the components are boiled and fully dissolved in the culture medium during the preparation, and the pH value is adjusted to 7.2, and the culture medium is autoclaved at 121 ℃ for 15 minutes.
As a preferable technical scheme of the invention, the identification medium in the step 2 comprises 3.2 g of polyvalent peptone, 2 g of hydrolyzed casein, 2 g of potassium sulfate, 0.3 g of magnesium chloride, 2.2 g of agar, 2 ml of glycerin and 200 ml of distilled water, and the components are boiled and fully dissolved in the preparation process, the PH value is regulated to 7.2, the mixture is autoclaved at 121 ℃ for 15 minutes, and the dissolved cetyltrimethylammonium bromide, sodium chain run and cephalosporin are added when the mixture is cooled to about 50 ℃.
As a preferable technical scheme of the invention, the colony irradiated by the ultraviolet detection equipment in the step 4 is a milky white, smooth and clean-edged colony.
As a preferred embodiment of the present invention, the culturing of the fluorogenic colonies in the step 5 is performed at 30℃for 48 hours.
The utility model provides a kiwi fruit pollen ulcerative disease detect reagent box, includes the fixed frame, the fixed frame is the I shape setting, all seted up the movable groove on the left and right sides wall of fixed frame, the left and right sides of fixed frame all is provided with the fly leaf, the fly leaf is the spill setting, the both sides lateral wall of fly leaf inserts the activity inslot respectively and with the swing joint between the movable groove, fixedly connected with first spring on the movable groove inner chamber lateral wall, first spring other end and fly leaf fixed connection, bilateral symmetry has inserted two kit casings in the fixed frame, fixedly connected with rectangle frame on the top surface of kit casing, bilateral symmetry swing joint has two apron in the rectangle frame, all seted up the gas pocket on the apron, all inserted the plug in the gas pocket.
As a preferable technical scheme of the invention, four circular grooves are symmetrically arranged on the inner side wall of the rectangular frame in a left-right mode, rotating shafts are movably connected in the circular grooves, the other ends of the two rotating shafts on the same side are fixedly connected with the same cover plate, a second spring is sleeved on the rotating shafts, one end of the second spring is fixedly connected with the inner wall of the circular groove, and the other end of the second spring is fixedly connected with the side wall of the cover plate.
The embodiment of the invention provides a kiwi fruit pollen canker detection method and a kit thereof, which have the following beneficial effects:
1. according to the invention, the detection sample of the kiwi fruit pollen canker is cultivated on two sides through the cultivation medium and the identification medium, on one hand, the once detection operation on the cultivated colonies can be realized through the ultraviolet detection equipment, on the other hand, the re-detection on the cultivated colonies can be realized through the gram staining method, and the kiwi fruit pollen canker can be determined only through the detection twice, so that the detection error caused by single detection measures is effectively avoided, and compared with the detection method of the gene level in the prior art, the detection method has the advantages of good operability and low cost;
2. according to the invention, the kit shell, the fixed frame, the movable plate and the cover plate are arranged, so that the culture medium and the identification medium can be respectively contained through the two kit shells during detection, the cover plate is used for sealing the inside of the kit shell, the interference of external factors to detection is reduced, the movable plate can be used for firmly placing the kit shell, the stability during detection is improved, the kit shells can be separated during detection, and the fixed frame is used for operating the kit shells, so that the mutual interference is reduced and the kit shell is convenient to use.
Drawings
The accompanying drawings are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate the invention and together with the embodiments of the invention, serve to explain the invention. In the drawings:
FIG. 1 is a flow chart of the method for detecting kiwi pollen canker of the invention;
FIG. 2 is a schematic overall perspective view of the kiwi pollen canker detection kit of the invention;
FIG. 3 is a schematic perspective view of the kit housing of the kiwi fruit pollen ulcer disease detection kit of the present invention;
FIG. 4 is a schematic diagram showing the internal perspective structure of a fixed frame of the kiwi fruit pollen ulcer disease detection kit of the present invention;
FIG. 5 is a schematic diagram showing a three-dimensional structure of a cover plate of the kiwi fruit pollen ulcer disease detection kit of the present invention;
in the figure: 1. a fixed frame; 2. a movable groove; 3. a movable plate; 4. a first spring; 5. a kit housing; 6. a rectangular frame; 7. a cover plate; 8. air holes; 9. a rubber plug; 10. a circular groove; 11. a rotating shaft; 12. and a second spring.
Detailed Description
The preferred embodiments of the present invention will be described below with reference to the accompanying drawings, it being understood that the preferred embodiments described herein are for illustration and explanation of the present invention only, and are not intended to limit the present invention.
Examples: as shown in fig. 1-5, the method for detecting kiwi fruit pollen canker comprises the following steps:
s1: collecting and processing samples of kiwi fruit tree infection parts, and crushing the samples after collecting the kiwi fruit tree infection parts;
s2: preparing a culture medium, wherein the culture medium comprises two kinds of culture medium and an identification medium, the culture medium comprises 4 g of peptone, 0.3 g of dipotassium hydrogen phosphate, 0.3 g of magnesium sulfate, 3 g of agar, 2 ml of glycerol and 200 ml of distilled water, the components are fully dissolved in boiling mode when being prepared, the pH value is regulated to 7.2, the culture medium is autoclaved at 121 ℃ for 15 minutes, the identification medium comprises 3.2 g of polyvalent peptone, 2 g of hydrolyzed casein, 2 g of potassium sulfate, 0.3 g of magnesium chloride, 2.2 g of agar, 2 ml of glycerol and 200 ml of distilled water, the components are fully dissolved in boiling mode when being prepared, the pH value is regulated to 7.2, the pH value is regulated to 121 ℃ for 15 minutes, and the dissolved cetyltrimethylammonium bromide, sodium chain sulfonate and cephalosporin are added when being cooled to about 50 ℃;
s3: filling the extracted kiwi fruit tree infection part sample into a culture medium for culturing strains;
s4: observing the bacterial colony cultured on the culture medium, and carrying out irradiation identification on the bacterial colony on the culture medium through ultraviolet detection equipment, wherein the bacterial colony irradiated by the ultraviolet detection equipment is a milky white, smooth and clean-edged bacterial colony;
s5: extracting the bacterial strain of the fluorogenic bacteria irradiated by the ultraviolet detection equipment, adding the extracted bacterial strain to an identification culture medium for culturing, and culturing the fluorogenic bacteria colony at 30 ℃ for 48 hours;
s6: performing a gram staining procedure on the identification medium and observing the results;
s7: if red is observed, the kiwi fruit is infected with kiwi fruit pollen canker, otherwise, the kiwi fruit is not infected with kiwi fruit pollen canker.
The kiwi fruit pollen canker detection kit comprises a fixed frame 1, wherein the fixed frame 1 is in an I-shaped structure, movable grooves 2 are formed in the left side wall and the right side wall of the fixed frame 1, movable plates 3 are arranged on the left side and the right side of the fixed frame 1, the movable plates 3 are in concave structures, the side walls on the two sides of the movable plates 3 are respectively inserted into the movable grooves 2 and are movably connected with the movable grooves 2, a first spring 4 is fixedly connected with the other end of the first spring 4 and the movable plates 3, two kit shells 5 are symmetrically inserted in the fixed frame 1 left and right, a rectangular frame 6 is fixedly connected with the top surface of the kit shells 5, two cover plates 7 are symmetrically and horizontally movably connected in the rectangular frame 6, air holes 8 are formed in the cover plates 7, rubber plugs 9 are inserted into the air holes 8, two culture mediums can be contained, and the kiwi fruit pollen canker can be conveniently detected twice through one kit;
four circular grooves 10 are symmetrically formed in the inner side wall of the rectangular frame 6, rotating shafts 11 are movably connected in the circular grooves 10, the other ends of the two rotating shafts 11 on the same side are fixedly connected with the same cover plate 7, a second spring 12 is sleeved on the rotating shafts 11, one end of the second spring 12 is fixedly connected with the inner wall of the circular groove 10, the other end of the second spring 12 is fixedly connected with the side wall of the cover plate 7, the cover plate 7 can automatically reset, the inner cavity of the kit shell 5 can be automatically sealed after strains are conveniently filled, the interference of the outside on detection results is reduced, and the use effect is good.
The foregoing description is only a preferred embodiment of the present invention, and the present invention is not limited thereto, but it is to be understood that modifications and equivalents of some of the technical features described in the foregoing embodiments may be made by those skilled in the art, although the present invention has been described in detail with reference to the foregoing embodiments. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (8)
1. The kiwi fruit pollen canker detection method comprises the following steps of:
s1: collecting and processing samples of kiwi fruit tree infection parts;
s2: preparing a culture medium, wherein the culture medium comprises a cultivation medium and an identification medium;
s3: filling the extracted kiwi fruit tree infection part sample into a culture medium for culturing strains;
s4: observing the colonies cultured on the culture medium, and carrying out irradiation identification on the colonies on the culture medium through ultraviolet detection equipment;
s5: extracting the fluorescent bacteria after irradiation of the ultraviolet detection equipment, and filling the extracted bacteria on an identification culture medium for culture;
s6: performing a gram staining procedure on the identification medium and observing the results;
s7: if red is observed, the kiwi fruit is infected with kiwi fruit pollen canker, otherwise, the kiwi fruit is not infected with kiwi fruit pollen canker.
2. The method for detecting kiwi fruit pollinosis according to claim 1, wherein in the step 1, the sample is crushed after the sample is collected from the kiwi fruit infected part.
3. The method for detecting kiwi fruit pollinosis according to claim 1, wherein the culture medium in the step 2 comprises 4 g of peptone, 0.3 g of dipotassium hydrogen phosphate, 0.3 g of magnesium sulfate, 3 g of agar, 2 ml of glycerin and 200 ml of distilled water, and the components are boiled and dissolved sufficiently during the preparation, and the pH value is adjusted to 7.2 and the temperature is adjusted to 121 ℃ for 15 minutes.
4. The method for detecting kiwi fruit pollinosis according to claim 1, wherein the identification medium in the step 2 comprises 3.2 g of polyvalent peptone, 2 g of hydrolyzed casein, 2 g of potassium sulfate, 0.3 g of magnesium chloride, 2.2 g of agar, 2 ml of glycerin and 200 ml of distilled water, and the components are boiled and fully dissolved during the preparation, the PH value is adjusted to 7.2, the sterilization is carried out at 121 ℃ for 15 minutes, and the dissolved cetyltrimethylammonium bromide, sodium chain run and cephalosporin are added when the medium is cooled to about 50 ℃.
5. The method for detecting kiwi fruit pollinosis according to claim 1, wherein the colony irradiated by the ultraviolet detection device in the step 4 is a milky white, smooth and clean-edged colony.
6. The method for detecting kiwi fruit pollen canker according to claim 1, wherein the culturing of the fluorogenic colonies in the step 5 is performed at 30℃for 48 hours.
7. The kiwi fruit pollen ulcer disease detection kit according to any one of claims 1 to 6, comprising a fixed frame (1), the fixed frame (1) is an I-shaped setting, movable slots (2) are all formed in the left and right side walls of the fixed frame (1), movable plates (3) are arranged on the left and right sides of the fixed frame (1), the movable plates (3) are concave, the side walls on the two sides of the movable plates (3) are respectively inserted into the movable slots (2) and are movably connected with the movable slots (2), a first spring (4) is fixedly connected to the side walls of the inner cavity of the movable slots (2), the other ends of the first springs (4) are fixedly connected with the movable plates (3), two kit shells (5) are symmetrically inserted in the left and right sides of the fixed frame (1), rectangular frames (6) are fixedly connected to the top surfaces of the kit shells (5), two cover plates (7) are symmetrically arranged in the left and right sides of the rectangular frames (6), and are respectively inserted into air holes (8) and are formed in the cover plates (7).
8. The kiwi fruit pollen ulcer disease detection kit according to claim 7, wherein four circular grooves (10) are symmetrically formed in the inner side wall of the rectangular frame (6), rotating shafts (11) are movably connected in the circular grooves (10), the other ends of the rotating shafts (11) on the same side are fixedly connected with the same cover plate (7), a second spring (12) is sleeved on the rotating shafts (11), one end of the second spring (12) is fixedly connected with the inner wall of the circular groove (10), and the other end of the second spring (12) is fixedly connected with the side wall of the cover plate (7).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311475243.6A CN117551733A (en) | 2023-11-08 | 2023-11-08 | Kiwi fruit pollen canker detection method and kit thereof |
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