CN106191205A - A kind of ash arrhizus bacteria method for quick drug-fast to various medicaments - Google Patents
A kind of ash arrhizus bacteria method for quick drug-fast to various medicaments Download PDFInfo
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- CN106191205A CN106191205A CN201610826193.5A CN201610826193A CN106191205A CN 106191205 A CN106191205 A CN 106191205A CN 201610826193 A CN201610826193 A CN 201610826193A CN 106191205 A CN106191205 A CN 106191205A
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- ash arrhizus
- arrhizus bacteria
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- 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
- C12Q1/18—Testing for antimicrobial activity of a material
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Abstract
The invention discloses a kind of ash arrhizus bacteria method for quick drug-fast to various medicaments, by first preparing ash arrhizus bacteria detection kit, then carry out field sampling to detect on the spot, whether can determine that the Drug resistance of various medicaments is used for quickly detecting by the method whether bacterial strain develops immunity to drugs by normal growth finally according to pathogen；Overcome traditional pathogen liquefaction resistance technology and need Field sampling, laboratory is isolated and purified, it is inoculated into the time and effort consuming on the plant tissue of pastille culture medium or spraying agent and the limited defect of disposable amount detection, it is time saving and energy saving, easy to operate, without sampling to have, various medicaments can be detected the most simultaneously and advantage, practical value and replicability are notable fast and accurately.
Description
Technical field
The present invention relates to pathogen liquefaction resistance technical field, particularly relate to a kind of ash arrhizus bacteria to various medicaments drug resistance
The method for quick of property.
Background technology
Phytopathogen Drug resistance, refers to that pathogen, for a long time under single medicament or similar medicament selection pressure, is led to
Cross heredity, variation, the adaptability that this is obtained.Along with the development of science and technology, the drug-fast research of pathogen becomes a focus and asks
Topic, modern Molecular Detection means are maked rapid progress, and can be detected the drug resistance of antibacterial by the molecular diagnostic techniques of nucleic acid level
Property, but for genome controlling the indefinite pathogen in the mutational site of resistance or not possessing Drug resistance Molecular Detection condition
Unit, traditional pathogen liquefaction resistance technology can only be used.
Traditional pathogen liquefaction resistance technology, mainly mycelial growth method, spore germination method, need Field sampling,
Laboratory is isolated and purified, is inoculated on the plant tissue of pastille culture medium or spraying agent, and this process is time-consuming, laborious, and
Disposable amount detection is limited.
Summary of the invention
Goal of the invention: in order to overcome defect present in prior art, the present invention proposes a kind of time saving and energy saving, operation side
Just, without sampling, can detect the most simultaneously various medicaments and fast and accurately ash arrhizus bacteria drug-fast quickly to various medicaments
Detection method.
Technical scheme: in order to solve above-mentioned technical problem, the technical solution adopted in the present invention is:
A kind of ash arrhizus bacteria method for quick drug-fast to various medicaments, it is characterised in that comprise the following steps:
(1) ash arrhizus bacteria detection kit is prepared；
(2) sampling detection: field sampling detects on the spot；
(3) the drug-fast judgement of medicament: whether can determine whether bacterial strain develops immunity to drugs by normal growth according to pathogen；
Wherein said prepare ash arrhizus bacteria detection kit concretely comprise the following steps determine for survey medicament differentiation metering, determine confession
Survey the kinds of culture medium required for medicament, determine the therapeutic amount of one-time detection, select culture plate, configuration culture medium, finally add
Work becomes ash arrhizus bacteria detection kit.
According to the quantity for surveying medicament, described culture plate selects in 6 holes, 12 holes, 24 holes, 48 hole sterilizing Tissue Culture Plates
A kind of.
Being more highly preferred to, the preparation of described culture medium prepares different culture medium according to the kind for survey medicament.
Further, the preparation of described culture medium is specially and adds for surveying medicament mother solution in the medium, is configured to institute
The concentration needed, joins in culture plate by culture medium on aseptic working platform, adds a cover after completing, and seals, and 4 DEG C of Refrigerator stores are standby
With, one month effect duration.
Be more highly preferred to, described field sampling and the drug-fast concrete operations judged of medicament as, find the cause of disease bacterium time
Sample with aseptic independent packaging cotton swab or toothpick, in direct inoculation to described detection kit, add a cover, seal and preserve, take back reality
Testing indoor 23 DEG C of constant temperature culture 2d, normal growth is resistant strain, is otherwise sensitive strain.
Beneficial effect: a kind of ash arrhizus bacteria method for quick drug-fast to various medicaments that the present invention provides, passes through
First prepare ash arrhizus bacteria detection kit, then carry out field sampling and detect on the spot, whether can be just finally according to pathogen
It is frequently grown and determines that the Drug resistance of various medicaments is used for quickly detecting by the method whether bacterial strain develops immunity to drugs；Overcome traditional
Pathogen liquefaction resistance technology needs Field sampling, and laboratory is isolated and purified, is inoculated into pastille culture medium or spraying agent
Time and effort consuming on plant tissue and the limited defect of disposable amount detection, it is time saving and energy saving, easy to operate, without adopting to have
Sample, can detect various medicaments the most simultaneously and advantage, practical value and replicability are notable fast and accurately.
Accompanying drawing explanation
Fig. 1 is 6 hole sterilizing Tissue Culture Plate schematic diagrams of the embodiment of the present invention 1；
Fig. 2 is 12 hole sterilizing Tissue Culture Plate schematic diagrams of the embodiment of the present invention 2.
Detailed description of the invention
Below in conjunction with embodiment, the present invention is described in further detail:
Embodiment 1:
A) test site: white rabbit town, Jurong City of Jiangsu Province peasant household Fructus Fragariae Ananssae roc
B) test period: in April, 2016
C) test objective: detect 50 strain Botrytis cinerea pathogenic bacterias to carbendazim, procymidone, diethofencarb, pyraclostrobin, phonetic bacterium
The Detection of insecticide resistance of 5 kinds of medicaments of ester.
D) implementation:
(1) ash arrhizus bacteria detection kit is prepared: made by the laboratory of Jiangsu Hilly Ground Zhenjiang Agriculture Science Research Institute
Standby ash arrhizus bacteria detection kit: first determine the differentiation metering for surveying medicament and kinds of culture medium (being specifically shown in Table 1), choose 6
Hole sterilizing Tissue Culture Plate (see Fig. 1), is prepared as pastille culture medium by sterilising medium, joins in culture plate, and each hole adds
A kind of pastille culture medium, using PDA culture medium as blank, wherein Fluoxastrobin and pyraclostrobin are for preventing bypass oxidation,
Culture medium all contains SHAM 100mg L-1, the detection kit prepared is sealed, at 4 DEG C, Refrigerator store is standby.
The differentiation metering of table 1 medicament and kinds of culture medium
(2) sampling detection: take 50 mentioned reagent boxes, loads in box, brings test field into；
(3) the drug-fast judgement of medicament: in strawberry greenhouse, with independent packaging sterilized cotton swabs or toothpick, sending out gently
Sick and produce and on the strawberry fruit of mould layer, dip pathogen spore, in point each hole in test kit, seal, every strain
Pathogen one test kit of inoculation；Postvaccinal test kit is taken back laboratory, 23 DEG C of constant temperature culture 2d, check result, according to
Whether pathogen can determine whether bacterial strain develops immunity to drugs by normal growth, normal growth for resistant strain, it is impossible to normally give birth to
Long for sensitive strain.
E) result of the test is shown in Table 2.
The table 2 Botrytis cinerea pathogenic bacteria liquefaction resistance to 5 kinds of medicaments
| Medicament title | Fastness frequency (%) |
| Carbendazim | 90 |
| Procymidone | 92 |
| Diethofencarb | 82 |
| Fluoxastrobin | 80 |
| Pyraclostrobin | 80 |
Embodiment 2:
A) test site: Lao Fang vineyard, Jurong City of Jiangsu Province
B) test period: in May, 2016
C) test objective: detect 50 strain Botrytis cinereas to carbendazim, procymidone, diethofencarb, Prochloraz, pyrazoles ether bacterium
Ester, Fluoxastrobin, Boscalid, fluopyram, fluxapyroxad, pyrimethanil, the Detection of insecticide resistance of 11 kinds of medicaments of CGA-173506.
D) implementation:
(1) ash arrhizus bacteria detection kit is prepared: made by the laboratory of Jiangsu Hilly Ground Zhenjiang Agriculture Science Research Institute
Standby ash arrhizus bacteria detection kit: first determine the differentiation metering for surveying medicament and kinds of culture medium (being specifically shown in Table 3), choose 12
Hole sterilizing Tissue Culture Plate (see Fig. 2), is prepared as pastille culture medium by sterilising medium, joins in culture plate, and each hole adds
A kind of pastille culture medium, using PDA culture medium as blank, wherein Fluoxastrobin and pyraclostrobin are for preventing bypass oxidation,
Culture medium all contains SHAM 100mg L-1, the detection kit prepared is sealed, at 4 DEG C, Refrigerator store is standby.
The differentiation metering of table 3 medicament and kinds of culture medium
| Medicament title | Distinguish metering (ppm) | Kinds of culture medium |
| Carbendazim | 10 | PDA |
| Procymidone | 0.5 | PDA |
| Diethofencarb | 0.5 | PDA |
| Prochloraz | 0.5 | PDA |
| Fluoxastrobin | 10 | MEA |
| Pyraclostrobin | 10 | MEA |
| Boscalid | 75 | YBA |
| Fluopyram | 10 | YBA |
| Fluxapyroxad | 5 | YBA |
| Pyrimethanil | 5 | CzA |
| CGA-173506 | 0.5 | CzA |
(2) sampling detection: take 50 mentioned reagent boxes, loads in box, brings test field into；
(3) the drug-fast judgement of medicament: in vineyard, uses independent packaging sterilizing toothpick, gently in morbidity and produce
Dip pathogen spore on the grape leave of mould layer or carpopodium, shake off in point each hole in test kit or by pathogenic bacteria spore
In each hole, seal, every pathogen strain bacterium one test kit of inoculation；Postvaccinal test kit is taken back laboratory, 23 DEG C
Constant temperature culture 2d, checks result, whether can determine whether bacterial strain develops immunity to drugs by normal growth according to pathogen, normal growth
For resistant strain, it is impossible to normal growth for sensitive strain.
E) result of the test is shown in Table 4.
Table 4 Botrytis cinerea liquefaction resistance to 11 kinds of medicaments
It should be pointed out that, that above detailed description of the invention is merely to illustrate the present invention rather than limits the scope of the present invention,
After having read the present invention, the amendment of the various equivalent form of values of the present invention is all fallen within and weighs appended by the application by those skilled in the art
Profit requires limited range.
Claims (5)
1. an ash arrhizus bacteria method for quick drug-fast to various medicaments, it is characterised in that comprise the following steps:
(1) ash arrhizus bacteria detection kit is prepared；
(2) sampling detection: field sampling detects on the spot；
(3) the drug-fast judgement of medicament: whether can determine whether bacterial strain develops immunity to drugs by normal growth according to pathogen；
Wherein said prepare ash arrhizus bacteria detection kit concretely comprise the following steps determine for survey medicament differentiation metering, determine for survey medicine
Kinds of culture medium required for agent, determine the therapeutic amount of one-time detection, select culture plate, configuration culture medium, be finally processed into
For ash arrhizus bacteria detection kit.
Ash arrhizus bacteria the most according to claim 1 method for quick drug-fast to various medicaments, it is characterised in that: institute
State culture plate and select the one in 6 holes, 12 holes, 24 holes, 48 hole sterilizing Tissue Culture Plates.
Ash arrhizus bacteria the most according to claim 1 method for quick drug-fast to various medicaments, it is characterised in that: institute
The preparation stating culture medium prepares different culture medium according to the kind for survey medicament.
Ash arrhizus bacteria the most according to claim 3 method for quick drug-fast to various medicaments, it is characterised in that: institute
The preparation stating culture medium adds the most in the medium for surveying medicament mother solution, is configured to required concentration, in aseptic work
Joining in culture plate by culture medium on platform, add a cover after completing, seal, 4 DEG C of Refrigerator stores are standby, one month effect duration.
Ash arrhizus bacteria the most according to claim 1 method for quick drug-fast to various medicaments, it is characterised in that: institute
State field sampling and the drug-fast concrete operations judged of medicament as, find the cause of disease bacterium time with aseptic independent packaging cotton swab or tooth
Sign sampling, in direct inoculation to described detection kit, add a cover, seal and preserve, take back 23 DEG C of constant temperature culture 2d in laboratory, just
It is frequently grown and is resistant strain, be otherwise sensitive strain.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610826193.5A CN106191205A (en) | 2016-09-14 | 2016-09-14 | A kind of ash arrhizus bacteria method for quick drug-fast to various medicaments |
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| CN201610826193.5A CN106191205A (en) | 2016-09-14 | 2016-09-14 | A kind of ash arrhizus bacteria method for quick drug-fast to various medicaments |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109338004A (en) * | 2018-11-12 | 2019-02-15 | 上海市农业科学院 | A kind of detection ash arrhizus bacteria is to the combination of the primer of Boscalid resistance, kit and method |
Citations (3)
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|---|---|---|---|---|
| CN102453742A (en) * | 2010-10-20 | 2012-05-16 | 华中农业大学 | Detection kit for Sclerotinia sclerotiorum strain drug-resistance and detection method of Sclerotinia sclerotiorum strain drug-resistance |
| CN102925565A (en) * | 2012-10-19 | 2013-02-13 | 中国农业大学 | Method for identifying nucleotide point mutation of phytophthora sojae beta-microtubulin gene and use thereof for diagnosing zoxamide resistance |
| CN103789409A (en) * | 2013-11-15 | 2014-05-14 | 中国农业大学 | Molecular detection method for identifying insecticide resistance of Botrytis cinerea on zoxamide |
-
2016
- 2016-09-14 CN CN201610826193.5A patent/CN106191205A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102453742A (en) * | 2010-10-20 | 2012-05-16 | 华中农业大学 | Detection kit for Sclerotinia sclerotiorum strain drug-resistance and detection method of Sclerotinia sclerotiorum strain drug-resistance |
| CN102925565A (en) * | 2012-10-19 | 2013-02-13 | 中国农业大学 | Method for identifying nucleotide point mutation of phytophthora sojae beta-microtubulin gene and use thereof for diagnosing zoxamide resistance |
| CN103789409A (en) * | 2013-11-15 | 2014-05-14 | 中国农业大学 | Molecular detection method for identifying insecticide resistance of Botrytis cinerea on zoxamide |
Non-Patent Citations (2)
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109338004A (en) * | 2018-11-12 | 2019-02-15 | 上海市农业科学院 | A kind of detection ash arrhizus bacteria is to the combination of the primer of Boscalid resistance, kit and method |
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