CN110376341A - A kind of measuring method of graw mold of tomato protective agents photolysis - Google Patents
A kind of measuring method of graw mold of tomato protective agents photolysis Download PDFInfo
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- CN110376341A CN110376341A CN201910593031.5A CN201910593031A CN110376341A CN 110376341 A CN110376341 A CN 110376341A CN 201910593031 A CN201910593031 A CN 201910593031A CN 110376341 A CN110376341 A CN 110376341A
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Abstract
The invention discloses a kind of measuring methods of graw mold of tomato protective agents photolysis, this method is that fungicide drug and agar medium are prepared by mixing into culture plate, it carries out lighting process and causes drug that photodegradation occurs, and drug photolysis causes the activity of light processing product to change, and shows the photolysis of drug to the reaction of light processing product by testing botrytis cinerea.Methods && steps of implementation is as follows: 1) being equipped with agar medium；2) compounding pharmaceutical plate；3) light processing prodrug Activity determination；4) light processing；5) pharmaceutical activity detects after light processing；6) photolysis result is obtained.The invention has the advantages that 1) overall technology is simple, operated without using high-precision instrument and equipment or the serological technique of complexity；2) it can reflect the relationship of drug photodissociation and drug effect；3) it can get the information that non-photolysis causes pharmaceutical activity to change.
Description
Technical field
The present invention relates to the detection technique of fungicide photolysis, specifically a kind of graw mold of tomato protective agents photolysis
Measuring method.
Background technique
Graw mold of tomato is the important disease of bulk vegetable tomato, and plant disease epidemic can bring heavy losses to tomato production,
The prevention and treatment of the current disease depends on pesticide control.The protective agents of pesticide control are once applied to field, it will usually
Natural degradation occurs, and photodecomposition caused by illumination acts on is the most prominent.Therefore the control efficiency of photolysis and drug
And medicament residue is in close relations.Obviously, the photolysis rule of drug is understood to the preventive and therapeutic effect for giving full play to drug, and
It is significant to reduce medicament residue.
The detection technique of the photolysis of drug is currently mostly to utilize physical and chemical means and serology means.Physical and chemical means are
Using the chemical property and physical features of drug, drug is shown and identified in modern comfort instrument, the means rapid sensitive,
But need modern large-scale precision instrument.Serology means are to convert drug to antigen and thus prepare antibody, utilize antigen-
Drug shows and identified by the idiosyncrasy of antibody, and the means are also more sensitive, but need to prepare specific antibody.Current technology
Means have a common feature, and detection process only tracks the original molecular structure of drug, is not directed to drug to controlling object
Bioactivity, as a result can not obtain drug degradants to the activity of controlling object be enhancing or weaken for information about, not
It can reflect the relationship of drug degradation and drug effect.
Whether the photodegradation of drug, its action activity to controlling object can be directly affected, action activity can pass through shadow
It rings the growth and development of controlling object and is shown, but, also favorably do not use the bioactivity means of drug as drug so far
The detection technique of photolysis.
Summary of the invention
The purpose of the present invention is the photolysises using drug, and the drug effectiveness of light processing product to be caused to change,
A kind of measuring method of graw mold of tomato protective agents photolysis is provided.
The technical scheme to solve the above technical problems is that
A kind of measuring method of graw mold of tomato protective agents photolysis, is that fungicide drug is mixed in agar culture
In base plate, and carry out lighting process, the drug in agar plate illumination effect issue the third contact of a total solar or lunar eclipse decompose, light processing product to kind
The action activity of solanum cinerea bacterium changes, and shows drug to the reaction of light processing product by testing botrytis cinerea
The step of photolysis of object, drug photolysis measuring method, is as follows:
1. being equipped with agar medium: with the culture medium containing agar according to a conventional method, conventional sterilant is spare.
2. compounding pharmaceutical plate: under aseptic condition, the culture medium that drug is equipped with step 1 in the proper ratio is sufficiently mixed
Close, and use Nostoc commune Vanch ware inverted plate, acquisition Drug plates；The culture medium flat plate conduct of same operation processing but non-drug containing
Compare plate.
3. light processing prodrug Activity determination: on the Drug plates and control plate that step 2 is equipped with, being quantitatively implanted into often
The botrytis cinerea that rule culture prepares；It is cultivated under 23 DEG C of appropriate temperature conditions, measures botrytis cinerea on plate
Increment calculates and obtains drug to inhibiting rate I before the light processing of botrytis cinerea0。
4. light processing: sealing the Drug plates of step 2 with common sealed membrane, be then placed in the Drug plates of the sealing
Lighting process is carried out under the illumination condition of setting, dark control CK processing is made in the double-deck masking foil encapsulating of same Drug plates.
5. pharmaceutical activity detects after light processing: the light processing plate of step 4 is gone back to after the light processing of setting time t
Mouth is sealed off under aseptic condition, then according to the operation of step 3 light processing prodrug Activity determination, carries out pharmaceutical activity after light processing
Detection；It calculates and obtains the inhibiting rate that drug handles inhibiting rate It1 after the light processing of botrytis cinerea and dark control CK
It2。
6. obtaining photolysis result: the I obtained according to step 30The It1 and It2 obtained with step 5 is calculated and is obtained medicine
Object light solution exercising result.
The invention has the advantages that
1. overall technology is simple, operated without using the serological technique of large-scale high-precision instrument and equipment or complexity.
2. detection process, to the action activity of controlling object, can obtain the work of photolytic product dependent on drug and photolytic product
Property be enhancing, weaken for information about, can reflect the relationship of drug photodissociation and drug effectiveness.
3. can get the information that non-photolysis causes pharmaceutical activity to change.
Specific embodiment
The present invention will be further described below with reference to examples.
A kind of measuring method of graw mold of tomato protective agents photolysis of the present invention.It is that fungicide drug is mixed in fine jade
Lighting process is carried out in rouge culture medium, light processing causes the drug in agar medium that photodegradation occurs, and photodecomposition causes
Drug changes to the action activity of botrytis cinerea, therefore the life by detection light processing product to botrytis cinerea
Object action activity can obtain the result of photolysis.
Used medium of the present invention is the agar medium of routine culture botrytis cinerea energy normal growth, conventional high temperature
It sterilizes spare.The culture medium of preparation is mixed in the proper ratio with drug to be determined, " ratio appropriate " refers to medicine here
After object is mixed with culture medium, the concentration dose of drug is in the concentration dose of botrytis cinerea response sensitivity.By drug and training
The mixture inverted plate for supporting base, is made drug containing tablet, and the plate changed into after using sterile water to mix with culture medium is flat as compareing
Plate.
Pharmaceutical activity detection after preparing drug containing tablet, before taking the leading lighting process of part drug containing tablet.Take routine culture
The botrytis cinerea of preparation is quantitatively implanted into drug containing tablet and its control plate, trains under the conditions of 23 DEG C of suitable growth temperature
Support reasonable time.When measurement be mycelium morphology factor when, close to the time for covering with plate needs be with bacterium colony mycelia it is appropriate, it is logical
It is often 3 days；When measurement be the germination rate of spore when, sufficiently sprouted with spore the time of needs be it is appropriate, usually 12 hours.
It cultivates to after the time, culture plate is taken out, measure the increment of botrytis cinerea on plate, calculate drug to tomato gray mould
Germ inhibiting rate.
Drug inhibition rate=(control plated growth amount-Drug plates increment)/control plated growth amount.
The Drug inhibition rate being calculated is the pharmaceutical activity I before lighting process0。
While coming into effect the detection of the pharmaceutical activity before lighting process, by same drug containing tablet first with common sealing
Film sealing, is subsequently placed under the illumination condition of setting and carries out lighting process.Separately take the double-deck tinfoil paper paper bag of same Drug plates
Envelope is protected from light, and is placed under same illumination condition and is made dark control CK processing.Light processing duration t is set by need of work.
After the lighting process of t time, plate is gone back under aseptic condition and is sealed off, it is living according to the drug before lighting process
Property detection operation, carry out lighting process after pharmaceutical activity detection.After testing result obtains the illumination t time, the medicine of lighting process
Object inhibiting rate It1；And the Drug inhibition rate It2 of corresponding dark control CK processing.Comparison is found, after t time-triggered protocol, fungicide
The inhibitory activity of drug changes, this phenomenon is referred to as decay of activity by the present invention, and attenuation can be calculated as follows:
Pharmaceutical activity attenuation It3:It3=I after light processing0-It1；
Dark CK pharmaceutical activity attenuation It4:It4=I after light processing0-It2；
Dark CK pharmaceutical activity attenuation It4 after light processing, is actually that non-photolysis leads to pharmaceutical activity attenuation；
And pharmaceutical activity attenuation It3 after light processing, actually photolysis and non-photolysis summation cause pharmaceutical activity to be decayed
Amount, therefore, photolysis leads to pharmaceutical activity attenuation It5 are as follows:
It5=It3-It4；
Non- photolysis is led to the pharmaceutical activity I before pharmaceutical activity attenuation It4 and light processing by the present invention0The ratio between, claim
Make decay of activity rate P1 caused by non-photolysis, then:
P1=It4/I0；
Photolysis is led to the pharmaceutical activity I before pharmaceutical activity attenuation It5 and light processing by the present invention0The ratio between, referred to as
Decay of activity rate P2 caused by photolysis, then:
P2=It5/I0。
Embodiment 1
Using a kind of measuring method of graw mold of tomato protective agents photolysis of the present invention, the illumination in routine experimentation room
Condition makees material with ash arrhizus bacteria bacterial strain Bc-5, measures the photolysis of fungicide iprodione, implements operation as follows:
1. being equipped with agar medium: being surveyed using the potato dextrose agar (PDA) for being suitable for botrytis cinerea growth
Culture medium on probation, in parts by weight: potato 200g, glucose 20g, agar 20g, water 900mL are prepared and conventional high temperature sterilizing is standby
With.
2. compounding pharmaceutical plate: the mycelia of known graw mold of tomato bacteria strain Bc-5 grows the inhibition to iprodione sensibility
Middle concentration EC50For 1 μ g/mL.Under aseptic condition, iprodione medical fluid and the PDA culture medium that step 1 is equipped with are sufficiently mixed, prepared
At ratio containing iprodione be 1 μ g/mL pastille culture medium, and use Nostoc commune Vanch ware inverted plate, acquisition Drug plates, by sterile water
The culture medium being equipped with step 1 is sufficiently mixed and inverted plate, obtains control plate.
3. light processing prodrug Activity determination: on the Drug plates and its control plate that step 2 is equipped with, being quantitatively implanted into
One piece of mycelia bacteria cake of the graw mold of tomato bacteria strain Bc-5 that routine culture prepares, after cultivating 3 days under the conditions of 23 DEG C of temperature, takes
The colony diameter of botrytis cinerea on plate is measured out.It calculates and obtains drug to inhibiting rate before the light processing of botrytis cinerea
I0。
4. light processing: sealing the Drug plates of step 2 with common sealed membrane, be then placed in the Drug plates of the sealing
In routine experimentation room on the bright desktop of nearly window, under the illumination condition that Natural light intensity is 1000Lux at noon, natural light is carried out
According to processing, same Drug plates encapsulate and make at dark control CK under same illumination condition with the double-deck masking foil
Reason.
5. pharmaceutical activity detects after light processing: the light processing plate of step 4, respectively by 7 days, 14 days, 21 days natures
It after lighting process, goes back under aseptic condition and seals off mouth, then according to the operation of step 3 light processing prodrug Activity determination, carry out
Drug detects the inhibitory activity of botrytis cinerea after light processing；The inhibiting rate It1 for obtaining lighting process drug is calculated,
And the inhibiting rate It2 of dark CK processing drug.
6. obtaining photolysis result: the I obtained by step 30And It1 and It2 that step 5 obtains, it calculates and obtains light
After processing 7 days, 14 days, 21 days, photolysis leads to decay of activity rate caused by pharmaceutical activity attenuation It5, non-photolysis
Decay of activity rate P2 caused by P1 and photolysis, as shown in table 1.
Iprodione inhibitory activity variation before the light processing of 1. embodiment 1 of table and after light processing
Embodiment 2
Using a kind of measuring method of graw mold of tomato protective agents photolysis of the present invention, in warm indoor lighting conditions,
Make material with ash arrhizus bacteria bacterial strain Bc-5, measures the photolysis of fungicide iprodione.It is grasped by step 1 to the step 6 of embodiment 1
Make, the light processing condition of different only steps 4 is not the illumination item that general room edge of window noon Natural light intensity is 1000Lux
Part, but noon Natural light intensity is the illumination condition of 11000Lux in greenhouse, measures the result such as table 2 of iprodione photolysis
It is shown.
Iprodione inhibitory activity variation before the light processing of 2. embodiment 2 of table and after light processing
Claims (1)
1. a kind of measuring method of graw mold of tomato protective agents photolysis, which is characterized in that fungicide drug to be mixed in
In agar medium plate, and lighting process is carried out, the drug in agar plate issues the third contact of a total solar or lunar eclipse in illumination effect and decomposes, light processing
Product changes to the action activity of botrytis cinerea, the reaction by test botrytis cinerea to light processing product
Show the photolysis of drug, the step of drug photolysis measuring method is as follows:
1) it is equipped with agar medium: with the culture medium containing agar according to a conventional method, conventional sterilant is spare；
2) compounding pharmaceutical plate: under aseptic condition, the culture medium that drug is equipped with step 1) in the proper ratio being sufficiently mixed,
And with Nostoc commune Vanch ware inverted plate, obtain Drug plates, the culture medium flat plate of same operation processing but non-drug containing as pair
According to plate；
3) light processing prodrug Activity determination: on the Drug plates and control plate that step 2) is equipped with, quantitatively it is implanted into routine
Cultivate the botrytis cinerea prepared；It is cultivated under 23 DEG C of appropriate temperature conditions, measures the life of botrytis cinerea on plate
Long amount calculates and obtains drug to inhibiting rate I before the light processing of botrytis cinerea0；
4) light processing: the Drug plates of step 2) are sealed with common sealed membrane, then the Drug plates of the sealing are placed in and are set
Lighting process is carried out under fixed illumination condition, dark control CK processing is made in the double-deck masking foil encapsulating of same Drug plates；
5) pharmaceutical activity detects after light processing: the light processing plate of step 4) is gone back to sterile after the light processing of setting time t
Under the conditions of seal off mouth, then according to the operation of step 3) light processing prodrug Activity determination, pharmaceutical activity is examined after carrying out light processing
It surveys；It calculates and obtains the inhibiting rate that drug handles inhibiting rate It1 after the light processing of botrytis cinerea and dark control CK
It2；
6) photolysis result is obtained: the I obtained according to step 3)0The It1 and It2 obtained with step 5) is calculated and is obtained drug light
Solve exercising result.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103798291A (en) * | 2014-02-18 | 2014-05-21 | 中国水稻研究所 | Compound pesticide containing triadimefon and copper oxychloride |
| CN104904723A (en) * | 2014-03-11 | 2015-09-16 | 江苏龙灯化学有限公司 | Sterilization composition |
| CN204694630U (en) * | 2015-06-18 | 2015-10-07 | 淮阴工学院 | A kind of device of Portable ultraviolet light degrading pesticide residues |
| CN106508944A (en) * | 2016-08-31 | 2017-03-22 | 柳州市惠农化工有限公司 | Pesticidal composition containing 2-methylsulfonyl-5-(2,4-dichlorophenyl)-1,3,4-oxadiazole and prochloraz |
| CN107219205A (en) * | 2017-06-05 | 2017-09-29 | 河海大学 | A kind of pesticide degradation bacteria activity test method based on Fluorescent Staining Observation |
| CN107410382A (en) * | 2017-09-13 | 2017-12-01 | 湖南省农业生物资源利用研究所 | A kind of bactericide for tobacco black shank and its preparation method and application |
| CN109105379A (en) * | 2018-09-24 | 2019-01-01 | 青岛农业大学 | A kind of preparation and application of chemical bactericide pesticide that preventing and treating blueberry fruit rot |
-
2019
- 2019-06-24 CN CN201910593031.5A patent/CN110376341B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103798291A (en) * | 2014-02-18 | 2014-05-21 | 中国水稻研究所 | Compound pesticide containing triadimefon and copper oxychloride |
| CN104904723A (en) * | 2014-03-11 | 2015-09-16 | 江苏龙灯化学有限公司 | Sterilization composition |
| CN204694630U (en) * | 2015-06-18 | 2015-10-07 | 淮阴工学院 | A kind of device of Portable ultraviolet light degrading pesticide residues |
| CN106508944A (en) * | 2016-08-31 | 2017-03-22 | 柳州市惠农化工有限公司 | Pesticidal composition containing 2-methylsulfonyl-5-(2,4-dichlorophenyl)-1,3,4-oxadiazole and prochloraz |
| CN107219205A (en) * | 2017-06-05 | 2017-09-29 | 河海大学 | A kind of pesticide degradation bacteria activity test method based on Fluorescent Staining Observation |
| CN107410382A (en) * | 2017-09-13 | 2017-12-01 | 湖南省农业生物资源利用研究所 | A kind of bactericide for tobacco black shank and its preparation method and application |
| CN109105379A (en) * | 2018-09-24 | 2019-01-01 | 青岛农业大学 | A kind of preparation and application of chemical bactericide pesticide that preventing and treating blueberry fruit rot |
Non-Patent Citations (2)
| Title |
|---|
| 施宏梅等: "雪白弯颈霉C41 发酵液稳定性及粗提物的抑菌活性", 《农药》 * |
| 陈召亮: "季铵盐类化合物对几种蔬菜致病菌的生物活性及应用前景探讨", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
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