CN107267591A - To the method for fine horse jujube alternaria Toxicity Determination - Google Patents
To the method for fine horse jujube alternaria Toxicity Determination Download PDFInfo
- Publication number
- CN107267591A CN107267591A CN201610205776.6A CN201610205776A CN107267591A CN 107267591 A CN107267591 A CN 107267591A CN 201610205776 A CN201610205776 A CN 201610205776A CN 107267591 A CN107267591 A CN 107267591A
- Authority
- CN
- China
- Prior art keywords
- fine horse
- virulence
- horse jujube
- alternaria
- value
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Toxicology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention is the method to fine horse jujube alternaria Toxicity Determination, it is intended to which the medicament of South Sinkiang fine horse jujube black spot can effectively be prevented and treated by filtering out, and reduces the blindness of peasant's medication, so as to improve disease control level, strengthens jujube garden economic benefit.Toxicity Determination uses spore germination method, determines that concentration of sterilant uses scope first on the basis of trial test, then determines the indoor virulence of bactericide.Result of the test shows 25% pyraclostrobin EC50 value virulence highests, and its EC50 value is 0.0373mg/L.Secondly it is 50% fluorine silicon Fluoxastrobin AS, 80% carbendazim WP and 10% Difenoconazole WG, its EC50 value is respectively 16.0752mg/L, 43.8137mg/L, 63.198mg/L.50% penta azoles miaow AS virulence are worst, and its EC50 value is 7007.383mg/L.25% pyraclostrobin EC, 50% fluorine silicon Fluoxastrobin AS, 10% Difenoconazole WG and 80% carbendazim WP medicament are relatively strong to fine horse jujube alternaria virulence, and application can be further tested in production.
Description
Technical field
The present invention relates to the method for toxicity test, more specifically for, it is more particularly to black to fine horse jujube
The method of pinta bacterium Toxicity Determination.
Background technology
Jujube tree (Zizyphus jujube Mill.), is Rhamnaceae (Rhamnaceae) jujube,
It is Chinese distinctive Fruit Tree Resources and the advantage fruit tree species shown unique characteristics.Jujube tree is with its adaptability
By force, fruit maturation is early, yield is high, manageability and ecological benefits and economic benefit etc. are significantly excellent
Point, occupies critical role in Chinese agriculture production.Recently as the Production and Construction Corps of Xinjiang
The adjustment of agricultural structure, jujube is widely applied plantation.Because single variety fine horse jujube exists
South Sinkiang large area plantation, occurs in that increasing new expression, wherein fine horse jujube black spot is jujube
The main Common Diseases in producing region, endanger most serious.
Through Morphological Identification, South Sinkiang fine horse jujube black spot is mainly by alternaric bacteria (Alternaria
Alternate caused by) infecting.Alternaric bacteria species is various, the novel species of published in recent years, Combination nova,
Newname and infraspecies taxonomy unit have 47.It is pathogenic between alternaric bacteria is not of the same race to have very big difference
Different, in South Sinkiang, alternaric bacteria only endangers jujube fruit, yet there are no the feelings of harm jujube leaf, jujube flower and branch
Condition, and above-mentioned jujube histoorgan can be caused harm in Shandong, the Shanxi Jujube producing region bacterium.It is right
In the disease caused by alternaric bacteria, the prevention and controls used are the synthesis based on chemical prevention
Prevention and controls.Although biological control and the utilization of microbial antibacterial active material are to suppress disease
One important channel, but practical application is less.It is black that winter jujube was once just done in 2000 by Shandong Province
The research of pinta Prevention Technique, effective chemical agent has been filtered out by Toxicity Determination,
These medicaments also play good prevention effect in production.
At present, the synthesis based on chemical prevention is used to the preventing and treating of South Sinkiang fine horse jujube black spot
Control strategy.But country's agricultural chemicals type of merchandize is various, most people None- identified is so as to abuse
Agricultural chemicals, not only causes environmental pollution but also prevention effect is poor.In order to reduce orchard worker in jujube garden medication
Blindness, the author uses spore germination method, determines in the market common 10 indoors
The virulence of bactericide and its mixed agent is planted, suppression big to fine horse jujube alternaria virulence to filter out
The good bactericide of bacterium effect, reference is provided for the upper scientific and reasonable medication of production.
It is to gather fine horse jujube disease fruit from Xinjiang Awat County for examination strain, according to Koch's Postulates to disease
Opportunistic pathogen is separated, identified and carried out Pathogenicity, and pathogen is chain through Morphological Identification
Lattice spore belongs to (Alternaria) fungi, and strain and Saving specimen are in plant science institute of Tarim University
Plant pathology laboratory.
It is pyraclostrobin (25% missible oil, BASF Co., Ltd), phenylate for examination bactericide
Methyl cyclic-azole (10% water-dispersible granules, Switzerland first just reaches crop protection Co., Ltd), many bacterium
The false monospore bar of clever (80% wettable powder, Zhengzhou Han Xiang chemical products Co., Ltd), fluorescence
Bacteria powder (300,000,000,000/g, Jianou City Wang Da agrochemical Co., Ltd), 3 120 (4% aqua,
Zhejiang 31 change bio tech ltd), fluorine silicon-Fluoxastrobin (50% aqua, rising sun agriculture
Industry Science and Technology Development Co., Ltd.), (50% missible oil, rising sun chemical industry (Shandong) is limited for ethirimol
Company), fluorine ring-phenylate azoles (65% aqua, rising sun agricultural science and technology development corporation, Ltd.), penta
Azoles-miaow (50% aqua, rising sun agricultural science and technology development corporation, Ltd.), streptomycin sulphate (72%
Soluble powder, Beijing middle peasant's Hong Lu developments in science and technology Co., Ltd).
The content of the invention
The purpose of the present invention is the method to fine horse jujube alternaria Toxicity Determination, passes through research
The Water Consumption Characteristics of jujube under condition of drip irrigation, analyze the water consumption of jujube different bearing stage and its right
The influence of yield, inquires into the Soil Moisture and orchard Water productivity for being adapted to jujube growth,
Determine the Optimal soil moisture index of each growing stage, formulate to economize on water, high-quality, be efficiently mesh
Target drip irrigation jujube garden irrigation program, creates jujube garden efficient water use technical system under condition of drip irrigation,
Theoretical foundation and technical support are provided for South Sinkiang local special fruit tree industry sustainable development.
The foregoing method to fine horse jujube alternaria Toxicity Determination, it is characterised in that：
For a long time, largely easily germ is developed immunity to drugs using single medicament, 2 kinds of medicament science are mixed
With can effectively delay drug-fast to produce and improve prevention effect.
The foregoing method to fine horse jujube alternaria Toxicity Determination, it is characterised in that：
The different ratio of carbendazim and pyraclostrobin can preferably suppress fine horse jujube alternaria spore
Sprout.
The foregoing method to fine horse jujube alternaria Toxicity Determination, it is characterised in that：
In 10 kinds of medicaments for examination, pyraclostrobin is to the virulence highest of fine horse jujube black spot, its EC50
It is worth for 0.0373mg/L；Secondly it is fluorine silicon-Fluoxastrobin, many bacterium and clever Difenoconazole, its EC50
Value respectively 16.0752,43.8137,63.198mg/L；Remaining medicament virulence is relatively low, fluorine
Ring-phenylate azoles EC50 values are that 165.9195mg/L, pseudomonas fluorescens EC50 values are
The sweet phenol EC50 values of 237.0493mg/L, second are that 524.4414mg/L, three 120EC50 values are
736.4497mg/L, streptomycin sulphate EC50 values are 1032.048mg/L, wherein penta azoles-miaow
Virulence is worst, and its EC50 value is 7007.383mg/L.
The beneficial effects of the invention are as follows because this experiment is that in vitro is studied indoors
, and prevention effect of the medicament in field is influenceed by several factors, it is therefore, malicious indoors
The higher medicament of power in production prevention effect performance how, in addition it is also necessary to further research.Chemistry
Preventing and treating is a kind of method in the disease comprehensive regulation, to improve plant disease management level, realizes agriculture
Industry sustainable development, should carry out the comprehensive regulation to disease.However, many jujube agricultures are black to fine horse jujube
The pathogenic factor and occurrence regularity of pinta lack understanding, cause the evil of being critically ill to aggravate.From now on should be from
Strengthen studying in terms of Variety distribution, orchard management, disease monitoring and combine chemical prevention, so that
Fine horse jujube black spot is avoided in Southern Xinjiang expansion and prevalence.
Embodiment
The present invention relates to the method to fine horse jujube alternaria Toxicity Determination, embodiment
It is as follows：
The preparation of germ Spore cultivation and spore suspension
Strains tested is placed in 25 DEG C of constant temperature dark culturing casees in PDA culture medium and cultivated, mirror
When inspection there are a large amount of conidiums generations, water elution bacterium colony spore is distilled with 5mL, and fill spore
Divide and suspend and dilute, per 80~100, visual field conidium under 10 × 10 power microscopes.
Toxicity Determination
Each medicament is respectively set to 5 concentration gradients on the basis of trial test, each agriculture is drawn
The μ L of medicine dilution 50 and equivalent spore suspension, mixing are dripped in clean concave slide, are placed in
In the moisturizing culture dish of wet filter paper, heat and moisture preserving culture is carried out in 25 DEG C of constant temperature dark culturing casees.
The concentration gradient of pyraclostrobin and fluorine silicon-Fluoxastrobin is set to 0.5,5,12.5,25,100mg/L；
Difenoconazole, carbendazim, penta azoles-miaow, 3 120, fluorine ring-phenylate azoles, streptomycin sulphate
Concentration gradient is 5,12.5,25,100,300mg/L；Ethirimol, Pseudomonas bar
Bacteria concentration gradient is 1,5,10,50,200mg/L；The carbendazim of same multiple will be diluted
Carried out with pyraclostrobin by A (6: 1), B (3: 1), C (1: 1), D (1: 3), E (1: 6) volume ratio
Mixture, concentration gradient is set to 0.005,0.05,0.5,5,50,500mg/L, the above is each
Processing is repeated 3 times, using the distilled water without agricultural chemicals as blank control.When control group spore germination
When rate reaches more than 80%, each processing spore germination rate is counted.
Fungicide compounding agent effect assessment
Mixed agent action effect is evaluated according to co-toxicity coefficient (CTC), each medicament toxicity index (TI),
The actual toxicity index of built agent (ATI), theoretical toxicity index (TTI) and co-toxicity coefficient (CTC) are calculated
Method is with reference to SUN.
TI=(EC50 of the EC50/ reagent agents of standard agent) × 100
ATI=(EC50 of the alone EC50/ built agent medicaments of standard agent) × 100
TTI=TI (A) × A in mixture percentage+TI (B) × B shared hundred in built agent
Divide ratio
CTC=[the actual toxicity index of mixture (ATI)/mixture theoretical toxicity index (TTI)] × 100
If the notable synergistic of CTC >=170,120≤CTC ＜ 170, slightly synergy, 70≤CTC ＜ 120
Summation action, CTC ＜ 70 are antagonism.
The statistics of data and analysis
Each processing checks that spore sum will reach 100, and statistics takes being averaged for 3 repetitions
Value, is calculated as follows spore correction germination rate, the correction bacteriostasis rate of each processing.
Correct germination rate=(treatment group germination rate/control group germination rate) × 100%
Correct bacteriostasis rate=1- correction germination rates
To correct the corresponding niqueMin of bacteriostasis rate as ordinate, pair of the series concentration set is tested
Numerical value is abscissa, makees regression straight line, obtains virulence regression equation and EC50 values.
Analysis to result
Mixed agent Evaluation of Germicidal Efficacy
Because carbendazim and pyraclostrobin are of a relatively high to fine horse jujube alternaria virulence, and two
Person's bactericidal action mechanism is different, so both progress Compounding trials of selection, its different blended is with combination
To the toxicity test result of fine horse jujube alternaria.Proportioning combination A, B, C, E EC50 values
It is smaller, illustrate that this 4 kinds of compounding methods have preferable fungistatic effect；Mixture combination D its
EC50 values are higher, are 60.032mg/L, show that its fungistatic effect is poor.Combine A, B and E
Co-toxicity coefficient be respectively 698.3147,5131.721,322.2999, its CTC ＞ 170, table
Now acted on for notable synergistic；It is 154.32,70≤CTC ＜ 120, performance to combine C co-toxicity coefficients
To slightly have synergy；And D co-toxicity coefficients are combined less than 70, show as antagonism.
Inhibition of 10 kinds of reagent agents to fine horse jujube alternaria spore germination
Carried out in 25 DEG C of constant temperature dark culturing casees after heat and moisture preserving culture 12h, control group germ
Spore germination rate all reaches more than 80%, counts each processing spore germination rate.For 10 kinds of medicines of examination
Sprouting of the agent to fine horse jujube alternaria spore has certain inhibitory action, and every kind of medicament is with dense
The reduction of gradient is spent, germ spore germination rate gradually rises, but all in all, 25% pyrazoles ether
Bacterium ester EC fungistatic effects preferably, from high concentration to low concentration in the range of, spore is averagely sprouted
Rate is controlled 11.33%~30.67%, and the average germination rate of its control group spore is 88.33%；And
50% penta azoles-miaow AS fungistatic effects are worst, and its spore germination rate reaches during highest drug concentration
69.33%, spore germination rate is up to more than 77% under remaining reagent agent concentration.
Medicament mixture combines the inhibition to fine horse jujube alternaria spore germination
The different ratio of carbendazim and pyraclostrobin can preferably suppress fine horse jujube alternaria spore
The sprouting of son, especially combines B, when medicament maximum concentration is 500mg/L, spore germination rate
Only 2%, when medicament least concentration is 0.005mg/L, spore germination rate also only has 30.67%,
And control group spore germination rate reaches 80.00%；And combine D fungistatic effects and show relatively poor,
Except under medicament high concentration its spore germination rate it is relatively low in addition to, spore germination rate under remaining concentration
Reach more than 66%.
Summary to result
Virulence of 10 kinds of bactericide to fine horse jujube alternaria is determined using spore germination method, it is indoor
Result of the test shows that 10 kinds of medicaments have certain fungistatic effect to germ spore, and with medicament
Its bacteriostasis of the rise of concentration progressively strengthens.Shown according to EC50 value result of calculations：Pyrazoles
Kresoxim-methyl virulence is most strong, next to that fluorine silicon-Fluoxastrobin, Difenoconazole and carbendazim, remaining medicine
Agent virulence is relatively low.It is therefore proposed that pyraclostrobin, fluorine silicon-Fluoxastrobin, Difenoconazole and many
Bacterium spirit is producing enterprising step progress drug effect checking test.
For a long time, largely easily germ is made to develop immunity to drugs using single medicament, by 2 kinds of Pharmacies
Drug-fast generation can effectively be delayed and improve prevention effect by learning to use with.To alleviate target organismses
Carbendazim and pyraclostrobin 2 have been selected in resistance, expansion preventing and treating spectrum and raising drug effect, this experiment
Kind virulence is of a relatively high, and the different medicament of the mechanism of action carries out mixture, and experiment shows carbendazim
Mixture is carried out by A, B and E with pyraclostrobin, co-toxicity coefficient is all higher than 170, not only bright
It is aobvious to improve bactericidal effect, it is also beneficial to prevent or delays the drug-fast generation of germ, for exploitation
Prevent and treat fine horse jujube black spot mixture preparation and provide important theoretical foundation.
Alternaric bacteria belongs to Deuteromycotina fungi, and vegetative propagation can produce a large amount of conidiums,
Therefore rod method is mainly propagated, infected and endangered by conidium.This research is from chain lattice
The conidial angle of spore is studied, and has been filtered out stronger to rod method conidium toxicity
Chemical agent so that carrying out chemical prevention for the upper large area of production provides theoretical direction.
Claims (3)
1. a kind of method of fine horse jujube alternaria Toxicity Determination, it is characterised in that：For a long time, largely easily germ is made to develop immunity to drugs using single medicament, 2 kinds of medicament science, which are used with, can effectively delay drug-fast generation and improve prevention effect.
2. the method for fine horse jujube alternaria Toxicity Determination as claimed in claim 1, it is characterised in that：The different ratio of carbendazim and pyraclostrobin can preferably suppress the sprouting of fine horse jujube alternaria spore.
3. the method for fine horse jujube alternaria Toxicity Determination as claimed in claim 1, it is characterised in that：In 10 kinds of medicaments for examination, pyraclostrobin is to the virulence highest of fine horse jujube black spot, and its EC50 value is 0.0373mg/L；Secondly it is fluorine silicon-Fluoxastrobin, many bacterium and clever Difenoconazole, its EC50 value respectively 16.0752,43.8137,63.198mg/L；Remaining medicament virulence is relatively low, fluorine ring-phenylate azoles EC50 values are that 165.9195mg/L, pseudomonas fluorescens EC50 values are that 237.0493mg/L, the sweet phenol EC50 values of second are that 524.4414mg/L, three 120EC50 values are that 736.4497mg/L, streptomycin sulphate EC50 values are 1032.048mg/L, the virulence of wherein penta azoles-miaow is worst, and its EC50 value is 7007.383mg/L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610205776.6A CN107267591A (en) | 2016-04-06 | 2016-04-06 | To the method for fine horse jujube alternaria Toxicity Determination |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610205776.6A CN107267591A (en) | 2016-04-06 | 2016-04-06 | To the method for fine horse jujube alternaria Toxicity Determination |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107267591A true CN107267591A (en) | 2017-10-20 |
Family
ID=60051730
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610205776.6A Pending CN107267591A (en) | 2016-04-06 | 2016-04-06 | To the method for fine horse jujube alternaria Toxicity Determination |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107267591A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102484998A (en) * | 2010-12-02 | 2012-06-06 | 江苏丘陵地区镇江农业科学研究所 | Broad-spectrum combined bactericide |
| WO2013027863A1 (en) * | 2011-08-24 | 2013-02-28 | Sumitomo Chemical Company, Limited | Composition and method for controlling plant diseases |
| CN102972398A (en) * | 2012-12-22 | 2013-03-20 | 云南农业大学 | Application of compound to prevention and treatment of plant fungal diseases caused by alternaria alternata |
-
2016
- 2016-04-06 CN CN201610205776.6A patent/CN107267591A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102484998A (en) * | 2010-12-02 | 2012-06-06 | 江苏丘陵地区镇江农业科学研究所 | Broad-spectrum combined bactericide |
| WO2013027863A1 (en) * | 2011-08-24 | 2013-02-28 | Sumitomo Chemical Company, Limited | Composition and method for controlling plant diseases |
| CN102972398A (en) * | 2012-12-22 | 2013-03-20 | 云南农业大学 | Application of compound to prevention and treatment of plant fungal diseases caused by alternaria alternata |
Non-Patent Citations (1)
| Title |
|---|
| 陈小飞等: "10种杀菌剂及其混配剂对骏枣黑斑病菌室内毒力测定", 《中国农学通报》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104872205B (en) | The mixture and its application method prevented and treated for soil-borne disease of tobacco | |
| CN103210925B (en) | Sterilization composition containing pyrazole naphthalene bacteria amine | |
| CN103493818A (en) | Sterilization composition containing metconazole and JS399-19 | |
| CN103271050A (en) | Compound bactericide | |
| CN108294014A (en) | A kind of seed coat agent and the preparation method and application thereof of prevention root rot | |
| CN103081920B (en) | Sterilizing composition containing penconazole | |
| CN104770373B (en) | A kind of containing copper 8-hydroxyquinolinate with the bactericidal composition of metconazole | |
| CN107242242A (en) | Bactericidal composition comprising folpet and prothioconazoles | |
| CN103975929A (en) | Compound preparation for preventing anthracnose of grapes | |
| CN105076212B (en) | A kind of bactericidal composition containing bacillus subtilis and 2-cyano-3-amino-3-phenylancryic acetate | |
| CN103651396B (en) | Sterilization composition | |
| CN107267591A (en) | To the method for fine horse jujube alternaria Toxicity Determination | |
| CN104920359B (en) | Plant essential oil for preventing and controlling rice blast efficiently | |
| CN104430384B (en) | A kind of bactericidal composition | |
| CN103798262A (en) | Sterilization composition containing thiophanate-methyl and resin acid copper salt | |
| CN107593738A (en) | A kind of composition pesticide containing captan and metrafenone | |
| CN107711864A (en) | Agricultural bactericidal composition and purposes for industrial crops | |
| CN108432787A (en) | A kind of bactericidal composition containing thiophanate methyl Yu the gloomy copper of thiophene | |
| CN109418273A (en) | A kind of bactericidal composition | |
| CN106879639A (en) | A kind of composition pesticide of the flavones of fruit containing ground | |
| CN106719744A (en) | Bactericidal composition and its application | |
| CN103651397B (en) | Compound bactericide | |
| CN103814949A (en) | Triadimefon and cuprous oxide containing compound pesticide | |
| CN104756996A (en) | Sterilization composition containing quinolone copper and ipconazole | |
| CN103636643B (en) | Thiophanate-methyl-containing sterilization composition |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20171020 |
|
| RJ01 | Rejection of invention patent application after publication |