CN104267152B - A kind of drug-fast method of detection barnyard grass fast - Google Patents
A kind of drug-fast method of detection barnyard grass fast Download PDFInfo
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- CN104267152B CN104267152B CN201410517531.8A CN201410517531A CN104267152B CN 104267152 B CN104267152 B CN 104267152B CN 201410517531 A CN201410517531 A CN 201410517531A CN 104267152 B CN104267152 B CN 104267152B
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Abstract
The invention discloses and a kind of detect the drug-fast method of barnyard grass fast, comprise the following steps: (1) collects barnyard grass seed: collect the barnyard grass seed of herbicide sensitive as material; (2) cultivation of barnyard grass: after barnyard grass seed breaking dormancy step (1) collected, first use potassium nitrate Steeping and budding, then the barnyard grass of germination being put into nutrient solution, to be cultured to 2 leaves stand-by; (3) process of barnyard grass: the barnyard grass being cultured to 2 leaves in step (2) is bonded on plastic plate, the herbicide liquid putting into different gradient concentration again soaks 30 minutes, then take out nature to dry, put into nutrient solution and continue cultivation 15 days, assessment barnyard grass is in leaf color, young leaves growth and the change of root system development, investigation barnyard grass mortality ratio, obtains the examination concentration of herbicide, can detect whether barnyard grass creates the resistance to the action of a drug to herbicide fast.The inventive method can disposable detection large batch of barnyard grass sample, and has time short, advantage accurately and reliably.
Description
Technical field
The present invention relates to technical field of plant protection, particularly a kind of quick detection barnyard grass method that whether herbicide is developed immunity to drugs.
Background technology
Barnyard grass (
echinochloacrusgalli) be one of worldwide ten large malignant weeds, Ye Shi China chief crop endangers one of the most serious weeds.Particularly serious to Rice Production harm, not only hazard area is large, and causes the paddy rice underproduction also the most remarkable.
Current chemical control remains the main approach preventing and kill off barnyard grass.Because in barnyard grass farmland worldwide, distribution area is wide, harm is large.Therefore, the exploitation major part of herbicide is with barnyard grass for mainly to prevent and kill off target, and the barnyard grass agent that removes of various dissimilar, different mechanism of action is developed in succession.
But along with the increase of tenure of use and the increase of Selective Pressure, barnyard grass all creates the resistance to the action of a drug to the long-term a large amount of herbicide used of part.Especially in the Dongting Lake Rice Areas in Hunan Province, because Direct-seeding Rice developing history is long, the frequency of utilization of herbicide is high, and Selective Pressure is large, and barnyard grass development of drug resistance is rapid.From 1991, dichloro quinolinic acid used at this area's spread, and to 2002, the barnyard grass in paddy field of this area created the serious resistance to the action of a drug to dichloro quinolinic acid.Dichloro quinolinic acid was stopped using afterwards, gradually substitute by the penoxsuam of triazolopyrimidine class.Within nearly 2 years, according to local farmers reaction, the preventive effect of penoxsuam to barnyard grass sharply declines, and double increase using dosage is also difficult to gather effect.In addition, Jiangxi, Anhui, Jiangsu, Zhejiang, Shanghai etc. are economized and are all found that barnyard grass is to the drug-resistant organisms type of dichloro quinolinic acid.The barnyard grass of Liaoning and Guangdong Dao Qu also creates the obvious resistance to the action of a drug to butachlor and benthiocarb.
The drug-fast detection method of current barnyard grass is a lot, the most frequently used method is greenhouse whole strain determination of plant method, basic step is, first from long-term single used herbicide and suspect have drug-fast weed seed to adopt back, be seeded in the basin in greenhouse, conventional dispenser process is carried out before seedling or after seedling after broadcasting, medicament arranges variable concentrations gradient, then the emergence rate of weeds under various dose is investigated, germinate and mortality ratio, leaf area, the indexs such as dry weight, and compare with blank, use different herbicide concentration process resistances and responsive seed, and make regression curve according to dosage and anti-effect relationship and calculate ED
50, simple and easy to do to determine resistant multiple's the method technology, plant in enormous quantities carries out simultaneously, reproducible, is often utilized.But the large problem that the method exists is that the whole test period is oversize, and spended time is many, and workload is very big.Other method many in addition, as target enzymes activation measurement, chloroplast resistant determination method, fluorescence signal detection and target resistance Molecular Detection method etc.But the detecting instrument that some methods are used is expensive, as chloroplast resistant determination method and fluorescence signal detection.The test reagent also having Part Methods to relate to is expensive, needs technical professional just can complete, as target resistance Molecular Detection method.
Summary of the invention
The technical problem to be solved in the present invention overcomes the deficiencies in the prior art, there is provided a kind of and detect the drug-fast method of barnyard grass fast, the method is simple and reliable, can detect the Detection of insecticide resistance of a large amount of barnyard grass sample to cauline leaf treating agent (as dichloro quinolinic acid, cyhalofop-butyl and penoxsuam etc.) fast.
For solving the problems of the technologies described above, the technical scheme that the present invention proposes is:
A kind of drug-fast method of detection barnyard grass fast, comprises the following steps:
(1) barnyard grass seed is collected: collect the barnyard grass seed of herbicide sensitive as material;
(2) cultivation of barnyard grass: the barnyard grass seed potassium nitrate Steeping and budding that step (1) is collected, then the barnyard grass of germination being put into nutrient solution, to be cultured to 2 leaves stand-by;
(3) process of barnyard grass: the barnyard grass being cultured to 2 leaves in step (2) is bonded on plastic plate, the herbicide liquid putting into different gradient concentration soaks 30 minutes, then take out nature to dry, put into nutrient solution again and continue cultivation 15 days, assessment barnyard grass is in leaf color, young leaves growth and the change of root system development, investigation barnyard grass mortality ratio, obtains the examination concentration of herbicide liquid.
Preferably, in described step (2), the cultivation of barnyard grass specifically: after the barnyard grass seed breaking dormancy after collection, first use 0.25% potassium nitrate soak at room temperature 1 day, then filter out unnecessary moisture, be placed in incubator and cultivate, after it germinates, put into nutrient solution continue to cultivate, when being cultured to for 1 leaf phase, barnyard grass seedling 10 strain is affixed on a long 15cm, on the plastic plate of wide 7cm, then plastic plate is positioned over nutrient solution is housed cup in continue to grow to 2 leaves.
Preferably, described nutrient solution is that nutrient solution sowed by the fertilizer ten thousand of concentration 0.526ml/ml, allows plastic plate tiltedly be put in cup when barnyard grass is cultivated, to ensure that plant root immerses in cup Middle nutrition liquid.
Preferably, the examination concentration of described herbicide liquid is barnyard grass mortality ratio herbicide concentration corresponding when being 95%.
Compared with prior art, the invention has the advantages that:
One of the present invention detects the drug-fast method of barnyard grass fast, can detect whether barnyard grass creates the resistance to the action of a drug to herbicide fast, in enormous quantities and exactly, field barnyard grass can being grasped in time dynamic to the resistance to the action of a drug of herbicide, providing theoretical foundation for using herbicide scientifically and rationally.The method is suitable for the monitoring for resistance of grassy weed barnyard grass.
Embodiment
For the ease of understanding the present invention, hereafter will do to describe more comprehensively, meticulously to the present invention in conjunction with preferred embodiment, but protection scope of the present invention is not limited to following specific embodiment.
An embodiment for the drug-fast method of quick detection barnyard grass, comprises the following steps:
(1) barnyard grass seed is collected: collect the barnyard grass population seed to herbicide sensitive, this sensitive population is determined by pot-culture method;
(2) cultivation of barnyard grass: after the barnyard grass seed breaking dormancy that step (1) is collected, first use 0.25% potassium nitrate soak at room temperature 1 day, then unnecessary moisture is filtered out, be placed in incubator and cultivate, after it germinates, the concentration put into through groping to determine is that the fertilizer ten thousand of 0.526ml/ml is sowed and (is set to normal concentration, under this concentration, the growth of barnyard grass is best) continue in nutrient solution to cultivate, when being cultured to for 1 leaf phase, barnyard grass seedling 10 strain is affixed on a long 15cm, on the plastic plate of wide 7cm, again plastic plate is tiltedly positioned over and is equipped with fertile ten thousand and sows continued growth in the cup of nutrient solution, plant root is kept to immerse in cup Middle nutrition liquid during cultivation, be cultured to after 2 leaves until barnyard grass, stand-by,
(3) process of barnyard grass: the herbicide liquid barnyard grass being cultured to 2 leaves in step (2) being put into different gradient concentration soaks 30 minutes, then take out nature to dry, put into nutrient solution again and continue cultivation 15 days, assessment barnyard grass is in leaf color, young leaves growth and the change of root system development, investigation barnyard grass mortality ratio, herbicide concentration corresponding when acquisition barnyard grass mortality ratio is 95%, this concentration is the examination concentration of herbicide liquid.
embodiment one
barnyard grass is to the resistance to the action of a drug method for quick of dichloro quinolinic acid
First by after the barnyard grass seed breaking dormancy of collection, with 0.25% potassium nitrate soak at room temperature 1 day, filter out unnecessary moisture, be placed in again in incubator and cultivate, after it germinates, put into concentration is that nutrient solution continuation cultivation sowed by 0.526ml/ml fertilizer ten thousand, when being cultured to for 1 leaf phase, barnyard grass seedling 10 strain is affixed on a long 15cm, on the plastic plate of wide 7cm, again plastic plate is tiltedly positioned over and is equipped with fertile ten thousand and sows continued growth in the cup of nutrient solution, keep plant root to be immersed in the water during cultivation, be cultured to after 2 leaves until barnyard grass, stand-by.
Screen the screening of concentration:
With the field dosage of dichloro quinolinic acid for reference, first 6 are established to process 0,0.05,0.5,5,50,500,5000ga.i./hm with 10 times amount gradients
2, each process repetition 3 times, tiltedly puts into dichloro quinolinic acid liquid and soaks 30 minutes by the plastic plate being stained with 10 strain barnyard grasses, taking-up allows it naturally dry, put into nutrient solution again and continue cultivation 15 days, assessment barnyard grass in leaf color, young leaves growth and the change of root system development, investigation barnyard grass mortality ratio.Result shows: 6 process 0,0.05,0.5,5,50,500,5000ga.i./hm
2mortality ratio be respectively 0,0,0,35%, 65%, 100%, 100%.Determine that first time screens concentration range at 5-500ga.i./hm
2between.
According to above-mentioned scope 5-500ga.i./hm
2second time establishes 6 to process 0,31.25,62.5,125,250,500ga.i./hm
2, each process repetition 3 times, barnyard grass is cultivated and is processed the same.Result shows: 6 process 0,31.25,62.5,125,250,500ga.i./hm
2mortality ratio be respectively 0,41%, 72%, 100%, 100%, 100%.Determine that second time screens concentration range at 62.5-125ga.i./hm
2between.
According to above-mentioned scope 62.5-125ga.i./hm
2third time establishes 6 to process 0,50,70,90,110,130ga.i./hm
2, each process repetition 3 times, barnyard grass is cultivated and is processed the same.Result shows: 6 process 0,50,70,90,110,130ga.i./hm
2mortality ratio be respectively 0,67%, 89%, 98%, 100%, 100%.Determine that third time screens concentration range at 70-90ga.i./hm
2between.
According to above-mentioned scope 70-90ga.i./hm
26 are established for 4th time to process 0,60,70,80,90,100ga.i./hm
2, each process repetition 3 times, barnyard grass is cultivated the same, and the sticky 20 strain barnyard grasses of current every block plate, process the same.Result shows: 6 process 0,60,70,80,90,100ga.i./hm
2mortality ratio be respectively 0,75%, 90%, 95%, 100%, 100%.Determine that screening concentration the 4th time is 80ga.i./hm
2.
With the examination concentration 80ga.i./hm of the 4th time
2process the resistant multiple that the barnyard grass sample 1 of anti-dichloro quinolinic acid, sample 2 and sample 3(pot-culture method determine and be respectively 397.1,125.7 and 95.8) found that: 3 samples mortality ratio after screening concentration process is respectively 15%, 37% and 65%.Consistent with traditional pot-culture method result.
embodiment two
barnyard grass is to the resistance to the action of a drug method for quick of penoxsuam
First by after the barnyard grass seed breaking dormancy of collection, with 0.25% potassium nitrate soak at room temperature 1 day, filter out unnecessary moisture, be placed in again in incubator and cultivate, after it germinates, put into concentration is that nutrient solution continuation cultivation sowed by 0.526ml/ml fertilizer ten thousand, when being cultured to for 1 leaf phase, barnyard grass seedling 10 strain is affixed on a long 15cm, on the plastic plate of wide 7cm, again plastic plate is tiltedly positioned over and is equipped with fertile ten thousand and sows continued growth in the cup of nutrient solution, keep plant root to be immersed in the water during cultivation, be cultured to after 2 leaves until barnyard grass, stand-by.
Screen the screening of concentration:
With the field dosage of penoxsuam for reference, first 6 are established to process 0,0.0025,0.025,0.25,2.5,25,250ga.i./hm with 10 times amount gradients
2, each process repetition 3 times, tiltedly puts into penoxsuam liquid and soaks 30 minutes by the plastic plate being stained with 10 strain barnyard grasses, taking-up allows it naturally dry, put into nutrient solution again and continue cultivation 15 days, assessment barnyard grass in leaf color, young leaves growth and the change of root system development, investigation barnyard grass mortality ratio.Result shows: 6 process 0,0.0025,0.025,0.25,2.5,25,250ga.i./hm
2strain preventive effect be respectively 0,0,0,0,55%, 100%, 100%.Determine that primary examination concentration range is at 2.5-25ga.i./hm
2between.
According to above-mentioned scope 2.5-25ga.i./hm
2second time establishes 6 to process 0,3.125,6.25,12.5,25,50ga.i./hm
2, each process repetition 3 times, barnyard grass is cultivated and is processed the same.Result shows: 6 process 0,3.125,6.25,12.5,25,50ga.i./hm
2mortality ratio be respectively 0,51%, 63%, 90%, 100%, 100%.Determine that secondary examination concentration range is at 12.5-25ga.i./hm
2between.
According to above-mentioned scope 12.5-25ga.i./hm
2third time establishes 6 to process 0,5,10,15,20,25ga.i./hm
2, each process repetition 3 times, barnyard grass is cultivated and is processed the same.Result shows: 6 process 0,5,10,15,20,25ga.i./hm
2mortality ratio be respectively 0,61%, 87%, 91%, 100%, 100%.Determine that third time screens concentration range at 15-20ga.i./hm
2between.
According to above-mentioned scope 15-20ga.i./hm
26 are established for 4th time to process 0,14,16,18,20,22ga.i./hm
2, each process repetition 3 times, barnyard grass is cultivated the same, and the sticky 20 strain barnyard grasses of current every block plate, process the same.Result shows: 6 process 0,14,16,18,20,22ga.i./hm
2mortality ratio be respectively 0,85%, 89%, 92%, 95%, 100%.Determine that the examination concentration of the 4th time is 20ga.i./hm
2.
With the examination concentration 20ga.i./hm of the 4th time
2the resistant multiple that process resistance barnyard grass sample 1, sample 2 and sample 3(pot-culture method are determined is respectively 354.7,119.7 and 112.1) found that: 3 samples mortality ratio after screening concentration process is respectively 19.7%, 36.2% and 47.8%.Consistent with traditional pot-culture method result.
Claims (3)
1. detect the drug-fast method of barnyard grass fast, it is characterized in that, comprise the following steps:
(1) barnyard grass seed is collected: collect the barnyard grass seed of herbicide sensitive as material;
(2) cultivation of barnyard grass: the barnyard grass seed potassium nitrate Steeping and budding that step (1) is collected, then the barnyard grass of germination being put into nutrient solution, to be cultured to 2 leaves stand-by;
(3) process of barnyard grass: the barnyard grass being cultured to 2 leaves in step (2) is bonded on plastic plate, the herbicide liquid putting into different gradient concentration soaks 30 minutes, then take out nature to dry, put into nutrient solution again and continue cultivation 15 days, assessment barnyard grass is in leaf color, young leaves growth and the change of root system development, investigation barnyard grass mortality ratio, obtains the examination concentration of herbicide liquid;
Described herbicide is cauline leaf treating agent dichloro quinolinic acid, cyhalofop-butyl or penoxsuam, and described nutrient solution is concentration 0.526
fertilizer ten thousand sow nutrient solution;
The examination concentration of described acquisition herbicide liquid is specifically: first establish 6 process with the herbicide liquid of 10 times amount gradient concentrations, determines that primary examination concentration range be barnyard grass mortality ratio is concentration corresponding when being greater than 35%; Establish 6 process according to primary examination concentration range again, determine that secondary examination concentration range is the concentration corresponding when being greater than 72% of barnyard grass mortality ratio; Then establish 6 process according to secondary examination concentration range, determine that the examination concentration range of third time is the concentration corresponding when being greater than 89% of barnyard grass mortality ratio; Finally establish 6 process according to the examination concentration range of third time, screen the examination concentration that concentration is herbicide liquid for the 4th time corresponding when determining that barnyard grass mortality ratio is 95%.
2. the drug-fast method of quick detection barnyard grass according to claim 1, it is characterized in that, in described step (2), the cultivation of barnyard grass specifically: after the barnyard grass seed breaking dormancy after collection, first use 0.25% potassium nitrate soak at room temperature 1 day, then unnecessary moisture is filtered out, be placed in incubator and cultivate, after it germinates, put into nutrient solution continue to cultivate, when being cultured to for 1 leaf phase, barnyard grass seedling 10 strain is affixed on a long 15cm, on the plastic plate of wide 7cm, then plastic plate is positioned over nutrient solution is housed cup in continue to grow to 2 leaves.
3. the drug-fast method of quick detection barnyard grass according to claim 2, is characterized in that, allows plastic plate tiltedly be put in cup when barnyard grass is cultivated, to ensure that plant root immerses in cup Middle nutrition liquid.
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| CN105353094A (en) * | 2015-12-09 | 2016-02-24 | 山东省农业科学院植物保护研究所 | Quick detection method for resistant digitaria sanguinalis |
| CN106802337A (en) * | 2016-11-23 | 2017-06-06 | 北海高创电子信息孵化器有限公司 | A kind of method of work of herbaceous plant resistance to the action of a drug Information Collection System |
| CN109379920B (en) * | 2018-09-21 | 2021-11-02 | 湖南省植物保护研究所 | Method for rapidly breaking barnyard grass seed dormancy and improving germination rate |
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| WO1997020209A2 (en) * | 1995-12-01 | 1997-06-05 | E.I. Du Pont De Nemours And Company | Agricultural product microscreen method and apparatus |
| CN1544937A (en) * | 2003-11-13 | 2004-11-10 | 南京农业大学 | Method for identifying pesticide resistance of dentes foxtail to high performance cover grass |
| CN1544936A (en) * | 2003-11-13 | 2004-11-10 | 南京农业大学 | Method for identifying pesticide resistance of dentes foxtail to sethoxydim |
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