CN102577857B - Method for identifying herbicide resistance of cotton leaves by cotton ball positioning - Google Patents
Method for identifying herbicide resistance of cotton leaves by cotton ball positioning Download PDFInfo
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- CN102577857B CN102577857B CN 201210019682 CN201210019682A CN102577857B CN 102577857 B CN102577857 B CN 102577857B CN 201210019682 CN201210019682 CN 201210019682 CN 201210019682 A CN201210019682 A CN 201210019682A CN 102577857 B CN102577857 B CN 102577857B
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- 238000000034 method Methods 0.000 title claims abstract description 46
- 235000009161 Espostoa lanata Nutrition 0.000 title claims abstract description 45
- 240000001624 Espostoa lanata Species 0.000 title claims abstract description 44
- 229920000742 Cotton Polymers 0.000 title claims abstract description 29
- 239000004009 herbicide Substances 0.000 title claims abstract description 28
- 241000219146 Gossypium Species 0.000 title claims abstract description 21
- 230000002363 herbicidal effect Effects 0.000 title claims abstract description 20
- 239000000463 material Substances 0.000 claims abstract description 38
- 238000012545 processing Methods 0.000 claims abstract description 14
- 238000006243 chemical reaction Methods 0.000 claims abstract description 8
- 239000012224 working solution Substances 0.000 claims abstract description 8
- 230000006378 damage Effects 0.000 claims abstract description 4
- 241000196324 Embryophyta Species 0.000 claims abstract description 3
- 230000000875 corresponding effect Effects 0.000 claims abstract 2
- 240000002024 Gossypium herbaceum Species 0.000 claims description 20
- 235000004341 Gossypium herbaceum Nutrition 0.000 claims description 20
- 239000007788 liquid Substances 0.000 claims description 10
- 239000005562 Glyphosate Substances 0.000 claims description 9
- XDDAORKBJWWYJS-UHFFFAOYSA-N glyphosate Chemical group OC(=O)CNCP(O)(O)=O XDDAORKBJWWYJS-UHFFFAOYSA-N 0.000 claims description 9
- 229940097068 glyphosate Drugs 0.000 claims description 9
- 239000003814 drug Substances 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 230000029553 photosynthesis Effects 0.000 claims description 2
- 238000010672 photosynthesis Methods 0.000 claims description 2
- IAJOBQBIJHVGMQ-UHFFFAOYSA-N 2-amino-4-[hydroxy(methyl)phosphoryl]butanoic acid Chemical compound CP(O)(=O)CCC(N)C(O)=O IAJOBQBIJHVGMQ-UHFFFAOYSA-N 0.000 claims 1
- 239000005561 Glufosinate Substances 0.000 claims 1
- 238000011160 research Methods 0.000 abstract description 3
- 239000000575 pesticide Substances 0.000 abstract description 2
- 230000037039 plant physiology Effects 0.000 abstract description 2
- 239000000126 substance Substances 0.000 abstract description 2
- 238000002474 experimental method Methods 0.000 abstract 1
- 238000011156 evaluation Methods 0.000 description 13
- YZHUMGUJCQRKBT-UHFFFAOYSA-M sodium chlorate Chemical compound [Na+].[O-]Cl(=O)=O YZHUMGUJCQRKBT-UHFFFAOYSA-M 0.000 description 8
- 238000005507 spraying Methods 0.000 description 7
- 238000011835 investigation Methods 0.000 description 6
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- 238000009395 breeding Methods 0.000 description 3
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- 238000012797 qualification Methods 0.000 description 3
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- 238000005516 engineering process Methods 0.000 description 2
- 231100000518 lethal Toxicity 0.000 description 2
- 230000001665 lethal effect Effects 0.000 description 2
- 231100000572 poisoning Toxicity 0.000 description 2
- 230000000607 poisoning effect Effects 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000007639 printing Methods 0.000 description 2
- 230000007226 seed germination Effects 0.000 description 2
- 239000007921 spray Substances 0.000 description 2
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 description 2
- WTEJQBARPJSNLZ-UHFFFAOYSA-N C(C(=O)N)(=O)O.P Chemical compound C(C(=O)N)(=O)O.P WTEJQBARPJSNLZ-UHFFFAOYSA-N 0.000 description 1
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- 238000007429 general method Methods 0.000 description 1
- 238000005213 imbibition Methods 0.000 description 1
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Abstract
本发明涉及植物生理学和农药学，具体地，本发明涉及棉花叶片棉球定位鉴定除草剂抗性的方法。根据本发明的方法，包括步骤：1)材料处理叶片的选择：选择完好无损无受害现象的功能叶或其邻位叶；2)棉球定位处理：在每株的功能叶正面自叶柄处开始顺时针或逆时针方向设定作用位置，将大小基本一致的棉球蘸满设定浓度的除草剂工作液，按固定方向依序放置各种浓度药剂浸泡的棉球于叶片表面的相应作用位置；3)观察处理部位反应情况来确定其抗性水平。该方法可大大节省材料，减小实验规模，大大减少人为误差，结果精确可靠，对精确定义材料抗性水平提供了技术保障，也为材料当代单株进行深入的抗性研究提供保证，操作极其简单，容易掌握。
The invention relates to plant physiology and pesticide science, in particular, the invention relates to a method for identifying herbicide resistance by locating cotton balls on cotton leaves. According to the method of the present invention, comprise steps: 1) the selection of material processing blade: select the functional leaf or its adjacent leaf without damage phenomenon intact; Set the action position clockwise or counterclockwise, dip the cotton balls of basically the same size into the herbicide working solution of the set concentration, and place the cotton balls soaked in various concentrations of chemicals on the corresponding action positions on the leaf surface in sequence according to the fixed direction ; 3) Observing the reaction of the treatment site to determine its resistance level. This method can greatly save materials, reduce the scale of experiments, greatly reduce human errors, and the results are accurate and reliable. It provides technical support for accurately defining the resistance level of materials, and also provides guarantee for in-depth resistance research of contemporary single plants of materials. The operation is extremely Simple and easy to master.
Description
Technical field
The present invention relates to plant physiology and Pesticide Science, particularly, the present invention relates to the method for cotton leaf cotton balls location evaluation Herbicid resistant.
Background technology
at present, it is usually the seed germination survey method that the cotton Herbicid resistant is identified, different times variable concentrations weed killer herbicide spraying method, flower bud phase blade spot printing growing point method, the culture herbicide treatment methods such as cell-line etc., but the seed germination survey method requires grain weight enough large, and spraying rule, different times variable concentrations weed killer herbicide require each seedling amount larger in period, also can be because spraying inequality, the elegant experimental error that causes of liquid, in addition in flower bud phase blade spot printing growing point method, growing point cannot represent the level of growth of whole cotton plant, and the higher concentration processing can cause that growth rate of cotton plant point is controlled, severe deformities, cotton plant can not blossom and bear fruit, and above the whole bag of tricks all may be lethal with material, and, when at the same time lot of materials being identified, require experimental size larger, be not easy to realize.
Simultaneously, when limited or rare cotton material being carried out the Herbicid resistant evaluation, due to the material quantitative limitation, process according to above-mentioned each method and very likely can destroy material, can not guarantee to find resistance level or the lethal concentration of material.At this moment, effective method is cotton plant to be carried out the part identify, so both can find the resistance level of material, can preserve material again.The present inventor recognizes, the functional leaf of cotton can represent the upgrowth situation of cotton plant, can process the cotton plant functional leaf and carry out cotton to the evaluation of Herbicid resistant.
Summary of the invention
The present inventor explores cotton plant Herbicid resistant authentication method for a long time, find to utilize the weed killer herbicide of variable concentrations to process simultaneously the cotton plant functional leaf, but both fast accurate is found out the resistance level of cotton plant, can continue again to preserve, the limited or rare cotton material of breeding; The Herbicid resistant that both had been applicable to a large amount of cotton materials is identified, also is applicable to the Herbicid resistant evaluation of limited or rare material, i.e. cotton balls location survey method.
Therefore, the method that the purpose of this invention is to provide cotton leaf cotton balls location evaluation Herbicid resistant.
Method according to cotton leaf cotton balls of the present invention location evaluation Herbicid resistant comprises the following steps:
1) selection of material processed blade: select excellent functional leaf or its ortho position leaf without the phenomenon of being injured;
2) cotton balls localization process: in the functional leaf front of every strain from petiole clockwise or counterclockwise set active position, the cotton balls that size is basically identical dips in the weed killer herbicide working solution of full setting concentration, sequentially places the cotton balls of various concentration medicaments immersions in the respective action position of blade surface by fixed-direction;
3) observe the treatment sites response situation and determine its resistance level.
According to the method for cotton leaf cotton balls of the present invention location evaluation Herbicid resistant, the step that it comprises cotton balls location evaluation wherein, is treated to Local treatment.According to requirement of the present invention, the blade of herbicide treatment is chosen functional leaf or its ortho position leaf, be generally down three or fall four leaves, it is the strongest blade of photosynthesis, because conditioned response is responsive to external world to cross tender tissue, when using the low concentration glyphosate, very easily show symptom of chemical damage, can not represent the resistance level of material; And the old tissue metabolism is not vigorous excessively, processes rear blade and may not react, and the natural Physiology and biochemistry of some of leaf tissue changes also easily affects qualification result.
Method according to cotton leaf cotton balls of the present invention location evaluation Herbicid resistant, require higher to weather conditions, guarantee to process rear 6 hours without rain, and be preferably in and process during without dew the calm sunny morning, guarantee that liquid and the blade of accurate concentration fully acts on; Too strict to do not need requiring breeding time of cotton plant, namely seedling stage, flower bud phase, flowering and boll-setting period all can.
The evaluation of cotton balls positioning mode according to the present invention to the cotton Herbicid resistant, wherein, absorbent cotton will be placed according to fixing order, or clockwise, or counterclockwise; It is sufficient that the absorbent cotton dose is wanted, but will guarantee cotton balls to be placed in blade and do not have liquid to flow; The cotton balls liquor strength can not unrestrictedly increase, otherwise may cause death of seedling or growing point deformity, and is seriously controlled; The absorbent cotton size is suitable, is sure not to occur variable concentrations absorbent cotton adhesion phenomenon; Cotton balls will use absorbent cotton, guarantees water imbibition.
According to the specific embodiment of the present invention, the method for described cotton leaf cotton balls location evaluation Herbicid resistant specifically comprises: the 1) selection of material processed blade: select excellent functional leaf without the phenomenon of being injured, a strain a slice; 2) the cotton balls positioning mode is processed, begin clockwise direction in the functional leaf front of every strain from the petiole place and set 5 positions, dip in the weed killer herbicide working solution of full setting concentration with the tweezers cotton balls that size is basically identical (absorbent cotton), sequentially be positioned over the blade surface relevant position.3) investigation in the 4th day after processing is observed the treatment sites response situation and is determined its resistance level.
The present invention is important technology invention in the basic scientific research cause, and international and domestic for creating, it has the following advantages:
1, spraying processing meeting is because spraying inequality, the legal adopted material resistance level of Pyatyi poisoning, can because visually observing, estimate that injured area or the order of severity difference of being injured are forbidden and causes human error, the definition meetings such as planting percent to select the improper experimental error that causes because of control time; Whether the cotton balls positioning mode is pressed the concentration classification, react as standard definition material resistance take treatment sites, greatly reduces human error, and result is accurately reliable.
2, for limited or rare material, adopt the cotton balls positioning mode to identify, for continuing normal growth, material provides guarantee, provide technical guarantee to explication material resistance level, also for carrying out deep resistance research, material individual plant in the present age gives security.
3, for the screening of lot of materials to Herbicid resistant, adopt the cotton balls positioning mode to identify and can greatly save material, reduce experimental size.
4, described method is quick than other method, and the cycle is short, general method will be after processing investigation in 10-15 days, but result namely appearred in cotton balls location survey method in 4 days, this has greatly reduced qualification cycle, has effectively improved determination rates.
5, described method can arrange several variable concentrations simultaneously on a strain cotton plant, and single treatment can be determined the reaction interval of cotton plant, greatly saves material, shortens qualification time, improves determination rates, has shown that the method is quick, economizes material, advantage in batches.
When 6, adopting the cotton balls positioning mode to carry out the weed killer herbicide evaluation to lot of materials, not only can know each material resistance level, simultaneously can be at comparative material Resistant Difference under same level.
7, dosage is few, significantly reduces experimental cost, and is substantially pollution-free to environment simultaneously, and huge contribution is done in environmental protection.And, operate extremely simply, easily grasp.
Description of drawings
Fig. 1 cotton balls positioning mode is processed cotton leaf, and on cotton leaf, 1-5 weed killer herbicide concentration increases successively.
Fig. 2 cotton balls location survey method observed result is the paramount sense of high resistance from a-f, wherein, a. high resistance, blade is reactionless, 1 grade; B. anti-, 5 reactions, 1-4 does not react, 2 grades; C. anti-, 4-5 responds, and 1-3 is reactionless, 3 grades; D. not anti-, 3-5 responds, and 1-2 is reactionless, 4 grades; E. sense, 2-5 responds, and 1 is reactionless, 5 grades; F. high sense, 1-5 has significant reaction, 6 grades.
Embodiment
1, processing material blade selects
Material plantation, management etc. do not have specific requirement, and the material of general planting management or self-sow all can.Blade selects there is no specific (special) requirements from the cotton plant position, stem, vegetative shoot, fruit branch, secondary or three grades of branches all can, but require excellent functional leaf without the phenomenon of being injured, a strain a slice; , be not strict with on breeding time from cotton plant, seedling stage, flower bud phase, flowering and boll-setting period all can.Before processing, blade is carried out mark, but the mark time can not be oversize in advance, otherwise for herbicide treatment, leaf growth is too fast, blade can be partially old, and error appears in result.
2, liquid preparation
At first liquid preparation is mixed with stoste mother liquor (every 10 milliliter of 41% glyphosate stoste fully be dissolved in be mixed with mother liquor in 400 ml waters be 1% glyphosate solution) by a certain percentage, further be diluted to again working solution (respectively 1% glyphosate mother solution of 6 milliliters, 30 milliliters, 90 milliliters, 150 milliliters and 200 milliliters fully is dissolved in the water of 994 milliliters, 970 milliliters, 910 milliliters, 850 milliliters and 800 milliliters, is mixed with respectively 0.006%, 0.03%, 0.09%, 0.15% and 0.2% glyphosate working solution).Stoste, mother liquor preferably are stored in refrigerator, and mother liquor and working solution can not preserve for a long time, and working solution can not be exposed to the sun in the sun, also can not be placed in the air port, and is preferably In Shade.
3, cotton balls localization process
The cotton balls localization process is to begin clockwise direction in the functional leaf front of every strain from the petiole place to set several positions, will dip in the cotton balls (absorbent cotton) of full setting concentration glyphosate with tweezers, sequentially is positioned over the blade surface relevant position, sees accompanying drawing 1.Key is that the cotton balls size is basically identical, and the absorbent cotton dose is sufficient, but not too many in case flow on blade.In addition, use blunt nosed tweezers, in order to avoid the tip tweezers scratch blade.
4, investigation
Investigation in about the 4th day after processing, response situation clearly with material resistance grading evaluation, is seen accompanying drawing 2 according to response situation, a. high resistance, blade is reactionless, 1 grade; B. anti-, 5 reactions, 1-4 does not react, 2 grades; C. anti-, 4-5 responds, and 1-3 is reactionless, 3 grades; D. not anti-, 3-5 responds, and 1-2 is reactionless, 4 grades; E. sense, 2-5 responds, and 1 is reactionless, 5 grades; F. high sense, 1-5 has significant reaction, 6 grades.
When cotton is identified in the cotton balls location; every cotton plant is the processing capacity leaf all; have uniformity and representativeness; and every functional leaf is all processed ad-hoc location, and is pointed, and the dose of the accurate concentration of every treatment sites is enough sufficient; the liquid that can guarantee every processing position tissue and accurate concentration fully acts on; and inequality usually can appear spraying in spraying, every material repeats to spray the phenomenon that dose differs in size, the variable concentrations liquid is elegant, causes the method to have randomness, difficulty to control, and error is larger.In addition, investigate every processing position when the cotton balls positioning mode is processed four days and reaction definition resistance level whether occurs, it is qualitative survey, artificial subjective error is less, the investigation of Pyatyi poisoning method will be estimated injured area or extent of injury seriousness, it is quantitative interview, artificial subjective error is larger, may be subjected to macroscopic the impact, and the spray liquid uniformity is very large on the result impact, the planting percent investigation method can more difficult unified dose level difference causes when processing delay of response, serious different, thereby control time hold the improper experimental error that causes.
Identify just to the local dispenser of cotton plant cotton balls location, and dose is very little, although herbicide glyphosate and phosphine oxamate have certain conductivity, the excessive concentration weed killer herbicide in cotton balls location only otherwise on cotton plant just is unlikely to suppress fully or lethal cotton plant.Therefore, the cotton plant after this method authentication process can be grown by normal growth substantially.
Claims (5)
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| CN 201210019682 CN102577857B (en) | 2012-01-24 | 2012-01-24 | Method for identifying herbicide resistance of cotton leaves by cotton ball positioning |
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| CN103081731B (en) * | 2013-02-28 | 2014-12-31 | 中国农业科学院棉花研究所 | Method for identifying glyphosate resistant cotton rapidly |
| CN105379570A (en) * | 2015-11-16 | 2016-03-09 | 中国农业科学院棉花研究所 | Method for rapidly identifying glufosinate-ammonium resisting cotton |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102165885A (en) * | 2010-12-15 | 2011-08-31 | 中国农业科学院植物保护研究所 | Method for identifying glyphosate resistance of cotton |
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| US6818807B2 (en) * | 2001-08-06 | 2004-11-16 | Bayer Bioscience N.V. | Herbicide tolerant cotton plants having event EE-GH1 |
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| CN102165885A (en) * | 2010-12-15 | 2011-08-31 | 中国农业科学院植物保护研究所 | Method for identifying glyphosate resistance of cotton |
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| Title |
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| 作物的功能叶;鄂玉江;《新农业》;19841231(第15期);第15页 * |
| 转基因抗草甘膦棉花鉴定方法研究;王秀丽等;《山东农业科学》;20081231(第09期);第79-80页 * |
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