CN103411971A - Method for quickly evaluating glyphosate tolerance of cotton in field - Google Patents
Method for quickly evaluating glyphosate tolerance of cotton in field Download PDFInfo
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- CN103411971A CN103411971A CN2013103621945A CN201310362194A CN103411971A CN 103411971 A CN103411971 A CN 103411971A CN 2013103621945 A CN2013103621945 A CN 2013103621945A CN 201310362194 A CN201310362194 A CN 201310362194A CN 103411971 A CN103411971 A CN 103411971A
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Abstract
The invention provides a method for quickly evaluating glyphosate tolerance of cotton in a field. The method comprises the following steps: sowing cotton seeds to be tested and sowing cotton seeds sensitive to the glyphosate to serve as a contrast; spraying the glyphosate on to-be-tested cotton and contrast cotton at the seedling stage of cotton; starting from the first day after spraying the glyphosate, surveying the phytotoxicity occurrence of the contrast cotton every day by a grading eye survey method and calculating the phytotoxicity index of the contrast cotton; when the phytotoxicity index of the contrast cotton is equal to or more than 50, surveying the phytotoxicity occurrence of the to-be-tested cotton by the same grading eye survey method and calculating the phytotoxicity index of the to-be-tested cotton; and calculating the relative phytotoxicity index of the to-be-tested cotton according to the phytotoxicity index of the to-be-tested cotton and the phytotoxicity index, which is equal to or more than 50, of the contrast cotton, and evaluating the glyphosate tolerance of the to-be-tested cotton according to the relative phytotoxicity index obtained through calculation. The method has the advantages of small occupied space, short time, low cost, high accuracy and high repeatability.
Description
Technical field
The application relates to the plant protection technology field, more specifically, relates to the method for a kind of field Fast Evaluation cotton to glyphosate tolerant.
Background technology
Weed in cotton field harm is one of major obstacle of Cotton in China production, and annual hazard area accounts for 67% of the cotton planting total area, causes damage in 15% left and right, has seriously restricted high-quality, the High-efficient Production of cotton.Chemical weed control, as the important component part of cotton Growing method system, is prevented and kill off in process and is had irreplaceable effect at weed in cotton field.Yet, in actual production, alternative cotton field herbicide kind is single, the herbicide spectrum is narrower, broadcast front or broadcast the soil processing herbicide used before rear seedling and mainly contain alachlor, Acetochlor, pendimethalin, S-metolachlor, trefanocide etc., be used to preventing and kill off annual gramineous weed and part broad leaved weed; The emerge cauline leaf of rear use of cotton is processed herbicide, as the spirit of efficient pyrrole fluorine first standing grain, sethoxydim, smart quizalofopethyl, fenoxapropPethyl, efficient fluazifop, be mainly used in preventing and kill off grassy weed, and relative less for the herbicide kind of cotton field perennial broadleaf weed and nutgrass flatsedge, the current glyphosate that only has that has obtained registration.
As in a kind of wide spectrum, inhaling steriland herbicide, glyphosate can be prevented and kill off nearly all annual and perennial grassy weed, broad leaved weed and nutgrass flatsedge effectively, plays an important role in the preventing and kill off of weed in cotton field.But to the glyphosate sensitivity, when with glyphosate, carrying out chemical weed control, cotton also can suffer damage due to cotton, even dead.The plantation of resistance glyphosate cotton, will greatly improve range of application and the dirigibility of glyphosate, improves the preventive effect of weed in cotton field.
Since the U.S. in 1997 took the lead in planting transgenosis resistance glyphosate cotton, the cultivated area of global transgenosis resistance glyphosate cotton increased year by year, and 2012 Annual planting areas reach the fifty-five million hectare, accounted for 23% left and right of the total cultivated area of global transgene cotton.Although also there are larger gap in aggregate level and the developed country of China's transgenosis resistance glyphosate cotton research, but through research and effort for many years, at present separated and identified a plurality of glyphosate high resistance genes that have independent intellectual property right, and having obtained the monoclonal antibody glyphosate of a collection of stably express and the transgene cotton new material with the pest-resistant complex character of resistance glyphosate.Along with the quickening of China's Development of China's Urbanization, a large amount of transfers of rural laborer and the rising of labor cost, the demand of cotton production Glyphosate-Resistant kind is more and more stronger.Transgenosis resistance glyphosate cotton certainly will will become China and after Insect Resistant Cotton, be expected to the another kind of transgene cotton industrialization kind of promoting.In resistance glyphosate cotton variety Breeding Process, the evaluation cotton material is the key link of breeding success or failure to the tolerance of glyphosate, is also one of content of transgene cotton environmental safety evaluation.
At present the evaluation of cotton glyphosate resistance degree is mainly adopted to the plant bioassay method, namely in container or field sowing cotton, the glyphosate solution of a series of concentration of the rear use of emerging sprays cotton seedling, according to the death condition of cotton after dispenser and the impact of output is estimated its resistance, the method can be measured the resistance level of cotton to glyphosate accurately, but required time is longer, from sowing, results to showing that measurement result needs about 150 days, this is for many materials, large batch of mensuration, length consuming time, take up room large, efficiency is low.Therefore, set up a kind of quick, easy and estimate accurately and effectively the method for cotton to glyphosate tolerant, significant for the cultivation of resistance glyphosate cotton, popularization and production application from now on.
Summary of the invention
The application's purpose is to provide the method for a kind of field Fast Evaluation cotton to glyphosate tolerant.
Particularly, the application provides the method for a kind of field Fast Evaluation cotton to glyphosate tolerant, and described method comprises:
Sow cotton seeds to be measured, and sowing to the cotton seeds of glyphosate sensitivity in contrast;
Cotton in seedling stage, spray glyphosate to cotton to be measured and contrast cotton;
Spray after glyphosate the 1st day and start, adopt the classification ocular estimate, a situation arises and calculate the poisoning index of contrast cotton in the poisoning of investigation contrast every day cotton;
When the poisoning index of contrast cotton, while being equal to or greater than 50, adopts identical classification ocular estimate, investigate the poisoning of cotton to be measured a situation arises and calculate the poisoning index of cotton to be measured;
According to the poisoning exponential sum of cotton to be measured contrast cotton described, be equal to or greater than the relative poisoning index that 50 poisoning index calculates cotton to be measured, and according to the tolerance of the relative poisoning index assessment cotton to be measured calculated to glyphosate.
In some embodiments, to the cotton of glyphosate sensitivity, can be the whole dead cottons of cotton plant after after the glyphosate that sprays 61.5～123.0g a.i./mu in 3～6 leaf phases the 20th day.Preferably, described contrast cotton can be " CCRI 49 ".
In some embodiments, sowing cotton seeds to be measured with the method for contrast cotton seeds can be: field sowing, drilling, line-spacing 30～50cm, the rear final singling of emerging; Preferably, line-spacing can be 30cm, and the final singling spacing in the rows can be 5～6cm, and contrast cotton and each cotton to be measured can be planted respectively 170～200 strains.
In some embodiments, can grow to for 3～6 leaf phases in cotton, spray glyphosate to cotton to be measured and contrast cotton.Preferably, can grow to for 3～4 leaf phases in cotton, spray glyphosate to cotton to be measured and contrast cotton.
In some embodiments, the spraying concentration of glyphosate can be 61.5～123.0g a.i./mu.Preferably, described glyphosate can reach for the agriculture that contains 41% glyphosate isopropyl amine salt, and the described agriculture that contains 41% glyphosate isopropyl amine salt reaches by About Monsanto Chemicals production.
In some embodiments, described classification ocular estimate can be 5 grades of ocular estimates.
In some embodiments, the grade scale of described 5 grades of ocular estimates can be: 0 grade, growth rate of cotton plant is normal, without symptom of chemical damage; 1 grade, 0～25% blade produces poisoning; 2 grades, 25%～50% blade produces poisoning; 3 grades, 50%～75% blade produces poisoning; 4 grades, 75%～100% blade produces poisoning.
Wherein, value range " 0～25% ", " 25%～50% ", " 50%～75% " reach the lower-limit point that " 75%～100% " does not comprise this value range, but comprise upper change point, such as the scope of " 0～25% " does not comprise 0 lower-limit point, but comprise 25% upper change point.
In some embodiments, the computing method of the described relative poisoning index of described poisoning exponential sum can be respectively:
The highest level of poisoning index=∑ (the poisoning strain number of each poisoning rank * appropriate level) * total strain number of 100/(* classification ocular estimate);
The poisoning index of poisoning index=cotton to be measured * 50/ contrasts described 50 the poisoning index that is equal to or greater than of cotton relatively.
In some embodiments, the computing method of the described relative poisoning index of described poisoning exponential sum can be respectively:
Poisoning index=∑ (the poisoning strain number of each poisoning rank * appropriate level) * total strain number of 100/(* 4), wherein 4 are the highest level of classification ocular estimate;
The poisoning index of poisoning index=cotton to be measured * 50/ contrasts described 50 the poisoning index that is equal to or greater than of cotton relatively.
In some embodiments, according to the relative poisoning index assessment cotton calculated to the standard of the tolerance of glyphosate, can be: poisoning index 0～10.0 relatively be high-resistance glyphosate; Poisoning index 10.1～20.0, be resistance glyphosate relatively; Poisoning index 20.1～35.0, be glyphosate tolerant relatively; Relative poisoning index > 35.0, for to the glyphosate sensitivity.
Wherein, in this application, the concentration of glyphosate is the concentration of glyphosate in water.
Wherein, in this application, " classification ocular estimate " can be this area 5 grades of ocular estimates, 6 grades of ocular estimates and 11 grades of ocular estimates etc. commonly used, and these classification ocular estimates can adopt grade scale known in the art.
And, in this application, the poisoning of investigation contrast cotton a situation arises can be for investigating every day 1～2 time, and the time of investigation can be the random time in a day, when carrying out twice investigation, should be greater than 8 hours interval time.
Compared with prior art, the application has following benefit:
(1) take up room littlely, each expert evidence only need take up an area approximately 3 square metres;
(2) consuming time short, from sowing cotton seed to drawing evaluation result, only need about 50 days;
(3) easy and simple to handle, testing cost is low, in whole testing process, only uses a kind of medicament of glyphosate, and only needs range estimation, without instrument and equipment;
(4) accuracy is high and reproducible, and evaluation method of the present invention arranges responsive contrast cotton, has avoided the impact of the factors such as weather, soil fertility, but duplicate detection, high conformity as a result.
Therefore, the application's method is suitable especially applies at agriculture field.
Embodiment
Below by embodiment, describing the application's embodiment, one of ordinary skill in the art appreciates that these specific embodiments only show the enforcement technical scheme of selecting for the purpose that reaches the application, is not the restriction to technical scheme.According to the application's instruction, in conjunction with prior art, be obvious to the improvement of present techniques scheme, all belong to the scope of the application's protection.
Embodiment 1
Materials and methods
Material:
5 parts of transgenosis resistance glyphosate cotton materials, be numbered respectively KCM1, KCM2, KCM3, KCM4, KCM5; The non-Bt cotton kind is CCRI 44, CCRI 79; The hybrid cotton kind is CCRI 63; Responsive contrast cotton is CCRI 49, provides by the Chinese Academy of Agriculture Science and Technologys Cotton Research Institute's breeding chamber.
The glyphosate used reaches (Roundup) as agriculture, and it contains 41% glyphosate isopropyl amine salt, by About Monsanto Chemicals production.
Method:
In field, delimit 54 1m
2Experimental plot, 100cm * 100cm, sowing cotton, the long 100cm of row, line-spacing 30cm, drilling, the cotton rear final singling of emerging, spacing in the rows 5～6cm, every row 16～20 strains, cotton is planted respectively 3 row and all plants 6 communities with each cotton to be measured in contrast.Adopt split block design, the processing of each concentration repeats 3 times.
When contrasting cotton and cotton to be measured and grow to 3～4 leaf periods, the herbicide spraying agriculture reaches, and concentration is respectively 61.5g a.i./mu and 123.0g a.i./mu, amount of water 30L/ mu.Spraying equipment is Singapore profit agriculture HD400 knapsack hand sprayer, Fan spray head.
Spray glyphosate started on the 1st day, adopt 5 grades of ocular estimates, every morning 9:00 and afternoon about 5:00 respectively investigate the poisoning of 1 contrast cotton a situation arises, investigation 50 strains of every community, and calculate the poisoning index according to following computing method: poisoning index=∑ (the poisoning strain number of each poisoning rank * appropriate level) * total strain number of 100/(* 4), wherein 4 is the highest level of 5 grades of ocular estimates.
Wherein, the grade scale of 5 grades of ocular estimates is: 0 grade, growth rate of cotton plant is normal, without symptom of chemical damage; 1 grade, 0～25% blade produces poisoning; 2 grades, 25%～50% blade produces poisoning; 3 grades, 50%～75% blade produces poisoning; 4 grades, 75%～100% blade produces poisoning.
When the poisoning index of contrast cotton while being equal to or greater than 50, adopting 5 grades of identical ocular estimates to investigate the poisoning of each cotton to be measured, a situation arises that 50 strains are investigated in the ，Mei community.As above calculate the poisoning index of each cotton to be measured, and according to the relative poisoning index of following calculating cotton to be measured: described 50 the poisoning index that is equal to or greater than of the poisoning index of poisoning index=cotton to be measured * 50/ contrast cotton relatively.
Relative poisoning index and the evaluation of resistance result of cotton to be measured are presented in table 1.In order to check the accuracy of relative poisoning assessment of indices method, table 1 has also been listed the single-strain fresh weight of the cotton of measuring in 20 days after the dispenser and the death condition of cotton while measuring the same day.
After table 1 sprays the glyphosate of variable concentrations, single-strain fresh weight and the death condition of 20 days after the poisoning index of contrast cotton is equal to or greater than the relative poisoning index of the cotton to be measured detected and dispenser at 50 o'clock
Wherein, the numerical value in table 1 all provides with the form of mean value.
As can be seen from Table 1, according to the application's method, it is consistent in glyphosate concentration, being respectively resulting evaluation of resistance result in the test of 61.5g a.i./mu and 123.0g a.i./mu.Simultaneously, the single-strain fresh weight of the cotton of measuring in 20 days after the evaluation of resistance result obtained according to the application's method and dispenser and while measuring the same day death condition of cotton be consistent.Explanation thus, the application's evaluation method has high accuracy.
The application includes but not limited to above embodiment, and every any being equal to of carrying out under the principle of the application's spirit substitutes or local improvement, all will be considered as within the application's protection domain.
Claims (10)
1. the field Fast Evaluation cotton method to glyphosate tolerant, described method comprises:
Sow cotton seeds to be measured, and sowing to the cotton seeds of glyphosate sensitivity in contrast;
Cotton in seedling stage, spray glyphosate to cotton to be measured and contrast cotton;
Spray after glyphosate the 1st day and start, adopt the classification ocular estimate, a situation arises and calculate the poisoning index of contrast cotton in the poisoning of investigation contrast every day cotton;
When the poisoning index of contrast cotton, while being equal to or greater than 50, adopts identical classification ocular estimate, investigate the poisoning of cotton to be measured a situation arises and calculate the poisoning index of cotton to be measured;
According to the poisoning exponential sum of cotton to be measured contrast cotton described, be equal to or greater than the relative poisoning index that 50 poisoning index calculates cotton to be measured, and according to the tolerance of the relative poisoning index assessment cotton to be measured calculated to glyphosate.
2. method according to claim 1, wherein, be the whole cottons of death of cotton plant after the 20th day after 3～6 leaf phases sprayed the glyphosate of 61.5～123.0g a.i./mu to the contrast cotton of glyphosate sensitivity; Preferably, described contrast cotton is " CCRI 49 ".
3. method according to claim 1, wherein, the method for sowing cotton seeds to be measured and contrast cotton seeds is: field sowing, drilling, line-spacing 30～50cm, the rear final singling of emerging; Preferably, line-spacing is 30cm, and the final singling spacing in the rows is 5～6cm, and contrast cotton and each cotton to be measured are planted respectively 170～200 strains.
4. method according to claim 1, wherein, grew to for 3～6 leaf phases in cotton, sprays glyphosate to cotton to be measured and contrast cotton; Preferably, grew to for 3～4 leaf phases in cotton, spray glyphosate to cotton to be measured and contrast cotton.
5. method according to claim 1, wherein, the spraying concentration of glyphosate is 61.5～123.0g a.i./mu, preferably, described glyphosate is that the agriculture that contains 41% glyphosate isopropyl amine salt reaches.
6. method according to claim 1, wherein, described classification ocular estimate is 5 grades of ocular estimates.
7. method according to claim 6, wherein, the grade scale of described 5 grades of ocular estimates is: 0 grade, growth rate of cotton plant is normal, without symptom of chemical damage; 1 grade, 0～25% blade produces poisoning; 2 grades, 25%～50% blade produces poisoning; 3 grades, 50%～75% blade produces poisoning; 4 grades, 75%～100% blade produces poisoning.
8. method according to claim 1, wherein, the computing method of the described relative poisoning index of described poisoning exponential sum are respectively:
The highest level of poisoning index=∑ (the poisoning strain number of each poisoning rank * appropriate level) * total strain number of 100/(* classification ocular estimate);
The poisoning index of poisoning index=cotton to be measured * 50/ contrasts described 50 the poisoning index that is equal to or greater than of cotton relatively.
9. according to the described method of claim 6 or 7, wherein, the computing method of the described relative poisoning index of described poisoning exponential sum are respectively:
Poisoning index=∑ (the poisoning strain number of each poisoning rank * appropriate level) * total strain number of 100/(* 4), wherein 4 are the highest level of classification ocular estimate;
The poisoning index of poisoning index=cotton to be measured * 50/ contrasts described 50 the poisoning index that is equal to or greater than of cotton relatively.
10. method according to claim 1 wherein, according to the relative poisoning index assessment cotton to be measured calculated to the standard of the tolerance of glyphosate is: poisoning index 0～10.0 relatively is high-resistance glyphosate; Poisoning index 10.1～20.0, be resistance glyphosate relatively; Poisoning index 20.1～35.0, be glyphosate tolerant relatively; Relative poisoning index > 35.0, for to the glyphosate sensitivity.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108575645A (en) * | 2018-04-03 | 2018-09-28 | 中国农业科学院棉花研究所 | A kind of method of field Fast Evaluation cotton to herbicide tolerant |
| CN111308059A (en) * | 2020-03-25 | 2020-06-19 | 河南省农业科学院植物保护研究所 | Method for quantitatively evaluating functions and characteristics of pesticides |
| CN114586496A (en) * | 2022-02-17 | 2022-06-07 | 江西省棉花研究所 | Identification method for glyphosate resistance of cotton seeds |
| CN117814083A (en) * | 2024-02-02 | 2024-04-05 | 湖北省农业科学院经济作物研究所 | Method for screening glyphosate sensitive cotton single plants in field |
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| US20030024015A1 (en) * | 2001-02-14 | 2003-01-30 | Davis William H. | Production of genetically-controlled herbicide resistance in cotton plants in the absence of genetic engineering |
| 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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2013
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030024015A1 (en) * | 2001-02-14 | 2003-01-30 | Davis William H. | Production of genetically-controlled herbicide resistance in cotton plants in the absence of genetic engineering |
| 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 |
Non-Patent Citations (1)
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| 李儒海等: "转基因抗草甘膦棉花与常规棉花对草甘膦耐受性的比较研究", 《中国棉花》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108575645A (en) * | 2018-04-03 | 2018-09-28 | 中国农业科学院棉花研究所 | A kind of method of field Fast Evaluation cotton to herbicide tolerant |
| CN111308059A (en) * | 2020-03-25 | 2020-06-19 | 河南省农业科学院植物保护研究所 | Method for quantitatively evaluating functions and characteristics of pesticides |
| CN111308059B (en) * | 2020-03-25 | 2023-12-01 | 河南省农业科学院植物保护研究所 | Method for quantitatively evaluating functions and characteristics of pesticide |
| CN114586496A (en) * | 2022-02-17 | 2022-06-07 | 江西省棉花研究所 | Identification method for glyphosate resistance of cotton seeds |
| CN117814083A (en) * | 2024-02-02 | 2024-04-05 | 湖北省农业科学院经济作物研究所 | Method for screening glyphosate sensitive cotton single plants in field |
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