CN102165885A - Method for identifying glyphosate resistance of cotton - Google Patents
Method for identifying glyphosate resistance of cotton Download PDFInfo
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- CN102165885A CN102165885A CN 201010604559 CN201010604559A CN102165885A CN 102165885 A CN102165885 A CN 102165885A CN 201010604559 CN201010604559 CN 201010604559 CN 201010604559 A CN201010604559 A CN 201010604559A CN 102165885 A CN102165885 A CN 102165885A
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Abstract
The invention relates to a method for identifying the glyphosate resistance of cotton, and the method is used for identifying the glyphosate resistance/ tolerance of cotton materials. The method comprises the following steps: seeding cotton seeds in a container or a field; after a certain leaf age of grown cotton, respectively spraying 61.5ai/mu-82g ai/mu or 123g ai/mu-246g ai/mu phytocide glyphosate; and recording the death rate of the cotton, and recording the yield properties while harvesting, wherein the cotton which is sprayed with the 61.5g ai/mu-82g ai/mu glyphosate and dies is sensitive to the glyphosate, the cotton which is sprayed with 123g ai/mu-246g ai/mu glyphosate, does not die and achieves no yield reduction is resistant to the glyphosate, and the cotton which is sprayed with 123g ai/mu-246g ai/mu glyphosate, does not die and achieves yield reduction is tolerant to the glyphosate. The method provided by the invention is suitable for identifying and evaluating the glyphosate resistance/tolerance of cottons of routine variety, hybrid variety and transgenic glyphosate-resistant variety, can be used for accurately measuring the glyphosate resistance/tolerance of the cotton and has strong practicability.
Description
Technical field
The present invention relates to the plant protection technology field, specifically, relate to an authentication method that grows cotton anti-/ glyphosate tolerant.
Background technology
Weeds are one of limiting factors of cotton stable yields.More than 7,000 ten thousand mu of the annual cotton field crop smothering generation areas of China, the underproduction is lost more than 12,500,000,000 yuan.In China 5 big cotton regions, the advantage weeds have kind more than 60, and the main harm kind is lady's-grass, eleusine indica, barnyard grass grass, green foxtail, moleplant seed, reed, Amaranthus retroflexus, kitchen garden, piemarker, acalypha copperleaf, lamb's-quarters, Veronica, Eclipta prostrata, field thistle, corn sow thistle, common sow thistle, Mengshan lettuce, field bindweed, the rhizome of nutgrass flatsedge etc.The cotton field weeding is based on chemical control, main adopt to broadcast to spray before the seedling of back spray the selectivity cauline leaf behind closed soil type weed killer herbicide Acetochlor, isopropyl methoxalamine or the seedling and handle methods such as weed killer herbicide Quizalotop-ethyl, haloxyfop-r-methyl.But above-mentioned medicament all can not satisfy the requirement of effective weeding and cotton volume increase.Herbicides for before-seedling stage such as Acetochlor are shorter to the closed soil phase of weeds, go down in the cotton back 45 days left and right sides drug effects of emerging, and cause weeds to be unearthed in a large number, influences that cotton blooms and the cotton boll growth, and Acetochlor is emerged to cotton and grown in early days simultaneously also certain harm; Herbicide after seedling Quizalotop-ethyl etc. only can be prevented and treated the grassy weed in cotton field, can not prevent and kill off broadleaf weed and sedge weed; At present, still untappedly in the world go out the selective herbicide that cotton field seedling rear defence removes broadleaf weed.Therefore, be badly in need of on the Cotton Production broad weed-killing spectrum, to the low-cost weed killer herbicide product of cotton safety.
Herbicide glyphosate is a kind of amino acid bio synthetic inhibitor, and its main action target is 5-enol acetone shikimic acid-3-phosphonate ester synthetase (EPSPS).Because to the inhibition of EPSPS, make through its catalysis to stop to the transition process of 5-enol acetone shikimic acid-3-phosphate (EPSP) by phosphoenolpyruvate (PEP) and shikimic acid-3-phosphoric acid (S3P), thus the biosynthesis of inhibition ArAA.In plant corpus, EPSPS mainly is present in chloroplast and the plastid, after glyphosate enters above-mentioned two organelles, glyphosate preferentially combines with the avtive spot of EPSPS, make EPSPS that variation on the configuration take place, thereby suppress itself and the combining and subsequently biocatalytic reaction of phosphoenolpyruvate.Glyphosate belongs to the wide spectrum steriland herbicide, can prevent and kill off nearly all annual and perennial grass family, dicotyledon and shrub etc. effectively.It is safe to people and animals, environment, soil etc., is the desirable grass product that removes.But because glyphosate does not have the selectivity between crop and weeds, conventional cotton variety is handled the death of promptly being injured once glyphosate.
The resistance glyphosate cotton is the effective way that solves cotton field crop smothering problem.Resistance glyphosate cotton 1997 is in american goodsization, to 2007, U.S.'s resistance glyphosate sown areas of cotton account for more than 52% of sowing cotton seed area because the glyphosate broad weed-killing spectrum, need not mix execute other medicament and save farming this etc. advantage, the resistance glyphosate sown areas of cotton enlarge just rapidly.China has also begun the cultivation of resistance glyphosate new cotton variety at present.In resistance glyphosate cotton variety seed selection process, to the cotton material glyphosate anti-/ the tolerance evaluation be breeding key of success link also be one of content of estimating of transgene cotton environmental safety.But have following problem at present in cotton is identified glyphosate resistance: used medicament is nonstandard, and glyphosate application period and application dosage are indeterminate, and the apparatus that sprays glyphosate is lack of standardization, the evaluation criterion disunity.The problems referred to above cause its qualification result to produce very big systematic error, influence the use of glyphosate in follow-up breeding work and the variety popularization process.Therefore, set up cotton to glyphosate anti-/ the tolerance authenticate technology is an urgent demand of the cultivation of resistance glyphosate cotton, popularization and production application.
Summary of the invention
One object of the present invention is to provide the anti-/ glyphosate tolerant authentication method that grows cotton.
In order to realize purpose of the present invention, cotton of the present invention resists/the glyphosate tolerant authentication method, comprises the steps:
1) at container or field sowing cotton seeds, the back thinning of emerging;
2) with the glyphosate of 61.5g ai/ mu～82g ai/ mu with 123g ai/ mu～246g ai/ mu two groups of identical cotton seedlings are sprayed respectively, the leaf age of described cotton seedling is 3～10 leaf ages;
3) after the dispenser, write down cotton lethality and biomass in 28～42 days, record yield traits during results;
4) cotton that sprays the glyphosate death of 61.5g ai/ mu～82g ai/ mu is the glyphosate sensitivity, spraying the not dead and cotton not underproduction of the glyphosate of 123g ai/ mu～246g ai/ mu is resistance glyphosate, and the glyphosate that sprays 123g ai/ mu～246g ai/ mu is not dead but the cotton underproduction is a glyphosate tolerant.
Wherein, described container is diameter 10～15cm, high 8～10cm vinyon alms bowl, and 2～5 0.2～0.5cm diameter permeable holes are arranged at the bottom of alms bowl, and permeable hole is covered by seepy material, and preferred seepy material is an individual layer filter paper.
Wherein, described container culturing substrate is preferably the immature soil: yellow sand: triumphant mattress nutrition soil 1: 1: 1, and its content of organic matter 2.5～5%, pH is 7.0～7.5.
Wherein, the container culturing method of cotton seeds is soaked into soil for pouring water from container bottom, and the cotton seeds of 5 vernalization of sowing covers thickness and be 0.75～1.5 centimetre matrix then, and the every alms bowl of back thinning of emerging stays 2 plant.
Wherein, the sub-district area of described field sowing is 50～100m
2, be preferably 20～30m
2
Wherein, described cotton field sowing method is the capable sowing of width, and wide every trade is apart from being 90cm, and the narrow row line-spacing is 45cm, and density is 4000～6000 strains/mu bunch planting, 2～5 cotton seeds of every bunch planting kind, and 1 plant is stayed in the every cave of thinning, back of emerging.
Wherein, the described cotton leaf age that sprays glyphosate is preferably 5～6 leaf ages.
Wherein, described glyphosate is preferably farming and reaches (Roundup), and it contains 41% glyphosate isopropyl amine salt, and U.S. Monsanto Company produces, available from Beijing Agriculture Prodn. Means Company.
Wherein, be used for the cotton of container culturing, preferably carry out in the greenhouse, the device that sprays glyphosate is preferably the spray tower of packaged type 8002 Fan spray heads of pressure 2.75Kpa.
Wherein, carry out the field with the sprayer that pressure limiting valve and 8001～8003 Fan spray heads are housed and spray glyphosate.
This authentication method can be measured anti-/ resistance level and the resistance stability of cotton to glyphosate effectively, is applicable to cotton conventional variety, Hybrid and transformed variety, and has characteristics easy and simple to handle, precise and high efficiency to have very strong practicality.This method has been stipulated period, medicament, dosage, dispenser facility and the evaluation criterion that cotton is identified glyphosate tolerant.And in the existing cotton resistance glyphosate degree evaluation method, often on the target weed killer herbicide adopt homemade glyphosate, and it is big that homemade medicament has a content luffing, and the normal situation such as synergist that adds causes the dosage that sprays inaccurate in the medicament; On spraying period, only spray medicine usually in seedling stage; The spray pharmacy amount adopts 61.5g ai/ mu～82g ai/ mu list multiple dose, and the result who draws can not reflect cotton resistance stability and resistance level; The spraying apparatus evaporating, emitting, dripping or leaking of liquid or gas of adopting is serious, and spray value is less than normal.The problems referred to above cause its qualification result to produce very big systematic error, influence the accuracy of estimating, thereby influence the use of glyphosate in follow-up breeding work and the variety popularization process.Evaluation method of the present invention has been filled up the blank that Cotton in China is measured glyphosate tolerant.Scientific evaluation and acceleration breeding process to breeding material resistance in the cotton resistance glyphosate breeding of new variety process are significant.
Embodiment
Following examples are used to illustrate the present invention, but are not used for limiting the scope of the invention.
Embodiment 1
Select diameter 15cm, high 10cm vinyon alms bowl, 3 0.5cm diameter permeable holes are made a call in the alms bowl bottom; Permeable hole individual layer filter paper covers; The immature soil of packing into: yellow sand: 1: 1: 1 culture matrix of triumphant mattress nutrition soil, pour water from container bottom and to soak into soil.
Sow transgenosis resistance glyphosate cotton MON88913, acceptor cotton variety Coker312 and the conventional variety Ji cotton 998 of vernalization respectively, 5 seeds of every alms bowl sowing.After planting cover 1 centimetre of same matrix.The every alms bowl of thinning stayed 2 strains after cotton emerged.
Treat that cotton grows to 3～4,6～8 and 9～10 leaf periods, put in the spray tower of packaged type 8002 Fan spray heads of pressure 2.75Kpa that the herbicide spraying farming reaches (Roundup) aqua; Glyphosate concentration is respectively 0g ai/ mu, 61.5g ai/ mu, 82g ai/ mu, 123g ai/ mu, 164g ai/ mu and 246g ai/ mu, amount of water 30L/ mu.The design of the employing factor repeats 5 times.
After the dispenser 28 days, withered with growth rate of cotton plant point be death standard mensuration cotton lethality; Measure cotton individual plant bell number, Dan Lingchong and ginning outturn Isoquant proterties during results, and be converted into per mu yield.When spraying with 2～3 times of spray values of glyphosate dosage 61.5～82g ai/ mu, the not underproduction of cotton is for anti-, and when above-mentioned dosage sprayed, cotton growth point was not dead but the underproduction is anti-.28d cotton material lethality sees Table 1 after using glyphosate, and the output during results sees Table 2
Table 1 is used 28d cotton material lethality (%) behind the glyphosate
Annotate: 1. L represents to handle the cotton that growing point is survived through the matched doses glyphosate in the table, and D represents to handle through the matched doses glyphosate cotton of growing point death.2.MON88913 be transgenosis resistance glyphosate cotton, Coker312 is the acceptor cotton variety of MON88913, is U.S. Monsanto Company and produces; Ji cotton 998 is the common cotton flower variety, and Hebei Ji Feng kind industry Co., Ltd produces.
Table 2 is used output of cotton (unginned cotton .kg/ mu) behind the glyphosate
Annotate: numeral back same letter represents that two data are not remarkable at α=0.05 level difference in the table.
From the result of table 1 and table 2 as can be known, cotton variety ' Mon88913 ' is a resistance glyphosate, and Coker312 and Ji cotton 998 is to the glyphosate sensitivity.
Embodiment 2
1, select cotton material MON88913, Coker312 and Ji cotton 998 and Ji cotton 20 standby.Delimit 162 30m in the field
2The experimental plot, the capable sowing cotton of width, wide every trade is apart from being 90cm, the narrow row line-spacing is 45cm, density 4000 strain/mus, bunch planting, 4 seeds of every bunch planting kind.1 plant was stayed in the every cave of thinning after cotton emerged; Above-mentioned each 54 sub-district of cotton material sowing;
2, treat above-mentioned cotton material length to 3,6 and 9 leaf periods, respectively with the compression CO that pressure limiting valve and 8001～8003 Fan spray heads are housed
2Sub-district sprayer herbicide spraying farming reaches (Roundup) aqua; Wherein, " compression CO
2The sub-district sprayer " be to adopt compression CO
2As source of the gas, with the field trial sub-district sprayer unit of agricultural chemicals ejection.
3, farming reaches concentration and is respectively 0g ai/ mu, 61.5g ai/ mu, 82g ai/ mu, 123g ai/ mu, 164g ai/ mu and 246g ai/ mu, amount of water 30L/ mu.Adopt split plot design, 3 repetitions.
4, after the dispenser 28 days, withered with growth rate of cotton plant point be death standard mensuration cotton lethality; Measure cotton individual plant bell number, Dan Lingchong and ginning outturn Isoquant proterties during results, and be converted into per mu yield.When spraying with 2～3 times of spray values of glyphosate dosage 61.5～82g ai/ mu, the not underproduction of cotton is for anti-, and when above-mentioned dosage sprayed, cotton growth point was not dead but the underproduction is anti-.28d cotton material lethality sees Table 3 after using glyphosate, and the output during results sees Table 4.
Table 3 is used 28d cotton material lethality (%) behind the glyphosate
Annotate: 1. L represents to handle the cotton that growing point is survived through the matched doses glyphosate in the table, and D represents to handle through the matched doses glyphosate cotton of growing point death.2.MON88913 be transgenosis resistance glyphosate cotton, Coker312 is the acceptor cotton variety of MON88913, is U.S. Monsanto Company and produces; Ji cotton 20 and Ji cotton 998 are the common cotton flower variety, and Hebei Ji Feng kind industry Co., Ltd produces.
Table 4 is used output of cotton (unginned cotton .kg/ mu) behind the glyphosate
Annotate: numeral back same letter represents that two data are not remarkable at α=0.05 level difference in the table.
From the result of table 3 and table 4 as can be seen, MON88913 is a resistance glyphosate, and Ji cotton 20 is a glyphosate tolerant.Coker312 and Ji cotton 998 is to the glyphosate sensitivity.
The above only is a preferred implementation of the present invention; should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the technology of the present invention principle; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (7)
1. anti-/ glyphosate tolerant the authentication method that grows cotton is characterized in that, comprises the steps:
1) at container or field sowing cotton seeds, the back thinning of emerging;
2) spray two groups of identical cotton seedlings with the glyphosate of 61.5g ai/ mu～82g ai/ mu and 123g ai/ mu～246g ai/ mu respectively, the leaf age of wherein said cotton seedling is 3～10 leaf ages;
3) after the dispenser, 28～42 days record cotton lethality, record yield traits during results;
4) spray the cotton of glyphosate death of 61.5g ai/ mu～82g ai/ mu for to the glyphosate sensitivity, spraying the not dead and cotton not underproduction of the glyphosate of 123g ai/ mu～246g ai/ mu is resistance glyphosate, and the glyphosate that sprays 123g ai/ mu～246g ai/ mu is not dead but the cotton underproduction is a glyphosate tolerant.
2. authentication method as claimed in claim 1 is characterized in that, described container is diameter 10～15cm, high 8～10cm vinyon alms bowl, and 2～5 0.2～0.5cm diameter permeable holes are arranged at the bottom of alms bowl, and permeable hole is covered by seepy material.
3. authentication method as claimed in claim 1 is characterized in that, the matrix of described container sowing is the immature soil: yellow sand: triumphant mattress nutrition soil 1: 1: 1, pH is 7.0～7.5.
4. authentication method as claimed in claim 1, it is characterized in that the type of seeding of described cotton container is soaked into soil for pouring water from container bottom, the cotton seeds of 5 vernalization of sowing, cover thickness then and be 0.75～1.5 centimetre matrix, the every alms bowl of back thinning of emerging stays 2 plant.
5. authentication method as claimed in claim 1 is characterized in that, described cotton field sowing method is the capable sowing of width, wide every trade is apart from being 90cm, and the narrow row line-spacing is 45cm, and density is 4000～6000 strains/mu bunch planting, 2～5 cotton seeds of every bunch planting kind, 1 plant is stayed in the every cave of thinning, back of emerging.
6. authentication method as claimed in claim 1 is characterized in that, the described cotton leaf age that sprays glyphosate is 5～6 leaf ages.
7. authentication method as claimed in claim 1 is characterized in that, described glyphosate is that farming reaches, and it contains 41% glyphosate isopropyl amine salt.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102577857A (en) * | 2012-01-24 | 2012-07-18 | 中国农业科学院棉花研究所 | Method for identifying herbicide resistance of cotton leaves by cotton ball positioning |
| CN102726185A (en) * | 2012-06-07 | 2012-10-17 | 河南科技大学 | Cotton cultivation method |
| CN103202128A (en) * | 2013-03-19 | 2013-07-17 | 盐城市育新种业有限公司 | Indoor puritytesting method of glyphosate-resistant cotton seed |
| CN103411971A (en) * | 2013-08-19 | 2013-11-27 | 中国农业科学院棉花研究所 | Method for quickly evaluating glyphosate tolerance of cotton in field |
| CN104012398A (en) * | 2014-06-13 | 2014-09-03 | 中国农业科学院棉花研究所 | Method for screening high-glyphosate-resistant semi-wild cotton material |
| CN104381123A (en) * | 2014-10-11 | 2015-03-04 | 中国农业科学院棉花研究所 | Screening method of G. hirsutum race with high glyphosate material sensibility |
| 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 |
| CN110972871A (en) * | 2019-12-04 | 2020-04-10 | 新疆农业科学院植物保护研究所 | Novel method for evaluating survival competitive capacity of transgenic cotton wasteland in extreme arid region |
| CN114586496A (en) * | 2022-02-17 | 2022-06-07 | 江西省棉花研究所 | Identification method for glyphosate resistance of cotton seeds |
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Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102577857B (en) * | 2012-01-24 | 2013-11-06 | 中国农业科学院棉花研究所 | Method for identifying herbicide resistance of cotton leaves by cotton ball positioning |
| CN102577857A (en) * | 2012-01-24 | 2012-07-18 | 中国农业科学院棉花研究所 | Method for identifying herbicide resistance of cotton leaves by cotton ball positioning |
| CN102726185A (en) * | 2012-06-07 | 2012-10-17 | 河南科技大学 | Cotton cultivation method |
| CN102726185B (en) * | 2012-06-07 | 2015-07-15 | 河南科技大学 | Cotton cultivation method |
| CN103202128A (en) * | 2013-03-19 | 2013-07-17 | 盐城市育新种业有限公司 | Indoor puritytesting method of glyphosate-resistant cotton seed |
| CN103411971A (en) * | 2013-08-19 | 2013-11-27 | 中国农业科学院棉花研究所 | Method for quickly evaluating glyphosate tolerance of cotton in field |
| CN104012398B (en) * | 2014-06-13 | 2016-05-04 | 中国农业科学院棉花研究所 | The method of screening high-resistance glyphosate semi-wild cotton material |
| CN104012398A (en) * | 2014-06-13 | 2014-09-03 | 中国农业科学院棉花研究所 | Method for screening high-glyphosate-resistant semi-wild cotton material |
| CN104381123A (en) * | 2014-10-11 | 2015-03-04 | 中国农业科学院棉花研究所 | Screening method of G. hirsutum race with high glyphosate material sensibility |
| 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 |
| CN110972871A (en) * | 2019-12-04 | 2020-04-10 | 新疆农业科学院植物保护研究所 | Novel method for evaluating survival competitive capacity of transgenic cotton wasteland in extreme arid region |
| CN110972871B (en) * | 2019-12-04 | 2021-09-21 | 新疆农业科学院植物保护研究所 | Novel method for evaluating survival competitive capacity of transgenic cotton wasteland in extreme arid region |
| CN114586496A (en) * | 2022-02-17 | 2022-06-07 | 江西省棉花研究所 | Identification method for glyphosate resistance of cotton seeds |
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