CN103070068A - Herbicide-resistant rape directive breeding method based on acetolactate synthase (ALS) target esterase - Google Patents
Herbicide-resistant rape directive breeding method based on acetolactate synthase (ALS) target esterase Download PDFInfo
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Abstract
The invention provides a herbicide-resistant rape directive breeding method based on acetolactate synthase (ALS) target esterase, which belongs to a plant trait breeding method. The method comprises the following steps of selecting materials needed by production of rape or breeding of the rape, processing seeds with ethylmethane sulfonate (EMS), mutagenizing the seeds to be isolated and propagated to a M2 generation, and directionally screening herbicide-resistant mutant characters in a large group under the selection pressure of the herbicide adopting the ALS as the target esterase. In order to increase the probability for acquiring the mutant characters, the directive screening can be continuously conducted in multiple years, or conducted in multiple places in the same year or conducted in multiple places in multiple years until the needed mutant material is screened out.
Description
Technical field
The present invention relates to a kind of selection of Atrazine resistant Brassica napus, the selection, seed EMS processing method and the weed killer herbicide that are specifically related to the rape material select to depress the directed screening of resistance trait, belong to the plant trait selection.
Background technology
Farmland weed has a strong impact on yield of rape and quality, has become one of main biological epidemics of rape production, applies the Atrazine resistant Brassica napus kind, uses chemical herbicide to carry out control of weeds, not only fast, effectively, and also economical and practical.Compare with traditional rape, it is easy and simple to handle, efficient that Atrazine resistant Brassica napus has farming, and the grower only needs just can effectively kill weeds in field at 1～2 weed killer herbicide of seedling spraying, compares with manpower weeding and can save the labour, reduces production costs, and improves economic well-being of workers and staff.Since the last century the nineties, the Canada of North America, the U.S. etc. are large-scale commercial applications plantation Atrazine resistant Brassica napus, has obtained preferably through the agent benefit.Yet China's Atrazine resistant Brassica napus is not yet got permission the commercialization plantation so far, and mainly contain two reasons: (1) Atrazine resistant Brassica napus is generally genetically modified crops, enters to commercially produce to pass through strict safety evaluatio.(2) up to now, domestic rape anti-herbicide gene is mainly from abroad, and resistant gene is subjected to international intellectual property protection; in case commercialization plantation; need to pay expensive patent fee, this will increase substantially the rape production cost, run counter to the original intention of plantation Resistant Herbicide Crops.2004, we have found the rape new germ plasm M9 of the anti-imidazolinone herbicide of spontaneous mutation in the experimental field of rape and for many years crop rotation of soybean, through test for identification for many years, this mutant resistance trait is stable, the resistance effect is obvious, the concentration of anti-imidazolinone herbicide imazethapyr is more than 2 times of the effective herbicide concentration of imazethapyr, has practical value.Genetic research shows, the resistance of this mutant is dominant character, and by 1 pair of nuclear gene control, resistant gene is at F
2And BC
1Follow mendelian inheritance in the colony, use the conventional breeding method such as hybridize, backcross and just resistant gene can be imported target variety, for the Atrazine resistant Brassica napus kind that seed selection has independent intellectual property right and not limited by transgene rape environment release conditions is laid a good foundation.Imidazolinone herbicide is a class wide spectrum, the high activity weed killer herbicide that American Cyanamid Company succeeded in developing in phase early 1980s, be one of present most widely used weed killer herbicide, its broad weed-killing spectrum (comprising grass family and broad leaved weed), consumption be low, to mammal low toxicity and environmentally friendly.But this class weed killer herbicide both can do processing soil treatment agent also cauline leaf spray, weeding ratio is high, easy to use firmly gets users' welcome.Studies show that, acetolactate synthestase (acetolactate synthase, ALS) be the target enzyme of imidazolinone herbicide, imidazolinone herbicide enters the enzyme active sites path by forming compound blocking-up substrate with ALS, suppress ALS active, cause the branched-chain amino acid biosynthesis block, make plant tissue chlorosis, yellow, dead gradually at last.But imidazolinone herbicide is long residual herbicide, has the safety issue of pedo relict phase length and late stubble sensitive crop.This type of weed killer herbicide is the faintly acid characteristic, and existing middle condition has again the anion attitude in soil, and its residual life is typical pH dependent form.Because of soil pH value difference, its absorption, leaching, decomposition and residual differing greatly.In high pH soil, anion attitude large percentage, degraded is rapid, and along with soil pH reduces, middle condition increases, the corresponding increase of the amount of being adsorbed, degraded is slow, and residual life, prolong.Such weed killer herbicide only can use in ripe season area for these reasons, is the soybean herbicides special in China northeast soybean producing region, and the registration granting is failed in the area how ripe season in the Yangtze river basin.Therefore, with the rape resistant variety of the M9 seed selection of anti-imidazolinone herbicide rape new germ plasm or transformation, can not apply in China winter rape main producing region, greatly limit the using value of the anti-imidazolone proterties of rape.
In fact the at present existing five large classes of the weed killer herbicide of developing take ALS as the target enzyme, be sulfonylurea (SU), imidazolone type (IMI), sulfonamides (SCT), pyrimidine salicylic acid (PTB) and triazolo pyrimidine class (TP), the weed killer herbicide kind that registration is provided in the production is countless especially.Selectivity is strong because the ALS weed killer herbicide has, broad weed-killing spectrum, using dosage are low, to advantages such as mammalian toxicity are little, after the eighties in last century of early development out, just be widely applied.But meanwhile because a large amount of this class weed killer herbicide of continuously use so that the resistance biotype weeds quantity of anti-such weed killer herbicide sharply increases, has surpassed triazine herbicide at present, become the most serious class weed killer herbicide of resistant weed.According to incompletely statistics, more than 30 countries in the whole world find that the weeds biotype of anti-acetolactate synthestase (ALS) inhibitor class weed killer herbicide has 98 kinds, and wherein broadleaf weed is 68 kinds, 30 kinds of monocotyledon weeds.Studies show that most the change by the target site produces in the weeds biotype of anti-ALS inhibitor class weed killer herbicide, due to being substituted by an amino acid in 5 essential amino acids.122 (calculating with model plant arabidopsis amino acid position, lower same) alanine are replaced by valine, and imidazolinone herbicide is produced resistance.197 proline are replaced by histidine or threonine, arginine, leucine, isoleucine, serine, alanine or glutamine, sulfonylurea herbicide is produced very strong resistance, and some site also can produce cross resistance to triazolo pyrimidine class weed killer herbicide.205 alanine are replaced by valine, and all kinds of ALS target enzyme weed killer herbicides are all had to a certain degree resistance of wide spectrum.574 tryptophans are replaced by leucine, and sulfonylurea, imidazolone type and triazolo pyrimidine class weed killer herbicide are all had very strong cross resistance.653 serines are replaced by threonine or asparagine, and imidazolinone herbicide is had very strong resistance.Above-mentioned 5 essential amino acids have 3 amino acid (122 alanine, 197 proline, with 205 alanine) be positioned at the aminoterminal of acetolactate synthestase, 2 (574 tryptophans and 653 serines) are positioned at the c-terminus of enzyme in addition, and this 5 amino acid position in the acetolactate synthestase sequence of most known plants is quite constant.The ALS resistance that base mutation also will produce different modes occurs in other zone beyond the conservative functional domain of als gene in addition, is replaced by leucine such as 569 tryptophans, and sulfonylurea and imidazolinone herbicide are produced resistance.591 tryptophans are replaced by leucine, and all ALS weed killer herbicides are produced very strong resistance of wide spectrum.No matter be that weeds or crop als gene have its common ground in higher plant, all there are five discontinuous conservative functional domains, the origination point sudden change will cause the ALS resistance (Ren Xueling based on the target position change on these conservative functional domains, Wu Chao, Yang Huazheng, the resistance problem of the weed killer herbicide that point mutation causes, chemistry circular 2006, the 1st phase, 9-15; Zhang Chaoxian, Ni Hanwen, Wei Shouhui, Huang Hongjuan, Liu Yan, Cui Hailan, Sui mark peak, Zhang Meng, Guo Feng, the herbicide resistance progress, Scientia Agricultura Sinica, 2009,42(4): 1274-128).The research of point mutation resistance provides theoretical foundation for cultivating resistance crop, utilizes the high conservative of plant als gene and target site to change the characteristic that produces anti-ALS inhibitor class weed killer herbicide, provides new way for seed selection resists the crop varieties of such weed killer herbicide.It is reported and use ethylmethane sulfonate (EMS) mutagenesis and organize cultivation to breed the corn variety of anti-imidazolone type and anti-sulfonylurea herbicide, breed the rape variety of anti-imidazolinone herbicide with the mutagenesis of nitroso ureas ethyl ester and microspores culture, breed the rice varieties of anti-imidazolinone herbicide with EMS mutagenesis, breed the wheat breed (Shen Xiaoxia of anti-imidazolinone herbicide with Sodium azide and EMS mutagenesis, the Ni Changchun compiling, the past of imidazolinone herbicide tolerance crop, status quo and future (on), world pesticide, 2005,27(5): 9-13; Su Shaoquan, the new development of non-glyphosate herbicidal resistance crop, agricultural chemicals, 2009,48(12): 859-863).All find in addition the natural mutation of anti-imidazolidinone weedicide in the crops such as sunflower, beet, cotton, soybean, tomato, tobacco and rape, people are screening and cultivation resistant variety from the organism that weed killer herbicide is produced resistance owing to the nucleotide point mutation directly.
The biologist generally believes, weeds such as sustainable existence herbicide selective pressure, can be brought out relevant target enzyme and produce point mutation in its process of growth, thereby makes weeds and produce resistance.Therefore, the prevention weeds produce the major measure of resistance for reducing the weed killer herbicide selection pressure in the production, in plant growing process, avoid depending on unduly chemical herbicide as far as possible, same geographic area, the crop rotation Different Crop, the weed killer herbicide that selection has different target sites uses in turn, all is conducive to reduce selection pressure, reduces the possibility that resistant weed occurs.In the crop breeding process, then can take full advantage of this selection pressure on the contrary, accelerate the cultivation of antiweed kind, and the relevant target enzyme of applied chemistry mutagen direct mutagenesis produces point mutation, recycling weed killer herbicide selection pressure directed screening mutational site, the seed selection process (Ren Xueling that then can greatly accelerate crop antiweed kind, Wu Chao, Yang Huazheng, the resistance problem of the weed killer herbicide that point mutation causes, chemistry circular 2006, the 1st phase, 9-15).The present invention just is being based on a kind of new method of cultivating anti-ALS target enzyme weed killer herbicide rape that above-mentioned principle proposes.
Summary of the invention
Technical problem
The present invention can remedy the deficiency of existing patent or technology, and a kind of Atrazine resistant Brassica napus directive breeding method based on ALS target enzyme is provided.
Technical scheme
A kind of Atrazine resistant Brassica napus directive breeding method based on ALS target enzyme comprises step:
A, seed EMS process: to the required rape seed that carries out mutagenesis, carry out the EMS mutagenic treatment with the half lethal concentration of this seed;
B, mutagenic progeny expand numerous: the mutagenesis seed expands numerous to M in the isolation solarium
2In generation, results are enough carried out the seed of large group directed screening, and low temperature saves backup;
C, weed killer herbicide pressure selection: in order to the weed killer herbicide of ALS target enzyme exploitation to M
2Carry out the antiweed directed screening for population mixture, dosing is the recommendation concentration of this weed killer herbicide controlling weeds, and sprays continuously 2 times, carries out in 3～4 leaf phases of rape, sprays after 10 days the 2nd time again for the 1st time;
D, to EMS mutagenesis M
2In generation, carried out the antiweed directed screening: the plant that is not killed by weed killer herbicide behind the spray medicine be directed screening to the Atrazine resistant Brassica napus new material take ALS as the target enzyme;
E, resistant material checking are identified: to the Atrazine resistant Brassica napus new material individual plant take ALS as the target enzyme of screening acquisition, florescence bagging selfing, the results selfed seed, verify evaluation: treat that rape grows to the 3-4 leaf phase, again spray the tribenuron-methyl weeding agent, dosing is the recommendation concentration of tribenuron-methyl controlling weeds, effective dose 90ga.i./hm
2, the plant that spray is not killed by weed killer herbicide behind the medicine be directed screening to the Atrazine resistant Brassica napus new material take ALS as the target enzyme.
Said method is specifically used:
1) selects the rather assorted No. 15 male parent N131 of double-low hybrid rapeseed new varieties;
2) with the EMS of N131 partly till death 0.4% couple of rape N131 of concentration mass ratio seed carry out the EMS mutagenic treatment;
3) expand numerous to M to mutagenic progeny in the isolation solarium
2Generation, results M
2Seed low temperature saves backup;
4) plantation M
2For rape seed, plantation M
2For rape seed, every square chi of density of reserved seedlings is the 50-70 strain.Brassica campestris L seedling sprays the tribenuron-methyl weeding agent when growing to 3 leaves, 1 heart, dosing is the recommendation concentration of tribenuron-methyl controlling weeds, effective dose 90ga.i./hm
2, spray again again 1 time after 10 days; Select to process the individual plant of surviving, do not have symptom of chemical damage in the Brassica campestris L seedling, 5-6 leaf after date moves to the rapeseed breeding land for growing field crops with the dish seedling, and bagging selfing when rape is bloomed obtains selfed seed;
5) the resistant material checking is identified: the selfed seed that the plantation screening obtains, treat that rape grows to the 3-4 leaf phase, and again spray the tribenuron-methyl weeding agent, dosing is the recommendation concentration of tribenuron-methyl controlling weeds, effective dose 90ga.i./hm
2, the plant that spray is not killed by weed killer herbicide behind the medicine be directed screening to the Atrazine resistant Brassica napus new material take ALS as the target enzyme.
Beneficial effect
The present invention can improve the Breeding Efficiency that obtains anti-ALS target enzyme weed killer herbicide rape, for anti-weeding rapeseed breeding provides new way, new method greatly by said method.
A: although EMS is random to plant offspring's proterties mutagenesis, and probability is very little.No matter be that weeds or crop als gene have its common ground in higher plant, all there are five discontinuous conservative functional domains, the origination point sudden change will cause the ALS resistance (Ren Xueling based on the target position change on these conservative functional domains, Wu Chao, Yang Huazheng, the resistance problem of the weed killer herbicide that point mutation causes, chemistry circular 2006, the 1st phase, 9-15; Zhang Chaoxian, Ni Hanwen, Wei Shouhui, Huang Hongjuan, Liu Yan, Cui Hailan, Sui mark peak, Zhang Meng, Guo Feng, the herbicide resistance progress, Scientia Agricultura Sinica, 2009,42(4): 1274-128).The research of point mutation resistance provides theoretical foundation for cultivating resistance crop, utilizes the high conservative of plant als gene and target site to change the characteristic that produces anti-ALS inhibitor class weed killer herbicide, provides new way for seed selection resists the crop varieties of such weed killer herbicide.It is reported and use ethylmethane sulfonate (EMS) mutagenesis and organize cultivation to breed the corn variety of anti-imidazolone type and anti-sulfonylurea herbicide, breed the rape variety of anti-imidazolinone herbicide with the mutagenesis of nitroso ureas ethyl ester and microspores culture, breed the rice varieties of anti-imidazolinone herbicide with EMS mutagenesis, breed the wheat breed (Shen Xiaoxia of anti-imidazolinone herbicide with Sodium azide and EMS mutagenesis, the Ni Changchun compiling, the past of imidazolinone herbicide tolerance crop, status quo and future (on), world pesticide, 2005,27(5): 9-13; Su Shaoquan, the new development of non-glyphosate herbicidal resistance crop, agricultural chemicals, 2009,48(12): 859-863).But the present invention is directed to the characteristics that the rape seed seed is little, reproduction coefficient is high, being chosen in the Brassica Napus Seedling phase carries out the large group directed screening to mutagenic progeny, until the resistance trait that acquisition needs, the method is simple and easy to do, highly effective, and this method will be accelerated the seed selection process of Atrazine resistant Brassica napus greatly.
B: evidence, the EMS of N131 of the present invention partly till death concentration be that mass ratio 0.4% is processed rape N131 seed, plantation 6000 gram M
2Seed (approximately 1,000,000 young plants), processing has 5 strains survival in the dish seedling, although it is slack-off to grow, lobus cardiacus is dark green, does not have symptom of chemical damage.5-6 leaf after date moves to the rapeseed breeding land for growing field crops with the dish seedling, obtains selfed seed, identifies through the resistance checking, obtains altogether 4 resistance strains.
C: the rape seed seed is less, thousand kernel weight is generally about 4 grams, and reproduction coefficient is up to more than 1000 times, but the isolation booth pure keeping of a 6 * 50m expands numerous 30～50 kilograms of seeds, the seed sum enough carries out the large group directed screening up to more than 800～1,200 ten thousand.Carry out the Herbicid resistant screening in 3～4 leaf phases of rape, because the dish seedling is less, and 2 spray medicines, can under higher density, screen.As carrying out directed screening with 1～2 mu of ground every year, every square chi of density of reserved seedlings is 60 strains, 1 year orientable screening 36～720,000 seedling, as carrying out continuously 5 years or 5 square chi of soils? then the mutagenesis seedling of orientable screening can reach 180～3,600,000 strains, the directed screening of large group can have increased access to the probability of mutant character greatly like this, is the practicable method of seed selection Atrazine resistant Brassica napus.
Embodiment
Embodiment: the seed selection of anti-sulfonylurea herbicide tribenuron-methyl rape.
A:EMS concentration gradient test: autumn in 2007, selecting the rather assorted No. 15 male parent N131 of double-low hybrid rapeseed new varieties is material (Pu Huiming etc., the dual-purpose new varieties of the oily vegetables of cabbage type rape (B.napus) are assorted No. 15 seed selection rather, Jiangsu agricultural journal, 2010,26 (6): 1432～1434), with pH be 7.0, the phosphate buffer of 0.1M preparation 0.2%, 0.4%, 0.6%, the 0.8% and 1.0% EMS solution of totally 5 concentration, take clear water as contrast, carry out the test of EMS concentration gradient.The test seed soaks 12h with clear water first, blotting paper blots the surface of the seed moisture, carry out the EMS seed soaking, after processing 8h seed is washed 4h with flowing water, the seed proper alignment of handling well in the culture dish that is covered with filter paper, 100 in every ware, repeat 3 times, place in the artificial climate incubator, 25 degree illumination are germinateed, and add up germination rate after 3 days.Send out the corresponding decline of rate along with EMS concentration increases seed, wherein the germination rate processed of 0.4%EMS concentration is 53.13%, approaches partly till death concentration, the EMS of true bright N131 partly till death concentration be mass ratio 0.4%(table 1).
The germination rate of table 1 rape seed under EMS variable concentrations (mass ratio) is processed
| EMS | 0% | 0.2% | 0.4% | 0.6% | 0.8% | 1.0% |
| Germination rate | 100% | 72.95% | 53.13% | 42.97% | 6.64% | 0.0% |
The EMS mutagenic treatment of B:N131: rape seed carried out the EMS mutagenic treatment in front 1 day, with the 250 gram N131 seeds nylon net bag of packing into, put into the container that fills with clear water and soak 12h, nylon net bag is hung on eminence after moisture content drains substantially, place with pH be 7.0, the 0.4%EMS solution of 0.1M phosphate buffer preparation processes 8h, to process seed with flowing water flushing 4h after, after again nylon net bag being hung on eminence water being drained, immediately with whole seed broadcasting kinds in the 6 * 25m isolation booth with isolation condition.Because this moment, seed was in germination period, the part seed has revealed bud, should gently take during sowing and gently spread, and will in time water after broadcasting, in case return bud, guarantees once neat seedling.
The expansion of C, mutagenic progeny is numerous: spring rape in 2008 is bloomed rear with nylon net cover pure keeping, because mutagenesis (M in the present age
0) grain weight is less, needs again to expand numerous after the seed results, randomly draws a certain amount of M
1Seed adopts group's bulk method to be seeded in the 6 * 50m isolation booth with isolation condition in the autumn in 2008, and spring rape was bloomed rear with nylon net cover pure keeping, summer harvest M in 2009 in 2009
2For more than 60 jin in seed, be stored in the low temperature storehouse for subsequent use.
D, large group directed screening: 2009 and continuous 2 years of autumn in 2010 are to M
2Seed carries out the evaluation and screening of resistance trait with the sulfonylurea herbicide tribenuron-methyl.Autumn sowing kind 1500 gram M in 2009
2Seed is planted 1.2 mu, the " Invest, Then Investigate " density of emerging, and 40.5 strain/square chis, the total seedling number of colony is 29.17 ten thousand strains.Autumn sowing kind 4500 gram M in 2010
2Seed is planted 1.5 mu, the " Invest, Then Investigate " density of emerging, and 84.5 strain/square chis, the total seedling number of colony is 76.09 ten thousand strains.Brassica campestris L seedling sprays the tribenuron-methyl weeding agent when growing to 3 leaves, 1 heart, dosing is the recommendation concentration of tribenuron-methyl controlling weeds, effective dose 90ga.i./hm
2Because planting density is higher, in order to prevent drain spray, increases simultaneously the weed killer herbicide selection pressure, sprays again 1 time after 10 days again.29.17 ten thousand strains in 2009 are processed the dish seedling and are all killed by the tribenuron-methyl weeding agent, fail to screen resistant plant.In 76.09 ten thousand strains processing dish seedling 5 strains survival was arranged in 2010, although it is slack-off to grow, lobus cardiacus is dark green, does not have symptom of chemical damage.5-6 leaf after date moves to the rapeseed breeding land for growing field crops with the dish seedling, bagging selfing when spring rape was bloomed in 2011, and the summer harvest obtains selfed seed.
The checking of E, resistant material is identified: autumn sowing in 2011,5 strain selfed seeds to the Screening and Identification acquisition, take original parent N131 as contrast, plant into 6 strains, each strain kind 2 row, every row 20-30 seedling, treat that rape grows to the 3-4 leaf phase, again spray the tribenuron-methyl weeding agent, dosing is the recommendation concentration of tribenuron-methyl controlling weeds, effective dose 15g a.i./hm
2Beginning in the 7th day behind the spray medicine, each is observed phytotoxicity reaction in week, 1 strain is wherein arranged and to impinging upon behind the spray medicine 7 days phytotoxicity reaction is just arranged, and dish seedling lobus cardiacus begins flavescence, and gradually rots, last death, this young plant should be under the high-density planting condition due to the medicament drain spray.1 strain shows resistance to a certain degree, the drug effect sluggish, and last dish seedling can be survived, but the dish seedling is lopsided, and the bull seedling that formation is grown thickly is numbered M343 then.3 strains show stronger resistance, and under the recommendation concentration of tribenuron-methyl weeding agent control of weeds, spray medicine posterior lobe look deepens, without any symptom of chemical damage, still can normal growth, and number then and be respectively M339, M341 and M342.Autumn sowing in 2012 is carried out Resistance Identification to M339, M341, M342 and four strains of M343 again, and the result is highly consistent with the prior year, and wherein M342 growth potential is strong, yielding ability good, has practical value.
Claims (3)
1. Atrazine resistant Brassica napus directive breeding method based on ALS target enzyme comprises step:
A, seed EMS process: to the required rape seed that carries out mutagenesis, carry out the EMS mutagenic treatment with the half lethal concentration of this seed;
B, mutagenic progeny expand numerous: the mutagenesis seed expands numerous to M in the isolation solarium
2In generation, results are enough carried out the seed of large group directed screening, and low temperature saves backup;
C, weed killer herbicide pressure selection: in order to the weed killer herbicide of ALS target enzyme exploitation to M
2Carry out the antiweed directed screening for population mixture, dosing is the recommendation concentration of this weed killer herbicide controlling weeds, and sprays continuously 2 times, carries out in 3～4 leaf phases of rape, sprays after 10 days the 2nd time again for the 1st time;
D, to EMS mutagenesis M
2In generation, carried out the antiweed directed screening: the plant that is not killed by weed killer herbicide behind the spray medicine be directed screening to the Atrazine resistant Brassica napus new material take ALS as the target enzyme;
E, resistant material checking are identified: to the Atrazine resistant Brassica napus new material individual plant take ALS as the target enzyme that screening obtains, and florescence bagging selfing, the results selfed seed is verified evaluation.
2. method according to claim 1 is characterized in that:
1) selects the rather assorted No. 15 male parent N131 of double-low hybrid rapeseed new varieties;
2) with the EMS of N131 partly till death 0.4% couple of rape N131 of concentration mass ratio seed carry out the EMS mutagenic treatment;
3) expand numerous to M to mutagenic progeny in the isolation solarium
2Generation, results M
2Seed low temperature saves backup;
4) plantation M
2For rape seed, Brassica campestris L seedling sprays the tribenuron-methyl weeding agent when growing to 3 leaves, 1 heart, and dosing is the recommendation concentration of tribenuron-methyl controlling weeds, effective dose 90ga.i./hm
2, spray again again 1 time after 10 days; Select to process the individual plant of surviving, do not have symptom of chemical damage in the Brassica campestris L seedling, 5-6 leaf after date moves to the rapeseed breeding land for growing field crops with the dish seedling, and bagging selfing when rape is bloomed obtains selfed seed;
5) the resistant material checking is identified: the selfed seed that the plantation screening obtains, treat that rape grows to the 3-4 leaf phase, and again spray the tribenuron-methyl weeding agent, dosing is the recommendation concentration of tribenuron-methyl controlling weeds, effective dose 90ga.i./hm
2, the plant that spray is not killed by weed killer herbicide behind the medicine be directed screening to the Atrazine resistant Brassica napus new material take ALS as the target enzyme.
3. method according to claim 2 is characterized in that:
Plantation M
2For rape seed, every square chi of density of reserved seedlings is the 50-70 strain.
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| CN105734071A (en) * | 2016-04-22 | 2016-07-06 | 江苏省农业科学院 | ALS mutant gene and application of protein thereof to herbicide resistance |
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| CN104642104A (en) * | 2015-02-10 | 2015-05-27 | 山东省林业科学研究院 | Method for constructing alfalfa EMS mutant library |
| CN105104166A (en) * | 2015-07-23 | 2015-12-02 | 上海市农业科学院 | Method used for breeding imidazolinone herbicide-resistant japonica rice |
| CN105104166B (en) * | 2015-07-23 | 2021-05-07 | 上海市农业科学院 | Method for breeding imidazolinone herbicide-resistant japonica rice |
| CN105734071A (en) * | 2016-04-22 | 2016-07-06 | 江苏省农业科学院 | ALS mutant gene and application of protein thereof to herbicide resistance |
| CN107460195A (en) * | 2017-08-16 | 2017-12-12 | 江苏省农业科学院 | Cabbage type rape als gene promoter and application |
| CN108707592A (en) * | 2018-05-23 | 2018-10-26 | 北京市农林科学院 | CLALS albumen, its encoding gene and their applications in predicting watermelon Herbicid resistant |
| WO2019223676A1 (en) * | 2018-05-23 | 2019-11-28 | 北京市农林科学院 | Clals protein and encoding gene of the protein, and uses thereof in prediction of herbicide resistance of watermelon |
| CN108707592B (en) * | 2018-05-23 | 2022-06-28 | 北京市农林科学院 | CLALS protein, encoding gene thereof and application of CLALS protein and encoding gene thereof in prediction of herbicide resistance of watermelons |
| CN113166775A (en) * | 2018-09-25 | 2021-07-23 | 科沃施种子欧洲股份两合公司 | Method for controlling weeds, sugar beets and other weeds |
| CN110157828A (en) * | 2019-04-15 | 2019-08-23 | 上海市农业科学院 | The application of Brassica Napus sulfonylurea herbicide resistant mutants and its method |
| CN110679480A (en) * | 2019-11-22 | 2020-01-14 | 江苏省农业科学院 | Breeding method of high-resistance sulfonylurea herbicide rape |
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