CN1251597C - Chemical crossbred agent composition, its using method and application - Google Patents
Chemical crossbred agent composition, its using method and application Download PDFInfo
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Abstract
The present invention relates to a rape chemical hybridization agent composition and an application thereof. The composition (SX-1) contains the effective components of 2-[3-(4-methoxy group-6-methyl-1, 3, 5-triazine-2-group)-3-methyl carbamido group sulfuryl] methyl benzoate, a wet unfolding agent, a dispersion agent, a stiffening agent, a disintegrating agent, soluble starch, etc., and the composition (SX-1) is a rape chemical hybridization agent with high efficiency, low toxicity, low residues and low prices. The present invention also relates to the application method of the composition and the application of the 2-[3-(4-methoxy group-6-methyl-1, 3, 5-triazine-2-group)-3-methyl carbamido group sulfuryl] methyl benzoate in the rape chemical hybridization. The composition has a thorough effect of male killing, and has a basically consistent male killing effect with varieties in different gene types; the sterile plant rate can reach 95% to 100%. By utilizing the SX-1, the rape male sterility can be induced, and the rape chemical hybrids can be obtained. By utilizing the SX-1 to assistantly produce seeds for CM, the micropowder of the CM can be reduced in the prior period, and the purity of the seeds produced can be improved.
Description
[technical field]
The present invention relates to a kind of chemical hybridizing agent composite, its using method and it application in rape.
[background technology]
Utilizing hybrid vigour is to improve crop yield, quality and resistance one very valid approach, also is the focus of all kinds of crop breedings researchs.Hybrid vigour has obtained extensive use on crops such as corn, Chinese sorghum, paddy rice and rape, and has obtained remarkable achievement.
At present, the method and approach that carries out rape heterosis utilization research both at home and abroad mainly contains 5 kinds, i.e. cytoplasm (CMS) male sterile, cell nucleus (GMS) male sterile, chemical hybridizing agent (CHA) do not induce male sterile, selfing diligent and be (SI), the blunt system of selfing etc.Wherein in rape heterosis utilization research, what real achieving success had also been carried out profound demonstration of large tracts of land and application mainly is CMS, GMS, CHS approach.
Cytoplasmic male sterility (CMS) is the male sterile line of nucleo_cytoplasmic interaction, the sterile cytoplasm source of pressing male sterile line is different, it can be divided into trailing plants crawl cytoplasmic sterility type (Ogu sterile cytoplasm), nap cytoplasmic sterility type (nap cms), Pori's horse cytoplasmic sterility type (Pol cms), Shan 2A cytoplasmic sterility type (Shan 2A cms) and other cytoplasmic sterility types again.Wherein the Ogu sterile cytoplasm exist recover to be that difficulty is looked for, three subject matters such as minus green and shortage nectary, though and the recovery of nap sterile cytoplasm system is many, its sterility instability is very responsive to temperature, pollinia appears in temperature when being higher than 20 ℃, so they all are difficult to utilize aborning.Utilize maximum cytoplasmic male sterile line systems still Pol sterile cytoplasm and Shan 2A sterile cytoplasm on China produces at present.Its outstanding feature is to be easy to find their maintenance line and to recover system and realize three series mating, and shortcoming is that male sterile line also exists very unstable problem, and promptly trace-pollen can appear in the part flower on plant top under the early spring cryogenic conditions, and influences seed production purity.GMS is cell and male sterile line, is characterized in that the male sterile pollen abortion is thorough, and good stability.Though easily look for but recessive cytoblast sterile recovers system, and maintenance line is difficult to find, needs during the production of hybrid seeds in field and male sterile line are capable, to pull out 50% educated strain.More owing to examining sterile type class in addition, hereditary more complicated, so seed selection is relatively more difficult.That has utilized in China produces at present has three classes: promptly two dominant genic male steriles, double recessive nuclear is sterile and by recessiveness overlapping and recessive epistasis make the nuclear sterile (three recessive cytoblast steriles) controlled mutually.Wherein two dominant genic male steriles and three recessive cytoblast steriles all can utilize temporary maintainer line to obtain the nuclear complete sterile line, realize " three line method " production of hybrid seeds, thus overcome double recessive nuclear sterile in " two is method " production of hybrid seeds, need to dial remove 50% trouble that can educate strain.But the recovery source electrode of two dominant genic male steriles is few, is difficult to obtain good hybrid combination, though and three recessive cytoblast sterile recovery sources are extensive, its temporary maintainer line only could separate in the test cross offspring and obtains, so the maintenance line source is narrow.
CHA compares with approach such as CMS, GMS, and the outstanding feature of CHA approach is to avoid some bad kytoplasm effect that a certain cytoplasmic dependence is caused; Its combination selection is relatively freer, can directly utilize rape conventional variety (being) as hybrid strain, need not cultivate special male sterile line and recover system, thereby choose the strong advantage combination of good quality and high output easily, application speed is fast, and can guarantee on the basis of stable yields, to obtain high yield, not influence the solid problem of the first generation of hybrid and do not exist because of recovery rate; Secondly, the male sterile that chemical emasculation causes can not heredity, and its second generation can not isolated male sterile plant, this again for utilize the combination of advantage extra-heavy two generation kind possibility is provided, also conventional breeding and the population improvement for crop provides an important means.But CHA pathway key technology is that the chemical hybridizing agent and the ripe seeding technique of a cover of high effect nontoxic must be arranged.
The target of CHA exploitation is to obtain can be used for the male sterilant that commerce is produced hybrid seed, and the strategy of therefore external exploitation CHA is not chemicals or the technology of selling, but sells seed.CHA exploitation at crop, mainly be cereal crops such as the big wheat of cultivated area, paddy rice.For rape only people such as Bangas.s (J.Agric.Sci.camb, 1983,101:453-455) utilize the male sterile of ethephon-induction mustard type rape, hybrid rate reaches 17%～79%.And other does not appear in the newspapers.
In recent years, domesticly obtaining remarkable progress aspect the utilization research of rape chemical hybridization breeding, occupy first place in the world, a plurality of crossbreed such as assorted No. 2, No. 3, No. 5, No. 1, the assorted oil in Hunan, excellent No. 1 of Fu, Chongqing assorted 05 are used for producing successively to have selected another name for Sichuan Province, change and kill nearly 70,000 hm of hybrid cultivated area
2The efficient chemical hybridizing agent of publishing at present that is applicable to rape has MG
1, MG
2, MG
3, MG
4, GA
3And EN.Wherein with MG
1(No. 1, male gametocide), MG
2(Kms-1), MG
4(the cooperation reagent of No. 1+gibberellin of male gametocide), GA
3(gibberellin), EN (composition the unknown) are comparatively desirable.But MG
1And MG
4Gametocidal effect be subjected to rape development degree environmental influence, it is thorough inadequately to kill hero, poor stability.It contains toxic component arsenic in addition, although the content of arsenic in edible oil does not exceed standard (0.1mg/kg), use all the year round and also can cause the dangerous of people, and the complete sterility strain rate that it produced 60%-80% only.MG
2After variable concentrations is handled certain poisoning is arranged all, it is heavier and be difficult to pollination to cause closing flower bud strain and dead strain phenomenon.GA
3Cost an arm and a leg and in the chemical hybridization production of hybrid seeds, do not have actual application value.Hybridization agent EN only Liu gives in to (northwest agricultural journals such as rosy clouds, 1999,8 (4): 60-62) made Preliminary report, its gametocidal effect sterile plant rate can reach 100%, sterile degree is more than 95%, but to fertility influence at whole florescence of rape how it, and to its susceptibility how the different genotype kind, and problems such as toxic residue, pollution do not appear in the newspapers as yet, do not see on producing yet and use.Therefore the novel rape C HA that seeks exploitation high-efficiency low-toxicity low-residual is very necessary.2-[3-(4-methoxyl group-6-methyl isophthalic acid; 3; 5-triazine-2-yl)-and 3-methyl ureido sulfuryl] methyl benzoate is the compound of our a kind of high-efficiency low-toxicity of finding in the screening study process of rape chemical hybridizing agent; past only is used for this compound weed killer herbicide (sulfonylurea); in existing weed killer herbicide; the content of this compound is 10% or 75%; and other composition; as wet spreader, dispersant, thickener, anticaking agent, defoamer, filler etc. totally 90% or 25%, the concrete content of these compositions is ominous.
[summary of the invention]
[problem that will solve]
The purpose of this invention is to provide a kind of rape chemical hybridizing agent composite.
Another object of the present invention provides the using method of described chemical hybridizing agent.
Another object of the present invention is described compound 2-[3-(4-methoxyl group-6-methyl isophthalic acid, 3,5-triazine-2-yl)-3-methyl ureido sulfuryl] application of methyl benzoate in the rape chemical emasculation.
[technical scheme]
2-[3-(4-methoxyl group-6-methyl isophthalic acid, 3,5-triazine-2-yl)-3-methyl ureido sulfuryl] methyl benzoate is a kind of high-efficiency low-toxicity compound that we find in the screening study process of rape chemical hybridizing agent.Describing below that to contain with this compound be the chemical hybridizing agent composite (be called for short SX-1) of active ingredient, 2-[3-(4-methoxyl group-6-methyl isophthalic acid, 3,5-triazine-2-yl)-3-methyl ureido sulfuryl] chemical constitution of methyl benzoate is as follows:
The physicochemical property of this compound is as follows: white solid, and 41 ℃ of fusing points, the vapour pressure under 25 ℃ is 2.7 * 10
-7Millimetres of mercury, the solvability in following solvent are respectively (milligram/kilogram solvent): acetone 43.8, acetonitrile 54.2, carbon tetrachloride 3.12, ethyl acetate 17.5, methyl alcohol 3.39, water (PH4) 28, water (PH5) 50, water (PH6) 280.
The toxicity of this compound: the oral acute lethal dose of 50＞5000mg/kg of rat, rabbit is acute through the skin lethal dose of 50＞2000mg/kg.
Said composition also contains wet spreader, dispersant, thickener, disintegrant, soluble starch and/or filler.
Preferably, said composition (in described composition total weight) composed as follows:
Compound 2-[3-(4-methoxyl group-6-methyl isophthalic acid, 3,5-triazine-2-yl)-3-methyl ureido sulfuryl] methyl benzoate 10-75%
Wet spreader dodecyl sulphate potassium or sodium 2.0-4.5%
Dispersant beta-naphthalenesulfonic-acid potassium or sodium acid carboxylic compound 4.0-6.0%
Thickener hydroxy cellulose sodium 0.3-0.5%
Disintegrant PTPP or sodium 2.0-5.0%
Soluble starch 3.0-5.0%
Filler supplies 100%.
Particularly preferably, said composition (in described composition total weight) composed as follows:
2-[3-(4-methoxyl group-6-methyl isophthalic acid, 3,5-triazine-2-yl)-3-methyl ureido sulfuryl] methyl benzoate: 70%
Wet spreader lauryl sodium sulfate and/or potassium 3.5%
Dispersant beta-naphthalenesulfonic-acid potassium and/or sodium acid carboxylic compound 5.0%
Thickener hydroxy cellulose sodium 0.5%
Disintegrant PTPP and/or sodium 4.0%
Soluble starch: 5.0%
Filler supplies 100%.
The invention still further relates to the using method of described rape chemical hybridizing agent composite; the method is characterized in that the monokaryotic stage of growing at the rape pollen mother cell; be the maximum bud length 1.5-2mm of rape master inflorescence; implement to kill male to female parent; spraying medicine concentration is 1.8 mg/litre-2.0 a mg/litre 2-[3-(4-methoxyl group-6-methyl isophthalic acid; 3; 5-triazine-2-yl)-and 3-methyl ureido sulfuryl] methyl benzoate; divide and use for 2 times; it is 3 milliliters that dose is accepted in each every strain; to father and mother's one's own profession ratio is 1: 1 female parent, every 667m
2Spray 17.5-20kg, i.e. monokaryotic stage dispenser for the first time 1.0-1.2 mg/litre, the 0.8-0.9 of dispenser for the second time mg/litre after 7-10 days, twice dispenser total concentration dosage is no more than 1.9～2.0 mg/litre, and twice every strain accepted dose and amounted to about 6 milliliters.
Rape chemical hybridizing agent composite of the present invention is used for the elimination of the early stage micro mist of CMS production of hybrid seeds process male sterile line, it is characterized in that utilizing the matched breeding technology of the auxiliary CMS of described composition, being about to SX-1 evenly sprays in the female parent of CMS seed farm capable, best period is the monokaryotic stage that rape cell is grown, be the maximum bud length of rape master inflorescence 1.5-2mm, best spraying medicine concentration is 0.5-0.7mg/L, and it is about 3ml that dose is accepted in every strain, every approximately 667m
2Spray 17.5-20kg.
At last, 2-[3-(4-methoxyl group-6-methyl isophthalic acid, 3,5-triazine-2-yl)-3-methyl ureido sulfuryl] application of methyl benzoate in the rape chemical emasculation.
The auxiliary agent of described composition (SX-1) is described now: because auxiliary agent is at the androcidal action of performance SX-1, prolong and kill the male time, be increased in the deposition on the target, reduce drift and improve other spraying proterties or biologically active, and reduce cost and alleviate the environmental pollution aspect and have important and positive effect.We are miscible with a spot of soluble starch, algae powder, ethanol, washing powder or sucrose respectively with SX-1, and result of study shows: washing powder not only has the gametocidal effect of strengthening SX-1, but also can alleviate poisoning.And other material is little to its influence.There are some researches show that surfactant has dispersion, adhesion, froth breaking, hydrotropy and activation, it is added on promotes adhesion, moistening, diffusion and blade to absorb in the flushing liquor, therefore can be used as the good auxiliary agent of agricultural chemicals.Petroleum oil, non-ionic surface active agent also can be used as the synergist of sulfonylurea herbicide, can improve the activity of chlorimuronethyl as the ammonium ion in the amine chemical fertilizer.Studies show that further the non-ionic surface active agent of hydrophil lipophil balance (HLB)＞10 can significantly improve the activity of sulfonylurea herbicide, can make the activity of rimsulfuron 25 (rimsulfuron) improve 10 times when the 12-17 as: HLB.Above-mentioned various surfactant also has effect same to SX-1, can help to improve its drug effect earlier with the SX-1 non-ionic surface active agent that adds spouting liquid 0.15～0.25% more soluble in water when using.
It is strong to inhale conductivity in the characteristics of SX-1 and mechanism of action: the SX-1, and active ingredient is active high, so consumption is minimum, and it can decompose rapidly by chemical hydrolysis in soil, about 60 days longevities of residure.The root leaf of SX-1 by rape absorbs, and two-way conduction in its body, but by root upwards the amount of conduction greater than the leaf portion amount of conduction downwards.(J.Bio.Chem., 1984,295:8753-7 such as La Rossa; Plant physiol.1984,75:827-31; Science (Washington), 1984, find that 223:1145-51) SX-1 acts on the ALS (acetolactate synthestase also is the acetohydroxy acid synthetase AHAS) in the plant corpus, and does not have second action site again.Because ALS is suppressed, the biosynthesis of branched-chain amino acid is hindered, and mainly makes valine, leucine, the biosynthesis of isoleucine is obstructed, and causes the accumulation of substrate α-butanone, block cell interkinesis DNA's is synthetic, and mitosis is stopped, and the cell cycle rests on G
1, G
2Stage.Because the sensitive part of crop is colored, using of low dosage, suppress and hindered the synthetic and normal normal growth that divides and influence stamen of flower pesticide cell DNA, played and killed male effect, but concerning insensitive position, do not play inhibition as yet, and poisoning even death can take place in over administration.Think that thus ALS may be the action target of CHA.
The action target of chemical hybridizing agent SX-1:
Researchs such as La Rossa think that ALS is sulfonylurea herbicide unique action site on plant, and the active ingredient of chemical hybridizing agent SX-1 is the sulfonylurea material.Therefore ALS also may be a better action target of chemical hybridizing agent.We select the identical salicylic another kind of Compound P yrithiobacsodium of pyrimidine that belongs to of effect rake mark of action target and SX-1 for use
[28], rape A74, K407 and wheat are handled, found that when its during in the above concentration of treatment of 2500mg/L, also can cause the male sterile of rape and wheat.Although owing to inhale factor such as conductivity in the effect of medicine and itself, it makes rape leaf withered spot occur in male killing, and wheat leaf blade is withered and be difficult to heading.But this is that the action target of CHA provides strong proof for ALS still.From the progress of present chemical hybridizing agent, the action target position of known CHA is not clear, does not also see the design of mentioning its MOLECULE DESIGN.Just because of do not understand the mechanism of action and the target of compound, often cause CHA to be difficult to screening, active compound is eliminated.Therefore, the discovery of this target of ALS is very useful to screening and the initiative of ALS.In recent years the He Cheng chemical high effective substance that acts on this target of CHA is very many, mainly contains sulfonylurea, imidazolone type, sulfonamides, thiazole pyrimidine sulfonyl amine, pyrazoles pyrimidine sulphonamide and pyrimidine salicylic acid
[28]SX-1 belongs to sulfonylurea, the material similar with it compare also have that some have efficiently, low toxicity and compound that residual life is grown.As: 2-[3-(4-methoxyl group-6-methyl isophthalic acid; 3; 5-triazine-2-yl) ureido sulfuryl] methyl benzoate and 2-{[(4; 6-dimethyl pyrimidine-2-yl) amino carbonyl] amino-sulfonyl } methyl benzoate etc. (referring to Su Shaoquan. the classification of herbicide action target and use. agricultural chemicals; 1998; 37 (11): 1-5.); though see that from the angle of agricultural chemicals residual life is shorter better; but male for killing of indefinite inflorescence rape, the hybridization agent of selecting 45-50 days residual lives for use with minimum consumption guarantee the whole florescence of rape 100% sterile be very necessary and significant.
Since the eighties in 20th century, aspect wheat breeding, domestic breeding research is subjected to bigger attention to CHA especially, but used CHA mostly is the new CHA that has abroad abandoned as the PGR of commercial production of hybrid seeds research etc. or external exploitation, is difficult to be used for produce but price is higher.Therefore utilize the method for combinatorial chemistry (promptly to adopt similar reaction condition, disposable synchronously synthetic thousands of kinds of molecules that structure is different, form a compound library and carry out high efficiency screening) and the rapid screening technology by to discovery, initiative and the screening of the high effective substance that acts on this target of ALS (as changing substituting group on the sulfonylureas bridge etc.), be expected to obtain efficient wheat male gametocide.Same this target of ALS also is very useful to the research of the CHA of other crop.
Seeding technique:
The discovery of chemical hybridizing agent SX-1, and, mainly be to kill the male production of hybrid seeds for using SX-1 to carry out rape to the research of SX-1 gametocidal effect on rape, and optimize ripe seeding technique theory support is provided for obtaining a cover.For the very low SX-1 of dosage, improve pesticide application technology such as protective spray-on process, wiping dose regimen, and develop novel medical instruments, all be to utilize SX-1 to carry out male sterilant production of hybrid seeds good problem to study.Think that according to research for many years carrying out rape with SX-1 kills the male production of hybrid seeds and need carry out following work:
1, safety is isolated: the chemical hybridization production of hybrid seeds should be carried out in isolated area, does not plant rape and brassicaceous vegetable (production of hybrid seeds is except male parent) in the scope about 1500m at least around the isolated area.Between breeding time, if spontaneous rape plant occurs and should in time root out in the seed farm or around the seed farm.
2, suitable row ratio: reasonably row is than being a key factor that improves hybrid seed yield and quality.Studies have shown that good in flower synchronization, under the condition of pollen amount abundance, father and mother's one's own profession is than adopting 1: 2,1: 3 outcrossing seed-setting rate difference is little, but from hybrid seed yield and the requirement of production of hybrid seeds benefit, adopts 1: 3 row than being both economical rational.
3, this flower synchronization of father and mother: can this florescence of father and mother meet well, is directly connected to the height of hybrid seeding output.Studies have shown that SX-1 can make the florescence of rape postpone 2-4 days, therefore when selecting strong advantage combination, the preferably maternal florescence was than male parent advance flowering period 3-4 days.Can adopt male parent to play the measure of tongue as not meeting, postpone its florescence.
4, the maternal hero that kills: get hold of the period that sprays and the mode of SX-1, carry out twice spray of low dosage medicine.When showing flower bud, monokaryon initial stage rape carries out the dispenser first time, secondary dispenser after 8-10 days, and twice spraying medicine concentration sum is less than 2.0 mg/litre, accepts dose and amounts to about 6 milliliters.For fear of medicine is sprayed on the male parent, can adopt simple and easy protection spray medicine frame, male parent is separated.As because of the period of spraying improper, its flower bud tongue can being plucked of micro mist appears in initial bloom stage, and be sterile fully until female parent.
5, roguing: catch the crucial growthdevelopmental stage that can manifest parent typical case proterties, the assorted strain that does not meet parent's characteristic feature is removed.As the phase of surviving the winter, elongation can carry out mutually with seedling according to Ye Se.
6, supplementary pollination: supplementary pollination is the ancillary method that improves hybrid seed yield, generally at first flower to full-bloom stage fine or cloudy calm morning 10 up to 12 o'clock, paternal pollen caught up with fallen on the maternal column cap with the feather duster or the pole of dialling that is tied with cloth.If the isolated area scope is big, can in isolated area, put honeybee, every 0.3-0.4hm
2Dispose a hive bee, be placed in assigned position when containing flower, male parent is in time withdrawn after spending eventually.
[beneficial effect]
Through the continuous test in 6 years in 1997～2002 years, we induce male sterile effect to SX-1 under different spraying medicine concentrations, dosage, period and number of times, and it has carried out multi-level omnibearing research to different rape varieties and rape floral organ, pollen vigor, rape growth and solid many-sides such as influence.And from its field seeding technique with utilize aspect such as the auxiliary rape C MS production of hybrid seeds of CHA to make good try, think by consulting a large amount of interrelated datas simultaneously: compare with other chemical hybridizing agent of rape, SX-1 is the chemical hybridizing agent of a kind of comparatively desirable new and effective, low toxicity, low-residual, low price, using the every strain of 1.0-2.0 mg/litre concentration accepts all can induce male sterile under 3 milliliters of left and right sides doses, gametocidal effect is thorough, and sterile plant rate can reach 95%-100% and not have poisoning or have only slight poisoning; Gynoecium is normally solid or normal substantially.Secondly to different genotype kind gametocidal effect basically identical.Just because rape belongs to indefinite inflorescence; in dispenser period, number of times, dose, concentration, effectively kill on male interval and the dispenser uniformity and there are differences; so normal flower can appear in some plant before spending eventually in 1.0-2.2 mg/litre concentration range, thereby cause that sterile degree differs between different disposal.But studies have shown that this problem can be overcome by insecticide-applying way and pesticide application technology.
The assessment of SX-1 seed production purity:
Genotype and CHA, environment and CHA are depended in effective utilization of CHA and diversification thereof exploitation, and genotype and environment and CHA make the size of effect mutually.Therefore, obtain higher hybrid seed yield, need be according to different kinds and specific dosage and time of environmental selection.SX-1 is a kind of excellent in chemical hybridization agent, but rape is an indefinite inflorescence, and SX-1 in use also can influence seed production purity because of the factors such as longer blooming period that abnormal climate causes that dispenser period, concentration and rape seed later stage occur.Therefore the assessment to the SX-1 seed production purity is the important indicator of reflection SX-1 practical value.Test selects for use K407,20Z71 and Y37 for maternal, earlier with 1.0 mg/litre again with 42.5 kilograms/667m of 2 concentration of 0.9 mg/litre
2SX-1 it sprayed by stages carry out the production of hybrid seeds, test shows with SX-1 and sprays made several hybrids that its purity all between 90%～98%, meets the requirement of country-level crossbreed fully.
SX-1 assists the CMS production of hybrid seeds:
The hybrid vigour of rape is extremely remarkable, and utilizing cytoplasmic male sterility (CMS) at present is the main path that rape heterosis utilizes.By bred Qin oil No. two, assorted oil 59, assorted No. 1 of China, assorted No. 2 of China, yellow assorted No. 1, No. 7, Qin You of this approach, rather a series of CMS hybrids such as assorted No. 1 are applied to produce.But the CMS sterility is subject to the influence of environment, is prone to micro mist in early spring under the cryogenic conditions, and reduces seed production purity.Official's spring cloud etc. (chemical hybridizing agent is induced the research of rapeseed male sterility mechanism. and Botany Gazette, 1997,23 (50): 513-518.) studies show that: chemical hybridizing agent can induce rape flower pesticide tapetum to depart from normal development, produces abortive pollen.Zhang Xuekun etc. (chemical hybridizing agent is to the influence of rape cytoplasmic sterile line Pohle horse fertility. Agricultural University Of Southwest's journal, 1999,21 (2): 140-143.) utilize No. one, male gametocide to handle Pohle horse cytoplasmic sterile line result and show: No. one, male gametocide can both be induced Pohle horse cytoplasmic sterile line rape effectively in pollen mother cell period to the mature pollen phase, can make the abortion once more of its trace-pollen, reduce trace-pollen effectively, thereby obviously improved sterile degree and seed production purity.2001～2002 years, we adopted SX-1 to assist the CMS production of hybrid seeds, through test of many times, with the SX-117.5-20 kilogram/667m of 0.5-0.7 mg/litre concentration
2In the rape monokaryon initial stage female parent of CMS seed farm is handled, reduce and eliminate the micro mist in maternal early stage in the CMS production of hybrid seeds process effectively, improve rape seed purity, special very remarkable for the bigger regional result of use of China's south winter rape production of hybrid seeds micro mist amount.We adopted SX-1 to handle to the female parent of No. 7 seed farms of more than the 250 mu of Qin You in Shan south in 2002, compared with contrast (not spraying medicine), and its seed production purity has improved 4-5 percentage point, and hybrid rate is to have reached the first order seed standard more than 93%.
[embodiment]
To further specify the present invention by non-restrictive example below.
Embodiment 1
This embodiment studies the influence of using dosage to gametocidal effect.Test consisting of of used chemical hybridizing agent composite:
2-[3-(4-methoxyl group-6-methyl isophthalic acid, 3,5-triazine-2-yl)-3-methyl ureido sulfuryl] methyl benzoate: 70%
Wet spreader: lauryl sodium sulfate 3.5%
Dispersant: sodium acid carboxylic compound 5.0%
Thickener: hydroxy cellulose sodium 0.5%
Disintegrant: PTPP 4.0%
Soluble starch: 5.0%
Filler supplies 100%
All to be solvent with water be made into finite concentration (with active ingredient weight note) with said composition to each embodiment tests.Confession formation testing dish is cabbage type rape (B.napus) kind (being) K407, Shan 3B (the confession formation testing vegetable kind among the embodiment 2,3,5,6 is identical with it).The standard that the standard of perfection employing official spring cloud of gametocidal effect provides (referring to Fu Tingdong etc., " breeding of cross-bred rape and utilization ", Hubei science tech publishing house, 1995).The spray pharmacy amount is the key factor that influences rape colony gametocidal effect, and each individual plant is accepted dose and crossed that I haven't seen you for ages and reduce gametocidal effect under finite concentration, and dose too much can produce poisoning again.Working concentration scope and the dosage of SX-1 on rape tested in 1997.This embodiment establishes 0,0.5,1.5,3.0,4.5,10.0 altogether, a 15.0mg/L7 concentration is handled; 40,42.5 and 45 kilograms/667 meters
2Three spray dosage, 3.36 meters m of every processing
2, density of reserved seedlings 1.2 ten thousand strains/667 meters
2Evidence: 6 concentration all can be induced rapeseed male sterility, but SX-1 evenly sprays less than 45 kilograms/667 meters under 0.5,1.5 mg/litre concentration
2Can not produce poisoning or slight poisoning is only arranged, 3.0 mg/litre or above concentration are handled and can be produced serious poisoning.Spraying concentration is SX-145 kilogram/667 meter of 1.5 mg/litre
2Induce male sterile effect better, can obtain 100% complete sterility strain.Therefore determine that SX-1 as the using dosage scope of rape male gametocide is: concentration between 0.5-3.0mg/L spray value less than 45 kilograms/667 meters
2
Table 1 SX-1 variable concentrations is handled the influence to the rape fertility
| Material | Handle (mg/L) | Processing period | Handle strain number (strain) | Initial bloom stage complete sterility rate (%) | Full-bloom stage complete sterility rate (%) | Complete sterility rate (%) before spending eventually | Moderate poisoning strain rate (%) |
| Shan 3B | 0 1.0 1.3 1.6 1.9 2.2 | The monokaryon later stage | 62 63 67 67 71 74 | 0 85.7 88.1 94.0 100 100 | 0 95.2 97.3 98.5 100 100 | 0 80.95 85.1 85.1 94.4 100 | 0 0 1.5 1.5 5.6 13.5 |
| K407 | 0 1.0 1.3 1.6 1.9 2.2 | The monokaryon initial stage | 75 75 89 87 87 90 | 0 96.0 100 100 100 100 | 0 97.3 100 100 100 100 | 0 72 79.8 93.1 95.4 100 | 0 0 0 2.3 3.4 12.2 |
| K407 | 0 1.0 1.3 1.6 1.9 2.2 | The monokaryon later stage | 60 88 90 92 90 98 | 0 92.1 95.6 95.6 100 100 | 0 96.6 96.7 100 100 100 | 0 83.0 92.2 95.6 97.8 100 | 0 0 1.1 3.5 4.8 14.7 |
Select 42.5 kilograms/667 meters on this basis for use
2Continuous 4 years of formulation rate to 0,1.0,1.3,1.6,1.9,6 concentration research experiment of 2.2mg/L (table 1), 4.48 meters of every processing
2, stay seedling 1.2 ten thousand strains/667 meters
2The result shows: in 1.0-2.2 mg/litre scope with handle concentration to increase gametocidal effect thorough more, wherein 1.0,1.3,1.6mg/L three handles all has the minority plant micro mist to occur early stage, and to continue mostly 4 days right and lefts change into sterile fully, 1.0 mg/litre of K407 handle and three processing of 1.0,1.3,1.6 mg/litre of Shan 3B in have that all there is micro mist at the whole florescences of plant until normally educating extremely individually.Once just can reach 100% complete sterility up to whole flower 2.2 mg/litre is only handled, solid abnormal moderate poisoning strain rate is higher but main inflorescence shortens, and reaches more than 12%; And 1.9 mg/litre are handled and generally can be reached 100% complete sterility strain rate, and have 95% plant still to keep sterile to flower eventually, and suffered poisoning is also lighter, and about 5% half sterile strain only is to spend individually before spending eventually near normal, to the purity influence of crossbreed not quite.Therefore consider that from the fertility and the suffered poisoning situation of rape at whole florescence 1.9 mg/litre processing gametocidal effect is better.
Embodiment 2
This embodiment studies the influence of spraying period to gametocidal effect.From the male lasting period consideration of killing of SX-1, adopt the SX-1 of 1.9 mg/litre to begin dispenser in pollen mother cells to the monokaryon initial stage, processing in per afterwards 5 days, up to the pollen maturation phase, the every processing of totally 4 phases is 4.48 meters
2, stay seedling 1.2 ten thousand strains/667 meters
2Result of the test is listed in table 2.
Table 2 SX-1 different disposal period is to the influence of rape fertility
| Material | Handle (mg/L) | Processing period | Handle strain number (strain) | Initial bloom stage complete sterility rate (%) | Full-bloom stage complete sterility rate (%) | Complete sterility rate (%) before spending eventually | Moderate poisoning strain rate (%) |
| K407 | 1.9 1.9 1.9 1.9 | Two nuclear pollen maturation phases phase of monokaryon initial stage monokaryotic stage | 95 93 86 95 | 100 98.9 65.1 0 | 100 100 99 100 | 100 100 99 100 | 4.2 4.3 1.2 1.05 |
| Shan 3B | 1.9 1.9 1.9 1.9 | Two nuclear pollen maturation phases phase of monokaryon initial stage monokaryotic stage | 80 74 82 73 | 100 98.6 68.3 0 | 100 100 100 100 | 92.8 93.2 100 98.6 | 6.0 5.7 1.2 0 |
The result shows: rape sprays medicine from pollen mother cells to the pollen maturation phase all have gametocidal effect, but with monokaryon initial stage and monokaryotic stage pesticide supplying effect the best, two kinds all reach the complete sterility strain more than 98%, wherein K407 still keeps 100% sterile before spend eventually, and the sterile plant rate of Shan 3B is also greater than 92%.Form from plant colony; when the sub-district reaches squaring period, the maximum bud length of main inflorescence effect of spraying pesticide in the time of 1.5～2.0 millimeters of 50% individual plant is better, and pollen maturation phase pesticide supplying effect is the poorest; mainly be that the initial bloom stage drug effect is failed performance, cause occurring the normal florescence about 10 days.Therefore SX-1 induces male sterile suitable time to arrive monokaryotic stage for the monokaryon initial stage.
Embodiment 3
This embodiment studies the influence of spray medicine number of times to gametocidal effect.Rape is an indefinite inflorescence, in growth course its flower bud differentiation successively different, so the dosage of dispenser and number of times directly influence SX-1 rapeseed male sterility are induced effect, duration and poisoning weight.Therefore, establishing monokaryotic stage only handles once with 1.9 mg/litre and compares; The monokaryon initial stage sprays for the first time with the SX-1 of 1.0 mg/litre and 1.6 mg/litre earlier, after 8-10 days respectively the SX-1 with 0.6,0.9,1.0 mg/litre spray for the second time.Spray dosage at every turn and be 42.5 kilograms/667 meters
2, totally 7 processing, result of the test is listed in table 3.
The different spraying times of table 3 SX-1 variable concentrations are to the influence of fertility
| Material | First concentration of treatment (mg/litre) | Concentration of treatment (mg/litre) for the second time | Handle strain number (strain) | Initial bloom stage complete sterility rate (%) | Full-bloom stage complete sterility rate (%) | Complete sterility rate (%) before spending eventually | Moderate poisoning strain rate (%) |
| K407 | 1.0 1.0 1.0 1.6 1.6 1.6 1.9 | 0.6 0.9 1.0 0.6 0.9 1.0 0 | 86 87 90 90 81 87 87 | 98.8 100 100 100 100 100 100 | 100 100 100 100 100 100 100 | 94.2 99.0 100 100 100 100 94.3 | 0 1.1 2.2 8.9 12.3 12.6 4.5 |
| Shan 3B | 1.0 1.0 1.0 1.6 1.6 1.6 1.9 | 0.6 0.9 1.0 0.6 0.9 1.0 0 | 84 76 71 76 69 69 74 | 96.4 98.7 100 100 100 100 100 | 97.6 100 100 100 100 100 100 | 84.5 96.1 98.6 100 100 100 94.6 | 0 2.6 4.2 8.0 11.6 10.1 5.4 |
The result shows: the contrast that sprays 1 time with 1.9 mg/litre can obtain 100% sterile good result, but still has 5.4% sterile strain micro mist flower and fertile flower to occur in the later stage.Spray twice processing, when spraying medicine concentration close more than or equal to 1.9 mg/litre the time can obtain 100% complete sterility strain rate.Wherein K407 still has the complete sterility rate more than 99% before spending eventually, and Shan 3B also has the complete sterility rate more than 96%, poor effect when twice spraying medicine concentration sum is 1.6 mg/litre.It is light that the processing that twice spraying medicine concentration sum is 1.9 mg/litre is killed male longer duration and poisoning than the processing that once sprays 1.9 mg/litre.But because twice dispenser meeting produces the accumulation of medicine in the plant body, thus when twice spraying medicine concentration sum during greater than 2.2 mg/litre poisoning heavier, close the increase of flower bud strain and dead plant rate.Therefore the dose summation of twice spray medicine should be controlled at 1.9 mg/litre between the 2.0 mg/litre concentration.
Embodiment 4
This embodiment studies SX-1 to the genotypic influence of different rapes.A kind of good CHA not only requires its gametocidal effect thoroughly to the not influence of the fertility of gynoecium, and require its to different genotype to make effect mutually little, promptly to the gametocidal effect no significant difference of different genotype kind.In SX-1 being carried out the research in 6 years, altogether the kind of 32 separate sources, different genotype has been carried out killing male test, and be the 42.5 kilograms/667m of SX-1 of 1.9 mg/litre with concentration
29 kinds are handled simultaneously, and result of the test is listed in table 4.
Table 4 SX-1 is to the influence of different rape variety fertility
| Kind | Handle (m g/) | Processing period | Handle strain number (strain) | Initial bloom stage complete sterility rate (%) | Full-bloom stage complete sterility strain (%) | Complete sterility rate (%) before spending eventually | Moderate poisoning strain rate (%) |
| K407 92P137 8906B 20NH705 20NH701 Shan 3B 20O30 20O424 20O368 | 1.9 1.9 1.9 1.9 1.9 1.9 1.9 1.9 1.9 | The monokaryotic stage monokaryon initial stage monokaryotic stage monokaryon initial stage monokaryotic stage monokaryon initial stage monokaryotic stage monokaryon initial stage at monokaryon initial stage | 87 90 95 93 87 71 60 73 73 | 100 97.8 100 94.9 100 100 100 95.9 100 | 100 98.9 100 100 100 100 100 100 100 | 95.4 88.9 92.6 94.6 100 94.4 90.0 95.9 89.0 | 3.4 4.4 3.2 2.2 1.1 5.6 6.3 5.5 5.5 |
The result shows: SX-1 is little to the gametocidal effect difference of different genotype rape, and the complete sterility strain rate of each kind from first flower to the Sheng flower is all greater than 95%.But because different cultivars genotype differences causes the male gametocide amount of required SX-1 slightly different.As the kind NH of Korea S
701100% induced male sterile after treatment, and can be retained to whole flower.K
407, NH
705, Shan 3B, 20O
424Though induced sterilely Deng 100%, had only the sterile rate about 95% before spend eventually.And in the test of SX-1 spraying times, K407 all can obtain 100% sterile good result until whole flower as adopting twice dispenser.As seen SX-1 all has gametocidal effect preferably to different genotype kind (being), if dispenser period, concentration and number of times are suitable, it is reached or near 100% gametocidal effect.
Embodiment 5
This embodiment studies the influence of SX-1 to rape floral organ, pollen vigor.SX-1 is an important evidence of determining the SX-1 gametocidal effect to the observation of rape floral organ, pollen effect of vigor.The male gametocide gametocidal effect standard of perfection that proposes according to official's spring cloud etc. and the week the lady in the moon of Wuhan University (referring to Fu Tingdong etc., " breeding of cross-bred rape and utilization ", Hubei science tech publishing house, 1995) the pollen vigor observation caliber of Ti Chuing, mainly clear water is handled (CK) and with the K407 of SX-1 after monokaryotic stage processing once of 1.9 mg/litre, the floral organ of Shan 3B and pollen vigor are observed, respectively at the rape initial bloom stage, full-bloom stage and whole florescences three sub-sampling, adopt every class spends 50 to measure the corolla diameter at every turn, the petal size, filigree, flower pesticide and gynoecium length, ask its mean value, and adopting the methylene blue decoration method to measure pollen viability, result of the test is listed in table 5.
Table 5 SX-1 is to the influence of rape floral organ and pollen vigor
| Material | Classification | Corolla diameter mm | Petal | The long mm of filigree | The long mm of flower pesticide | The long mm of gynoecium | The pollen vigor is identified | Solid situation * | ||||||
| The bagging selfing | Artificial different friendship | |||||||||||||
| Wide mm | Long mm | Investigation pollen number (grain) | Debility pollen number (grain) | Debility pollen rate (%) | Effective angle fruit (individual) | Kernal number (grain) | Effective angle fruit (individual) | Kernal number (grain) | ||||||
| K407 | The complete sterility strain | 15.28 | 6.18 | 11.47 | 5.08 | 2.04 | 8.57 | 1034 | 1031 | 99.7 | 0 | 0 | 67 | 927 |
| Half sterile strain | 16.25 | 6.31 | 12.49 | 7.36 | 2.03 | 6.89 | 803 | 542 | 67.5 | 38 | 143 | 76 | 983 | |
| Normal strain (CK) | 18.69 | 7.20 | 14.42 | 7.92 | 2.06 | 7.27 | 1855 | 128 | 6.9 | 52 | 546 | 78 | 1013 | |
| Shan 3B | The complete sterility strain | 15.88 | 5.76 | 12.31 | 6.21 | 2.08 | 7.44 | 862 | 855 | 99.19 | 0 | 0 | 77 | 1353 |
| Half sterile strain | 16.26 | 6.36 | 12.85 | 6.98 | 2.14 | 7.76 | 834 | 491 | 58.87 | 43 | 182 | 75 | 1147 | |
| Normal strain (CK) | 18.15 | 6.06 | 13.12 | 7.52 | 2.12 | 8.45 | 1079 | 53 | 4.91 | 55 | 806 | 75 | 1261 | |
Annotate: 1, the pollen vigor is identified: carry out according to the methylene blue decoration method that week the lady in the moon of biology department of Wuhan University (1964) proposes; Floral organ is measured: every class flower is adopted 50 each indexs of measurement of firm opening and is asked its mean value.2: pollen is measured in solid situation * bagging selfing debility and solid situation; The activity and the pollination ability thereof of gynoecium measured in artificial different friendship.10 inflorescences are respectively made in selfing and different friendship.
The result shows: the male sterile plant of the acquisition that compositions-treated is induced through containing SX-1 shows as the complete sterility strain at first flower to full-bloom stage, indivedual plant show as partly sterile before spending eventually, the feature of comparing this three classes plant with the normal strain of contrast is respectively, the complete sterility strain: staminody is needle-like, filigree significantly shortens or is dried not plump tall and thin, the flower pesticide WUHUAFEN, and it is not plump to wrinkle, look light or be empty not plump, lopsided, dead abortive pollen, and the pollen vigor is below 0.9%.Corolla is slightly less than normal flower, and gynoecium length increases, and the bagging selfing is shaky, but outcrossing is normal.Half sterile strain: floral organ is slightly littler than normal flower, and staminody has pollen grain in the flower pesticide, and pollen part deformity is identified its vitality through methylene blue dyeing, the not plump death of wrinkling of about pollen of about 50%.The bagging self-fruitful rate is low, and outcrossing is normal.Normal strain: floral organ is normal, and dead pollen rate is below 7%.
Embodiment 6
This embodiment studies SX-1 rape is grown and solid influence.Present embodiment selects for use K407 and two rape varieties of Shan 3B with 42.5 kilograms/667 meters of 6 concentration of 0,1.0,1.3,1.6,1.9,2.2 mg/litre
2SX-1 with the compression manual sprayer it is handled, every processing 4 row, per 3 days of the back of blooming is observed once until whole flower each fertility and developmental state thereof of handling rape, the normal strain that initial bloom stage is handled with 0 mg/litre is contrast, the complete sterility strain that the growing way unanimity is chosen in other processing mark of listing, 10 strains are selected in every processing, in the species test of rape maturing stage, calculate its single plant yield, result of the test is listed in table 6.
Table 6 is grown and solid situation with the rape that SX-1 handles
| Material | Handle (mg/litre) | Plant height (cm) | Effective branch position (cm) | Main inflorescence effective length (cm) | Complete stool effective angle fruit number (individual) | Every fruit grain number (grain) | Thousand kernel weight (g) | Single plant yield (g) |
| Shan 3B | ck 1.0 1.3 1.6 1.9 2.2 | 136.60 134.60 136.14 133.38 132.75 132.38 | 45.63 54.56 60.86 53.88 47.50 53.13 | 40.00 35.88 39.00 33.13 39.13 33.5 | 286.94 268.89 259.83 273.00 284.07 255.64 | 26.85 25.98 24.48 24.14 21.70 20.14 | 2.93 2.75 3.14 2.96 2.98 3.00 | 20.96 19.21 19.97 19.51 18.37 15.45 |
| K407 | Ck 1.0 1.3 1.6 1.9 2.2 | 168.57 173.14 170.13 164.75 171.13 159.29 | 62.33 60.14 61.50 63.00 62.13 62.43 | 71.70 65.14 65.13 61.38 57.63 58.57 | 319.36 329.50 307.01 296.33 286.76 302.12 | 18.99 16.27 16.14 16.88 15.64 13.38 | 3.12 3.22 3.24 3.20 3.18 3.05 | 18.92 17.26 16.05 16.01 14.26 12.33 |
Annotate: CK belongs to normal strain contrast, and all the other are treated to the solid situation of sterile strain spontaneous pollination behind the hero extremely.
The result shows: the growth to the rape elongation when the concentration of treatment of SX-1 〉=1.6 mg/litre has certain inhibitory action, and increasing inhibitory action with concentration strengthens, the whole growing of rape prolongs, and is subjected to the obvious cripetura of main inflorescence of poisoning strain, but divides branch growth normal to development later stage.Maturing stage except that the poisoning strain, each plant height of handling with contrast between no significant difference, and slightly improve at effective branch position of Shan 3B.Main inflorescence effective length reduces, and sees 1.9 mg/litre are normal and single plant yield that 2.2 mg/litre are handled is minimum from the plant of handling with the SX-1 variable concentrations, differs greatly with CK, and its reason is that concentration is higher, suppress greatly due to.Prove also by artificial different friendship and natural crossing that in addition SX-1 handles the sterile strain that is obtained solid normal (except the poisoning strain).Therefore under suitable concentration, utilize SX-1 to induce the male sterile of rape can not produce ill effect.
Claims (5)
1; a kind of using method of rape chemical hybridizing agent composite; said composition contains 2-[3-(4-methoxyl group-6-methyl isophthalic acid; 3; 5-triazine-2-yl)-and 3-methyl ureido sulfuryl] methyl benzoate; it is characterized in that; with the water-soluble formation soup of said composition; when the maximum bud length 1.5-2mm of rape master inflorescence, female parent to be implemented to kill male, spraying medicine concentration is 2-[3-(the 4-methoxyl group-6-methyl isophthalic acid of 1.8 mg/litre-2.0 mg/litre; 3; 5-triazine-2-yl)-and 3-methyl ureido sulfuryl] methyl benzoate, divide and use for 2 times, it is 3 milliliters that dose is accepted in each every strain; to father and mother's one's own profession ratio is 1: 1 female parent, every 667m
2Spray the 17.5-20kg soup, the 1.0-1.2 of dispenser for the first time mg/litre, the 0.8-0.9 of dispenser for the second time mg/litre after 7-10 days, twice dispenser total concentration is no more than 2.0 mg/litre, and twice every strain receiving amount amounts to 6 milliliters.
2, using method as claimed in claim 1 is characterized in that, described composition also contains wet spreader, dispersant, thickener, disintegrant, soluble starch and/or filler.
3, using method as claimed in claim 2 is characterized in that, described composition is made up of following material by weight percentage:
2-[3-(4-methoxyl group-6-methyl isophthalic acid, 3,5-triazine-2-yl)-3-methyl urea 10-75% base sulfonyl] methyl benzoate
Dodecyl sulphate potassium or sodium 2.0-4.5%
Beta-naphthalenesulfonic-acid potassium or sodium acid carboxylic compound 4.0-6.0%
Hydroxy cellulose sodium 0.3-0.5%
PTPP or sodium 2.0-5.0%
Soluble starch 3.0-5.0%
Filler supplies 100%.
4, using method as claimed in claim 3 is characterized in that, described composition is made up of following material by weight percentage:
2-[3-(4-methoxyl group-6-methyl isophthalic acid, 3,5-triazine-2-yl)-3-methyl urea 70% basic sulfonyl] methyl benzoate
Dodecyl sulphate potassium or sodium 3.5%
Beta-naphthalenesulfonic-acid potassium or sodium acid carboxylic compound 5.0%
Hydroxy cellulose sodium 0.5%
PTPP or sodium 4.0%
Soluble starch 5.0%
Filler supplies 100%.
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| CN101595836B (en) * | 2009-07-02 | 2011-11-09 | 西北农林科技大学 | Method for preservation of pure state of hybrid plants by chemical emasculation |
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| CN100345484C (en) * | 2006-05-29 | 2007-10-31 | 西北农林科技大学 | Application of compound in preparation of chemical hybridization agent for brassica plants |
| CN101473838B (en) * | 2009-02-11 | 2012-05-09 | 西北农林科技大学 | Chemical anthericide for cabbage stalk and formulating method thereof |
| CN101595837B (en) * | 2009-07-13 | 2012-08-29 | 陕西省杂交油菜研究中心 | Ultra high-oil rapeseed germplasm and breed selection method of high-oil hybrid thereof |
| CN101884335A (en) * | 2010-07-17 | 2010-11-17 | 陕西合和种业科技有限公司 | Chemical male sterilant composition of cruciferae crops and application method thereof |
| CN101911873B (en) * | 2010-07-26 | 2012-06-27 | 山东省农业科学院蔬菜研究所 | Application of ethylmethylsulfone in preparation of chemical anthericide for crops, and breeding |
| CN103081798B (en) * | 2012-11-12 | 2015-04-22 | 陕西省杂交油菜研究中心 | Method for improving rape hybrid by chemically inducing male sterility |
| CN103539539B (en) * | 2013-10-12 | 2015-03-11 | 华中农业大学 | Viola x wittrockiana chemical hybridizing agent composition and its preparation method and use |
| CN106332774A (en) * | 2016-11-02 | 2017-01-18 | 陕西省杂交油菜研究中心 | Efficient breeding method adopting multi-female-parent one-male-parent mode to prepare oilseed rape hybrid |
| CN106718840A (en) * | 2016-12-16 | 2017-05-31 | 咸阳市农业科学研究院 | A kind of Guanzhong area cabbage type rape cytoplasmic sterility cross-breeding method |
| CN106577263B (en) * | 2016-12-19 | 2019-04-05 | 陕西省杂交油菜研究中心 | A kind of rape chemical hybridizing agent accuracy pesticide applying method |
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