CN101133715A - Tobacco NaN3 mutagenesis idioplasm innovation and breed selecting and cultivating technique - Google Patents
Tobacco NaN3 mutagenesis idioplasm innovation and breed selecting and cultivating technique Download PDFInfo
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- CN101133715A CN101133715A CNA2007100779665A CN200710077966A CN101133715A CN 101133715 A CN101133715 A CN 101133715A CN A2007100779665 A CNA2007100779665 A CN A2007100779665A CN 200710077966 A CN200710077966 A CN 200710077966A CN 101133715 A CN101133715 A CN 101133715A
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Abstract
The present invention discloses a technique for making tobacco NaN3 mutation germplasm innovation and selecting-breeding variety. Said technique includes the following steps: (1), mutagenic material selection, selecting air-dried seed of tobacco cultivated variety with good comprehensiveness and strong adaptability; (2), preparing NaN3 solution; (3), seed treatment, using NaN3 solution whose concentration is 4mmol/L to treat seed for 6h at 25deg.C; (4), planting and management of M1; (5), planting and selection of M2; and (6), M3 and next-treatment. Said method can greatly raise mutation frequency, and its beeding effect is high.
Description
Technical field
The present invention relates to methods for the treatment of seed, specifically, relate to tobacco germplasm innovation and breeding technology.
Technical background
Since 1948, Gustafsson etc. once handled barley with mustard gas and obtain mutant; Nilan in 1967 handles barley seed with dithyl sulfate and has bred of short stem, high yielding variety, and research of chemical mutagen after this and application just progressively grow up.Compare with physical mutagen, the characteristics of chemical mutagen have: 1. induced mutation rate is higher, and chromosome aberration is less; 2. light to handling material damage, some privileged site that the chemical mutagen that has is only limited to DNA morphs; 3. most of effective chemical mutagen are big to biological damage than physical mutagen, cause that easily vitality and fertility descend.Aspect the new varieties that cultivate plants by mutagenesis, the technical scheme application has been arranged patent, for example Chinese patent application part ZL02136506.7 number " breeding method of natural multi-colour lawn " carries out direct mutagenesis of seed or stripped mutagenic treatment with sodium azide, cultivated multi-colour lawn; 01139096.4 number " method of biotechnology improvement rice quality " handles seed rice with chemical mutagen EMS+NEU, improved rice quality; 200410053947.5 number " a kind of selection that has leaf color marker corn " handles corn seed with sodium azide or ethylmethane sulfonate, mutagenesis obtains having the corn of leaf color marker; 200510043191.0 number " a kind of preparation method of plant with yellow seedlings in red pepper seedling stage " soaks pepper seed with nitrous acid solution, obtains plant with yellow seedlings etc.But up to now, the technology that adopts the mutagenesis method to handle tobacco seed is not appeared in the newspapers as yet.
Summary of the invention
The object of the present invention is to provide tobacco NaN
3Mutagenesis idioplasm innovation and breeding technology are selected NaN
3As the mutagen of practicality, and the mutagen that this induced mutation rate is high and safe are applied to the tobacco bred seed selection efficiently.
The inventor recognizes by long-term experimental study, compares with physical mutagenesis, and chemical mutagen has more danger to human body, must selection not affect the active principle of operator ' s health as mutagens.By screening, find NaN
3As a kind of vegeto-animal respiration inhibitor, DNA base in duplicating is replaced, having higher mutation frequency, have efficient, nontoxic, low price and advantage such as safe in utilization again, is the high and a kind of mutagen of safety of induced mutation rate at present few in number.
Tobacco NaN provided by the invention
3Mutagenesis idioplasm innovation and breeding technology are complete technical systems, and it comprises:
(1) mutant materials is selected
Select that comprehensive proterties is better, high-quality is suitable produces, and adaptability is stronger, the conventional kind or crossbreed of excellence cultivation that need improve indivedual bad proterties;
(2) preparation NaN
3Solution
4.11ml 0.2M sodium hydrogen phosphate and 15.89ml 0.1M citric acid are mixed, be mixed with citric acid-disodium phosphate soln of pH=3.Wherein the 0.1M citric acid is with the citric acid 21.01g preparation 1000ml aqueous solution of a water of crystallization; 0.2M sodium hydrogen phosphate is with the 35.01g phosphate dihydrate disodium hydrogen preparation 1000ml aqueous solution; Again with 0.26g NaN
3Be dissolved in the citric acid-disodium phosphate soln of pH=3 among the 1L, be mixed with the NaN of 4mmol/L
3Solution.
(3) seed treatment
The dry seed of selected materials with clear water preimpregnation 24h, is used the NaN of 4mmol/L in the drier that fills 50% glycerine, one aqueous solution behind the balance 7d then in temperature is 25 ℃ insulating box
3Solution is handled 6h down at 25 ℃, the seed of handling washes 8h in flowing water after, obtains M
1For seed; The seed number of handling should be more than 10000.
(4) plantation of mutagenesis tobacco, selection and management
M with above-mentioned processing
1Sow, grow seedlings, transplant, cultivate, open pollination for seed, and sowing obtains mutagenesis two generations (M
2) seed, again sow, grow seedlings, transplant, cultivate, the pollination of blooming, and sowing obtains seed of future generation, so repeatedly plantation and to second, third, the plant in the 4th generation selects, after strain is stable, all surpasses or approach the material that contrasts for continuous 2 years excellent strain comparative test Comprehensive Traits, can submit regional testing to, enter the variety certification program.
In the said process, a mutagenesis generation (M
1) plantation with the management main points be: M
1Should accomplish as far as possible to sow in positive season of ground main cultivation of preliminary election kind for seed, avoid making M because changing sowing time and ecological condition
1For plant change because of conditions such as sunshine and temperature in growth course, making grows normally is affected; M
1The whole ground in generation, sow, grow seedlings, transplanting and field management should carry out according to the suitableeest mode in locality, makes every effort to meticulous, to guarantee the higher survival rate of seedling; A field period bud picking is not pinched; But, M
1The planting density in generation can increase about 50% than the density of the common employing in locality, in order to dwindle M
1The planting scale in generation is saved cost and labour; M
1Open pollination of generation according to the few grain of strain method sowing, and is gathered in the crops the seed of the different fruit branch joint of same strain position, as far as possible to take full advantage of the sudden change chimera; Seed with same kind mutagenic progeny mixes then.
In the said process, mutagenesis two generations (M
2) plantation with selecting main points be: M
2In generation, adopted the mode of classifying type miscegenation, the i.e. M of same kind usually
2For mixed seeding, and plant 1 row contrast every 3-5 is capable, in order to observe possible sudden change; M
2In generation, also should accomplish to plant the positive season sowing in ground at the main of preliminary election kind, and sow, grow seedlings, transplanting and whole field management all adopt the suitableeest mode in locality to carry out; A field period bud picking is not pinched; For take the disease resistance seed selection as purpose M
2In generation, can plant in the sick garden of the evaluation of purpose disease or disease district occurred frequently, and contrast every capable 1 row of planting of 3-5, in order to depress the raising Breeding Efficiency in certain selection; For according to the M that selects in the breeding objective
2For plant, should be at the bagging in squaring period, independent sowing, and by a part strict numbering, put down in writing main feature; Plant for not middle choosing can mix sowing, also can according to circumstances abandon sowing.
In the said process, mutagenesis three generations (M
3) and later medical record be: M
3In generation, also should accomplish to plant the positive season sowing in ground at preliminary election kind main, sows, grows seedlings, transplanting and whole field management all adopt the suitableeest local mode to carry out, and add the contrast of 1 row every the capable mutant material of 3-5; M
2The individual plant kind that generation is selected becomes M
3For strain, every strain 150-200 strain; Be main purpose M for the disease resistance seed selection
2Generation, can continue to plant in corresponding sick garden or disease district occurred frequently, every strain 500-800 strain, and plant 1 row contrast every 3-5 is capable is in order to the degree of stability of investigating mutant character with proceed to select; Should compare with control material during selection, be defined as mutating strain series after, select earlier excellent mutating strain series, therefrom select again excellent sudden change individual plant; The individual plant of selecting from high stability excellent mutating strain series changes M over to after the mixed receipts of plant division
4Carry out the test of output pilot study and resistance and Quality Identification.According to result of the test, M
4Select again best strain in the strain of participating in the experiment in generation, and from best strain, select excellent strain and carry out numerous kind; For sudden change shape unstabilized mutating strain series still, after the roguing at M
4Generation even M
5In generation, still plant by strain, continues to select; For the M that mixes sowing
2For material, its M
3In generation, is still with reference to M
2In generation, carried out miscegenation, the individual plant of middle choosing, its M
4In generation, be similar to M
2M for single sowing
3For processing method, postpone according to this; In the ordinary course of things, the tobacco mutagenic progeny is passed through M
3The selection in generation, most M
4Stable for strain, so M
4In generation, should be carried out seminal propagation work simultaneously except carrying out the output pilot study, be M
5In generation, participate in the major clique rating test seed preparation be provided; For continuous 2 years excellent strain comparative test Comprehensive Traits all above or approach the material of contrast, can submit regional testing to, enter the variety certification program.
The inventor points out: population size suitably is the necessary condition that ensures mutagenesis idioplasm innovation and breed breeding success, and therefore, in the seed treatment stage, the seed number of processing should be more than 10,000; M
1In generation, should increase planting density, M
1The plantation colony in generation should be 5,000-8 usually, 000 strain; Tobacco M
2Generation is the abundantest generation of variation type, also is the generation of the key selected, M
2The population size in generation should be at 8,000-10,000 strain; M
3Generation is that the tobacco mutant is identified and the generation of continuing to select, should be at M
2Cultivate on the basis of the population size in generation, select elite plant strain.
Tobacco NaN of the present invention
3Mutagenesis idioplasm innovation and breeding technology advantage are: 1) clear and definite tobacco NaN
3The optimal dose of mutagenesis and method for treating seeds have improved mutagenic frequency; 2) the breeding technique method integrates parsimony and practicality, under try one's best little population size and input, and maximized seed selection effect and the efficient of having improved.
Embodiment
Embodiment carries out tobacco NaN according to following steps
3Mutagenesis idioplasm innovation and breed breeding:
The first step, mutant materials are selected: select former cigarette presentation quality and inherent chemical quality better, and anti-multiple diseases, but relatively poor, the pitch of performance tobacco leaf identity is greatly when plant in Guizhou Province, higher your cigarette 11 kind dry seeds of plant height are as mutant materials.Be intended to shorten by mutagenic treatment seed selection pitch, plant height suitably reduces, the mutant that while tobacco leaf identity makes moderate progress.
In second step, preparation NaN3 solution: earlier 4.11ml 0.2M sodium hydrogen phosphate and 15.89 ml0.1M citric acids are mixed, wherein the 0.1M citric acid is with the citric acid 21.01g preparation 1000ml aqueous solution of a water of crystallization; 0.2M sodium hydrogen phosphate is with the 35.01g phosphate dihydrate disodium hydrogen preparation 1000ml aqueous solution; Again with 0.26g NaN
3Be dissolved in citric acid-disodium phosphate soln of 1L pH=3, be mixed with the NaN of 4mmol/L
3Solution.
The 3rd step, seed treatment:, in temperature is 25 ℃ insulating box,, use the NaN of 4mmol/L then with clear water preimpregnation 24 hours with about 10,000 your cigarette 11 dry seeds balance after 7 days in the drier that fills 50% glycerine, one aqueous solution
3Solution was handled 6 hours down at 25 ℃, the seed of handling washes 8 hours in flowing water after, obtained M
1For seed.
The 4th step, M
1The plantation in generation and management: M
1The sowing in generation, grow seedlings, transplanting and field management all carry out according to the suitableeest mode of the Fuquan City, state, the south of Guizhou Province at place, experimental field, Guizhou Province Tabacco Scientific Research Institute.But field period is not pinched, and Transplanting Density is 1450 strain/mus, 1100 strain/mus commonly used, raising 32%.M
1Generation plantation 5000 strains, open pollination after the few grain of strain method sowing, all mixes institute's sowing.
The 5th step, M
2The plantation in generation and selection: M
2In generation, sowed, grow seedlings, transplant in the proving ground, Guizhou Province Tabacco Scientific Research Institute, and its method comprises that later field management all carries out according to the suitableeest method of locality.During transplanting, the capable mutant material of every 3-5 is planted the contrast of 1 row, plants altogether 8,000 strains.The pitch of centering choosing shortens, the M that plant height reduces
2For 18 independent baggings of mutant strain, reserve seed for planting.
The 6th step, M
3Generation and later processing: M
3In generation, sowed, grow seedlings, transplant in the proving ground, Guizhou Province Tabacco Scientific Research Institute, and its method comprises that later field management all carries out according to the suitableeest method of locality.M
2The individual plant kind that generation is selected becomes to plant into respectively 18 strains, every strain 150 strains, and the capable mutant material of every 3-5 is planted the contrast of 1 row, selects with proceeding in order to the degree of stability of investigating mutant character; Choose earlier M
3-1-M
3Totally 7 major cliques such as-7 grades, successively therefrom choose again 12 excellent variation individual plants.After reserving seed for planting, above-mentioned individual plant plant division bagging changes M over to
4Carry out output preliminary experiment and resistance and quality approval test.According to result of the test, from 12 strains, screen 6 best strains, and therefrom select 6 excellent strains and carry out numerous kind.As finding above-mentioned 6 M through the genetic stability test
4Stable for strain, can carry out simultaneously seminal propagation work, be M
5In generation, participate in the major clique rating test seed preparation be provided.Through excellent strain comparative test in 2 years, finding had 1 part of mutant material to have objective trait in the aforementioned stable material, and Comprehensive Traits all surpasses or approaches the material that contrasts.
Claims (5)
1. tobacco NaN
3The technology of mutagenesis idioplasm innovation and breed breeding, its feature comprises:
(1) mutant materials is selected: the selection Comprehensive Traits is better, high-quality is suitable produces, and adaptability is stronger, need to cultivate conventional kind and cenospecies to the excellence that indivedual bad proterties improve;
(2) preparation NaN
3Solution: earlier 4.11ml 0.2M sodium hydrogen phosphate and 15.89ml 0.1M citric acid are mixed, wherein the 0.1M citric acid is with the citric acid 21.01g preparation 1000ml aqueous solution of a water of crystallization; 0.2M sodium hydrogen phosphate is with the 35.01g phosphate dihydrate disodium hydrogen preparation 1000ml aqueous solution; Again with 0.26g NaN
3Be dissolved in citric acid-disodium phosphate soln of 1L pH=3, be mixed with the NaN of 4mmol/L
3Solution;
(3) seed treatment: the dry seed of selected materials with clear water preimpregnation 24h, is used the NaN of 4mmol/L in the drier that fills 50% glycerine, one aqueous solution behind the balance 7d then in temperature is 25 ℃ insulating box
3Solution is handled 6h down at 25 ℃, the seed of handling washes 8h in flowing water after, obtains M
1For seed;
(4) plantation of mutagenesis tobacco, selection and management: with the M of above-mentioned processing
1For the seed pollination of sowing, grow seedlings, transplant, cultivate, bloom, and sowing obtains mutagenesis two generations seed, the pollination of again sowing, grow seedlings, transplant, cultivate, bloom, and sowing obtains seed of future generation, so repeatedly plantation and to second, third, the plant in the 4th generation selects, after strain is stable, all surpasses or approach the material that contrasts for continuous 2 years excellent strain comparative test Comprehensive Traits, can submit regional testing to, enter the variety certification program.
2. technology as claimed in claim 1 is characterized in that the plantation of a mutagenesis generation and management main points are: M
1Should accomplish as far as possible to sow in positive season of ground main cultivation of preliminary election kind for seed, avoid making M because changing sowing time and ecological condition
1For plant change because of conditions such as sunshine and temperature in growth course, making grows normally is affected; M
1The whole ground in generation, sow, grow seedlings, transplanting and field management should carry out according to the suitableeest mode in locality, makes every effort to meticulous, to guarantee the higher survival rate of seedling; A field period bud picking is not pinched; M
1The planting density in generation increases about 50% than the density of local common employing, in order to dwindle M
1The planting scale in generation is saved cost and labour; M
1Open pollination of generation according to the few grain of strain method sowing, and is gathered in the crops the seed of the different fruit branch joint of same strain position, as far as possible to take full advantage of the sudden change chimera; Seed with same kind mutagenic progeny mixes then.
3. technology as claimed in claim 1 is characterized in that the plantation in two generations of mutagenesis and the main points of selection are: M
2In generation, adopted the mode of classifying type miscegenation, the i.e. M of same kind usually
2For mixed seeding, and plant 1 row contrast every 3-5 is capable, in order to observe possible sudden change; M
2In generation, should be planted the positive season sowing in ground at the main of preliminary election kind, and sow, grow seedlings, transplanting and whole field management all adopt the suitableeest mode in locality to carry out; A field period bud picking is not pinched; For take the disease resistance seed selection as purpose M
2In generation, can plant in the sick garden of the evaluation of purpose disease or disease district occurred frequently, and contrast every capable 1 row of planting of 3-5, in order to depress the raising Breeding Efficiency in certain selection; For according to the M that selects in the breeding objective
2For plant, should be at the bagging in squaring period, independent sowing, and by a part strict numbering, put down in writing main feature; Plant for not middle choosing can mix sowing, also can according to circumstances abandon sowing.
4. technology as claimed in claim 1 is characterized in that mutagenesis three generations and later medical record are: M
3In generation, also should be planted the positive season sowing in ground at preliminary election kind main, sows, grows seedlings, transplanting and whole field management all adopt the suitableeest local mode to carry out, and add the contrast of 1 row every the capable mutant material of 3-5; M
2The individual plant kind that generation is selected becomes M
3For strain, every strain 150-200 strain; Be main purpose M for the disease resistance seed selection
2Generation, can continue to plant in corresponding sick garden or disease district occurred frequently, every strain 500-800 strain, and plant 1 row contrast every 3-5 is capable is in order to the degree of stability of investigating mutant character with proceed to select; Should compare with control material during selection, be defined as mutating strain series after, select earlier excellent mutating strain series, therefrom select again excellent sudden change individual plant; The individual plant of selecting from high stability excellent mutating strain series changes M over to after the mixed receipts of plant division
4Carry out the test of output pilot study and resistance and Quality Identification; According to result of the test, M
4Select again best strain in the strain of participating in the experiment in generation, and from best strain, select excellent strain and carry out numerous kind; For sudden change shape unstabilized mutating strain series still, after the roguing at M
4Generation even M
5In generation, still plant by strain, continues to select; For the M that mixes sowing
2For material, its M
3In generation, is still with reference to M
2In generation, carried out miscegenation, the individual plant of middle choosing, its M
4In generation, be similar to M
2M for single sowing
3For processing method, postpone according to this; In the ordinary course of things, the tobacco mutagenic progeny is passed through M
3The selection in generation, most M
4Stable for strain, so M
4In generation, should be carried out seminal propagation work simultaneously except carrying out the output pilot study, be M
5In generation, participate in the major clique rating test seed preparation be provided; For continuous 2 years excellent strain comparative test Comprehensive Traits all above or approach the material of contrast, can submit regional testing to, enter the variety certification program.
5. according to the described technology of claims 1 to 3, it is characterized in that the seed treatment stage, the seed number of processing should be more than 10000; M
1The plantation colony in generation should be the 5000-8000 strain usually; M
2The population size in generation should be in the 8000-10000 strain.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007100779665A CN101133715A (en) | 2007-10-09 | 2007-10-09 | Tobacco NaN3 mutagenesis idioplasm innovation and breed selecting and cultivating technique |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007100779665A CN101133715A (en) | 2007-10-09 | 2007-10-09 | Tobacco NaN3 mutagenesis idioplasm innovation and breed selecting and cultivating technique |
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| Publication Number | Publication Date |
|---|---|
| CN101133715A true CN101133715A (en) | 2008-03-05 |
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|---|---|---|---|
| CNA2007100779665A Pending CN101133715A (en) | 2007-10-09 | 2007-10-09 | Tobacco NaN3 mutagenesis idioplasm innovation and breed selecting and cultivating technique |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102047844A (en) * | 2010-12-13 | 2011-05-11 | 湖北省烟草科研所 | Effective mutagenesis method for tobacco pollen |
| CN104542265A (en) * | 2013-10-25 | 2015-04-29 | 北京林业大学 | Mutation breeding method for cunninghamia lanceolata |
| CN106718864A (en) * | 2017-03-01 | 2017-05-31 | 浙江省萧山棉麻研究所 | A kind of method of the germplasm innovation of tawny daylily |
| CN113575158A (en) * | 2021-08-13 | 2021-11-02 | 泰安市泰山林业科学研究院 | Mutagenesis method of new variety of pyrus betulaefolia variation based on tissue culture system |
-
2007
- 2007-10-09 CN CNA2007100779665A patent/CN101133715A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102047844A (en) * | 2010-12-13 | 2011-05-11 | 湖北省烟草科研所 | Effective mutagenesis method for tobacco pollen |
| CN104542265A (en) * | 2013-10-25 | 2015-04-29 | 北京林业大学 | Mutation breeding method for cunninghamia lanceolata |
| CN106718864A (en) * | 2017-03-01 | 2017-05-31 | 浙江省萧山棉麻研究所 | A kind of method of the germplasm innovation of tawny daylily |
| CN113575158A (en) * | 2021-08-13 | 2021-11-02 | 泰安市泰山林业科学研究院 | Mutagenesis method of new variety of pyrus betulaefolia variation based on tissue culture system |
| CN113575158B (en) * | 2021-08-13 | 2022-07-15 | 泰安市泰山林业科学研究院 | Mutagenesis method of new variation variety of pyrus betulaefolia |
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Open date: 20080305 |