CN102763506A - Method for increasing purity of plant variety - Google Patents
Method for increasing purity of plant variety Download PDFInfo
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- CN102763506A CN102763506A CN2012102140873A CN201210214087A CN102763506A CN 102763506 A CN102763506 A CN 102763506A CN 2012102140873 A CN2012102140873 A CN 2012102140873A CN 201210214087 A CN201210214087 A CN 201210214087A CN 102763506 A CN102763506 A CN 102763506A
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Abstract
The present invention relates to a method for increasing purity of a plant variety. The method comprises step of: providing a plurality of plant seeds, wherein at least a portion of the plant seeds express an acetolactate synthase mutant and have herbicide resistance; and cultivating the seeds to obtain plants with herbicide resistance. Before the seed cultivation or in the cultivation process, at least one of the following treatments is carried out: before cultivation, a herbicide is utilized to treat the seeds, in order to remove seeds without herbicide resistance; and during the cultivation, herbicide is utilized to treat seedlings, so as to remove the seedlings without herbicide resistance. The method can improve and guarantee purity of hybrid varieties in the field and endow the hybrid seeds and seedlings with herbicide resistance activity.
Description
Technical field
The present invention relates to the plant genetics and breeding field.Particularly, the present invention relates to improve the method for plant variety purity method, the method for preparing the plant hybridization seed and raising and guarantee plant field Hybrid purity.
Background technology
Crossbreeding is the effective way of the universally acknowledged comprehensive productivity of raising crops.Crossbreeding can be hybridized strain and kind the different ecological type, that affiliation is far away, and the hybrid seed of generation shows the characteristic that surmounts the parent, i.e. hybrid vigour.In recent decades, crossbreeding becomes the main breeding method of many crops as a kind ofly improving crop yield, improvement crop quality, improving pest-resistant, disease-resistant, the degeneration-resistant means of crop by extensive utilization.
The key of production high-purity hybrid seed is effectively to control crop self-pollination fertilization.At present, the method for control crop self-pollination fertilization comprises the utilization of manual detasseling method, chemical emasculation method and male sterile line.Manual detasseling method is to remove plant spermary official through mechanical or manual operation; The chemical emasculation method is to utilize chemical reagent that the inhibitory action that plant spermary official grows is reached the castration effect; Two kinds of methods all need a large amount of labours; And castration is not thorough, therefore is difficult to effectively control the crop self-pollination, thereby influences hybrid seed purity.And the use of male sterile line not only can be omitted the artificial emasculation step, can also reduce the hybrid seeding cost, is the main path of crop hybrid breeding.
The male sterile line that is used for crossbreeding at present comprises that cytoplasmic sterility and cell nucleus are sterile.Cytoplasmic sterility (being Three-line Hybrid) needs male sterile line (female parent), recovers system's (male parent) and maintenance line (male sterile line is maintained).The cell nucleus male sterile line is by recessiveness or dominant male sterile gene control.Mostly the cell nucleus male sterile line that adopts at present is photo-thermo-sensitive genetic male sterile line; Its fertility receives the temperature and the illumination effect of environment, and promptly under certain temperature or illumination condition, mutant shows as sterile; But the change of temperature or illumination condition can cause fertility restorer, makes male sterile line be able to keep.Because the instability of environmental factor can directly influence the stability of male sterile line, cause male sterile line to produce selfed seed, therefore, in the unsettled time of environmental condition, use photo-thermo-sensitive genetic male sterile line can be difficult to control the purity of hybrid seed, thereby production of hybrid seeds risk is increased.In addition, cytoplasmic male sterile line and cell nucleus male sterile line all can produce false hybrid seed because of the pollen of the non-male parent accepting to drift about in the environment.The generation of false hybrid seed influences seed quality because of reducing hybrid seed purity on the one hand, causes direct economic loss; Reduce grain yield because of hybrid seed purity is low on the other hand, cause indirect economic loss.Therefore, the method for a kind of can effectively removing the " false hybrid seed ", raising plant variety purity is significant to improving grain-production, yet present method still remains to be improved.
Summary of the invention
The present invention is intended to solve at least one of technical problem that exists in the prior art.For this reason, one object of the present invention is to propose a kind of method that improves plant variety purity.
According to an aspect of the present invention, the invention provides a kind of method that improves plant variety purity.According to embodiments of the invention, this method comprises: the seed of a plurality of plants is provided, and at least a portion of these a plurality of seeds is expressed the acetolactate synthestase mutant and is had Herbicid resistant; Seed is cultivated; So that obtain having the plant of Herbicid resistant; Wherein, before seed is cultivated or in the cultivating process, carry out following processing one of at least: before cultivating; Utilize weed killer herbicide that a plurality of seeds are handled, so that remove the seed that does not have Herbicid resistant; With in cultivating process, utilize weed killer herbicide that rice shoot is handled, so that remove the rice shoot that does not have Herbicid resistant.Thus; Can remove " false hybrid seed " effectively; Thereby can improve and ensure the purity of the plant variety of utilizing hybrid seed production; And then can effectively promote crossbreeding and hybrid vigour in the breeding of crops and other plant with produce the application in the for example grain-production, improve grain yield.Wherein, the expression way that is adopted in this article " false hybrid seed ", be meant self-fertility that the male sterile line plant causes because of fertility instability or because of the non-recovery of accepting to drift about in the environment be the hybrid seed that the pollen of plant (male parent) produces.
The inventor finds that the acetolactate synthestase mutant is to weed killer herbicide, and particularly imidazolinone herbicide is insensitive.Therefore; The seed of above-mentioned " express the acetolactate synthestase mutant and have Herbicid resistant " has the imidazolinone herbicide resistance, thus; In the method for raising plant variety purity of the present invention; Can utilize imidazolinone herbicide that seed is handled before seed is cultivated or in the cultivating process, to remove seed or the rice shoot that does not have Herbicid resistant.According to embodiments of the invention, in the method for raising plant variety purity of the present invention, weed killer herbicide is an imidazolinone herbicide, and the acetolactate synthestase mutant has the amino acid sequence shown in any that is selected from SEQ ID NO:1-8.Further, according to embodiments of the invention, in the method for raising plant variety purity of the present invention, the acetolactate synthestase mutant is by the nucleic acid coding with the nucleotide sequence shown in any that is selected from SEQ ID NO:9-16.Wherein, employed in this article term " nucleic acid " can be any at least a polymer that comprises ribonucleotide and deoxyribonucleotide, include but not limited to through modification or without the nucleic acid of modifying.
According to embodiments of the invention; In the method for raising plant variety purity of the present invention; The seed that a plurality of plants are provided is that to be plant with the male sterile line plant that does not have Herbicid resistant hybridize realizes through the recovery that will have Herbicid resistant; Wherein, to be plant obtain one of at least through following in recovery:
(a) making recovery is that plant comprises the nucleic acid with the nucleotide sequence shown in any that is selected from SEQ ID NO:9-16;
(b) making recovery is that plant is expressed the protein with the amino acid sequence shown in any that is selected from SEQ ID NO:1-8.
Wherein, Protein with the amino acid sequence shown in any that is selected from SEQ ID NO:1-8 is foregoing acetolactate synthestase mutant; And as previously mentioned, it is coded by the nucleic acid with the nucleotide sequence shown in any that is selected from SEQ IDNO:9-16.
Particularly, according to embodiments of the invention, in the method for raising plant variety purity of the present invention, recovery is that plant is through hybridizing, backcrossing and vegetative at least a acquisition.Thus; Can make recovery effectively is that plant comprises the nucleic acid with the nucleotide sequence shown in any that is selected from SEQ ID NO:9-16; Or expression has the protein of the amino acid sequence shown in any that is selected from SEQ ID NO:1-8, promptly foregoing acetolactate synthestase mutant.
According to embodiments of the invention, in the method for raising plant variety purity of the present invention, may further include recovering is that plant is identified.Those skilled in the art will appreciate that to recovering be that the method that plant is identified does not receive special restriction.According to concrete examples more of the present invention, in the method for raising plant variety purity of the present invention, evaluation may further include: measure recovery is whether plant comprises the nucleic acid with the nucleotide sequence shown in any that is selected from SEQ ID NO:9-16; And/or measure recovery is whether plant expresses the acetolactate synthestase mutant.
According to embodiments of the invention; In the method for raising plant variety purity of the present invention; Before cultivating; Utilize weed killer herbicide that a plurality of seeds are handled, may further include so that remove the seed that does not have Herbicid resistant: utilize at least a of imidazolinone herbicide of following concentration range that hybrid seed is carried out immersion treatment: the weed eradication cigarette of 0.1-2.5 weight %; 0.012-0.2 the Imazethapyr of weight %; 0.012-0.2 hundred ridges of weight % are logical; 0.016-0.2 the weed eradication quinoline of weight %.Thus, seed can be removed effectively, thereby the purity of the plant variety of being planted can be effectively improved with Herbicid resistant.
According to embodiments of the invention; In the method for raising plant variety purity of the present invention; In cultivating process, utilize weed killer herbicide that rice shoot is handled, further do not comprise so that remove the rice shoot do not have Herbicid resistant: the imidazolinone herbicide that utilizes following concentration range is with the processing of spraying of hybrid seed rice shoot; Wherein, rice shoot is preferably the two leaf stage rice shoot: the weed eradication cigarette of 0.1-2.5 weight %; 0.012-0.2 the Imazethapyr of weight %; 0.012-0.2 hundred ridges of weight % are logical; 0.016-0.2 the weed eradication quinoline of weight %.Thus, rice shoot can be effectively removed, thereby the purity of the plant variety of being planted can be effectively improved with Herbicid resistant.
According to embodiments of the invention, the floristics that the method for raising plant variety purity of the present invention can be used does not receive special restriction.According to concrete examples more of the present invention, the plant that method of the present invention can be used can be preferably at least a of paddy rice, wheat, rape, corn, cotton, millet and clover for being selected from least a of crops and herbage.
According to a further aspect in the invention, the present invention also provides a kind of method for preparing the plant hybridization seed.According to embodiments of the invention; This method comprises: the recovery that will have Herbicid resistant is that plant hybridizes with the male sterile line plant that does not have Herbicid resistant; So that obtain the plant hybridization seed; Wherein, recovery is that plant is expressed the acetolactate synthestase mutant, and the male sterile line plant is not expressed said acetolactate synthestase mutant.According to embodiments of the invention; Utilize this method can prepare hybrid seed efficiently, further utilize weed killer herbicide that hybrid seed is handled, can remove not " the false hybrid seed " of antiweed effectively; Keep hybrid seed with Herbicid resistant; Thereby can greatly improve the purity of hybrid seed, and then can effectively promote crossbreeding and hybrid vigour in the breeding of crops and other plant with produce the application in the for example grain-production, improve grain-production power.The inventor is surprised to find, the plant hybridization seed that utilizes the method for preparing the plant hybridization seed of the present invention to prepare, and purity is high, quality is very good, and has the imidazolinone herbicide resistance.
According to embodiments of the invention, the floristics that the method for preparing the plant hybridization seed of the present invention can be used does not receive special restriction, recovers system and male sterile line cross-breeding as long as can utilize.According to concrete examples more of the present invention, what the plant that method of the present invention can be used can be for crops and herbage is at least a, is preferably at least a of paddy rice, wheat, rape, corn, cotton, millet and clover.Thus, can be effectively to paddy rice, wheat, corn and rape cross-breeding, and the hybrid seed quality that obtains is good, purity is high.
According to embodiments of the invention, in the method for preparing the plant hybridization seed of the present invention, weed killer herbicide is an imidazolinone herbicide, and the acetolactate synthestase mutant has the amino acid sequence shown in any that is selected from SEQ ID NO:1-8.Further, according to embodiments of the invention, in the method for preparing the plant hybridization seed of the present invention, the acetolactate synthestase mutant is by the nucleic acid coding with the nucleotide sequence shown in any that is selected from SEQ ID NO:9-16.Wherein, employed in this article term " nucleic acid " can be any at least a polymer that comprises ribonucleotide and deoxyribonucleotide, include but not limited to through modification or without the nucleic acid of modifying.
According to embodiments of the invention, in the method for preparing the plant hybridization seed of the present invention, to be plant obtain one of at least through following in recovery:
(a) making recovery is that plant comprises the nucleic acid with the nucleotide sequence shown in any that is selected from SEQ ID NO:9-16;
(b) making recovery is that plant is expressed the protein with the amino acid sequence shown in any that is selected from SEQ ID NO:1-8.
Wherein, Protein with the amino acid sequence shown in any that is selected from SEQ ID NO:1-8 is foregoing acetolactate synthestase mutant; And as previously mentioned, it is coded by the nucleic acid with the nucleotide sequence shown in any that is selected from SEQ ID NO:9-16.
Particularly, according to embodiments of the invention, in the method for preparing the plant hybridization seed of the present invention, recovery be plant through hybridization, backcross and vegetative at least a acquisition.Thus; Can make recovery effectively is that plant comprises the nucleic acid with the nucleotide sequence shown in any that is selected from SEQ ID NO:9-16; Or expression has the protein of the amino acid sequence shown in any that is selected from SEQ ID NO:1-8, promptly foregoing acetolactate synthestase mutant.
According to embodiments of the invention, in the method for preparing the plant hybridization seed of the present invention, may further include recovering is that plant is identified.Those skilled in the art will appreciate that to recovering be that the method that plant is identified does not receive special restriction.According to concrete examples more of the present invention, in the method for raising plant variety purity of the present invention, evaluation may further include: measure recovery is whether plant comprises the nucleic acid with the nucleotide sequence shown in any that is selected from SEQ ID NO:9-16; And/or measure recovery is whether plant expresses the acetolactate synthestase mutant.
According to a further aspect in the invention, the present invention also provides a kind of method that improves and ensure plant field Hybrid purity.According to embodiments of the invention, this method comprises: according to the method for preparing the plant hybridization seed of the present invention, obtain the plant hybridization seed; And said seed cultivated; So that obtain having the plant of Herbicid resistant; Before seed is cultivated or in the cultivating process; Carry out following processing one of at least: before cultivating, utilize weed killer herbicide that a plurality of seeds are handled, so that remove the seed that does not have Herbicid resistant; With in cultivating process, utilize weed killer herbicide that rice shoot is handled, so that remove the rice shoot that does not have Herbicid resistant.Thus; Can remove " false hybrid seed " effectively; Thereby can improve and ensure the purity of plant field Hybrid, and then can effectively promote crossbreeding and hybrid vigour in the breeding of crops and other plant with produce the application in the for example grain-production, improve grain yield.
According to embodiments of the invention; In the method for raising of the present invention and guarantee plant field Hybrid purity; Before cultivating; Utilize weed killer herbicide that a plurality of seeds are handled, may further include so that remove the seed that does not have Herbicid resistant: utilize at least a of imidazolinone herbicide of following concentration range that hybrid seed is carried out immersion treatment: the weed eradication cigarette of 0.1-2.5 weight %; 0.012-0.2 the Imazethapyr of weight %; 0.012-0.2 hundred ridges of weight % are logical; 0.016-0.2 the weed eradication quinoline of weight %.Thus, seed can be removed effectively, thereby plant field Hybrid purity can be effectively improved and ensure with Herbicid resistant.
According to embodiments of the invention; In the method for raising of the present invention and guarantee plant field Hybrid purity; In cultivating process, utilize weed killer herbicide that rice shoot is handled, further do not comprise so that remove the rice shoot do not have Herbicid resistant: the imidazolinone herbicide that utilizes following concentration range is with the processing of spraying of hybrid seed rice shoot; Wherein, rice shoot is preferably the two leaf stage rice shoot: the weed eradication cigarette of 0.1-2.5 weight %; 0.012-0.2 the Imazethapyr of weight %; 0.012-0.2 hundred ridges of weight % are logical; 0.016-0.2 the weed eradication quinoline of weight %.Thus, rice shoot can be effectively removed, thereby plant field Hybrid purity can be effectively improved and ensure with Herbicid resistant.
According to embodiments of the invention, the floristics that the method for raising of the present invention and guarantee plant field Hybrid purity can be used does not receive special restriction.According to concrete examples more of the present invention, the plant that method of the present invention can be used can be preferably at least a of paddy rice, wheat, rape, corn, cotton, millet and clover for being selected from least a of crops and herbage.
Need to prove, the method for raising plant variety purity of the present invention, the present inventor accomplishes through the arduous creative work and the work of optimization.
Additional aspect of the present invention and advantage part in the following description provide, and part will become obviously from the following description, or recognize through practice of the present invention.
Description of drawings
Above-mentioned and/or additional aspect of the present invention and advantage obviously with are easily understood becoming the description of embodiment from combining figs, wherein:
Fig. 1 has shown according to one embodiment of the invention, No. 4 germination after 0.036 weight % Imazethapyr is handled of common peaceful spring;
Fig. 2 has shown according to one embodiment of the invention, the germination of No. 4 wheat seeds of peaceful spring of the antiweed that isozygotys after 0.036 weight % Imazethapyr is handled; And
Fig. 3 shown according to one embodiment of the invention, the germination of seed after the processing of 0.036 weight % Imazethapyr that obtains after the filial generation selfing of No. 4 wheat seeds of peaceful spring of the antiweed that isozygotys and common peaceful spring No. 4 wheat seeds of antiweed not.
Embodiment
Describe embodiments of the invention below in detail, the example of said embodiment is shown in the drawings, and wherein identical from start to finish or similar label is represented identical or similar elements or the element with identical or similar functions.Be exemplary through the embodiment that is described with reference to the drawings below, only be used to explain the present invention, and can not be interpreted as limitation of the present invention.
Embodiment 1: the anti-imidazolinone herbicide mutant heredity of paddy rice compartment analysis
At first; (dna sequence analysis shows that acetolactate synthestase (ALS) gene of this plant exists with the heterozygote state to make yellow China of the anti-weed eradication cigarette of strain aqua weed killer herbicide account for the rice mutant plant; The ALS mutant gene is undergone mutation in the 1642nd and 1643 sites; Become AT by TG, cause the 548th of corresponding encoded albumen to become methionine, promptly with respect to the wild type acetolactate synthestase by tryptophan; Its acetolactate synthestase has the Trp548Cys sudden change) self-fertility, so that obtain selfed seed.Then, utilize weed eradication cigarette aqua (available from Shandong Cynda Chemical Co., Ltd) that the selfed seed of above-mentioned acquisition is steeped kind of experiment and rice shoot sprinkling experiment respectively, concrete steps are following:
1. bubble kind of experiment
Adopt weed eradication cigarette aqua (0.12 weight %, 0.36 weight % respectively; 0.6 weight %; 2.5 weight %); Steep kind of an experiment according to following steps: the selfed seed of above-mentioned acquisition is soaked 24 hours in weed eradication cigarette aqua after, place in 32 ℃ of incubators and carry out vernalization, after 24 hours; In 25 ℃ of incubators, sprout, observe budlet after 5 days and sprout and growing state.
Add up the budlet length of each seed, the result shows that the seed budlet length of antiweed reaches 1.5-2cm, and the seed budlet length of antiweed does not stop at about 0.2cm.Then, calculate the resistance individuality of each concentration weed eradication cigarette aqua processing and the number of non-resistance individuality, and resistance and the non-resistance ratio that separates, the result sees the following form 1.The data show of table 1; Self-fruitful seed shows as resistance to the weed killer herbicide in the experimental concentration scope: non-resistance separation ratio is near 3:1; The antiweed The Characters is a dominant inheritance; Accord with expectation show and can distinguish resistance seed and non-resistance seed through seed being carried out the weed killer herbicide immersion treatment, and the weed killer herbicide concentration range can be 0.12-2.5%.
Random choose 30 strain phenotypes are the budlet difference extracting DNA of resistance, and utilize high-resolution solubility curve (HRM) method to detect its genotype.Testing result shows that wherein 12 strains are the homozygous mutation body, and 18 strains are heterozygote, and homozygote: heterozygote is near 1:2.This result shows that further resistance seed (comprising homozygote and heterozygote) and non-resistance seed can be distinguished through the weed killer herbicide immersion treatment, and non-resistance seed can utilize the weed killer herbicide immersion treatment that it is removed to herbicide sensitive.
2. rice shoot sprays experiment
The weed eradication cigarette aqua of picked at random 0.36 weight % carries out the field rice shoot to above-mentioned selfed seed and sprays experiment, particularly, and to processings of spraying of 2 leaf phase seedlings, wherein, 87 milliliters of every square metre of sprayings, 2-3 week of back handled in spraying, adds up the rice shoot survival.The result shows that the growth of 62 strain seedlings is normal, and 20 strain seedling flavescence are also withered gradually; Show 62 strain seedling tool Herbicid resistants; 20 strain seedlings are not had a Herbicid resistant, and seedling is to the resistance of weed killer herbicide: non-resistance separation is than near 3:1 (as shown in table 1 below), the dominant inheritance of accord with expectation.Above result shows, can carry out herbicide sprays through the rice shoot to seed and handle and distinguish resistance seed and non-resistance seed, thus, utilizes the antiweed characteristic can ensure the purity of crop hybrid seed in the use of field.
With 30 strain phenotypes is the rice shoot difference extracting DNA of resistance, and utilizes high-resolution solubility curve (HRM) method to detect its genotype.Testing result shows that wherein 11 strains are the homozygous mutation body, and 19 strains are heterozygote, and homozygote: heterozygote is near 1:2.This result further shows, can handle through herbicide sprays and remove non-resistance seedling.
Table 1 weed eradication cigarette aqua soaks the self-fertility seed or sprays the two leaf stage rice shoot
| Weed killer herbicide concentration | Resistance is individual | Non-resistance individuality | Separate ratio |
| a1x | 62 | 18 | 3.44 |
| a3x | 61 | 19 | 3.21 |
Annotate:
aInstitute's weed eradication cigarette aqua that refers to soaks the experiment of self-fertility seed;
bInstitute's weed eradication cigarette aqua that refers to sprays the experiment of two leaf stage rice shoot.
Embodiment 2: utilize imidazolinone herbicide to carry out kind of the experiment of rice paddy seed bubble
Imidazolinone herbicide at Chinese production and selling comprises imidazoles nicotinic acid (weed eradication cigarette) at present, AC 263222 (hundred ridges are logical), imazethapyr (Imazethapyr), imazaquin (weed eradication quinoline).Utilize above-mentioned herbicide solution that embodiment 1 described paddy rice heterozygote self-fertility seed was carried out immersion treatment 24 hours; Take out seed; And in 32 ℃ of incubators, carried out vernalization 24 hours, and in 25 ℃ of incubators, sprout then, observe budlet after 5 days and sprout and growing state.
Add up the budlet length of each seed, the result shows that the budlet length of antiweed reaches 1.5-2cm, and the seed budlet length of antiweed does not stop at about 0.1-0.2cm.Then, the number of resistance individuality and non-resistance individuality in the seed of calculating after each weed killer herbicide immersion treatment, and resistance and the non-resistance ratio that separates, the result sees the following form 2.The data show of table 2; Self-fruitful seed all shows as resistance to these weed killer herbicides: non-resistance separation ratio is near 3:1; Show that these weed killer herbicides all can be used for immersion treatment and remove non-resistance seed; Thus, utilize the antiweed characteristic can ensure the purity of crop hybrid seed in the use of field.
Four kinds of imidazolinone herbicides of table 2 soak the self-fertility seed
| Weed killer herbicide | Resistance is individual | Non-resistance individuality | Separate ratio |
| 0.36 weight % weed eradication cigarette | 58 | 22 | 2.64 |
| 0.036% weight, hundred ridges are logical | 61 | 21 | 2.90 |
Embodiment 3: utilize imidazolinone herbicide to carry out rice seedling and spray experiment
Utilize respectively that weed eradication cigarette, hundred ridges are logical, Imazethapyr and weed eradication quinoline carry out the field rice shoot to embodiment 1 described paddy rice heterozygote self-fertility seed and spray experiment; Particularly; To the processing of spraying of 2 leaf phase seedlings, 87 milliliters of every square metre of sprayings, 2-3 week of back handled in spraying; Statistics rice shoot survival, the result sees the following form 3.Can be known that by table 3 rice shoot is to the resistance of various weed killer herbicides: non-resistance separation is than all near 3:1, the dominant inheritance of accord with expectation.This result further shows, can handle through herbicide sprays and remove non-resistance seedling, thus, utilizes the antiweed characteristic can ensure the purity of crop hybrid seed in the use of field.
Four kinds of imidazolinone herbicides of table 3 spray the two leaf stage rice shoot
| Weed killer herbicide | Resistance is individual | Non-resistance individuality | Separate ratio |
| 0.36 weight % weed eradication cigarette | 58 | 18 | 3.22 |
| 0.036 weight % hundred ridges are logical | 61 | 21 | 2.90 |
| 0.036 weight % Imazethapyr | 59 | 20 | 2.95 |
| 0.048 weight % weed eradication quinoline | 57 | 21 | 2.71 |
Embodiment 4: soak the wheat seed experiment with imidazolinone herbicide
Utilize 0.036 weight % Imazethapyr; Respectively to No. 4 (not antiweed) wheat seeds of common peaceful spring, No. 4 wheat seeds of peaceful spring of homozygote antiweed, and the seed that obtains after the filial generation selfing of No. 4 wheat seeds of peaceful spring of the antiweed that isozygotys and No. 4 (not antiweed) wheat seeds of common peaceful spring carries out the weed killer herbicide immersion treatment and is not with the Herbicid resistant seed germination to investigate whether can stop through herbicide treatment.Take out seed after 24 hours, and in 25 ℃ of incubators, carried out vernalization 24 hours, in 25 ℃ of incubators, sprout then, observe budlet after 5 days and sprout and growing state, and take a picture, the result sees Fig. 1-Fig. 3.As shown in the figure; After 0.036 weight % Imazethapyr immersion treatment; The sprouting and the growth of seedling of No. 4 (not antiweed) wheat seeds of common peaceful spring all receive obvious inhibition (Fig. 1); But the sprouting of homozygote antiweed wheat seed and growth of seedling normal (Fig. 2), and the seed that obtains after the filial generation selfing of No. 4 wheat seeds of peaceful spring of the antiweed that isozygotys and No. 4 (not antiweed) wheat seeds of common peaceful spring are because of the seed of existing antiweed; The not seed of antiweed is also arranged; Then the sprouting and the growth of seedling of a part of seed are normal after 0.036 weight % Imazethapyr is handled, and another part is suppressed, and the about 3:1 of seed rate (Fig. 3) normal and that be suppressed grows.Thus, further show, can distinguish resistance seed and non-resistance seed through seed being carried out the weed killer herbicide immersion treatment.
In the description of this specification, the description of reference term " embodiment ", " some embodiment ", " example ", " concrete example " or " some examples " etc. means the concrete characteristic, structure, material or the characteristics that combine this embodiment or example to describe and is contained at least one embodiment of the present invention or the example.In this manual, the schematic statement to above-mentioned term not necessarily refers to identical embodiment or example.And concrete characteristic, structure, material or the characteristics of description can combine with suitable manner in any one or more embodiment or example.
Although illustrated and described embodiments of the invention; Those having ordinary skill in the art will appreciate that: under the situation that does not break away from principle of the present invention and aim, can carry out multiple variation, modification, replacement and modification to these embodiment, scope of the present invention is limited claim and equivalent thereof.
| a5x | 59 | 21 | 2.80 |
| a10x | 58 | 22 | 2.64 |
| b3x | 62 | 20 | 3.1 |
| 0.036% weight Imazethapyr | 59 | 20 | 2.95 |
| 0.048% weight weed eradication quinoline | 57 | 21 | 2.71 |
Claims (10)
1. a method that improves plant variety purity is characterized in that, comprising:
The seed of a plurality of said plants is provided, and at least a portion of said a plurality of seeds is expressed the acetolactate synthestase mutant and is had Herbicid resistant;
Said seed is cultivated, so that obtain having the plant of Herbicid resistant,
Wherein,
Before seed is cultivated or in the cultivating process, carry out following processing one of at least:
Before cultivating, utilize weed killer herbicide that said a plurality of seeds are handled, so that remove the seed that does not have Herbicid resistant; With
In cultivating process, utilize weed killer herbicide that rice shoot is handled, so that remove the rice shoot that does not have Herbicid resistant.
2. method according to claim 1 is characterized in that, said weed killer herbicide is an imidazolinone herbicide, and said acetolactate synthestase mutant has the amino acid sequence shown in any that is selected from SEQ ID NO:1-8,
Randomly, said acetolactate synthestase mutant is by the nucleic acid coding with the nucleotide sequence shown in any that is selected from SEQ ID NO:9-16.
3. method according to claim 2 is characterized in that, the seed that a plurality of said plants are provided is that to be plant with the male sterile line plant that does not have Herbicid resistant hybridize realizes through the recovery that will have Herbicid resistant,
Wherein,
To be plant obtain one of at least through following in said recovery:
(a) making said recovery is that plant comprises the nucleic acid with the nucleotide sequence shown in any that is selected from SEQ ID NO:9-16;
(b) making said recovery is that plant is expressed the protein with the amino acid sequence shown in any that is selected from SEQ ID NO:1-8.
4. method according to claim 3 is characterized in that, said recovery is plant through hybridization, backcrosses and vegetative at least a acquisition,
Randomly, further comprise to said recovery being that plant is identified,
Randomly, said evaluation further comprises:
Measuring said recovery is whether plant comprises the nucleic acid with the nucleotide sequence shown in any that is selected from SEQ ID NO:9-16; And/or
Measuring said recovery is whether plant expresses said acetolactate synthestase mutant.
5. method according to claim 1 is characterized in that, before cultivating, utilizes weed killer herbicide that said a plurality of seeds are handled, and further comprises so that remove the seed that does not have Herbicid resistant:
Utilize at least a of imidazolinone herbicide of following concentration range that said hybrid seed is carried out immersion treatment:
0.1-2.5 the weed eradication cigarette of weight %; 0.012-0.2 the Imazethapyr of weight %; 0.012-0.2 hundred ridges of weight % are logical; 0.016-0.2 the weed eradication quinoline of weight %,
Randomly, in cultivating process, utilize weed killer herbicide that rice shoot is handled, further comprise so that remove the rice shoot that does not have Herbicid resistant:
The imidazolinone herbicide that utilizes following concentration range is with the processing of spraying of said hybrid seed rice shoot, and wherein, said rice shoot is preferably the two leaf stage rice shoot:
0.1-2.5 the weed eradication cigarette of weight %; 0.012-0.2 the Imazethapyr of weight %; 0.012-0.2 hundred ridges of weight % are logical; 0.016-0.2 the weed eradication quinoline of weight %.
6. method according to claim 1 is characterized in that said plant is be selected from crops and herbage at least a, is preferably at least a of paddy rice, wheat, rape, corn, cotton, millet and clover.
7. a method for preparing the plant hybridization seed is characterized in that, comprising:
The recovery that will have Herbicid resistant is that plant hybridizes with the male sterile line plant that does not have Herbicid resistant, so that obtain said plant hybridization seed,
Wherein, said recovery is that plant is expressed the acetolactate synthestase mutant, and said male sterile line plant is not expressed said acetolactate synthestase mutant,
Randomly, said plant is at least a of crops and herbage, is preferably at least a of paddy rice, wheat, rape, corn, cotton, millet and clover.
8. method according to claim 7 is characterized in that, said weed killer herbicide is for being selected from imidazolinone herbicide, and said acetolactate synthestase mutant has the amino acid sequence shown in any that is selected from SEQ ID NO:1-8,
Randomly, said acetolactate synthestase mutant is by the nucleic acid coding with the nucleotide sequence shown in any that is selected from SEQ ID NO:9-16,
Randomly, to be plant obtain one of at least through following in said recovery:
(a) making said recovery is that plant comprises the nucleic acid with the nucleotide sequence shown in any that is selected from SEQ ID NO:9-16;
(b) making said recovery is that plant is expressed the protein with the amino acid sequence shown in any that is selected from SEQ ID NO:1-8,
Randomly, said recovery is plant through hybridization, backcrosses and vegetative at least a acquisition,
Randomly, further comprise to said recovery being that plant is identified,
Randomly, said evaluation further comprises:
Measuring said recovery is whether plant comprises the nucleic acid with the nucleotide sequence shown in any that is selected from SEQ ID NO:9-16; And/or
Measuring said recovery is whether plant expresses said acetolactate synthestase mutant.
9. a method that improves and ensure plant field Hybrid purity is characterized in that, comprising:
According to claim 7 or 8 described methods, obtain the plant hybridization seed;
And
Said seed is cultivated, so that obtain having the plant of Herbicid resistant,
Before seed is cultivated or in the cultivating process, carry out following processing one of at least:
Before cultivating, utilize weed killer herbicide that said a plurality of seeds are handled, so that remove the seed that does not have Herbicid resistant; With
In cultivating process, utilize weed killer herbicide that rice shoot is handled, so that remove the rice shoot that does not have Herbicid resistant.
10. method according to claim 9 is characterized in that, before cultivating, utilizes weed killer herbicide that said a plurality of seeds are handled, and further comprises so that remove the seed that does not have Herbicid resistant:
Utilize at least a of imidazolinone herbicide of following concentration range that said hybrid seed is carried out immersion treatment:
0.1-2.5 the weed eradication cigarette of weight %; 0.012-0.2 the Imazethapyr of weight %; 0.012-0.2 hundred ridges of weight % are logical; 0.016-0.2 the weed eradication quinoline of weight %;
Randomly, in cultivating process, utilize weed killer herbicide that rice shoot is handled, further comprise so that remove the rice shoot that does not have Herbicid resistant:
The imidazolinone herbicide that utilizes following concentration range is with the processing of spraying of said hybrid seed rice shoot, and wherein, said rice shoot is preferably the two leaf stage rice shoot:
0.1-2.5 the weed eradication cigarette of weight %; 0.012-0.2 the Imazethapyr of weight %; 0.012-0.2 hundred ridges of weight % are logical; 0.016-0.2 the weed eradication quinoline of weight %,
Randomly, said plant is at least a of crops and herbage, is preferably at least a of paddy rice, wheat, rape, corn, cotton, millet and clover.
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| CN105755024A (en) * | 2016-04-12 | 2016-07-13 | 江苏省农业科学院 | ALS mutation gene as well as protein and application thereof |
| CN110291986A (en) * | 2017-09-28 | 2019-10-01 | 彭基明 | Rice paddy seed product containing anti-herbicide gene of field planting and preparation method thereof |
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