CN103222423A - Rice hybrid breeding material as well as cultivation method and application thereof - Google Patents
Rice hybrid breeding material as well as cultivation method and application thereof Download PDFInfo
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- CN103222423A CN103222423A CN2013101781526A CN201310178152A CN103222423A CN 103222423 A CN103222423 A CN 103222423A CN 2013101781526 A CN2013101781526 A CN 2013101781526A CN 201310178152 A CN201310178152 A CN 201310178152A CN 103222423 A CN103222423 A CN 103222423A
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Abstract
The invention discloses a rice hybrid breeding material as well as a cultivation method and application thereof. A female parent material with a herbicide sensitive gene and a male parent material with the herbicide sensitive gene are obtained by specific promoter drive and antisense inhibit expression methods; the herbicide sensitive gene is just expressed in stamen; and the herbicide sensitive gene is just expressed in pistil; the female parent material and the male parent material are mixed in application; mechanized cropping is carried out; the female parent emasculation and male parent female removal are achieved by spraying the herbicide before heading; and the high-quality F1 hybrid seed is produced by one-way mating between parents. By adopting the method disclosed by the invention, a sterile line and a restorer line can be freely obtained; high-purity hybrid is obtained; mixing of the self-fruitfulness seed is effectively removed; especially, serious decline of hybrid purity caused by fertility recovery of the photo-thermo-sensitive sterile line under an adverse light temperature condition is avoided; the breeding process of the parents of the hybrid rice is simplified; manpower and material resources are saved; and the production efficiency is improved.
Description
Technical field
The present invention relates to paddy rice production of hybrid seeds field, specifically a kind of paddy rice cross breeding breeding material and breeding method and application.
Background technology
Hybrid rice is that China and even global grain security have been made huge contribution.But the existing rice heterosis utilization and the production of hybrid seeds, combo are not free, and many, the complex procedures of seed production link, too much depend on intensive human and material resources, and production efficiency is low.Particularly in recent years, along with China's economic development, the rural area person between twenty and fifty go down town and work, rural laborer's shortage, and hybrid rice seed production faces serious challenge.Realize the hybrid rice mechanization mixed seeding production of hybrid seeds,, improve seed production quality and efficient, promote further applying of hybrid rice reducing the hybrid rice breeding cost, all significant.
Plant tissue Idiotype promotor is the specific dna fragmentation of a class, has the ability that gene is expressed in particular organization, cell that drives.According to the difference of expression characterization, organizing specific type promotor can be divided into types such as root-specific promoter, chlorenchyma specific promoter, stem specific promoter, stamen (pollen) specific promoter, gynoecium specific promoter.Good organizing specific promotor only has driving force in specific tissue or cell, and does not have driving force in other cells.Therefore, normal using-system specific promoter is finished the genetic manipulation in the specific cells and can the position beyond the target cell not exerted an influence in plant genetic engineering.Present research has had been found that some can be for the rice tissue specific promoter of gene engineering use, concerning the process that reproductive development is correlated with, comprise stamen (pollen) specific expression promoters such as Osg6B promotor, OsMADS58 promotor, OsACE1 promotor, RTS promotor.Gynoecium (column cap) the specific promoter research that derives from paddy rice is less, does not also have the report of using.But the clone obtains in tobacco, rape and potato gynoecium specific promoter such as STS14 promotor, SLG promotor, SRK promotor and SLR1 promotor still have the characteristics of gynoecium specifically expressing in paddy rice.The intensity of these female/stamen specific promoters also is enough to drive the herbicide sensitive that the artificial DNA fragment causes privileged site.
Cyhalofop-butyl is the insensitive weed killer herbicide of a kind of paddy rice, has another name called a thousand pieces of gold or cyanogen fluorine diclofop-methyl, is mainly used in the control grassy weed.Cyhalofop-butyl is interior suction conductivity weed killer herbicide, by blade and the leaf sheath absorption of plant, phloem conduction, resistance acetyl-CoA carboxylase activity is disturbed and is stopped fatty acid metabolism and synthesizes, and suppresses the plant cell division growth, destroy fat structures such as cell membrane system, finally cause cell death.Use cyhalofop-butyl and have fewer environmental impacts, zoopery shows no overt toxicity, is fine herbicide commonly used in the Rice Production.
The problem that exists the impurity of seeds of this self-fertility of father and mother to cause purity of hybrid to descend in the tradition production of hybrid seeds technology, as two being photo-thermo-sensitive genetic male sterile line fertility restorer under disadvantageous light and temperature condition in the production of hybrid seeds, self-fertility has a strong impact on purity of hybrid.
Summary of the invention
For overcoming the prior art deficiency, the object of the invention is to provide a kind of paddy rice cross breeding breeding material and breeding method and application, is intended to solve breeding cost height in the present production, and complex steps, purity are difficult to problems such as assurance.
One of purpose of the present invention provides a kind of paddy rice cross breeding breeding material, and the maternal material (male sterile line) in this material is for having herbicide sensitive gene, and herbicide sensitive gene is only expressed in stamen; Male parent material (recover system) is for having herbicide sensitive gene, and herbicide sensitive gene is only expressed in gynoecium.
Crossbreeding material of the present invention is to use the means that specific promoter drives and Antisense Suppression is expressed to obtain, and concrete breeding method is realized in the following way:
The herbicide resistance gene antisense fragment that the stamen specific promoter is driven changes the maternal kind of paddy rice over to, special the expressing at the stamen position of herbicide resistance gene is suppressed, realize that the stamen specific promoter drives the purpose of herbicide sensitive gene, the maternal material of stamen part to herbicide sensitive only arranged;
The herbicide resistance gene antisense fragment that the gynoecium specific promoter is driven changes paddy rice male parent kind over to, special the expressing at the gynoecium position of herbicide resistance gene is suppressed, realize that the gynoecium specific promoter drives the purpose of herbicide sensitive gene, obtain only to have the male parent material of gynoecium part herbicide sensitive.
Wherein herbicide sensitive gene is other acceptable sensitive gene in any one or the crop breeding in cyhalofop-butyl sensitive gene, the Bentazon sensitive gene etc.; The promotor that drives herbicide sensitive gene is any one or other the special promotor of acceptable floral organ in STS14 gynoecium specific expression promoter, the Osg6B stamen specific expression promoter.
The maternal kind of wherein said paddy rice (male sterile line) is acceptable other kind on kind NS629S or the thremmatology, and male parent kind (recovering system) is acceptable other kind on kind RH003 or the thremmatology.
Another object of the present invention provides a kind of hybrid rice mixed seeding producing method for seed.
This producing method for seed is realized in the following way:
That (1) cultivates as stated above has a herbicide sensitive gene, and the herbicide sensitive gene maternal material of only expressing in stamen and have herbicide sensitive gene, and the male parent material that herbicide sensitive gene is only expressed in gynoecium carries out mixed seeding; Herbicide spraying before heading, thus realize maternal the castration, male parent is gone female, and the seed that obtains is crossbreed, and it is feasible that mechanized harvest is become;
(2) at isolation field piece separately plantation above-mentioned maternal material (male sterile line) or male parent material (recovering system), under spraying herbicide condition not, breeding maternal (male sterile line) or male parent (recovering system).
Wherein weed killer herbicide is corresponding cyhalofop-butyl, Bentazon, pyrazosulfuron etc.
Compared with prior art, the invention has the advantages that:
1, helps freely matching of father and mother's basis.
2, by herbicide spraying, realize maternal the castration automatically, male parent is gone female, has alleviated workload greatly, has improved production efficiency.
3, having avoided mixing of this self-fertility of father and mother seed effectively, particularly is to have prevented that the purity of hybrid that maternal self-fertility causes from descending, and significantly improves purity of hybrid.
4, maternal (male sterile line) breeding is simple, only needs can obtain by selfing, need not use maintenance line, thereby simplify reproductive process, has reduced production cost.
5, production procedure is simple, is easy to control, because herbicide spraying stepfather female parent all can not self-fertility, all offsprings are crossbreed, therefore are convenient to mechanized operation, realize the hybrid rice mixed seeding mechanization production of hybrid seeds, enhance productivity, reduce breeding cost, save human and material resources.
Description of drawings
This cultivation of father and mother flow chart among Fig. 1 the present invention.
Producing method for seed realization flow figure among Fig. 2 the present invention.
Embodiment
Below in conjunction with drawings and the embodiments, the present invention is further detailed.
Osg6B stamen specific promoter, STS14 gynoecium specific promoter are known promotor in following examples; Herbicide sensitive gene also is a known; Cyhalofop-butyl, Bentazon, pyrazosulfuron etc. are known weed killer herbicide; Rice varieties NS629S and RH003 are existing conventional variety, are provided by the Academy of Agri-Science and Technology Anhui Province paddy rice;
In following examples, unless otherwise noted, used technological means is a conventional means well known to those skilled in the art among the embodiment.
The breeding method of embodiment 1 paddy rice cross breeding breeding material
With rice varieties NS629S and RH003 is example;
(1) maternal material breeding method
Clone's Osg6B stamen specific promoter also drives the cyhalofop-butyl sensitive gene or the antisense fragment rice transformation kind NS629S of Bentazon sensitive gene, select in the transgenosis filial generation that single copy inserts, economical character is good, and only in the stamen part to the high plant of cyhalofop-butyl or Bentazon susceptibility as maternal material;
(2) male parent material breeding method
The antisense fragment that clone's STS14 gynoecium specific promoter drives cyhalofop-butyl sensitive gene or Bentazon sensitive gene imports rice varieties RH003, select in the transgenosis filial generation that single copy inserts, economical character is good, and only in the gynoecium part to the high plant of cyhalofop-butyl or Bentazon susceptibility as the male parent material.
Embodiment 2, the mechanization mixed seeding production of hybrid seeds (Hefei ,Anhui plantation)
(1) will invent this material of acquisition father and mother seed and be mixed, its mass ratio is the 1:6(male parent: female parent), in sowing on 10-May 15 May, adopt conventional watery ricenursery mode to cultivate strong seedling.The automatic rice transplanter of Kubo field SPW-48C is adopted in rice transplanting, and average row is apart from 30cm, spacing in the rows 18cm.Field planting adopts conventional water and fertilizer management, sprays corresponding weed killer herbicide (as cyhalofop-butyl, aqua mass fraction 0.25%, every mu sprays 50L-60L) in preceding about 7 days in heading, kills male (female parent), goes female (male parent); By grain header results seed.Through sampling Detection, the seed of surveying is crossbreed, and no this self-fertility of father and mother mixes.
According to the kind of weed killer herbicide, the concentration of control herbicide spraying, spray time and position, should accomplish effective castration (female), avoid producing poisoning again, reduce crossbreed output.
(2) plant female parent (male sterile line) separately at the isolation field piece, under herbicide spraying condition not, conventional water and fertilizer management, under the condition of Pest management, breeding maternal (male sterile line).The breeding of male parent roughly the same.
Should be appreciated that above embodiment only is used for explaining the present invention, and is not limited to the present invention.Without departing from the spirit and substance of the case in the present invention, some modification that is equal to that those skilled in the art are done method of the present invention, crop varieties, step or condition etc. or replacement, conspicuous improvement all belong to protection scope of the present invention.
Claims (8)
1. a paddy rice cross breeding breeding material is characterized in that, maternal material has herbicide sensitive gene, and herbicide sensitive gene is only expressed in stamen; The male parent material has herbicide sensitive gene, and herbicide sensitive gene is only expressed in gynoecium.
2. the breeding method of the described paddy rice cross breeding breeding material of claim 1 is characterized in that,
Described maternal material breeding method is: herbicide resistance gene antisense fragment or other similar techniques that the stamen specific promoter is driven change maternal kind over to, special the expressing at the stamen position of herbicide resistance gene is suppressed, realize that the stamen specific promoter drives the purpose of herbicide sensitive gene, the maternal material of stamen part to herbicide sensitive only arranged;
The breeding method of described male parent material is: herbicide resistance gene antisense fragment or other similar techniques that the gynoecium specific promoter drives change the male parent kind over to, special the expressing at the gynoecium position of herbicide resistance gene is suppressed, realize that the gynoecium specific promoter drives the purpose of herbicide sensitive gene, obtain only to have the male parent material of gynoecium part herbicide sensitive.
3. the breeding method of a kind of paddy rice cross breeding breeding material according to claim 2 is characterized in that, described herbicide sensitive gene is any one in cyhalofop-butyl sensitive gene, the Bentazon sensitive gene etc.; The promotor of described driving herbicide sensitive gene is STS14 gynoecium specific expression promoter and Osg6B stamen specific expression promoter.
4. the breeding method of a kind of paddy rice cross breeding breeding material according to claim 2 is characterized in that, described weed killer herbicide is a wild type paddy rice non-sensitive type weed killer herbicide, is in cyhalofop-butyl, Bentazon, the pyrazosulfuron any one.
5. a hybrid rice mixed seeding producing method for seed is characterized in that, may further comprise the steps:
(1) cultivate and to have herbicide sensitive gene, and the herbicide sensitive gene maternal material of only in stamen, expressing; Cultivation has herbicide sensitive gene, and the herbicide sensitive gene male parent material of only expressing in gynoecium;
(2) maternal material of the paddy rice of cultivating in the step (1) and male parent material are carried out mixed seeding; Herbicide spraying before heading, thus realize maternal the castration, and male parent is gone female, by this unidirectional mating of father and mother, obtains the F1 crossbreed, carries out mechanized harvest.
6. a kind of hybrid rice mixed seeding producing method for seed according to claim 5, it is characterized in that, described maternal material is that herbicide resistance gene antisense fragment or other similar techniques that the stamen specific promoter drives are changed among the rice varieties NS629S, special the expressing at the stamen position of herbicide resistance gene is suppressed, realize that the stamen specific promoter drives the purpose of herbicide sensitive gene, the maternal material of stamen part to herbicide sensitive only arranged, described male parent material is that gynoecium specific promoter driving herbicide resistance gene antisense fragment or other similar techniques are changed among the rice varieties RH003, make special the expressing of herbicide resistance gene be suppressed at the gynoecium position, realize that the gynoecium specific promoter drives the purpose of herbicide sensitive gene, the male parent material of gynoecium part to herbicide sensitive only arranged.
7. a kind of hybrid rice mixed seeding producing method for seed according to claim 6 is characterized in that described herbicide sensitive gene is any one in cyhalofop-butyl sensitive gene, the Bentazon sensitive gene; The promotor of described driving herbicide sensitive gene is STS14 gynoecium specific expression promoter and Osg6B stamen specific expression promoter.
8. a kind of hybrid rice mixed seeding producing method for seed according to claim 7 is characterized in that, described weed killer herbicide is a wild type paddy rice non-sensitive type weed killer herbicide, is in cyhalofop-butyl, Bentazon, the pyrazosulfuron any one.
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Cited By (4)
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| CN104206268A (en) * | 2014-09-09 | 2014-12-17 | 扬州大学 | Novel method for sensitively cultivating efficient rice intelligent sterile line by using herbicides |
| CN104429935A (en) * | 2014-12-12 | 2015-03-25 | 扬州大学 | New method for increasing seed reproduction yield of rice intelligent sterile line |
| CN104946652A (en) * | 2015-07-07 | 2015-09-30 | 安徽省农业科学院水稻研究所 | Plant pistil specific expression promoter OsPisl and derivate thereof |
| CN106258930A (en) * | 2016-07-15 | 2017-01-04 | 安徽省农业科学院水稻研究所 | A kind of hybrid rice seed breeding process of entire mechanization |
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| CN104206268A (en) * | 2014-09-09 | 2014-12-17 | 扬州大学 | Novel method for sensitively cultivating efficient rice intelligent sterile line by using herbicides |
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| CN104946652B (en) * | 2015-07-07 | 2017-08-25 | 安徽省农业科学院水稻研究所 | Plant gynoecium specific expression promoter OsPis1 and its derivative |
| CN106258930A (en) * | 2016-07-15 | 2017-01-04 | 安徽省农业科学院水稻研究所 | A kind of hybrid rice seed breeding process of entire mechanization |
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