CN109479716A - A method of improving subspecies Tetraploid Rice new germ plasm induction frequency - Google Patents
A method of improving subspecies Tetraploid Rice new germ plasm induction frequency Download PDFInfo
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- CN109479716A CN109479716A CN201811587774.3A CN201811587774A CN109479716A CN 109479716 A CN109479716 A CN 109479716A CN 201811587774 A CN201811587774 A CN 201811587774A CN 109479716 A CN109479716 A CN 109479716A
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- subspecies
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- new germ
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
Abstract
The invention proposes a kind of methods for improving subspecies Tetraploid Rice new germ plasm induction frequency.By means of the inductive technology of rice chromosome group polyploidization, pass through the sexual hybridization of cultivated rice subspecies first, obtain the inter subspecific hybrid group (F1) that hereditary basis is in heterozygous state, the allometrosis (F2) with Multi-genotype is then obtained by selfing mode, and then induce and formulate subspecies Tetraploid Rice new germ plasm, thus improve the induced efficiency of subspecies Tetraploid Rice new germ plasm.In same primary test, if inducing the concentration (0.1%) of medicament and induction time (72h) being particular value, used test material is heterozygous genotypes (heterozygote i.e. genetically), then the probability of successful induced chromosome group polyploidization is bigger, genome polyploidizing induction is carried out to Multi-genotype material simultaneously in an induction experiment, successful probability is induced to greatly increase.
Description
Technical field
The present invention relates to a kind of methods for improving subspecies Tetraploid Rice new germ plasm induction frequency.
Background technique
Researcher has opened the research prelude of China's polyploid rice early 1950s.Due to many odjective causes and
Research purpose expected from researchers, potential value has not been reached yet about the research of polyploid rice at present in subjective reason
Still the concern of most of rice research persons is not caused.The material quantity (or material type) in the research of autopolyploid
Limitation is mainly manifested in induction and research by more than 60 years, at present the number of our autopolyploid rice materials obtained
Amount has not exceeded 2000 parts, and the ecotype or genotype of material are fairly limited.It can be seen that researcher is homologous in acquisition at present
Still have certain technical limitation in the inductive technology and abductive approach of Tetraploid Rice new germ plasm, induced efficiency up for
It further increases.
The course of China's polyploid rice research is made a general survey of, how major technology bottleneck is still expeditiously by various bases
Because the diploid rice induction and cultivation of type are polyploid rice new germ plasm, i.e., in the induction of polyploid rice germ plasm resource such as
What overcomes the randomness in inductive technology and reaches directional induction.Once being obtained in the technology of induction polyploid rice new germ plasm
Breakthrough, then essence variation will occur for the research of polyploid rice, rice genetic improvement thus will be made horizontal and water
Rice genetics research rises to new technology platform.
Subspecies Tetraploid Rice has special germplasm feature and important potential value in polyploid rice.It is luring
In the technical research for leading subspecies Tetraploid Rice new germ plasm, researcher mainly uses seed bud revulsion or seed bud clone to induce
Method, induced efficiency are relatively low (usually less than 1%).The main reason is that: material is tested in the induction of subspecies Tetraploid Rice
The genotype of material induces between the concentration and induction time of medicament there is certain correlation, in actual operation researcher
It is difficult to search out optimal 3 combinations of factors.In same primary test, if the concentration (0.1%) and induction time of induction medicament
(72h) is particular value, and test material is a kind of genotype (homozygote genetically), then succeed induced chromosome group polyploidization
Probability is smaller.
Summary of the invention
The present invention proposes a kind of method for improving subspecies Tetraploid Rice new germ plasm induction frequency, to solve existing method
The low technical problem of induced efficiency.
The technical scheme of the present invention is realized as follows:
Technical solution 1: a method of subspecies Tetraploid Rice new germ plasm induction frequency is improved, is included the following steps:
The first step obtains diploid indica and japonica subspecies by sexual hybridization method using diploid long-grained nonglutinous rice and japonica rice seed as starting material
Species hybrid first generation group；
Second step obtains diploid Xian round-grained rice by selfing mode based on diploid subspecies indica and japonica hybrid first generation group
Inter subspecific hybrid second generation group；
Third step is using concentration at seed bud initial stage using diploid subspecies indica and japonica hybrid second generation group as test material
0.1% colchicine solution carries out induction processing, obtains the seed that there is seed bud to expand feature；
4th step induces skill by clone based on the diploid subspecies indica and japonica hybrid second generation group that seed bud is expanded
Art obtains the callus of dedifferentiation, obtains hybrid generation plant using differentiation technique again；
5th step, hybrid generation plant previous step obtained according to the mode of appearance of plant and the morphological feature of polyploid rice into
Row primary election.
Technical solution 2: true using genome identification technology by experimental material of the plant tip of a root after the 5th step
Determine the caryotype of new germ plasm.
Technical solution 3: further doing following improvement on the basis of in technical solution 2, after determining caryotype, with
Between indica and japonica subspecies based on Tetraploid Hybrids offspring, new germ plasm is screened and purified according to rice breeding method, is lost
Pass the subspecies Tetraploid Rice new germ plasm of base fixed.
Further, further progress is improved on the basis of three above scheme, and diploid long-grained nonglutinous rice and japonica rice select respectively
With IR36 and new rice No. 10, to obtain technical solution 4-6.
Using above-mentioned technical proposal, the invention has the benefit that
The induced efficiency of Tetraploid Rice new germ plasm between indica and japonica subspecies can be significantly improved.In induction subspecies Tetraploid Rice novel species
In the technical research of matter, the method that researcher mainly uses is exactly asexual using kind as the seed bud revulsion or seed bud of donor material
It is revulsion, induced efficiency is relatively low (usually less than 1%).The relatively low basic reason of the induced efficiency of traditional revulsion is just
It is the material (homozygote i.e. genetically) relatively simple for genotype of donor material used by researcher.What this method used
Donor material is the material (heterozygote i.e. genetically) of genotype heterozygosis.Therefore, this induction new method that we are proposed
Facilitate the induction frequency that subspecies Tetraploid Rice new germ plasm is effectively improved in unit test period.
It is by the Biological Principles that this method effectively improves the induction frequency of subspecies Tetraploid Rice new germ plasm: in Asia
There is certain between the genotype of test material, the concentration and induction time of induction medicament in the induction of inter-species Tetraploid Rice
Correlation, researcher is difficult to search out optimal 3 combinations of factors in actual operation.In same primary test, if induction
The concentration (0.1%) and induction time (72h) of medicament are particular value, and test material is a kind of genotype (homozygote genetically),
Then the probability of successful induced chromosome group polyploidization is smaller.Based on this technical limitation, it is proposed that new technology
Method, if the concentration (0.1%) of induction medicament and induction time (72h) are particular value, is used that is, in same primary test
Test material be heterozygous genotypes (heterozygote i.e. genetically), then succeed induced chromosome group polyploidization probability compare
Greatly, i.e., genome polyploidizing induction is carried out to Multi-genotype material simultaneously in an induction experiment, induced successfully
Probability greatly increase.
This method will for polyploid other types of in oryza plant polyploid rice Germplasm Creation (autopolyploid,
Polyploid between autoallopolyploid and species) research of inductive technology provides enlightenment, thus promote oryza plant evolve and
Species upgrading.
Based on it is above three aspect technological improvement and technological merit, this method by using heterozygous genotypes (i.e. genetically
Heterozygote) be donor tests material carry out induction experiment, eventually lead to Tetraploid Rice novel species between significantly improving indica and japonica subspecies
The induced efficiency of matter.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this
Some embodiments of invention without any creative labor, may be used also for those of ordinary skill in the art
To obtain other drawings based on these drawings.
Fig. 1 is a kind of flow diagram of specific embodiment of the present invention.
Specific embodiment
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Site preparation description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts every other
Embodiment shall fall within the protection scope of the present invention.
As shown in Figure 1, with diploid rice variety " IR36(2) ", japonica rice variety " new rice No. 10 (2) " and its Hybrids F1 and
The test of Tetraploid Rice new germ plasm can be summarized as the inventive technique between the induction indica and japonica subspecies that hybrid F2 is carried out by material
Example.It is first that hybrid parent carries out sexual hybridization with " IR36(2) " and " new rice No. 10 (2) " in this research system,
Thus to obtain the subspecies indica and japonica hybrid first generation (F1).Pass through clone inductive technology (dedifferentiation, genome by material of F1
Double and break up again) tetraploid indica-japonica hybrid offspring is obtained, its caryotype is determined by means of genome identification technology
(Oryza sativa,2n=4x=48).Then, it is successively pressed based on tetraploid indica-japonica hybrid offspring (F1, F2, F3 and F4)
New germ plasm screening is carried out according to rice conventional breeding methods, thus to obtain the strain that hereditary basis is stable.Finally, between indica and japonica subspecies
The mass of 1000 kernel of Tetraploid Rice new lines is identified.Mass of 1000 kernel identification the result shows that, Tetraploid Rice is new between indica and japonica subspecies
The mass of 1000 kernel of strain is that (51.6g) increases more than 1 times than the mass of 1000 kernel (24.9g) of its diploid parent material (F1).
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Within mind and principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.
Claims (4)
1. a kind of method for improving subspecies Tetraploid Rice new germ plasm induction frequency, which comprises the steps of:
The first step obtains diploid indica and japonica subspecies by sexual hybridization method using diploid long-grained nonglutinous rice and japonica rice seed as starting material
Species hybrid first generation group；
Second step obtains diploid Xian round-grained rice by selfing mode based on diploid subspecies indica and japonica hybrid first generation group
Inter subspecific hybrid second generation group；
Third step is using concentration at seed bud initial stage using diploid subspecies indica and japonica hybrid second generation group as test material
0.1% colchicine solution carries out induction processing, obtains the seed that there is seed bud to expand feature；
4th step induces skill by clone based on the diploid subspecies indica and japonica hybrid second generation group that seed bud is expanded
Art obtains the callus of dedifferentiation, obtains hybrid generation plant using differentiation technique again；
5th step, hybrid generation plant previous step obtained according to the mode of appearance of plant and the morphological feature of polyploid rice into
Row primary election.
2. the method according to claim 1 for improving subspecies Tetraploid Rice new germ plasm induction frequency, which is characterized in that
After the 5th step, the genome of new germ plasm is determined using genome identification technology using the plant tip of a root as experimental material
Type.
3. the method according to claim 2 for improving subspecies Tetraploid Rice new germ plasm induction frequency, which is characterized in that
After determining caryotype, between indica and japonica subspecies based on Tetraploid Hybrids offspring, according to rice breeding method to new germ plasm
It is screened and is purified, obtain the stable subspecies Tetraploid Rice new germ plasm of hereditary basis.
4. the method according to claim 1-3 for improving subspecies Tetraploid Rice new germ plasm induction frequency,
It is characterized in that, diploid long-grained nonglutinous rice and japonica rice select IR36 and new rice No. 10 respectively.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111543310A (en) * | 2020-05-29 | 2020-08-18 | 东北师范大学 | Method for breeding by using rice indica subspecies cross/doubled tetraploid recombinant inbred line |
| CN112042529A (en) * | 2020-10-12 | 2020-12-08 | 武汉多倍体生物科技有限公司 | Breeding method of tetraploid rice |
| CN114190268A (en) * | 2021-10-21 | 2022-03-18 | 湖北大学 | Ploidy cyclic breeding method for polyploid rice |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1266615A (en) * | 2000-04-10 | 2000-09-20 | 湖北大学生命科学学院 | Method for breeding of new rice variety and hybridized rice by distant hybridization and double dominancy of polyploid |
| JP2004121032A (en) * | 2002-09-30 | 2004-04-22 | Genetic Id Kk | Method for discriminating individual of rice plant and base sequence comprising transposon |
| CN102726289A (en) * | 2012-06-12 | 2012-10-17 | 黄群策 | Induction method of autotetraploid rice |
-
2018
- 2018-12-25 CN CN201811587774.3A patent/CN109479716A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1266615A (en) * | 2000-04-10 | 2000-09-20 | 湖北大学生命科学学院 | Method for breeding of new rice variety and hybridized rice by distant hybridization and double dominancy of polyploid |
| JP2004121032A (en) * | 2002-09-30 | 2004-04-22 | Genetic Id Kk | Method for discriminating individual of rice plant and base sequence comprising transposon |
| CN102726289A (en) * | 2012-06-12 | 2012-10-17 | 黄群策 | Induction method of autotetraploid rice |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111543310A (en) * | 2020-05-29 | 2020-08-18 | 东北师范大学 | Method for breeding by using rice indica subspecies cross/doubled tetraploid recombinant inbred line |
| WO2021237924A1 (en) * | 2020-05-29 | 2021-12-02 | 东北师范大学 | Method for breeding using tetraploid recombinant inbred lines obtained by hybridization/doubling of rice indica and japonica subspecies |
| CN112042529A (en) * | 2020-10-12 | 2020-12-08 | 武汉多倍体生物科技有限公司 | Breeding method of tetraploid rice |
| CN114190268A (en) * | 2021-10-21 | 2022-03-18 | 湖北大学 | Ploidy cyclic breeding method for polyploid rice |
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