CN1092478C - Method for cultivating caryoplasmic hybrid wheat - Google Patents
Method for cultivating caryoplasmic hybrid wheat Download PDFInfo
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- CN1092478C CN1092478C CN98119847A CN98119847A CN1092478C CN 1092478 C CN1092478 C CN 1092478C CN 98119847 A CN98119847 A CN 98119847A CN 98119847 A CN98119847 A CN 98119847A CN 1092478 C CN1092478 C CN 1092478C
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Abstract
The present invention provides a method for cultivating caryoplasmic hybrid wheat. In order to overcome the hybridization incompatibility between certain wild species of aegilops as a female parent and the wheat, and shorten the period of nucleus substitution, the present invention adopts the stigmas of a young female parent and repeated pollination, and the stigmas of the young female parent after the pollination are treated in time by the solution of p-chlorophenoxyacetic acid to enhance the seed setting percentage for 25 to 38 times. In the present invention, the somatic chromosome number of the plants of BC1 and BC2 are checked to select the plats of 2n=42 in time, and the procedure of nucleus substitution can be completed by one or two times of replacing backcross the plants as a female parent; breeding cycle can be reduced for 3 to 4 years.
Description
Technical field
The present invention relates to a kind of method of cultivating caryoplasmic hybrid wheat.
Background technology
The nucleo-cytoplasmic hybrid method of breeding is a kind of emerging breeding technique, and its novelty and characteristics are to develop into from simple attention nuclear gene payes attention to cytoplasm, nucleo-cytoplasmic interaction effect simultaneously.Wheat caryoplasm hybrid breeding method can be concentrated the advantage of pure line breeding method and two kinds of methods of cytoplasmic male sterility heterosis breeding method, has very big yield potential, is expected to make the wheat breed improvement to make a breakthrough.
Cultivate the wheat nucleo-cytoplasmic hybrid and just the cell nucleus of wheat need be replaced or is transplanted in the cytoplasm of another species, breed wheat with allogeneic cytoplasm.For higher plant, though have the people to attempt adopting cell nucleus or organelle transplantation, induce technology such as arrhenokaryon is grown separately, hypochromatosis method to carry out caryoplasm displacement, these methods still can not practical application so far.The effective ways of cultivating at present the wheat nucleo-cytoplasmic hybrid are nucleus substitution backcross methods: promptly make original female parent with cytoplasm donor kind (wild relatives), the nuclear donor grow wheat is that male parent is hybridized and obtained distant hybrid.Make female parent with this distant hybrid again, nuclear parent wheat breed is that male parent is carried out backcrossing more than 6 times continuously, thereby obtains the nucleo-cytoplasmic hybrid of wheat nucleus and allogeneic cytoplasm combination.Beginning to finish to nucleus substitution from distant hybridization generally needs 8-10, and program is numerous and diverse, the cycle very long (Kihara is and prospect H.1982.WheatStudies:Retrospect, Kodensha, Ltd., ToKyo.131-162).This situation has hindered nucleo-cytoplasmic hybrid breeding technique progress of research.Different with general distant hybridization, must make female parent with wild species in order to obtain the cytoplasmic nucleo-cytoplasmic hybrid of wild species, need to utilize the wild species and the wheat hybridizing developing new germplasm source that were not studied as yet, this has just increased the difficulty of work.To cultivate nucleo-cytoplasmic hybrid is purpose, and overcomes the existing many reports of cross-incompatibility.Doing maternal hybridization with common wheat with the Triticum tauschii (Aegilops squarrosa) that comprises in this technical scheme is example, Japan's scholar's wood former equal (1973,1977) method of Cai Yonging is: earlier diplontic Triticum tauschii chromosome doubling is become autotetraploid, with it serves as that female parent is that wheat (AABB) hybridization obtains hybrid with two, making female parent with this hybrid again, is that male parent is backcrossed continuously with common wheat.Not only program is many, the cycle is long for this method, and difficulty is also very big.We had reported the earliest that adopting the rataria cultural method to overcome Triticum tauschii made part of test results maternal and the common wheat cross incompatibility in 1979, make Triticum tauschii and common wheat direct cross one go on foot successfully (Zhang Yan etc., 1979, Institute of Genetics, Academia Sinica's work annual report, 169-171 page or leaf; Zhang Yan etc., 1982, Acta Agronomica Sinica 8 (2): 137-140).Some authors have also carried out similar work but have not obtained the normal hybrid seed of endosperm development (the old filial piety of the Chinese Academy of Agricultural Sciences etc., 1987, wheat nucleo-cytoplasmic hybrid and cytoplasm engineering academic discussion abstract of a thesis selected works, the 6th page later on; Old filial piety etc., 1992, crop magazine (1): 11.The two will good fortune of academy of agricultural sciences, Shanxi Province crop genetic institute etc., 1987, wheat nucleo-cytoplasmic hybrid and cytoplasm engineering academic discussion abstract of a thesis selected works, the 2nd page).Because the hybrid seed that all authors obtain all seldom, so the hybrid first filial generation plant of turning out all has only the 1-3 strain, this is just for further research and utilization have caused difficulty.We test foundation raising cross-fertile rate, particularly raising for this reason the new method of embryo, the normal hybrid seed ripening rate of endosperm development.
As mentioned above, usually people carry out nucleus substitution backcross all be continue to use at random serve as maternally to backcross continuously by generation with the distant hybridization first filial generation, each selection of used female parent is from generation to generation paid attention to particularly more not noting from generation to generation in early days inadequately, ignore the chromosome structure of material therefor.Therefore, not only the cycle of nucleus substitution long, but also can cause that wheat and wild species interchromosomal exchange.We notice the part hybrid plant of the first filial generation of backcrossing of some wheats and goatweed, may be by the F of subtrahend not
1Oogamete and normal common wheat andro gamete are in conjunction with producing.Utilize the hybrid plant of this chromosome structure to do material and common wheat and backcross and obtain 2n=42 easily and have the genomic offspring of common wheat, shorten the nucleus substitution cycle effectively.
Summary of the invention
The method that the purpose of this invention is to provide a kind of new cultivation caryoplasmic hybrid wheat is to overcome the cycle that the wild species goatweed makes maternal and wheat hybridizing incompatibility and shortens nucleus substitution.
The method of cultivation caryoplasmic hybrid wheat of the present invention comprises:
1, choose sharp writing brush shape children of being of goatweed column cap in age, castrate, with the pollination of common wheat pollen, repeated to pollinate 1 time in first pollination back in 2-3 days at least;
2, last pollination is back 4-6 hour, on the maternal column cap of 1 to 5 dropping of p-chlorophenoxyacetic acid (PCPA) solution after awarding powder with 0.5-2.0% weight;
3, the pollination back peeled embryo, the normal hybrid seed of endosperm development in 15-20 days, a tip cut-off of band pappus was accounted for a section of seed length 1/2-1/4;
4, will be with the semina subdivision to be seeded on the medium, grow up to F
1Plant also is transplanted in the soil;
5, use F
1Do maternal backcrossing as male parent and obtain backcrossing the 1st generation BC with common wheat
1
6, check BC
1The plant chromosome number of somatic, choosing somatic chromosome number is the plant that common wheat somatic chromosome bar number and maternal wild species haploid cell chromosome bar are counted summation, does maternal backcrossing as male parent with common wheat with this plant and obtains BC
2
7, BC
2The plant somatic chromosome number checks, selects to have 42 chromosomal plant, and the morphology proterties is so far promptly finished nucleus substitution and obtained caryoplasmic hybrid wheat like the plant of male parent.
In the method for the invention, described goatweed can be a Triticum tauschii.
In the method for the invention, second day after on the maternal column cap after awarding powder with 1 to 5 dropping of p-chlorophenoxyacetic acid (PCPA) solution of 0.5-2.0% weight preferably repeats to drip 1 time.
In the method for the invention, the first pollination in step (1) preferably repeated to pollinate 1 time in back second day.
In the method for the invention, the medium described in the step (4) is preferably and does not contain 2,4-D, add in per 1000 ml solns 7.5 the gram agar amended B
5Medium.
That preferably will select in the method for the invention, has 42 chromosomal BC
2Plant is backcrossed as female parent and produces BC
3, and to BC
3The plant somatic chromosome number is checked, selects 2n=42, and the morphology proterties is like the plant of male parent, with this caryoplasmic hybrid wheat that has obtained as having finished nucleus substitution.
Open up the caryoplasmic hybrid wheat new germplasm source in order to utilize some wild species to do maternal hybridization with common wheat, the subject matter that presses for solution at present is to improve the normal hybrid seed ripening rate of distant hybridization ripening rate, particularly endosperm development.Method of the present invention adopts the children to make maternal column cap, repeat to pollinate and improved the hybridization success rate; Handle maternal column cap after the pollination with p-chlorophenoxyacetic acid solution in good time, promoted the normal development of after fertilization embryo and endosperm, make nomospermous ripening rate improve 25-38 doubly.Different with the general practice, when cultivating inoculation, this method does not peel rataria but with a seed excision part, does so not that the easy damaged embryo obviously improves survival rate and helps abolishing the seed dormancy phase, plant growing way stalwartness.
In the general kernel continued to use for a long time displacement backcross process, begin from the first filial generation that per generation does not make any distinction between to used hybrid plant or only select blindness big according to external form, nucleus substitution is backcrossed to be needed more than 6 generations.This technical scheme is to BC
1And BC
2Plant carries out the somatic chromosome number inspection, and is simple, can in time select the plant of 2n=42, is that female parent carries out one, twice displacement backcrossed, and just can finish the nucleus substitution program with it again, and breeding cycle can reduce 3-4.Need 8-10 if the past begins to finish to nucleus substitution from distant hybridization, adopt this technical scheme to finish so in 3-4.Cooperate in the experiment and utilize time that phytotron (case) quickens to cultivate from generation to generation caryoplasmic hybrid wheat 2-3 only.
Embodiment
Do maternal with Triticum tauschii (Aegilops Squarrosa) strain 011.Cultivation under field conditions (factors).Select 182 Xiao Hua and pull out tassel from Bao Yezhong on plant before tassel is extracted out naturally, earlier little floriglume shell top is cut off 1/3 put in order fringe, column cap is in the children and makes the stage be sharp writing brush shape at this moment.After the castration, award pollen with common wheat (Triticum aestivum) kind Kenya (Kenya).Pollinate and carry out during 9-10 the morning, the pollination bagging is isolated.The 2nd day repeats to pollinate 1 time with identical paternal pollen.Back 6 hours of the 2nd pollination, the p-chlorophenoxyacetic acid solution with 1.0% is with on the maternal column cap of dropper dropping after awarding powder, and 3 on every column cap repeats dropping point 1 time after 24 hours.Select out 180 Xiao Hua else in contrast, handle, just on the maternal column cap of p-chlorophenoxyacetic acid solution dropping point after awarding powder with distilled water replacement 1.0% with said method.Obtain 63 of embryo and the normal hybrid seeds of endosperm development with said method, 2 of the hybrid seeds that obtains from adjoining tree.
Pollinated back 18 days, the normal hybrid seed of profile is peeled from tassel,, a tip cut-off of planting the subband pappus is accounted for a section of seed length 1/3 with 0.1% mercuric chloride solution sterilization 5 minutes.Band semina subdivision is seeded in the B through revising
5On the medium (do not contain 2,4-D adds 7.5 gram agar in per 1000 ml solns).25 ℃-28 ℃ of culturing room's temperature, illumination every day 10 hours.
Hybrid first filial generation (F by cultured in vitro
1) vine growth and development is normal, has parents' feature, pollen mother cells chromosome pairing behavior inspection shows most of F
17 bivalents and 14 free univalents are arranged, hybrid F
1Caryotype should be ABDD.
Winter is at chamber planting then.With F
1Backcross with common wheat Kenya pollen for female parent and to obtain BC
1To 48 strain BC
1Plant makes root tip chromosomes and counts inspection, and 12 strains of therefrom selecting and remain have 49 chromosomal plant, and backcrossing with common wheat Kenya produces BC again
2, to BC
2Plant check the root tip chromosomes number select and remain have 42 chromosomal, and with this maternal BC of generation that backcrosses
3Winter greenhouse plantation BC
3, it is all similar to common wheat Kenya that its plant forms is learned proterties, pollen fertility, esterase isozyme zymogram, shows that nucleus substitution finishes.Later investigation proves, the BC of part 2n=42
2Plant has been finished the common wheat nucleus substitution.
Claims (6)
1, a kind of method of cultivating caryoplasmic hybrid wheat comprises:
(1) choose sharp writing brush shape children of being of goatweed column cap in age, castrate, with the pollination of common wheat pollen, repeated to pollinate 1 time in first pollination back in 2-3 days at least;
(2) last pollination is back 4-6 hour, on the maternal column cap of 1 to 5 dropping of p-chlorophenoxyacetic acid (PCPA) solution after awarding powder with 0.5-2.0% weight;
(3) the pollination back peeled embryo, the normal hybrid seed of endosperm development in 15-20 days, a tip cut-off of band pappus was accounted for a section of seed length 1/2-1/4;
(4) will be with the semina subdivision to be seeded on the medium, grow up to F
1Plant also is transplanted in the soil;
(5) use F
1Do maternal backcrossing as male parent and obtain backcrossing the 1st generation BC with common wheat
1
(6) check BC
1The plant chromosome number of somatic, choosing somatic chromosome number is the plant that common wheat somatic chromosome bar number and maternal wild species haploid cell chromosome bar are counted summation, does maternal backcrossing as male parent with common wheat with this plant and obtains BC
2
(7) BC
2The plant somatic chromosome number checks, selects to have 42 chromosomes, and the morphology proterties promptly obtains the plant of caryoplasmic hybrid wheat like the plant of male parent.
2, in accordance with the method for claim 1, it is characterized in that described goatweed is a Triticum tauschii.
3, in accordance with the method for claim 1, it is characterized in that, last pollination back 4-6 hour, second day after on the maternal column cap after awarding powder with 1 to 5 dropping of p-chlorophenoxyacetic acid (PCPA) solution of 0.5-2.0% weight repeats to drip 1 time.
4, in accordance with the method for claim 1, it is characterized in that the first pollination in step (1) repeated to pollinate 1 time in back second day.
5, in accordance with the method for claim 1, it is characterized in that the medium described in the step (4) is for containing 2,4-D adds the amended B of 7.5 gram agar in per 1000 ml solns
5Medium.
6, in accordance with the method for claim 1, it is characterized in that that will select has 42 chromosomal BC in step (7)
2Plant makes female parent and backcrosses, obtains BC
3Plant is checked its somatic chromosome number, selects 2n=42, and the morphology proterties promptly obtains caryoplasmic hybrid wheat like the plant of male parent.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN98119847A CN1092478C (en) | 1998-09-18 | 1998-09-18 | Method for cultivating caryoplasmic hybrid wheat |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN98119847A CN1092478C (en) | 1998-09-18 | 1998-09-18 | Method for cultivating caryoplasmic hybrid wheat |
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| CN1248394A CN1248394A (en) | 2000-03-29 |
| CN1092478C true CN1092478C (en) | 2002-10-16 |
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|---|---|---|---|
| CN98119847A Expired - Fee Related CN1092478C (en) | 1998-09-18 | 1998-09-18 | Method for cultivating caryoplasmic hybrid wheat |
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| CN107041292A (en) * | 2016-08-09 | 2017-08-15 | 东北师范大学 | The method for cultivating the new hexaploid wheat of homoeologous chromosomes restructuring |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1039946A (en) * | 1989-08-04 | 1990-02-28 | 张贵芝 | A kind of method of producing hybrid wheat |
| CN1082812A (en) * | 1992-07-23 | 1994-03-02 | 湖南农学院 | Utilize low temperature to castrate and prepare method little, hybrid seeds of barley |
| CN1104420A (en) * | 1994-09-20 | 1995-07-05 | 西北农业大学 | Method for producing hybrid wheat seeds |
-
1998
- 1998-09-18 CN CN98119847A patent/CN1092478C/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1039946A (en) * | 1989-08-04 | 1990-02-28 | 张贵芝 | A kind of method of producing hybrid wheat |
| CN1082812A (en) * | 1992-07-23 | 1994-03-02 | 湖南农学院 | Utilize low temperature to castrate and prepare method little, hybrid seeds of barley |
| CN1104420A (en) * | 1994-09-20 | 1995-07-05 | 西北农业大学 | Method for producing hybrid wheat seeds |
Non-Patent Citations (3)
| Title |
|---|
| 中国农业科学 1997.6.1 核质杂种春小麦小山村134选育及其产量因素分析与增产途径的研究 * |
| 陕西农业科学 1998.4.1 马三梅 张改生"异源细胞质诱导小麦单倍体的研究与进展" * |
| 陕西农业科学 1998.4.1 马三梅 张改生"异源细胞质诱导小麦单倍体的研究与进展";中国农业科学 1997.6.1 核质杂种春小麦小山村134选育及其产量因素分析与增产途径的研究 * |
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