CN109315285B - Method for rapidly and synchronously breeding cytoplasm sterile line and maintainer line of double-low cabbage type rape - Google Patents
Method for rapidly and synchronously breeding cytoplasm sterile line and maintainer line of double-low cabbage type rape Download PDFInfo
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- CN109315285B CN109315285B CN201811396974.0A CN201811396974A CN109315285B CN 109315285 B CN109315285 B CN 109315285B CN 201811396974 A CN201811396974 A CN 201811396974A CN 109315285 B CN109315285 B CN 109315285B
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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
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
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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
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
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Abstract
The invention discloses a method for rapidly and synchronously breeding a cytoplasmic sterile line and a maintainer line of double-low cabbage type rape, which comprises the following steps: 1. selecting a cabbage type rape maintainer line or sterile line containing cytoplasmic sterile genes as a female parent for hybridization or test crossing; 2. pollinating the hybrid F1 generation by rape double haploid induction line pollen; 3.f2 generation fertile single plant selfing; 4. cytoplasmic sterility and fertile separation in F3 generation strains, selecting single plants with high sterility degree and completely fertile single plants for sister hybridization; 5. the F4 generation formed a maintainer line and a sterile line with stable and consistent nuclear genes in pair relationship. The invention has the beneficial effects that: the maintainer line and the sterile line are bred quickly, the paired maintainer line and the sterile line can be bred in 3-4 generations, the efficiency is improved by 3-4 times compared with the traditional method, the period is shortened, and manpower and material resources are greatly reduced.
Description
Technical Field
The invention relates to a method for rapidly and synchronously breeding a cytoplasmic sterile line and a maintainer line of double-low cabbage type rape, belonging to the technical field of breeding of modern agricultural varieties and breeding parents.
Background
Cabbage type rape is the rape type with the largest planting area all over the world at present, and the cabbage type rape popularized at present mainly is double-low rape, low erucic acid (less than 1 percent) and low glucosinolate (less than 40 mu mol/g. cake). The cabbage type cytoplasm sterility system is one of the main ways of utilizing the rape heterosis. The currently common types of sterile cytoplasm of brassica napus include: cytoplasmic sterility in Policy horse (PolCMS), cytoplasmic sterility in radish (ogara CMS), cytoplasmic sterility in mustard (hau CMS), JA sterile cytoplasmic types. The breeding of the stable cabbage type rape cytoplasmic sterile line needs a longer period, and the investment of manpower and material resources is large. The conventional breeding method comprises the following steps: testing and crossing the material A which is basically stable in heredity (F6-F8 generation) or stable in heredity (high generation self-crossing line or DH line) with the stable cytoplasmic sterile line B, testing and crossing the progeny after testing and crossing to obtain all sterile lines C, proving that the test-crossing male parent A is a maintainer line (in short: the sterile state of the test-crossing female parent can be maintained), carrying out backcross or paired backcross on the test-crossing progeny C for multiple generations (8-10 generations), and breeding a pair of maintainer line A and new sterile line A, wherein the maintainer line A and sterile line A have the same nuclear gene, and only the difference of sterile and fertile cytoplasmic genes exists. The breeding process has long time, more than 15 generations are needed from breeding the maintainer line to forming the stable pair sterile line, more than 15 years are needed by 1 generation and 1 year, and a great amount of time, manpower and material resources are wasted. The conventional method is to replace the nuclear gene of the original sterile line by a maintainer line and sterile line multi-generation backcross method, so that the sterile line and the maintainer line achieve the aim of consistency of the nuclear gene.
Disclosure of Invention
The invention aims to overcome the defects of the prior art, provides a method for quickly and synchronously breeding a double-low cabbage type rape cytoplasmic sterile line and a maintainer line, can overcome the defects of the prior art, solves the problem of quickly and synchronously breeding the double-low cabbage type rape cytoplasmic sterile line and the maintainer line, and does not need to carry out backcross for multiple generations after the maintainer line is basically stable or stable so as to breed the corresponding sterile line and the maintainer line.
The purpose of the invention is realized by the following technical scheme: the fast and synchronous breeding process of double low rape cytoplasmic sterile line and maintainer line includes the following steps:
s1, crossing or testing crossing F1 generation: hybridizing or testing-crossing a known double-low cabbage type rape cytoplasm maintainer line or sterile line needing to be improved serving as a female parent and other double-low cabbage type rape materials with genetically stable excellent characters serving as male parents;
s2, cross or test cross F1 induction: emasculation of fertile plants in the F1 generation, pollination of rape double haploid induction line pollen, and induction of the F1 to generate F2 generation single plants homozygous for nuclear genes;
s3, F2 generation single plant breeding: through field character investigation, selecting a complete fertile and normal tetraploid individual plant with excellent male parent character in the step S1 for bagging selfing, and eliminating a high glucosinolate and high erucic acid quality selfing individual plant through a quality test;
s4, investigating and detecting strains of the F3 generation: f3 generation forms strains, the uniformity and consistency in the strains are identified at the seedling stage and the moss stage of the F3 generation, the strains with uniform shapes and plant types are selected, the sterile and fertile segregation conditions of stamens in the strains are investigated at the flowering stage, DNA is extracted from sterile and fertile single strains to carry out SNP gene chip detection, and the genetic similarity coefficient and the genetic relationship of the sterile single strains and the fertile single strains are analyzed;
s5, F3 generation strain breeding: in the F3 generation strain with regular shape and strain type, plants with sterile quality and fertile quality appear, a strain with genetic similarity coefficient of sterile single plants and fertile single plants more than 95 percent in S4 is selected, the completely fertile single plants in the strain are subjected to sister cross with sterile single plants with thorough sterility, and the completely fertile single plants subjected to sister cross in the F3 generation strain are bagged and selfed;
s6, identifying and forming a new double-low cabbage type rape maintainer line and a sterile line by the F4 generation: planting the S5 fully fertile single plant and sister cross sterile plant to observe the sterility, whether the sister cross sterile single plant is fully sterile or not, whether the progeny of the fertile single plant is fully fertile or not, if the F3 generation of sister cross sterile progeny is still fully sterile, the corresponding F3 generation of fully fertile progeny does not separate the sterile single plant, and the sterility and fertility form the pairing relation of the maintainer line and the sterile line with consistent nuclear genes; if the progeny of the F3 generation of the completely fertile single plant still separates sterile and fertile progeny, the step of S4 is repeated and the F3 generation breeding method is continuously adopted to carry out pairwise sister crossing of a plurality of single plants until the progeny of the completely fertile single plant can not separate sterile plants, and the sister crossing progeny is completely sterile.
The other genetic stable excellent character double-low cabbage type rape materials are any one of cabbage type rape cytoplasm maintainer lines, restorer lines or non-restorer non-maintainer materials; the excellent properties comprise one or more than two of properties of high oil content, high oleic acid, low glucosinolate, low erucic acid, prematurity, crack resistance, root swelling resistance, short stalk, red petals, purple petals, orange petals and the like.
The uniformity and consistency in the F3 generation seedling stage and moss stage identification strains. The SNP gene chip detection is to randomly select 10 sterile and completely fertile single plants to identify the genetic similarity coefficient among the single plants by utilizing a rape 60K SNP chip.
The morphological traits of the F3 strain comprise: leaf color, leaf type, plant height, plant type, branch angle, branch number, flowering phase, maturity phase and the like.
The principle of the invention for achieving the technical effect is as follows: the rape doubled haploid inducing line is utilized to induce the hybrid F1 generation mother body to generate doubled haploid (DH line), namely, the F1 generation egg cell nuclear gene is induced to be doubled to form a descendant with stable heredity, but the cytoplasm in the F1 generation egg cell contains sterile gene, the rape doubled haploid inducing line can not induce the cytoplasmic gene to be stable, so that the formed F2 generation single plant can be fertile, cytoplasmic sterile segregation can occur in the F3 generation strain with stable nuclear gene, but the nuclear gene heredity is consistent. Therefore, the genetic inheritance of the sterile and fertile individual plant cell nucleus genes in the F3 generation strain is consistent, except the genetic difference of sterility and fertility, the other characters are consistent in performance, and the genetic similarity of the genetic difference of nuclear DNA analysis can exceed 95%. The relation between the maintainer line and the sterile line can be realized by carrying out sister crossing on the fertile single plant and the sterile single plant. But the segregation of sterility and fertility still can occur in fertile plants to the F4 generation, because the cytoplasmic gene can also segregate, the method of brother crossing is needed to keep sterility until the sterile segregation of fertile plants does not occur any more and the filial generation of the brother crossing is sterile completely.
The invention has the beneficial effects that: 1. the maintainer line and the sterile line are bred quickly, the sterile line and the maintainer line are formed in one step, the maintainer line and the sterile line can be bred in pairs after 3-4 generations, and the efficiency is improved by more than 3-4 times compared with that of the traditional method; 2. The method is applicable to the synchronous breeding process of the cytoplasmic sterility of the double-low cabbage type rape Berlima, the cytoplasmic sterility of radish, the cytoplasmic sterility of mustard and the cytoplasmic sterility of JA type sterility and the maintainer line. 3. Compared with the existing method, the method has the advantages that the maintainer line is basically stable and then is backcrossed with the sterile line in a test mode, the maintainer line and the sterile line are not synchronously finished, the maintainer line is selected firstly, and then the sterile line is bred, so that the period of breeding is long, the investment of manpower and material resources is large, the breeding period is greatly shortened, and the investment cost of manpower and material resources is reduced.
Drawings
FIG. 1 is a schematic flow chart of the present invention.
FIG. 2 is a schematic flow chart of the rapid and synchronous breeding of the improved precocious double-low cabbage type rape Berima cytoplasmic male sterile line and the maintainer lines 4182A and 4182B of the present invention.
FIG. 3 is an electrophoresis chart of molecular markers for identifying 4182 genetic relationship of SSR molecular markers (primer H57).
FIG. 4 is a schematic flow chart of the rapid and synchronous breeding of the improved double low, high oil, early maturing cabbage type rape Berima cytoplasmic male sterile line and the maintainer lines 4173A and 4173B of the present invention.
FIG. 5 is an electrophoresis chart of molecular markers for identifying 4173 genetic relationship of SSR molecular markers (primer H53).
FIG. 6 is a schematic diagram of the procedure for rapid and synchronous breeding of the modified dwarf, precocious and double-low brassica napus radish cytoplasmic sterile line and maintainer lines 3772A and 3772B of the present invention.
FIG. 7 is an electrophoresis chart of molecular markers for identifying 3772 relatives of SSR molecular markers (primer H105).
Detailed Description
The technical solutions of the present invention are further described in detail below with reference to specific embodiments and drawings, but the scope of the present invention is not limited to the following.
Example 1:
as shown in figures 1 and 2, in order to improve the maturity of the existing double low cabbage rape cytoplasmic sterile A0464, the two early maturing and double low material provinces 9617 and Chinese 3F5 hybridize in 2016, F15 generation stable material (2 month low initial flower, mature before 4 month 30) is crossed with the maintainer line of the cotton A0464, F1 is obtained in 2016, F1 flowering period is removed in 3 month, Y3380 pollen pollination is carried out with the rape double haploid inducer line, the induced progeny seed (F2 generation) is obtained in 2017 month, the induced progeny (F2 generation) is bred in Szechuan Marc summer, the normal tetraploid, anther fertile, flowering early sterile single plant is selected for bagging selfing, F2 generation single plant (F3) is obtained in 2017 month, the bagging test is carried out on the bagging single plant, the crossing single plant is carried out, the high sterile line is selected, the anther fertile single plant is bred, the flowering early sterile single plant is self-bred in the sterile line, the sterile line of the No. 7 crossing No. 7, the No. 7 No. 4, the No. 4 sterile line, the sterile line is marked by the sterile line, the sterile line of the sterile line, the sterile line of the.
Example 2:
as shown in figure 4, in order to improve the oil content (lower than 40%) of the existing early-maturing and double-low cabbage type rape cytoplasmic sterile A0496, test crossing the stable material of the sterile line C142F 10 generation with double-low and high oil content (oil content is more than 51%) in 2016 year 3 month with the sterile line A0496, test crossing F1 in 2016 year 5 month, castration in 2017 month F1 flowering period (test crossing F1 semi-fertile), pollination with rape double haploid induction line Y60 pollen, obtaining induced progeny seed (F2 generation) in 2017 month 5 (F7 generation) in Marshall summer breeding additional generation, selecting normal tetraploid, anther fertile, early-fertile single strain bagging selfing, self-crossing the sterile line F2 generation single plant (F2 generation) in 2017 month, and testing the quality of bagging single-sterile line, and testing the sterile line F6373, and breeding of sterile line, and single-maturing single-sterile line NP 7 strain containing sterile gene 73, test shows that the sterile line of the sterile line A417-mature single-mature line A417 strain is equal and the sterile line 2-mature line, the sterile line is equal, the sterile line is obtained by the test, the test of the sterile line A-sterile line of the sterile line A-mature line, the sterile line 417 line, the stable marker DNA marker.
Example 3:
as shown in figure 6, the plant height and maturity of the existing double low cabbage type rape radish cytoplasmic sterile Rouginea A0068 are improved, the sterile Rouginea rape cytoplasmic sterile Rouginea 36 line F8 stable material (containing radish cytoplasmic restoring gene, early flowering at 2 months bottom, mature at 4 months bottom) and high CM of the sterile plant is tested and crossed with Rouginea 0068 in 2016 year 3, the test cross F1 is obtained in month 5, the sterile F1 flowering phase emasculation (test cross full fertile) is tested and crossed with sterile double haploid inducer Y3380 pollen of rape, the induced progeny seed (F2 generation) is obtained in month 2017, the induced progeny (F2 generation) is bred in March crossing summer in year 2017, normal tetraploid, anther fertile, early, single plant bag of stem, single plant is selected, the sterile rice rape transgenic rice, the sterile single rice stem rice cultivar rape cytoplasmic sterile rice is crossed with sterile rice stem rice, rice stem rice.
The foregoing is illustrative of the preferred embodiments of this invention, and it is to be understood that the invention is not limited to the precise form disclosed herein and that various other combinations, modifications, and environments may be resorted to, falling within the scope of the concept as disclosed herein, either as described above or as apparent to those skilled in the relevant art. And that modifications and variations may be effected by those skilled in the art without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (4)
1. The method for fast and synchronously breeding the cytoplasm sterile line and the maintainer line of the double low cabbage type rape is characterized by comprising the following steps:
s1, crossing or testing crossing F1 generation: hybridizing or testing-crossing a known double-low cabbage type rape cytoplasm maintainer line or sterile line needing to be improved serving as a female parent and other double-low cabbage type rape materials with genetically stable excellent characters serving as male parents;
s2, cross or test cross F1 induction: emasculation of fertile plants in the F1 generation, pollination of rape double haploid induction line pollen, and induction of the F1 to generate F2 generation single plants homozygous for nuclear genes;
s3, F2 generation single plant breeding: through field character investigation, selecting a complete fertile and normal tetraploid individual plant with excellent male parent character in the step S1 for bagging selfing, and eliminating a high glucosinolate and high erucic acid quality selfing individual plant through a quality test;
s4, investigation and detection of F3 generation strains: f3 generation forms strains, the uniformity and consistency in the strains are identified at the seedling stage and the moss stage of the F3 generation, the strains with uniform shapes and plant types are selected, the sterile and fertile segregation conditions of stamens in the strains are investigated at the flowering stage, DNA is extracted from sterile and fertile single strains to carry out SNP gene chip detection, and the genetic similarity coefficient and the genetic relationship of the sterile single strains and the fertile single strains are analyzed;
s5, F3 generation strain breeding: in the F3 generation strain with regular shape and strain type, plants with sterile quality and fertile quality appear, a strain with genetic similarity coefficient of sterile single plants and fertile single plants more than 95 percent in S4 is selected, the completely fertile single plants in the strain are subjected to sister cross with sterile single plants with thorough sterility, and the completely fertile single plants subjected to sister cross in the F3 generation strain are bagged and selfed;
s6, identifying and forming a new double-low cabbage type rape maintainer line and a sterile line by the F4 generation: planting the S5 fully fertile single plant and sister cross sterile plant to observe the sterility, whether the sister cross sterile single plant is fully sterile or not, whether the progeny of the fertile single plant is fully fertile or not, if the F3 generation of sister cross sterile progeny is still fully sterile, the corresponding F3 generation of fully fertile progeny does not separate the sterile single plant, and the sterility and fertility form the pairing relation of the maintainer line and the sterile line with consistent nuclear genes; if the progeny of the F3 generation of the completely fertile single plant still separates sterile and fertile progeny, the step of S4 is repeated and the F3 generation breeding method is continuously adopted to carry out pairwise sister crossing of a plurality of single plants until the progeny of the completely fertile single plant can not separate sterile plants, and the sister crossing progeny is completely sterile.
2. The method for rapid and synchronous breeding of cytoplasmic male sterile line and maintainer line of double low Brassica napus according to claim 1 wherein the other genetically stable and excellent trait material of double low Brassica napus is any one of cytoplasmic maintainer line, restorer line or non-restorer-non-maintainer line of Brassica napus; the excellent properties comprise one or more than two of properties of high oil content, high oleic acid, low glucosinolate, low erucic acid, precocity, crack resistance, root swelling resistance, short stalk, red petals, purple petals and orange petals.
3. The method for rapid and synchronous breeding of cytoplasmic sterile line and maintainer line of canola plants with double low cabbage type as claimed in claim 1, wherein the SNP gene chip detection is to randomly select 10 sterile and fully fertile individuals to identify the genetic similarity coefficient between the individuals by using 60K SNP chips of rape.
4. The method for rapid and synchronous breeding of cytoplasmic sterile line and maintainer line of brassica oleracea L.var.latifolia L.according to claim 1, wherein the morphological traits of the F3 strain line include leaf color, leaf type, plant height, plant type, branch angle, branch number, flowering phase and maturation phase.
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| CN114831021B (en) * | 2022-06-14 | 2022-12-20 | 江苏省农业科学院 | Ornamental rape variety cultivation method suitable for miniature bonsai |
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| CN1104418A (en) * | 1994-02-21 | 1995-07-05 | 江苏省农业科学院 | Breeding method for capitata double hybrid rape |
| CN1276150A (en) * | 1999-06-04 | 2000-12-13 | 华中农业大学 | Two-line male sterile method for assortively breeding ecological-type cytoplasm of rape |
| CN106069718A (en) * | 2016-06-23 | 2016-11-09 | 成都市农林科学院 | The method of Brassica campestris L dihaploid induction system selection-breeding cabbage type rape cytoplasmic male sterile line |
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| CN1104418A (en) * | 1994-02-21 | 1995-07-05 | 江苏省农业科学院 | Breeding method for capitata double hybrid rape |
| CN1276150A (en) * | 1999-06-04 | 2000-12-13 | 华中农业大学 | Two-line male sterile method for assortively breeding ecological-type cytoplasm of rape |
| CN106069718A (en) * | 2016-06-23 | 2016-11-09 | 成都市农林科学院 | The method of Brassica campestris L dihaploid induction system selection-breeding cabbage type rape cytoplasmic male sterile line |
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| Development of new restorer lines for CMS ogura system with the use of resynthesized oilseed rape (Brassica napus L.);Laurencja Szala等;《Breeding Science》;20160930;第66卷(第4期);全文 * |
| 大麦加倍单倍体(DH群体)的建立及其在遗传育种中的应用;汪军妹等;《大麦科学》;20020325(第1期);全文 * |
| 油菜双单倍体诱导系的发现及应用;付绍红等;《中国作物学会油料作物专业委员会第八次会员***暨学术年会综述与摘要集》;20181119;第182页第5段第2-7行 * |
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