CN114642168A - Method for obtaining radish cytoplasm male sterility restoring material and transferring cabbage type rape radish cytoplasm male sterility restoring line - Google Patents
Method for obtaining radish cytoplasm male sterility restoring material and transferring cabbage type rape radish cytoplasm male sterility restoring line Download PDFInfo
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- CN114642168A CN114642168A CN202111360793.4A CN202111360793A CN114642168A CN 114642168 A CN114642168 A CN 114642168A CN 202111360793 A CN202111360793 A CN 202111360793A CN 114642168 A CN114642168 A CN 114642168A
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- radish
- male sterility
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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
- A01H1/021—Methods of breeding using interspecific crosses, i.e. interspecies crosses
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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
- A01H1/022—Genic fertility modification, e.g. apomixis
- A01H1/023—Male sterility
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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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- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
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- Developmental Biology & Embryology (AREA)
- Environmental Sciences (AREA)
- Animal Husbandry (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention provides a method for obtaining a radish cytoplasmic male sterility restoring material and transferring a cabbage type rape radish cytoplasmic male sterility restoring line, which comprises the steps of selecting ladybug with radish cytoplasmic male sterility as a female parent, planting the ladybug in a village in a large scale, freely pollinating cruciferous vegetables such as radishes planted in villages in a natural state, harvesting natural fruiting seeds of the ladybug, planting and identifying, selecting fertile plants in the plants, freely pollinating, harvesting the natural fruiting seeds, continuously planting the natural fruiting seeds, and obtaining the radish cytoplasmic male sterility restoring material. And the second step of hybridizing the radish cytoplasm male sterility restoring material as female parent with excellent cabbage type rape as male parent and subsequent multiple generation backcross and selfing to breed cabbage type rape radish cytoplasm male sterility restoring line. The radish cytoplasmic male sterility restoring line is used as a female parent, and is hybridized, backcrossed and selfed with a cabbage type rape parent with breeding target characters as a male parent to breed the cabbage type rape restoring line with the target characters.
Description
Technical Field
The invention belongs to the technical field of hybrid rape variety breeding and seed production, and particularly relates to a method for obtaining radish cytoplasm male sterility restoring materials from the nature and breeding a cabbage type rape radish cytoplasm male sterility restoring line and a transfer method of the cabbage type rape radish cytoplasm male sterility restoring line.
Background
The radish cytoplasmic sterile line has the advantages of stable and thorough fertility, no environmental influence and the like, and has no restoring gene in the cabbage type rape, so that a maintainer line is easy to obtain and breed into a stable sterile line, but the breeding of the restoring line in the heterosis utilization of the radish cytoplasmic sterile line is difficult.
Pelletier et al in France introduced the restoring gene from Raphanus sativus into Brassica napus by protoplast fusion technique, achieving three-line mating. But still has the problems of unsatisfactory restoring capability, partial female sterility, linkage of higher glucosinolate and restoring genes and the like. In 2005, Primard-Brisset et al utilized gamma ray radiation mutagenesis to recombine DNA fragments to obtain a low-glucosinolate Brassica napus radish cytoplasm male sterility restorer line R2000, which enables radish cytoplasm male sterility hybrid rape to be widely used in Europe. However, foreign radish cytoplasmic male sterility restoring genes and related marking technologies are mostly protected by patents, and the application range of the restoring genes and the related marking technologies is limited.
Disclosure of Invention
The invention aims to obtain radish cytoplasm male sterility restoring material from nature and provide a method for breeding a cabbage type rape radish cytoplasm male sterility restoring line.
The invention has the advantages that the radish cytoplasm male sterility restoring material is found from the nature by adopting an effective method, the autonomy of radish cytoplasm sterility restoring genes is realized, and the excellent cabbage type rape radish cytoplasm male sterility restoring line is bred by continuously hybridizing, backcrossing and selfing with the excellent cabbage type rape for many years.
In order to realize the invention, the inventor combines the uninterrupted screening (searching) of the radish cytoplasm male sterility restoring material and the breeding research of the cabbage type rape radish cytoplasm male sterility restoring line for many years, and finally obtains the following high-efficiency breeding technical scheme:
the method comprises the steps of utilizing the ladybug sterile line with the cytoplasmic male sterility of radish as a female parent, freely pollinating radish planted in villagers in a natural state, harvesting natural seed and planting and identifying, screening fertile plants in the seed, and freely pollinating to obtain the cytoplasmic male sterility restoring material of the radish. Compared with the method for searching for the restoring gene by hybridizing the cabbage type rape and the radish, the method has the advantages that the radish cytoplasmic male sterility ladybug sterile line is used as a bridge, radish pollen randomly planted by villagers is freely pollinated, and the probability of searching for the restoring gene by hybridizing the rape and the radish is higher than that of purposefully. The reason is two: firstly, the ladybug is easier to naturally hybridize and seed with radish pollen than the cabbage rape, and can generate more filial generation; and secondly, the ladybug sterile line of the radish cytoplasm is used as a female parent, so that the radish cytoplasm male sterility restoring gene can be found only by finding a fertile plant in the filial generation, and the detection and restoration are much easier compared with the method of using the filial generation.
Further, the fertile plant is used as a female parent to be continuously hybridized, backcrossed and selfed with the cabbage type rape to breed the cabbage type rape radish cytoplasm male sterility restoring line. The method is characterized in that the fertile plants and the offspring fertile plants are used as female parents to ensure that cytoplasm is cytoplasmic male sterility of the radish, has a cytoplasmic male sterility restoring gene of the radish and simultaneously transmits the cytoplasmic male sterility restoring gene to the offspring.
Further, the bred cabbage type rape radish cytoplasm male sterility restoring line is used as a female parent, and is hybridized, backcrossed and selfed with a cabbage type rape parent with breeding target characters as a male parent, and the cabbage type rape radish cytoplasm male sterility restoring line with the target characters is transferred.
The bred cabbage type rape radish cytoplasm male sterility restoring line with breeding target characters and the cabbage type rape radish cytoplasm male sterility line are utilized to prepare a new variety (combination). The method has the advantages of easy breeding of radish cytoplasmic male sterile line, thorough and stable fertility, and high efficiency and safety of seed production. By the method, a new restorer line is easy to breed: the restoring line with sterile cytoplasm is used as female parent, and hybridized with target parent, and the filial generation is backcrossed or selfed to obtain the new restoring line with fertility-less separation and target character.
Detailed Description
The present invention is further described in the following description with reference to specific embodiments, many details are set forth in order to provide a thorough understanding of the present invention, but the present invention can be implemented in many other ways than those herein described, and it will be apparent to one skilled in the art that similar modifications can be made without departing from the spirit and scope of the invention, and therefore the scope of the invention is to be determined by the claims and not limited by the specific embodiments disclosed below. The following experimental methods are conventional methods and techniques in the art unless otherwise specified.
Example 1: screening (searching) radish cytoplasmic male sterility restoring material:
the invention comprises the acquisition of radish cytoplasmic male sterility restoring material, and the method for acquiring the restoring material comprises the following steps:
(1) the inventor utilizes the radish cytoplasmic cabbage sterile line donated by the forest tree spring teacher of Guizhou university to hybridize and continuously backcross with local village resident ladybug (local people called ladybug which is used as vegetable and feed) in Guizhou south county to breed the radish cytoplasmic ladybug sterile line.
(2) The radish cytoplasmic ladybug sterile line is distributed to villagers to be planted in multiple places, and is freely pollinated with radishes planted in the villagers in a natural state.
(3) And harvesting the freely pollinated natural seed setting, planting and identifying, and finding fertile plants in a planting and identifying garden in 2012.
(4) The fertile plants are marked by the hanging labels, are allowed to freely pollinate for natural fructification, and natural fructification seeds are harvested for continuous planting to obtain the radish cytoplasm male sterility restoring material (because cytoplasm is radish cytoplasm male sterility, the fertile plants inevitably have radish cytoplasm male sterility restoring genes).
Example 2: breeding (transformation) cabbage type rape radish cytoplasm male sterility restoring line RS116
The invention comprises transferring radish cytoplasm male sterility restoring gene into cabbage type rape to breed cabbage type rape radish cytoplasm male sterility restoring line, the method for breeding cabbage type rape radish cytoplasm male sterility restoring line comprises the following steps:
(1) the cytoplasmic male sterile restorer material (fertile strain) of radish obtained in example 1 is used as a female parent to be hybridized with an excellent cabbage type rape line 556(556 does not have the cytoplasmic sterile restorer gene of radish) as a male parent to obtain a hybrid F1And (4) seeds.
(2) Using the above F1Backcrossing the fertile plant as female parent with 556 to obtain BC1F1。
(3) Using the above BC1F1The fertile plant is used as female parent to be backcrossed with 556 continuously to obtain BC2F1This same procedure was repeated for multiple backcrosses to obtain BC6F1。
(4) Using the above BC6F1Bagging the fertile plants for selfing to obtain BC6F2。
(5) Using the above BC6F2Bagging the fertile plants for selfing to obtain BC6F3Selecting the plant line with the field number of 116 plants to obtain a plant line without sterile plant separation,
(6) and performing paired test cross identification on the 116 plant lines serving as male parents and the cabbage type rape radish cytoplasmic male sterile line, wherein the filial generation is all fertile, and the 116 plant lines are the radish cytoplasmic male sterile restoring line and are named as RS 116. The quality is identified, the erucic acid is less than or equal to 1 percent, and the glucosinolate is less than or equal to 30 mu mol/g.
Example 3: breeding (transformation) of high-quality high-oil-content lodging-resistant cabbage type rape radish cytoplasm male sterility restoring line RS211
The invention comprises a new cabbage type rape radish cytoplasm male sterility restoring line transferred according to breeding target requirements, and the method for transferring the new cabbage type rape radish cytoplasm male sterility restoring line comprises the following specific steps:
(1) the cabbage type rape radish cytoplasm male sterility restoring line RS116 obtained in the example 2 is used as a female parent and has high self-breeding quality and high oil content and lodging resistanceThe cabbage type rape line rape 211R (211R does not have radish cytoplasm sterile restoring gene) is taken as a male parent for hybridization to obtain a hybrid F1And (4) seeds.
(2) Using the above F1Backcrossing with YOUZHEN 211R as female parent to obtain BC1F1。
(3) Using the above BC1F1The fertile plant is used as female parent to be backcrossed with the excellent cabbage type rape to obtain BC2F1The same procedure was repeated for 2 additional backcrosses to obtain BC4F1。
(4) Using the above BC4F1Selecting fertile plants and bagging for selfing by combining agronomic characters to obtain BC4F2。
(5) Using the above BC4F2Selecting fertile plants and bagging for selfing by combining agronomic characters to obtain BC4F3Screening out the plant line without sterile plant separation,
(6) and (3) carrying out paired test cross identification on the strain line separated from the sterile plant serving as a male parent and the cabbage type rape radish cytoplasmic male sterile line.
Claims (5)
1. A method for obtaining a radish cytoplasm male sterility restoring material and transferring a cabbage type rape radish cytoplasm male sterility restoring line is characterized by comprising the following steps:
1) selecting a ladybug sterile line with the cytoplasmic male sterility characteristic of radish as a female parent, carrying out free pollination with radish in a natural state, harvesting naturally fruitful seeds, planting and identifying the seeds, screening fertile plants in the seeds, and carrying out free pollination to obtain a cytoplasmic male sterility restoring material of the radish;
2) selecting the obtained radish cytoplasm male sterility restoring material as a female parent, hybridizing with the cabbage type rape as a male parent, backcrossing and selfing to breed the cabbage type rape radish cytoplasm male sterility restoring line;
3) the cabbage type rape radish cytoplasm male sterility restoring line is used as a female parent, and is hybridized, backcrossed and selfed with a cabbage type rape parent with breeding target characters as a male parent to transform the cabbage type rape radish cytoplasm male sterility restoring line with the breeding target characters.
2. The method for obtaining cytoplasmic male sterility restorer line of radish and transferring the cytoplasmic male sterility restorer line of Brassica napus according to claim 1, wherein the eruca japonica sterile line with cytoplasmic male sterility of radish is non-heading cabbage sterile line or pakchoi sterile line.
3. The method for obtaining cytoplasmic male sterility restorer line of radish according to claim 1, wherein the radish providing pollen is selected from radish species reserved for long term in villages and radish species purchased.
4. The method for obtaining cytoplasmic male sterility restorer material of radish according to claim 1, wherein said brassica napus does not have a cytoplasmic male sterility restorer gene of radish, so as to ensure that said restorer gene is derived from the cytoplasmic male sterility restorer material of radish obtained in step 1).
5. The method for obtaining the radish cytoplasmic male sterility restoring material and the method for transforming the radish cytoplasmic male sterility restoring line for brassica napus according to claim 1, wherein in the backcross, a progeny fertile plant is selected as a female parent to keep the cytoplasm as the radish cytoplasmic male sterility characteristic, and the radish cytoplasmic male sterility restoring gene in the step 1) is transferred to progeny.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202111360793.4A CN114642168A (en) | 2021-11-17 | 2021-11-17 | Method for obtaining radish cytoplasm male sterility restoring material and transferring cabbage type rape radish cytoplasm male sterility restoring line |
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| CN202111360793.4A CN114642168A (en) | 2021-11-17 | 2021-11-17 | Method for obtaining radish cytoplasm male sterility restoring material and transferring cabbage type rape radish cytoplasm male sterility restoring line |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2932062A1 (en) * | 2008-06-04 | 2009-12-11 | Clause | Plant of Eruca genus, which is a carrier of a cytoplasmic male sterility, useful to produce hybrids, including in radish, carrot, celery, beetroot, onion, sunflower, and fennel |
| CN102640700A (en) * | 2012-05-04 | 2012-08-22 | 湖南省作物研究所 | Cabbage type rape Ogura cytoplasmic male sterility (OGU CMS) restorer and transformation method and application thereof |
| CN102696474A (en) * | 2012-05-15 | 2012-10-03 | 西北农林科技大学 | Breeding and application of cytoplasmic male sterility restoring line of brassica napus rapeseed and radish |
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2021
- 2021-11-17 CN CN202111360793.4A patent/CN114642168A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2932062A1 (en) * | 2008-06-04 | 2009-12-11 | Clause | Plant of Eruca genus, which is a carrier of a cytoplasmic male sterility, useful to produce hybrids, including in radish, carrot, celery, beetroot, onion, sunflower, and fennel |
| CN102640700A (en) * | 2012-05-04 | 2012-08-22 | 湖南省作物研究所 | Cabbage type rape Ogura cytoplasmic male sterility (OGU CMS) restorer and transformation method and application thereof |
| CN102696474A (en) * | 2012-05-15 | 2012-10-03 | 西北农林科技大学 | Breeding and application of cytoplasmic male sterility restoring line of brassica napus rapeseed and radish |
Non-Patent Citations (2)
| Title |
|---|
| 娄丽娜 等: "萝卜与芸薹属物种间的远缘杂交研究进展", 《江西农业学报》 * |
| 胡大有 等: "甘蓝型油菜与萝卜属间杂种的获得及分子鉴定", 《中国油料作物学报》 * |
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