CN105638434A - Japonica rice high-combining ability aromatic restorer line breeding method - Google Patents
Japonica rice high-combining ability aromatic restorer line breeding method Download PDFInfo
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- CN105638434A CN105638434A CN201511003895.5A CN201511003895A CN105638434A CN 105638434 A CN105638434 A CN 105638434A CN 201511003895 A CN201511003895 A CN 201511003895A CN 105638434 A CN105638434 A CN 105638434A
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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 relates to a japonica rice high-combining ability aromatic restorer line breeding method, and belongs to the field of biotechnology combined rice breeding. The japonica rice high-combining ability aromatic restorer line breeding method is provided to breed a restorer line used as aromatic three-line hybrid rice and improve the resource utilization rate. The method comprises the following steps: 1, extracting DNA of a sample material; 2, carrying out PCR amplification detection on the extracted DNA by using molecular markers InDel-Rfla and GRFM04 in patents; and 3, carrying out restoration gene and aroma gene detection through a biotechnology, carrying out test crossing and plantation with an aromatic three-line sterile line I with the genotype of aa as a female parent and a three-line restorer line II as a male parent to obtain hybrid F1 seeds, planting F1 plants, and determining and screening aromatic restoration variety III with the genotype of aa by adopting aroma, high quality, strong restoring force and high combining ability as selection standards.
Description
Technical field
The present invention relates to a kind of japonica rice high-combining ability odor type Breeding for restoration lines method, belong to the rice breeding field in conjunction with biotechnology.
Background technology
Oryza sativa L. is the first generalized grain crop of China, accounts for the 1/3 of View of World Rice total output. The long-term cultivated area of China Oryza sativa L. is more than 30,000,000 hectares, and wherein the cultivated area of indica Hybrid Rice reaches 1.733 ten thousand hectares, accounts for the 80% of long-grained nonglutinous rice area, accounts for more than the 50% of the Oryza sativa L. gross area; And japonica rice cultivated area is about 8,280,000 hectares, mainly based on conventional japonica rice, Japonica Hybrid ratio is less than 4%; Compared with indica Hybrid Rice, the development of Japonica Hybrid is very delayed. In recent years, along with the raising of living standards of the people and the continuous of Semen oryzae sativae consumption demand increase, wide space has been opened up in the development for having yield potential Japonica Hybrid. According to data, if the Annual planting area of Japonica Hybrid expands 50% to from account for japonica rice area 3%, just it is hopeful to increase production 3,500,000,000 kilograms of high quality paddies every year. Therefore, strengthen the breed breeding of Japonica Hybrid (particularly Three-line system), to ensureing China's grain security, promote that growth of agricultural efficiency, increasing peasant income are significant.
The cultivation of excellent parent is the basis of apolegamy Three-line system advantageous combination. The male sterile cytoplasm of current China Japonica Hybrid application is mainly BT type, next to that Yunnan I type, both sterile cytoplasms belong to the sterile type of gametocyte, have similar Rescued virus, and its fertility restorer is controlled by 1 pair of karyogene. In breeding, the sterile line of Japonica Hybrid be mostly by produce on promote bloom, the good conventional japonica rice of outcrossing habit is formed by backcross transformation, itself just has good economical character; And in restorer, owing to being generally not present corresponding Restorer varieties in japonica rice, its Restore gene is to be imported from long-grained nonglutinous rice by the method for artificial hybridization, inevitably also it is permeated with in long-grained nonglutinous rice while Restore gene some disadvantageous genetic constitutions introducing. Therefore, the key of the excellent Restoring Line of Japonica Rice of selection-breeding is that and carries out Restore gene identifying fast and accurately and selecting.
Practice is it has been proved that heterosis utilization is the effective way improving crop yield, and fragrant rice is the staple food that people like, one of quasi-aromatic hybrid rice breeding thus trend being increasingly becoming rice breeding. Fragrance is an important indicator of rice taste quality, in the world using rice aroma as an important quality index. Fragrant rice not only fragrance is unique, and mouthfeel is good, can strengthen the appetite of people, and its nutritive value is higher. Fragrant rice contains substantial amounts of protein, each seed amino acid, alkaloid, vitamin, amylase, lipase, maltase and calcium, phosphorus, ferrum and abundant zinc, selenium and other trace elements, the resistance of human body can be strengthened, improve metabolism, it it is the ancillary drug for the treatment of vitamin C deficiency, anaphylactic disease, acute and chronic diseases, blood capillary can also be perfected, hypertension and cardiovascular patient are had certain curative effect. Having dulcet rice, not only quality better, also have abundant nutritional health function, economic worth is very high. Therefore, fragrance has become one of rice important quality feature, and Quasi-aromatic Hybrid Rice breeding also becomes a focus and the developing direction of current rice breeding. Some rice breedings expert utilizes locality or introduces fragrant rice resource both at home and abroad, utilize various technical measures scent gene introductive crossing parental rice, successfully it is bred as Quasi-aromatic Hybrid Rice parent, i.e. aromatic indica CMS line or restorer, then hybridize combo with odor type sterile line and odor type restorer and obtain whole spice type hybrid rice variety, and by odor type sterile line and non-odor type restorer or hybridize combo by non-odor type sterile line and odor type restorer and obtain nature and refer and synthesize the Quasi-aromatic Hybrid Rice kind (the part grain of rice is savory) of type.
Although China is Rice Production the first big country, hybrid paddy rice develops quickly, and aromatic rice is various, but China's Hybrid Aromatic Rice kind is very low in international market popularity. Again owing to aromatic rice improvement is relatively backward, eating rice production fails always play a leading role, lack international influence, need on the basis of the fragrant rice of original tradition, accelerate breeding, select and a collection of meet high-quality odor type Hybrid domestic, international market, make China occupy a tiny space in the international market. Therefore, by animal nutrition, shortening the breeding cycle, in conjunction with field breeding, selection-breeding one japonica rice high-combining ability odor type restorer, and then cultivate new varieties, meeting market and consumption demand, quality of improving the people's livelihood, is one of the important content of modern rice breeding.
Summary of the invention
For Three-line Hybrid rice sterile line, restorer had different essential requirement, and high-quality high-combining ability hybrid rice restorer line breeding difficulty is big, there is the technical problem of the scent gene scarcity of resources of health care, in order to be able to select the restorer used as odor type Three-line Hybrid Oryza sativa L., improve resource utilization, the invention provides a kind of japonica rice high-combining ability odor type Breeding for restoration lines method.
For realizing object above, technical scheme step is as follows:
1. there is the selection-breeding of the odor type restorer kind III of high-combining ability, concrete Breeding Process is: carried out scent gene and Restore gene detection by animal nutrition after, using genotype be aa odor type CMS line I as female parent, it is that restorer II carries out test cross, plantation as male parent using three, obtain hybridization F1 seed, plantation F1 plant, to have fragrance, high-quality, restoring force is strong, combining ability is high for selection standard, measures, filters out the odor type recovery kind III that genotype is aa.
2. the feature of the present invention also resides in:
(1) have that fragrance, high-quality, restoring force be strong, combining ability high for selection standard particularly as follows: odor type refers to the overground part of Oryza sativa L., all savory including seed, blade, cane etc.; Restoring force refers to that by force determined variety source or material are as male parent, and the setting percentage performance of CMS line cross combination F1, and setting percentage is more than 70% is recovery, and setting percentage reaches more than 85% for strong restoring force; High-combining ability refers to the yield mean apparent measuring kind or material and multiple Testers hybridization F1, and what yield was high shows to measure kind or material fit power height, otherwise combining ability is low.
(2) being detected by biotechnology, this material has main effect Restore gene; Field is measured, the setting percentage performance of investigation cross combination F1, but have low containing Restore gene setting percentage, screening setting percentage reaches the corresponding male parent of combination of more than 85%, and this male parent system is target strain.
3., when step 1 carrying out scent gene and Restore gene detection by animal nutrition, first have to carry out DNA extraction, then carry out pcr amplification. During pcr amplification, select the related molecular marker GRFM04 (F:3 '-GTTAGGTTGCATTTACTGGGAG-5 ' of scent gene, R:3 '-GAATGATGCTCAAAGTGTCT-5 ') and the related molecular marker InDel-Rfla (F:3 '-CTGATGATCGAGGAGGAGGTA-5 ' of Restore gene, R:3 '-TAACGCGTCTTCCATCCTACT-5 ') carry out pcr amplification, finally carry out electrophoresis detection result.
(1) amplification reaction system is as follows:
(2) amplification reaction condition
The related molecular marker GRFM04 amplification reaction condition of scent gene is as follows:
94 DEG C of denaturation 5min; 94 DEG C of 1min, 55 DEG C of 1min, 72 DEG C of 2min, totally 35 circulations; Finally carry out 72 DEG C and extend 5min. 4 DEG C of preservations are stand-by.
The related molecular marker InDel-Rf1a amplification reaction condition of Restore gene is as follows:
94 DEG C of denaturation 5min; 94 DEG C of 30s, 55 DEG C of 30s, 72 DEG C of 1min, totally 35 circulations; Finally carry out 72 DEG C and extend 7min. 4 DEG C of preservations are stand-by.
The present invention is advantageous in that:
The whole spice type Quality Hybrid Japonica Rice not only yield being bred as with the inventive method is high, particularly except subterranean root, including seed, blade, cane etc., the aerial parts of whole plant is all savory, its rice is in digestion process, outwards give out pleasant fragrance, open steaming and decocting instrument, rice gives out one strong fragrance assailed the nostrils, and rice cold after do not bring back to life, not hardening, cold and hot all show moisten tasty and refreshing, quality is soft, high resilience, fragrant are sweet, glossy smooth, especially cold rear fragrance is richer.
Detailed description of the invention
1. a japonica rice high-combining ability odor type Breeding for restoration lines method, it is characterised in that:
Carried out scent gene and Restore gene detection by animal nutrition after, using genotype be aa odor type CMS line I as female parent, it is that restorer II carries out test cross, plantation as male parent using three, obtain hybridization F1 seed, plantation F1 plant, to have fragrance, high-quality, restoring force is strong, combining ability is high for selection standard, measures, filters out the odor type recovery kind III that genotype is aa.
2., when step 1 carrying out scent gene and Restore gene detection by animal nutrition, first have to carry out DNA extraction, then carry out pcr amplification. During pcr amplification, select the related molecular marker GRFM04 (F:3 '-GTTAGGTTGCATITACTGGGAG-5 ' of scent gene, R:3 '-GAATGATGCTCAAAGTGTCT-5 ') and the related molecular marker InDel-Rfla (F:3 '-CTGATGATCGAGGAGGAGGTA-5 ' of Restore gene, R:3 '-TAACGCGTCTTCCATCCTACT-5 ') carry out pcr amplification, finally carry out electrophoresis detection result.
(1) amplification reaction system is as follows:
(2) amplification reaction condition
The related molecular marker GRFM04 amplification reaction condition of scent gene is as follows:
94 DEG C of denaturation 5min; 94 DEG C of 1min, 55 DEG C of 1min, 72 DEG C of 2min, totally 35 circulations; Finally carry out 72 DEG C and extend 5min. 4 DEG C of preservations are stand-by.
The related molecular marker InDel-Rf1a amplification reaction condition of Restore gene is as follows:
94 DEG C of denaturation 5min; 94 DEG C of 30s, 55 DEG C of 30s, 72 DEG C of 1min, totally 35 circulations; Finally carry out 72 DEG C and extend 7min. 4 DEG C of preservations are stand-by.
Claims (2)
1. a japonica rice high-combining ability odor type Breeding for restoration lines method, it is characterised in that:
There is the selection-breeding of the odor type restorer kind III of high-combining ability, concrete Breeding Process is: carried out scent gene and Restore gene detection by animal nutrition after, using genotype be aa odor type CMS line I as female parent, it is that restorer II carries out test cross, plantation as male parent using three, obtain hybridization F1 seed, plantation F1 plant, to have fragrance, high-quality, restoring force is strong, combining ability is high for selection standard, measures, filters out the odor type recovery kind III that genotype is aa.
2. the feature of the present invention also resides in:
1) have that fragrance, high-quality, restoring force be strong, combining ability high for selection standard particularly as follows: odor type refers to the overground part of Oryza sativa L., all savory including seed, blade, cane etc.; Restoring force refers to that by force determined variety source or material are as male parent, and the setting percentage performance of CMS line cross combination F1, and setting percentage is more than 70% is recovery, and setting percentage reaches more than 85% for strong restoring force; High-combining ability refers to the yield mean apparent measuring kind or material and multiple Testers hybridization F1, and what yield was high shows to measure kind or material fit power height, otherwise combining ability is low.
2) being detected by biotechnology, this material has main effect Restore gene; Field is measured, the setting percentage performance of investigation cross combination F1, but have low containing Restore gene setting percentage, screening setting percentage reaches the corresponding male parent of combination of more than 85%, and this male parent system is target strain.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112586341A (en) * | 2020-12-04 | 2021-04-02 | 国家粳稻工程技术研究中心 | Preparation method of recovery line japonica rice in late flowering |
| CN112602589A (en) * | 2020-12-30 | 2021-04-06 | 江苏省农业科学院 | Method for rapidly and accurately breeding semi-waxy japonica rice restorer |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104823843A (en) * | 2015-05-06 | 2015-08-12 | 云南种业集团有限责任公司 | Breeding method of one-restore dual-purpose aromatic soft hybrid rice restorer line |
| CN104920202A (en) * | 2015-03-10 | 2015-09-23 | 文山壮族苗族自治州农业科学院 | Breeding method of three-line hybrid rice of aromatic purple rice |
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2015
- 2015-12-30 CN CN201511003895.5A patent/CN105638434A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104920202A (en) * | 2015-03-10 | 2015-09-23 | 文山壮族苗族自治州农业科学院 | Breeding method of three-line hybrid rice of aromatic purple rice |
| CN104823843A (en) * | 2015-05-06 | 2015-08-12 | 云南种业集团有限责任公司 | Breeding method of one-restore dual-purpose aromatic soft hybrid rice restorer line |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112586341A (en) * | 2020-12-04 | 2021-04-02 | 国家粳稻工程技术研究中心 | Preparation method of recovery line japonica rice in late flowering |
| CN112586341B (en) * | 2020-12-04 | 2022-02-15 | 国家粳稻工程技术研究中心 | Preparation method of recovery line japonica rice in late flowering |
| CN112602589A (en) * | 2020-12-30 | 2021-04-06 | 江苏省农业科学院 | Method for rapidly and accurately breeding semi-waxy japonica rice restorer |
| CN112602589B (en) * | 2020-12-30 | 2024-01-19 | 江苏省农业科学院 | Method for rapid and accurate breeding of semi-waxy japonica rice restorer |
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Effective date of registration: 20170616 Address after: 300457, No. 7, Ning Ning Road, Tianjin Development Zone Applicant after: CHINA NATIONAL JAPONICA RICE RESEARCH AND DEVELOPMENT CENTER Applicant after: Tianjin Tianlong Science & Technology Co Ltd Address before: Tianjin economic and Technological Development Zone, Tianjin City, 300457 Mu Ning Road No. 11 Applicant before: CHINA NATIONAL JAPONICA RICE RESEARCH AND DEVELOPMENT CENTER Applicant before: Tianjin Tianlong Seeds Science and Technology Co., Ltd. |
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Application publication date: 20160608 |