CN1762199A - Method for obtaining cytoplasm male sterility three lines from paddy TGMS(thermo-sensitive genic male sterile) line - Google Patents
Method for obtaining cytoplasm male sterility three lines from paddy TGMS(thermo-sensitive genic male sterile) line Download PDFInfo
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Abstract
The invention discloses a breeding method for a hybrid rice, particularly relating to a method of getting the cytoplasm male sterility three lines from the photo temperature sensing nucleon sterility line; comprising: choosing the photo temperature sensing nucleon sterility line and high quality rice cytoplasm male sterility keeping line for crossing, planting the filial generation in the high temperature and long day to charge off the photo temperature sensing nucleon sterility genes, separating the cytoplasm male sterility recovery line from the filial generation through the test-mating of cytoplasm sterility line, getting the rice cytoplasm male sterility line and recovery line respectively by carrying out the backcross and auto-copulation at the same time. In this method, the introducing the sound characteristics of the photo and temperature sensing nucleon sterility line such as the straight strain and high chapiter exposing rate to the keeping line, promotes the improvement of the cytoplasm male sterility line and establishes a mew way of generating the cytoplasm male sterility line, and at the same time overcomes the shortage of fertility of the rice photo and temperature sensing sterility line easy to be influenced by the light and temperature environment, which is favorable to increase the hybrid rice seed purity and the seed production.
Description
Technical field
The present invention relates to a kind of hybrid rice breeding method, more particularly, the present invention relates to a kind ofly obtain the method that cytoplasmic male sterilty three is from photo-thermo sensitive male nuclear sterile rice line.
Background technology
Along with the practical photo-thermo-sensitive genic male sterile line of paddy rice is trained breeding of short 64S, be that the long-grained nonglutinous rice double-linear hybrid rice of a collection of new seed selection of representative begins larger area application on producing with the two line system.Be rich in a lot of excellent genes in the display light temperature-sensitive sterile line, for example: the upright plant type relevant with super high-yielding, the stigma exposing ratio relevant with seed production, flowering habit is good etc.But because the fertility of photo-thermo-sensitive genic male sterile line is subject to the influence of light temperature environment, the purity and the output of hybrid rice seeds have been subjected to serious challenge, have increased difficulty also for the sustainable growth of hybrid rice.Thereby the stability of improvement photo-thermo-sensitive genic male sterile line is the important topic of two-line hybrid rice breeding, in contrast to this, the cytoplasm male sterility line stable fertility, three line method still is in the ascendance in hybrid rice breeding so far.
Selecting good maintenance line for use is the successful key of cultivating the paddy rice cytoplasm male sterility line.Yet male sterility of rice maintenance line resource is quite poor.The existing maintenance line great majority of China derive from the seed selection of Wenzhou, Zhejiang institute of agricultural sciences early precious Shan of Xian kind and sisters system thereof, and this is tangible illustration.Tracing it to its cause is, most rice varieties lack the paddy rice cytoplasm male sterility line is kept sterile ability, when shifting male sterility, fertility often occurs and recovers phenomenon by generation, finally causes shifting the sterility failure.Equally, also because of existing cytoplasmic male sterilty to recover gene, cause excellent genes contained in the photo-thermo-sensitive genic male sterile line to be difficult to the transfer of realization in the photo-thermo-sensitive genic male sterile line cell nucleus to cytoplasm male sterility line.Up to the present do not see the report that successfully produces maintenance line and male sterile line from the photoperiod-temperature sensitive genie male-sterile line material both at home and abroad as yet.
Summary of the invention
The objective of the invention is to overcome deficiency of the prior art, provide a kind of and obtain the method that cytoplasmic male sterilty three is from photo-thermo sensitive male nuclear sterile rice line.
The invention provides and a kind ofly obtain the method that cytoplasmic male sterilty three is, may further comprise the steps from photo-thermo sensitive male nuclear sterile rice line:
(1) selective light temperature-sensitive sterile line and good paddy rice cytoplasmic male sterilty maintenance line hybridization;
(2) the plantation filial generation is rejected the photoperiod-temperature sensitive genie male-sterile line gene under long day high temperature;
(3) isolate cytoplasmic male sterilty by the cytoplasmic sterile line test cross from filial generation and recover system;
(4) by backcross and selfing carry out synchronously, obtain paddy rice cytoplasm male sterility line and maintenance line respectively.
The present invention by with the cytoplasmic sterile line test cross, isolate the cytoplasmic male sterilty maintenance line from filial generation after, can utilize the maintenance line of synchronization gain to make recurrent parent, further cytoplasm male sterility line and the maintenance line of transformation Cheng Xin.
When the present invention backcrossed, microscopy abortive pollen rate was more than 99.7%, and abortive pollen loses based on allusion quotation.
The present invention recovers can also to utilize this male sterile restoring line to do the parent, further the new male sterile restoring line of seed selection after the system by isolating cytoplasmic male sterilty with the cytoplasmic sterile line test cross from filial generation.
Compared with prior art, the invention has the beneficial effects as follows:
The present invention successfully obtains maintenance line and male sterile line from the photoperiod-temperature sensitive genie male-sterile line material first, takes the lead in finishing from the conversion of paddy rice two line method caryon sterile line to the three line method cytoplasmic male sterile gene.Utilize new method provided by the invention, good characteristics such as the photo-thermo-sensitive genic male sterile line plant type is upright, stigma exposing ratio height are introduced maintenance line, have advanced the improvement of cytoplasm male sterility line, have created generation cytoplasm male sterility line new way.Also overcome the photo-thermo sensitive male nuclear sterile rice line fertility simultaneously and be subject to the shortcoming of light temperature environmental influence, thereby helped improving the purity of hybrid rice seeds, increased the output that seed is produced.
Embodiment
Below the present invention is further described by example.
May relate to term among the present invention is defined as follows:
Male sterile line: the normal and stamen pollen abortion of this strain gynoecium, can not self-fertility, fertility is controlled by plasmon or nuclear gene.Have the male sterile line of wild rice male sterile cytoplasm as wild abortion type sterile series, represent with A usually.Photo-thermo-sensitive genetic male sterile line then by nuclear gene control, is represented with S usually.
Male sterility maintainer line: the female stamen of this strain is grown normal, the energy self-fertility, and its offspring of back still keeps the male sterile characteristic to the male sterile line pollination, represents with B usually.Thereby can be called numerous kind by male sterile line and its corresponding maintenance line hybridization procreation male-sterile seed.
Male sterile restoring line: the female stamen of this strain is grown normal, and the first generation of hybrid fertility restorer that is produced to male sterile line pollination back is normal, has stronger advantage.Usually represent with R.By male sterile line and the normal and heterotic hybrid seed of tool of recovery system hybridization production fertility, be called the production of hybrid seeds.
As mentioned above, having comprised male sterile line, maintenance line and recovery in the application cell matter male sterile line production hybrid seed system is three systems, so be called " three line method ".And when using photo-thermo-sensitive genic male sterile line and producing hybrid seed, though caryon sterile line performance under long day, high temperature is sterile, but under short day, low temperature, can convert to and can educate, be that photo-thermo-sensitive genic male sterile line can raise up seed by changing light temperature environment, therefore, can be without maintenance line, so be called " two line method ".
Pollen types:
According to the display form of abortive pollen and to the staining reaction of 1% Wagner's reagent can be divided into that allusion quotation loses, circle loses, dye lose, normal four kinds.Allusion quotation loses that the pollen grain form is irregular, transparent not to dye, and circle loses that pollen grain is circular transparent not to dye, and dyes and loses pollen circle, slight dyeing; Normal pollen grain circle, dye brownish black.
The microscopy method: every strain picked at random spike of rice grain husk flower, take out flower pesticide and place on the slide, with the dyeing of 1% IKI liquid, under ordinary optical microscope, amplify 100 times of numbers of observing and adding up above-mentioned all kinds of pollen.
The genetic analysis of the described method of present embodiment:
The present invention produces the feasibility that cytoplasmic male sterilty three is by following genetic analysis proof from photo-thermo sensitive male nuclear sterile rice line, table 1 is for producing the analysis diagram that cytoplasmic male sterilty three is from photo-thermo sensitive male nuclear sterile rice line, the non-photoperiod-temperature sensitive genie male-sterile line gene of G1 (dominance) wherein, g1 photoperiod-temperature sensitive genie male-sterile line gene (recessiveness), the R2 cytoplasmic male sterilty recovers gene (dominance), and the r2 cytoplasmic male sterilty keeps gene (recessiveness); P (parent), F1 (1 generation of hybrid), F2 (2 generations of hybrid); S (G1G1r2r2) is a test cross kind genotype, and S is the cytoplasmic sterility gene.N is the cytoplasmic sterility gene.
| P | g1g1R 2R 2 | × | G1G1r2r2 | ||||
| (photoperiod-temperature sensitive genie male-sterile line recovery) | (non-photoperiod-temperature sensitive genie male-sterile line maintenance) | ||||||
| ↓ | |||||||
| F1 | G1g1R 2r 2 | (non-photoperiod-temperature sensitive genie male-sterile line recovery) | |||||
| ↓ | |||||||
| F2 | g1g1R2_ | g1g1r2r2 | G1_R2_ | G1_r2r2 | |||
| (photoperiod-temperature sensitive genie male-sterile line recovery) | (photo-thermo-sensitive genic male sterile line maintenance) | (non-photoperiod-temperature sensitive genie male-sterile line recovery) | (non-photoperiod-temperature sensitive genie male-sterile line maintenance) | ||||
| Under the long day high temperature | Sterile (eliminating) | Sterile (eliminating) | Can educate | Can educate | |||
| G1_R2_ | G1_r2r2 | ||||||
| ↓ | ↓ | ↓ | ↓ | ↓ | |||
| And S (G1G1r 2r 2) test cross | S(G1_R2_) | S(G1G1r2r2) | |||||
| Can educate | Sterile | ||||||
| Selfing and seed selection continuously | N or S (G 1G1R2R2) | N(G1G1r2r2) | |||||
| (recovery is R) | (maintenance line B) | ||||||
| And S (G1G1r2r2) | S(G1G1r2r2) | ||||||
| Continuous backcross and seed selection | (cytoplasm male sterility line A) | ||||||
Table 1 produces the analysis diagram that cytoplasmic male sterilty three is from photo-thermo sensitive male nuclear sterile rice line
Specific embodiment 1:
(1) selective light temperature-sensitive sterile line and good paddy rice cytoplasmic male sterilty maintenance line hybridization
By photo-thermo sensitive male nuclear sterile rice line is collected screening, find that most photo-thermo-sensitive genic male sterile lines lack cytoplasmic male sterilty and keep gene in the present embodiment.In other words, the paddy rice cytoplasm male sterility line has to a certain degree restorative to existing cytoplasm male sterility line, and the present invention isolates the fact that cytoplasmic male sterilty recovers system and also confirmed this point (table 1) fully from filial generation.
The photo-thermo sensitive male nuclear sterile rice line that present embodiment is cultivated with hybrid rice center, Hunan is trained short 64 and is done the blue or green B early of donor and cytoplasmic male sterilty maintenance line association hybridization, hybridizes F1 generation and F2 generation.From F2, select the plant and the cytoplasm male sterility line Xieqingzao A of the similar training of plant type short 64 to backcross.
(2) the plantation filial generation is rejected the photoperiod-temperature sensitive genie male-sterile line gene under long day high temperature.
At Hangzhou sowing backcross progeny, make them at the beginning of 4 months, select the high plant of ripening rate, supply further seed selection cytoplasmic male sterilty maintenance line and recover system in (long day hot environment in summer) booting in July, heading.Genetic analysis proved already that the photoperiod-temperature sensitive genie male-sterile line gene was controlled by recessive gene, and the plant that contains this gene can only be sterile in performance under long day high temperature, therefore, utilizes hot environment seed selection in long day of summer, can reject the photoperiod-temperature sensitive genie male-sterile line gene in the hybrid generation.
(3) by with the cytoplasmic sterile line test cross, isolate cytoplasmic male sterilty from filial generation and recover system.
Further utilize cytoplasmic sterile line Xieqingzao A (male sterile line of large-area applications) test cross to identify, as: identify that with 1% IKI liquid decoration method the almost whole pollen of pollen are blue, show normally and can educate; Generally more than 85%-90%, these tested plant can be considered and contain cytoplasmic male sterilty recovery gene the seed fertility.Present embodiment has restoring force to other cytoplasmic sterile lines after tested equally by how to recover system for obtaining cytoplasmic male sterilty after test cross and the selfing.By with the cytoplasmic sterile line test cross, microscopy abortive pollen rate is more than 99.7%, abortive pollen loses based on allusion quotation; Self-fruitful rate is lower than 0.1% under the isolation implant.Stigma exposing ratio is higher than 50%, and plant type is upright.
(4) by backcross and selfing carry out synchronously, obtain paddy rice cytoplasm male sterility line and maintenance line respectively.
Identify by cytoplasmic sterile line Xieqingzao A test cross, as: identify that with 1% IKI liquid decoration method pollen is irregular type, belong to allusion quotation and lose; Be round as pollen, but do not dye, then belong to circle and lose; These tested plant all are considered as containing cytoplasmic male sterilty and keep gene.The stable maintenance line of seed selection is at first wanted in male sterile line transformation in the past, and then the backcross transformation male sterile line, and breeding time is very long like this, and speed is slower.The present invention adopts the back-and-forth method of backcrossing synchronously (promptly implement to backcross and selfing is carried out synchronously), has not only accelerated sterile line breeding process (approximately than conventional method 3-4) soon, and paddy rice cytoplasm male sterility line and maintenance line are obtained respectively.Present embodiment through observe continuously backcross 1 to 4 generation (BC2F1 is to BC4F1) pollen fertility variation, the abortive pollen rate keeps high sterile degree all more than 99.5%, thereby successfully obtains paddy rice cytoplasm male sterility line and maintenance line.
When sterile line breeding, except that good and stable male sterility must be arranged; Also need possess good characteristics such as stigma exposing ratio height, flowering habit is good, plant type is upright.Therefore, pay attention to selecting plant type upright from filial generation, the stigma exposing ratio height, the good male sterile of flowering habit keeps material.
When having observed the variation of the different pollen fertilities from generation to generation backcrossed continuously, for making cytoplasmic male sterile gene very strong confining force is arranged, must be strict at pollen microscopic examination, microscopy abortive pollen rate is more than 99.7%, and abortive pollen loses based on allusion quotation.As find that the normal pollen rate of some material and dying loses the pollen rate and with increasing from generation to generation the tendency of increasing progressively arranged, then should in time eliminate, to alleviate workload.
The selection of male sterile line test cross kind
In view of maintenance line there are differences the sterile confining force of dissimilar CMS.Present embodiment be adopt on seed is produced large-area applications, sterility preferably Xieqingzao A as the test cross male sterile line.
The paddy rice cytoplasm male sterility line can utilize this male sterile line directly to survey combo and close after obtaining, the new combination of seed selection hybridization.
Paddy rice cytoplasmic male sterilty maintenance line also can utilize this male sterility maintainer line to do the parent after obtaining, the further new combination of configuration, male sterile line that seed selection is new and maintenance line.
The paddy rice cytoplasmic male sterilty recovers can also utilize this male sterile restoring line to do the parent after system's acquisition, the further new combination of configuration, and the male sterile restoring line that seed selection is new has characteristics such as plant type is upright, flowering habit is good.Its recovery source is different with existing recovery.
The selected photo-thermo-sensitive genic male sterile line of the present invention contains cytoplasmic male sterilty usually and recovers gene, and its photoperiod-temperature sensitive genie male-sterile line gene is rejected the photoperiod-temperature sensitive genie male-sterile line gene by environment plantation filial generation under long day high temperature.
The present invention adopts the back-and-forth method of backcrossing synchronously, accelerates the sterile line breeding process, and almost can obtain paddy rice cytoplasm male sterility line and corresponding maintenance line thereof simultaneously.
At last, it is also to be noted that what more than enumerate only is specific embodiments of the invention.Obviously, the invention is not restricted to above examples of implementation, many distortion can also be arranged.All distortion that those of ordinary skill in the art can directly derive or associate from content disclosed by the invention all should be thought protection scope of the present invention.
Claims (4)
1, a kind ofly obtain the method that cytoplasmic male sterilty three is, may further comprise the steps from photo-thermo sensitive male nuclear sterile rice line:
(1) selective light temperature-sensitive sterile line and good paddy rice cytoplasmic male sterilty maintenance line hybridization;
(2) the plantation filial generation is rejected the photoperiod-temperature sensitive genie male-sterile line gene under long day high temperature;
(3) isolate cytoplasmic male sterilty by the cytoplasmic sterile line test cross from filial generation and recover system;
(4) by backcross and selfing carry out synchronously, obtain paddy rice cytoplasm male sterility line and maintenance line respectively.
2, obtain the method that cytoplasmic male sterilty three is according to claim 1 is described, it is characterized in that, by with the cytoplasmic sterile line test cross, after filial generation is isolated the cytoplasmic male sterilty maintenance line, can utilize the maintenance line of synchronization gain to make recurrent parent, further cytoplasm male sterility line and the maintenance line of transformation Cheng Xin.
3, obtain the method that cytoplasmic male sterilty three is according to claim 1 is described, it is characterized in that when backcrossing, microscopy abortive pollen rate is more than 99.7%, abortive pollen loses based on allusion quotation.
4, obtain the method that cytoplasmic male sterilty three is according to claim 1 is described, it is characterized in that, recover after the system by isolating cytoplasmic male sterilty from filial generation with the cytoplasmic sterile line test cross, can also utilize this male sterile restoring line to do the parent, further the new male sterile restoring line of seed selection.
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1951172B (en) * | 2006-11-09 | 2010-05-12 | 国家杂交水稻工程技术研究中心 | A hybrid rice breeding method |
| CN101356894B (en) * | 2008-09-24 | 2012-07-04 | 湖南杂交水稻研究中心 | Method for breeding and using sporophyte type rice sterile line |
| CN103026964A (en) * | 2012-12-14 | 2013-04-10 | 西南科技大学 | Method for breeding hybrid rice new material through hybridizing two-line sterile line and three-line maintainer line |
| CN106900536A (en) * | 2015-12-22 | 2017-06-30 | 中国种子集团有限公司 | Photo-thermo sensitive male nuclear sterile rice line orients transformation into the method for protection cytoplasmic male sterile line |
| CN107435066A (en) * | 2017-07-13 | 2017-12-05 | 中国水稻研究所 | Stigma Extrusion of Rice main effect QTL and its localization method and application |
| CN109662026A (en) * | 2019-02-27 | 2019-04-23 | 江油市川江水稻研究所 | A method of utilizing cytoplasmic male sterile rice breeding rice photo-thermo-sensitive genic male sterile line |
| CN110547189A (en) * | 2019-10-16 | 2019-12-10 | 四川发生种业有限责任公司 | novel method for breeding rice cytoplasmic male sterile line by using rice cytoplasmic male sterile line |
| CN112314429A (en) * | 2020-10-29 | 2021-02-05 | 海南波莲水稻基因科技有限公司 | Breeding method of rice nuclear male sterility maintainer line |
| CN115777524A (en) * | 2021-12-17 | 2023-03-14 | 安徽省农业科学院水稻研究所 | Method for creating novel rice sterile line |
| CN116420613A (en) * | 2023-05-30 | 2023-07-14 | 福建华晋农业技术开发有限公司 | Rapid backcross stable cultivation method for three-line sterile line of rice |
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- 2005-08-31 CN CNB2005100605876A patent/CN100364384C/en not_active Expired - Fee Related
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1951172B (en) * | 2006-11-09 | 2010-05-12 | 国家杂交水稻工程技术研究中心 | A hybrid rice breeding method |
| CN101356894B (en) * | 2008-09-24 | 2012-07-04 | 湖南杂交水稻研究中心 | Method for breeding and using sporophyte type rice sterile line |
| CN103026964A (en) * | 2012-12-14 | 2013-04-10 | 西南科技大学 | Method for breeding hybrid rice new material through hybridizing two-line sterile line and three-line maintainer line |
| CN103026964B (en) * | 2012-12-14 | 2014-06-11 | 西南科技大学 | Method for breeding hybrid rice new material through hybridizing two-line sterile line and three-line maintainer line |
| CN106900536A (en) * | 2015-12-22 | 2017-06-30 | 中国种子集团有限公司 | Photo-thermo sensitive male nuclear sterile rice line orients transformation into the method for protection cytoplasmic male sterile line |
| CN106900536B (en) * | 2015-12-22 | 2022-03-08 | 中国种子集团有限公司 | Method for directionally transforming rice photo-thermo sensitive genic male sterile line into wide-range cytoplasmic male sterile line |
| CN107435066B (en) * | 2017-07-13 | 2021-02-26 | 中国水稻研究所 | Rice stigma exposure rate main effect QTL and positioning method and application thereof |
| CN107435066A (en) * | 2017-07-13 | 2017-12-05 | 中国水稻研究所 | Stigma Extrusion of Rice main effect QTL and its localization method and application |
| CN109662026A (en) * | 2019-02-27 | 2019-04-23 | 江油市川江水稻研究所 | A method of utilizing cytoplasmic male sterile rice breeding rice photo-thermo-sensitive genic male sterile line |
| CN110547189A (en) * | 2019-10-16 | 2019-12-10 | 四川发生种业有限责任公司 | novel method for breeding rice cytoplasmic male sterile line by using rice cytoplasmic male sterile line |
| CN112314429A (en) * | 2020-10-29 | 2021-02-05 | 海南波莲水稻基因科技有限公司 | Breeding method of rice nuclear male sterility maintainer line |
| CN112314429B (en) * | 2020-10-29 | 2023-03-28 | 海南波莲水稻基因科技有限公司 | Breeding method of rice nuclear male sterility maintainer line |
| CN115777524A (en) * | 2021-12-17 | 2023-03-14 | 安徽省农业科学院水稻研究所 | Method for creating novel rice sterile line |
| CN116420613A (en) * | 2023-05-30 | 2023-07-14 | 福建华晋农业技术开发有限公司 | Rapid backcross stable cultivation method for three-line sterile line of rice |
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