CN104082054B - A kind of method utilizing high temperature stress to identify the sterile pnca gene type of rape - Google Patents
A kind of method utilizing high temperature stress to identify the sterile pnca gene type of rape Download PDFInfo
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- CN104082054B CN104082054B CN201410231271.8A CN201410231271A CN104082054B CN 104082054 B CN104082054 B CN 104082054B CN 201410231271 A CN201410231271 A CN 201410231271A CN 104082054 B CN104082054 B CN 104082054B
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Abstract
The invention discloses a kind of method utilizing high temperature stress to identify the sterile pnca gene type of rape, it is by certain high temperature stress process, the genotype of sterile strain to be measured is differentiated according to the difference of process latter two sterile strain sterile stability, combine the advantage of test cross method and molecular labeling method, namely simple to operate, realize the present age detect, to laboratory condition and technical requirement low, reliable results.
Description
Technical field
The present invention relates to rape sterile pnca gene V-neck V territory, is exactly a kind of method utilizing high temperature stress to identify the sterile pnca gene type of rape.
Background technology
Recessive epistatically GMS is a kind of important channel that rape heterosis utilizes, this utilization ways obtained National Technical patent of invention (ZL97125803.1) in 2003, this patented method successfully realizes rapeseed plant recessive Genetic Sterility and to isozygoty dual purpose lines, interim maintainer and the production of hybrid seeds of restorer three series mating completely, first produce complete sterile line with the sterile strain of isozygotying in dual purpose lines and interim maintainer completely during the production of hybrid seeds, then produce crossbreed by complete sterile line and restorer.Therefore in line with genic sterile recessive epistatic interaction in system, have two kinds of sterile strains of genotype, in the dual purpose lines that namely isozygotys, sterile pnca gene type isozygotys, and is homozygous sterile strain; Complete sterile line genotype is heterozygosis, is the sterile strain of heterozygous.
The sterile strain of isozygotying in theory in dual purpose lines is homozygous sterile strain, and itself and complete temporary maintainer line test cross can produce the complete sterile line that sterile plant rate is 100%.Because rape is Cross Pollinated, very easily mix in reproductive process, in the dual purpose lines that therefore isozygotys, be very easily mixed into the sterile strain of heterozygous.These two kinds of sterile strains of genotype are difficult to distinguish in appearance, sterile for heterozygous strain cannot be extracted when producing complete sterile line, cause the complete sterile line sterile plant rate produced to decline, have a strong impact on seed production efficiency and purity.Therefore differentiate that these two kinds sterile strain types are extremely important on breed and production, the method for these two kinds of sterile strains of genotype of qualification at present has test cross method and molecular labeling method.
Test cross method uses interim maintainer and sterile strain test cross to be identified completely usually, and the Fertility segregation ratio according to test cross offspring differentiates sterile pnca gene type, if test cross offspring is all sterile strain, sterile strain to be identified is then homozygous sterile strain; If test cross offspring fertile plant and sterile strain segregation ratio are 1:1, sterile strain to be identified is then the sterile strain of heterozygous.This authentication method advantage is simple to operate, and shortcoming needs test cross, and needs just to know the genotype of sterile strain the second season (year), and spended time is long.
Molecular labeling method will first find and the closely linked molecular labeling in upper suppressor site, and then detect sterile strain with this molecular labeling, the presence or absence according to key band judges genotype.The method advantage is that detection speed is fast, can realize detecting the present age.Shortcoming requires higher to laboratory and operating personnel, only applies in minority unit at present.
Summary of the invention
The object of the invention is to solve the technical problem of two kinds of sterile pnca gene type qualifications in line with genic sterile recessive epistatic interaction in system,
Above-mentioned purpose is realized by following scheme:
Utilize high temperature stress to identify a method for the sterile pnca gene type of rape, it is characterized in that:
Comprise the following steps:
(1), field planting: the normal plantation in season of rape to be measured;
(2), transplant: rape is moved into the basin alms bowl with bottom outlet by squaring period, the strain of every basin one, and rape is planted into land for growing field crops together with basin alms bowl;
(3), high temperature stress: peduncle-growing period for rapeseed is when the clear and legible rape fertile plant of naked eyes and sterile strain, the sterile strain of band basin alms bowl is placed in phytotron or illumination box, control to process 14-16 hour under the illumination condition of 34-36 DEG C, remove basin alms bowl immediately, land for growing field crops is transplanted in sterile strain;
(4), fertility investigation: the flowers are in blossom decontrols and begin from sterile strain first, investigates the list flower fertility that every main inflorescence of sterile strain and top four primary branch main sequences are open day by day, extract immediately, continue one week of investigation after having investigated; By 7 grades of Fertility rating statistic laws, single flower fertility is divided into 0-6 level according to the number of its Fertile stamen, and wherein 0 grade is 6 stamen complete sterilities, and 1-6 level has 1-6 Fertile stamen respectively successively;
Fertility index M FI calculates: occur in joint account one week those days of 1 grade and above flower all by the average stamen number of average fertility index M FI=of investigation flower=(1 × 1 grade spend number+2 × 2 grades spend number+... spend number for+6 × 6 grades)/investigation flower number; If all investigated flowers are 0 grade in the week, then MFI is 0.
(5), genotype judges: work as MFI=0, then sterile strain to be measured is homozygous sterile strain; Work as MFI>1, then sterile strain to be measured is the sterile strain of heterozygous.
A kind of described method utilizing high temperature stress to identify the sterile pnca gene type of rape, is characterized in that: the illumination temperature of described step (3) is 35 DEG C, and the processing time is 15 hours.
Operation principle of the present invention is: research finds that the susceptibility of the sterile strain sterility of two types to temperature there are differences in rapeseed plant recessive epistatic interaction Genetic Sterility system, at a lower temperature, the sterile strain of two types all shows thoroughly sterile, be difficult to distinguish, and under certain high temperature stress, the sterility of the sterility strain more sterile than heterozygous of homozygous sterile strain is more stable, according to this phenomenon (principle), just can identify the genotype of the sterile strain of this two type.
Beneficial effect of the present invention is: by certain high temperature stress process, the genotype of sterile strain to be measured is differentiated according to the difference of process latter two sterile strain sterile stability, combine the advantage of test cross method and molecular labeling method, namely simple to operate, realize detecting the present age, to laboratory condition and technical requirement low, reliable results.
Embodiment
Utilize high temperature stress to identify a method for the sterile pnca gene type of rape, comprise the following steps:
(1), field planting: the normal plantation in season of rape to be measured;
(2), transplant: rape is moved into the plastic basin alms bowl with the diameter 10cm of bottom outlet by squaring period, the strain of every basin one, and rape is planted into land for growing field crops together with basin alms bowl;
(3), high temperature stress: peduncle-growing period for rapeseed is when the clear and legible rape fertile plant of naked eyes and sterile strain, the sterile strain of band basin alms bowl is placed on phytotron or illumination box, under 35 DEG C of illumination conditions, process 15 hours, remove basin alms bowl immediately, land for growing field crops is transplanted in sterile strain;
(4), fertility investigation: the flowers are in blossom decontrols and begin from sterile strain first, investigates the list flower fertility that every main inflorescence of sterile strain and top four primary branch main sequences are open day by day, extract immediately, continue one week of investigation after having investigated; By 7 grades of Fertility rating statistic laws, single flower fertility is divided into 0-6 level according to the number of its Fertile stamen, and wherein 0 grade is 6 stamen complete sterilities, and 1-6 level has 1-6 Fertile stamen respectively successively;
Fertility index M FI calculates: occur in joint account one week those days of 1 grade and above flower all by the average stamen number of average fertility index M FI=of investigation flower=(1 × 1 grade spend number+2 × 2 grades spend number+... spend number for+6 × 6 grades)/investigation flower number; If all investigated flowers are 0 grade in the week, then MFI is 0.
(5), genotype judges: work as MFI=0, then sterile strain to be measured is homozygous sterile strain; Work as MFI>1, then sterile strain to be measured is the sterile strain of heterozygous.
To the rape of qualification be needed in the normal plantation in season of large Tanaka.Moved in the plastic basin alms bowl with the diameter 10cm of bottom outlet squaring period at rape, the strain of every basin one, then rape is planted into large field together with basin alms bowl.By observing, 40 strains are chosen in the sterile strain of androecium abortion at rape peduncle-growing period for rapeseed, process 15 hours (phytotron, illumination box) under front 20 strains being placed on 35 DEG C of conditions, rear 20 strains process 15 hours as a control group at normal temperatures, remove basin alms bowl afterwards, transplant into land for growing field crops.From first of sterile strain, the flowers are in blossom decontrols and begin, and investigates every main inflorescence of sterile strain and the open list flower fertility of four, top primary branch main sequence day by day, extract immediately, continue one week of investigation after having investigated.Record in the week and occur that those days of 1 grade and above flower are all by the average fertility index (MFI) of investigation flower; If all investigated flowers are 0 grade in the week, then MFI is 0.The results are shown in Table 1.
table 1
Claims (1)
1. utilize high temperature stress to identify a method for the sterile pnca gene type of rape, it is characterized in that:
Comprise the following steps:
(1), field planting: the normal plantation in season of rape to be measured;
(2), transplant: rape is moved into the basin alms bowl with bottom outlet by squaring period, the strain of every basin one, and rape is planted into land for growing field crops together with basin alms bowl;
(3), high temperature stress: peduncle-growing period for rapeseed is when the clear and legible rape fertile plant of naked eyes and sterile strain, the sterile strain of band basin alms bowl is placed in phytotron or illumination box, control to process 14-16 hour under the illumination condition of 34-36 DEG C, remove basin alms bowl immediately, land for growing field crops is transplanted in sterile strain;
(4), fertility investigation: the flowers are in blossom decontrols and begin from sterile strain first, investigates the list flower fertility that every main inflorescence of sterile strain and top four primary branch main sequences are open day by day, extract immediately, continue one week of investigation after having investigated; By 7 grades of Fertility rating statistic laws, single flower fertility is divided into 0-6 level according to the number of its Fertile stamen, and wherein 0 grade is 6 stamen complete sterilities, and 1-6 level has 1-6 Fertile stamen respectively successively;
Fertility index M FI calculates: be designated as effectively investigate day for the same day having the flower of more than 1 grade and 1 grade classification to occur, investigated all effective investigation days in joint account one week the average stamen number of average fertility index M FI=of flower=(1 × 1 grade spend number+2 × 2 grades spend number+... spend number for+6 × 6 grades)/investigation flower number; If all investigated flowers are 0 grade in the week, then MFI is 0;
(5), genotype judges: work as MFI=0, then sterile strain to be measured is homozygous sterile strain; Work as MFI>1, then sterile strain to be measured is the sterile strain of heterozygous.
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Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4658084A (en) * | 1985-11-14 | 1987-04-14 | University Of Guelph | Hybridization using cytoplasmic male sterility and herbicide tolerance from nuclear genes |
| US4658085A (en) * | 1985-11-14 | 1987-04-14 | University Of Guelph | Hybridization using cytoplasmic male sterility, cytoplasmic herbicide tolerance, and herbicide tolerance from nuclear genes |
| EP0139674B1 (en) * | 1983-03-16 | 1987-11-04 | Institut National De La Recherche Agronomique | Method for the somatic hybridization of colza |
| CN1586160A (en) * | 2004-09-09 | 2005-03-02 | 中国农业科学院油料作物研究所 | Rape cytoplasmic male sterility + self incompatibility hybrid superiority using method |
| CN1778163A (en) * | 2004-11-22 | 2006-05-31 | 湛江海洋大学 | Cultivation of cryophilous plant hybrid species except rape by high-temperature sterile line |
| CN1868260A (en) * | 2006-06-05 | 2006-11-29 | 华中农业大学 | Method for breeding male sterile series of envelopped senvy cytoplasm |
| GB2429462A (en) * | 2005-08-23 | 2007-02-28 | Elsoms Seeds Ltd | Male sterile swede plants and F1 hybrids |
| CN101189952A (en) * | 2006-11-23 | 2008-06-04 | 国家杂交水稻工程技术研究中心天津分中心 | High yield low risk reproducing technology for temperature sensitive type genic male sterile rice in high altitude low yield field |
| CN101595839A (en) * | 2009-07-02 | 2009-12-09 | 西北农林科技大学 | Breeding and application of rape environmental sensitive genic male sterile line with self-incompatibility characteristic |
-
2014
- 2014-05-28 CN CN201410231271.8A patent/CN104082054B/en not_active Expired - Fee Related
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0139674B1 (en) * | 1983-03-16 | 1987-11-04 | Institut National De La Recherche Agronomique | Method for the somatic hybridization of colza |
| US4658084A (en) * | 1985-11-14 | 1987-04-14 | University Of Guelph | Hybridization using cytoplasmic male sterility and herbicide tolerance from nuclear genes |
| US4658085A (en) * | 1985-11-14 | 1987-04-14 | University Of Guelph | Hybridization using cytoplasmic male sterility, cytoplasmic herbicide tolerance, and herbicide tolerance from nuclear genes |
| CN1586160A (en) * | 2004-09-09 | 2005-03-02 | 中国农业科学院油料作物研究所 | Rape cytoplasmic male sterility + self incompatibility hybrid superiority using method |
| CN1778163A (en) * | 2004-11-22 | 2006-05-31 | 湛江海洋大学 | Cultivation of cryophilous plant hybrid species except rape by high-temperature sterile line |
| GB2429462A (en) * | 2005-08-23 | 2007-02-28 | Elsoms Seeds Ltd | Male sterile swede plants and F1 hybrids |
| CN1868260A (en) * | 2006-06-05 | 2006-11-29 | 华中农业大学 | Method for breeding male sterile series of envelopped senvy cytoplasm |
| CN101189952A (en) * | 2006-11-23 | 2008-06-04 | 国家杂交水稻工程技术研究中心天津分中心 | High yield low risk reproducing technology for temperature sensitive type genic male sterile rice in high altitude low yield field |
| CN101595839A (en) * | 2009-07-02 | 2009-12-09 | 西北农林科技大学 | Breeding and application of rape environmental sensitive genic male sterile line with self-incompatibility characteristic |
Non-Patent Citations (4)
| Title |
|---|
| 属间体细胞杂交创建甘蓝型油菜细胞质雄性不育系及其鉴定;胡琼等;《中国农业科学》;20040310;第37卷(第3期);第333-338页 * |
| 油菜品种资源细胞质育性类型及核不育基因型鉴定;文雁成等;《中国油料作物学报》;20030630;第25卷(第2期);第1-4页 * |
| 油菜温敏雄性不育系373S的选育;于澄宇等;《中国农学通报》;20070731;第23卷(第7期);第245-248页 * |
| 甘蓝型油菜新型细胞质雄性不育系NCa不育胞质类型的分子鉴定;危文亮等;《中国农业科学》;20051031;第38卷(第10期);第1965-1972页 * |
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