CN102640700A - Cabbage type rape Ogura cytoplasmic male sterility (OGU CMS) restorer and transformation method and application thereof - Google Patents
Cabbage type rape Ogura cytoplasmic male sterility (OGU CMS) restorer and transformation method and application thereof Download PDFInfo
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Abstract
The invention discloses a cabbage type rape Ogura cytoplasmic male sterility (OGU CMS) restorer and an obtained derivative restorer material after the cabbage type rape Ogu CMS restorer is transferred, wherein the preservation number of the Chinese microbial culture preservation management committee center for general microbiology is CGMCC No. 6001, and the plant name is Ogu CMS restorer CLR650. A transformation method of the cabbage type rape OgU CMS restorer comprises the following steps: using a cruciferae resource containing Ogu CMS exogenous restoring genes as a restoring gene donor parent, utilizing a grafting technology to overcome distant hybridization reproductive dysfunction, leading intergeneric radish restoring genes into cabbage type rapes, continuously culturing fertile plants with chlorosis, and obtaining the restorer CLR650 which can be fertile in selfing and test-crossing after screening, selfing and backcross screening for multiple generations. The restorer material is low in cost, good in safety and capable of being applied to a cabbage type rape Ogu CMS pollination system.
Description
Technical field
The invention belongs to rape heterosis and utilize research field, relate in particular to a kind of cabbage type rape and recover material and transfer method and application.
Background technology
The male sterile with radish cytoplasm (Ogu CMS) that Ogura (1968) creates has sterility and stablizes, is easy to distinguishing features such as maintenance, is acknowledged as heterosis utilization approach ideal in the whole rape crop.In the cabbage type rape application study, Bannerot (1974) utilizes backcross method to breed cabbage type rape Ogu CMS at first, and this sterile type is inharmonious because of matter nuclear, has negative characteristics such as leaf look low temperature minus green, nectary shortage.Pelletier (1983) carries out protoplast with cabbage type rape and radish and merges, and has bred Ogu-INRA CMS of new generation, has both kept original sterile stability, has overcome problems such as leaf look minus green again.Discovering thereafter can not find Ogu CMS and recovers gene in the whole Brassicas crop, the recovery gene exists only in the radish varieties in Japan or Europe.Heyn (1976) attempts through sexual hybridization radish being recovered gene and imports to cabbage type rape, obtains the Ogu CMS F that fertility is partly recovered
1The intergeneric cross kind, Pelletier (1983) merges the cybrid from the protoplast of cabbage type rape and radish, and the Ogu CMS that has obtained the cabbage type rape proterties recovers strain.Recover the radish chromosome segment of gene with certain-length owing to import in the cabbage type rape, and the material ripening rate is low, economical character is poor with recovering, with serial problem such as high-sulfur glycosides gene close linkage, the transformation of good homozygous recovery material is difficulty very.Primard-Brisset (2000) recovers gene through irradiation treatment reorganization, has reduced the radish fragment to recovering the influence of material, and it is one of technological that cabbage type rape Ogu CMS becomes rape heterosis utilizations of tool potentiality.
China introduces cabbage type rape Ogu CMS the end of the seventies in last century; Aspect the transformation of recovery material; Li Xufeng (2001) once imported to cabbage type rape Ogu CMS with the recovery gene among the blue beggar of radish mutation; And obtaining addition chromosome recovery materials A d-6, homozygous cabbage type rape Ogu CMS recovers material screening and never makes great progress.Patent protection is all implemented to the Ogu CMS recovery material of breeding in countries in the world at present, and creating new O gu CMS recovery material (source) has important function to the range of application that enlarges cabbage type rape Ogu CMS.
Summary of the invention
The technical problem that the present invention will solve is the deficiency that overcomes prior art; Provide the cabbage type rape Ogu CMS that a kind of cost is low, safety good, can be used in the cabbage type rape Ogu CMS pollination system to recover system; Also provide a kind of simple to operate, directly perceived, effect obviously, cabbage type rape Ogu CMS with important hereditary meaning recovers the transfer method of system, simultaneously corresponding this cabbage type rape Ogu CMS that provides recovers to tie up to the application in the cabbage type rape Ogu CMS pollination system.
For solving the problems of the technologies described above; The present invention has created and has a kind ofly recovered that the different cabbage type rape Ogu CMS in gene original parent source recovers system and by the series derivatives recovery material of this recoverys system through backcrossing, obtaining after the selfing transformation; The preserving number that said cabbage type rape Ogu CMS recovery ties up to China Committee for Culture Collection of Microorganisms common micro-organisms center is CGMCC No.6003, and it is CLR650 that its plant name is called Ogu CMS recovery.It is that CLR650 transfers to China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC) preservation on April 16th, 2012 that cabbage type rape Ogu CMS of the present invention recovers, and the address of depositary institution is positioned at Beijing Institute of Microorganism, Academia Sinica.
As a total technical conceive, the present invention also provides a kind of above-mentioned cabbage type rape Ogu CMS to recover the transfer method of system, may further comprise the steps:
(1) grafting imports the external source radish and recovers gene: with cabbage type rape Ogu CMS leaf look minus green male sterile line (Brassica Napus.L) and the Cruciferae resource that contains Ogu CMS external source recovery gene as the grafting parent; During grafting with the sterile strain of said Ogu CMS leaf look minus green male sterile line as scion; The educated strain of said Cruciferae resource is as stock; Scion becomes to live when blooming; Get the fertile flower powder of stock and the sterile flower of scion and hybridize, obtaining to import has the chromosomal cabbage type rape distant hybrid progeny of radish; Carry the recovery gene on the said radish chromosome; The method of this step can effectively overcome distant hybridization genitality obstacle, and on average tying the angle rate is about 16%, and average ripening rate reaches about 17%, the introduction method of other edge foreign gene far away relatively, and the grafting operation of this step has more directly, easier importing effect;
(2) screening cabbage type rape proterties: obtain the cabbage type rape distant hybrid progeny from step (1); Utilize the characteristic of separation of leaf look minus green proterties and leaf look minus green degree and plant forms performance dependency relation; Select the educated strain of leaf look minus green to continue to cultivate; After a plurality of generation screenings; Radish is recovered the assortment of genes to cabbage type rape chromosome; The Ogu CMS leaf look minus green that acquisition shows as the cabbage type rape proterties normally recovers strain (2N=38) (in general, the mosaic type that can obtain to show as the cabbage type rape proterties through 5～6 generations recovers strain, and the Ogu CMS leaf look minus green that can obtain the Wild cabbage type proterties through 8～9 generations normally recovers strain); In this step, utilize the distant hybridization of Ogu CMS leaf look minus green character screening to recover strain, got rid of the additional radish chromosome in the distant hybrid progeny quickly, realized the genomic reorganization of cabbage type rape (2N=38); But; Because the Ogu CMS leaf look minus green that obtains normally recovers a little less than the recovery capability of strain; Mostly fertility performance in the screening process is that partly sterile and mosaic type can be educated, and flower is relatively poor for white, fecundity, and natural solid only can the receipts of recovery individual plant planted 30～50; The recovery strain rate in inferior generation is generally about 10%, needs further to screen, isozygoty through follow-up step (3);
(3) purifying Ogu CMS recovers system: with Ogu-INRA CMS leaf look normal male sterile line (Brassica Napus.L) as female parent; The Ogu CMS leaf look minus green of cultivating with above-mentioned steps (2) normally recovers strain and hybridizes as male parent; From filial generation, obtaining the leaf look is that separation green, that pattern is yellow recovers strain; Fecundity be improved significantly, again through a plurality of generations give the screening of backcrossing certainly, obtaining selfing, test cross, to be the Ogu CMS that can the educate full recovery of isozygotying be CLR650; The efficient in operation of this step overcome the radish chromosome segment to recover the strain fertility separate the influence that causes (separate like self-fertility strain rate mostly be 50%, test cross can educate strain separate be mostly that 10%～20% fertility departs from), strengthened the reorganization probability between radish recovery gene and cabbage type rape genome.
The basic principle of the above-mentioned transfer method of the present invention is: to contain Cruciferae resource that Ogu CMS external source recovers gene as recovering the genetic donor parent; Utilize graft technology to overcome distant hybridization genitality obstacle earlier; Radish recovery gene imports in the cabbage type rape between will belonging to;, strengthen system of selection and combine with preferred through selfing, conventional breeding method such as backcross thereafter, overcome recover gene with radish fragment adverse effect that normal heredity is separated.Through this method, the present invention finally realized reorganization that radish recovers gene and isozygotied, and bred cabbage type rape Ogu CMS and isozygotied and recover material C LR650.
Above-mentioned transfer method; In the said step (3); Said from giving the offspring who separates that backcrosses all preferably to adopt the parallel back-and-forth method of microcommunity, many strains system to strengthen screening; This method can progressively improve the recovery rate of recovering strain, and part selfing strain system possibly break through Ogu CMS heterozygous and recover the obstacle that strain generally shows as the separation of 1:1 fertility, and finally reaches homozygotic state.
In the above-mentioned transfer method; The Cruciferae resource that the said Ogu of containing CMS external source is recovered gene is trailing plants-indigo plant (Raphanobrassica); Be the polyploid plant (2N=58) that radish (2N=20) and cabbage type rape (2N=38) natural crossing form, its original parent is that the source is different with the existing similar recovery of breeding.
Above-mentioned transfer method; In the said step (1); Significantly improved the affinity that cabbage type rape and trailing plants-indigo plant are carried out distant hybridization through grafting; Its average knot angle rate reaches 15%～17%, and average ripening rate reaches 16%～18%, imports to cabbage type rape (then generally adopting cell-fusion techniques to solve this difficult problem in the prior art) thereby can successfully the radish external source be recovered gene.
As a total technical conceive, the present invention also provides the cabbage type rape Ogu CMS that obtains after a kind of above-mentioned transformation to recover the application of system's (containing by the recovery material of deriving that obtains after its transformation) in cabbage type rape Ogu CMS pollination system.Relative other heterosis utilization method, clear superiorities such as that application of the present invention has is easy and simple to handle, production safety.
Compared with prior art; The invention has the advantages that: the grafting that the present invention adopts imports the transfer method that external source is recovered gene; Can be through the exchange of the physiologically substance between graft stock and scion; Overcome distant hybridization genitality obstacle effectively, improve the importing frequency of target gene, this importing to other external source beneficial gene also has important reference role.The present invention also utilizes the leaf look minus green characteristic of early stage cabbage type rape Ogu CMS; The cabbage type rape proterties of screening distant hybrid progeny; Have select directly perceived, effect obviously, operate easy technical advantage, to the influence of selecting effect a new technological approaches is provided for getting rid of additional radish chromosome.In addition, the homozygous recovery material C of the cabbage type rape Ogu CMS LR650 of the final seed selection of the present invention is to exist tangible molecular labeling difference with external similar recovery, and this has significance to genetic research that expansion Ogu CMS recovers material.
It is that CLR650 has transferred to China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC) preservation on April 16th, 2012 that cabbage type rape Ogu CMS of the present invention recovers; The address of depositary institution is positioned at Beijing Institute of Microorganism, Academia Sinica; Deposit number is CGMCC No.6003, and it is the classification called after cabbage type rape (Brassica napus) of CLR650 that cabbage type rape Ogu CMS recovers.
Description of drawings
Fig. 1 be in the embodiment of the invention to recovering gene linkage molecular labeling Preliminary detection comparison diagram, wherein, white portion is disclosed document results (2005) such as Primard-Brisset; The ash color part is the CLR650PCR amplification for cabbage type rape Ogu CMS recovery of the present invention.
Fig. 2 is the profile photo of original parent trailing plants-indigo plant (Raphanobrassica) 94-1 of using in the embodiment of the invention.
Fig. 3 is the profile photo of the original parent Ogu CMS leaf look minus green male sterile line H13A that uses in the embodiment of the invention.
Fig. 4 is the form photo that grafting imports grafting elongation when recovering gene in the embodiment of the invention.To be designated as with trailing plants-indigo plant be that stock, the sterile strain of Ogu CMS minus green are the grafting part of scion to arrow among the figure.
Fig. 5 is the form photo in grafting flowering stage when grafting import to be recovered gene in the embodiment of the invention.Arrow is designated as grafting part among the figure.
Fig. 6 is the fertility performance that Ogu CMS leaf look minus green recovers strain in the embodiment of the invention, and arrow is designated as and spends part in vain and can educate (flower pesticide tiny have a small amount of pollen) among this figure.
Fig. 7 is the fertility performance that Ogu CMS leaf look minus green recovers strain in the embodiment of the invention, and arrow is designated as and spends chimeric educating (fertile flower and sterile flower coexistence) in vain among this figure.
Fig. 8 is the fertility performance that Ogu CMS leaf look minus green recovers strain in the embodiment of the invention, and arrow is designated as to spend in vain normally and can educates (pollen amount is normal) among this figure.
Fig. 9 is CLR650 for the cabbage type rape Ogu CMS recovery that is in seedling stage in the embodiment of the invention.
Figure 10 is CLR650 for the cabbage type rape Ogu CMS recovery that is in flower, angle phase in the embodiment of the invention.
Figure 11 is the transformation schematic flow sheet of CLR650 for cabbage type rape Ogu CMS recovery in the embodiment of the invention.
Embodiment
Below in conjunction with Figure of description and specific embodiment the present invention is further described.
Embodiment:
A kind of is CLR650 like Fig. 9, cabbage type rape Ogu CMS recovery of the present invention shown in Figure 10; The preserving number that this cabbage type rape Ogu CMS recovery ties up to China Committee for Culture Collection of Microorganisms common micro-organisms center is CGMCCNo.6003, and it is CLR650 that its plant name is called Ogu CMS recovery.
The cabbage type rape Ogu CMS recovery of present embodiment is that the transfer method of CLR650 is shown in figure 11, specifically comprises the steps.
1. the external source radish recovers the importing of gene
1.1 material
Mainly comprise: (AACCRR 2n=58), gave (genetic background is not quite clear) in 1993 by the Fu Ting of a Hua Zhong Agriculture University academician to trailing plants-indigo plant (Raphanobrassica) 94-1 as shown in Figure 2; (Brassic Napus.L 2N=38) tells researcher Canada in 1979 by the Cui De of Hu'nan Prov. Crops Research Inst. and introduces cabbage type rape Ogu CMS as shown in Figure 3 (leaf look low temperature) minus green male sterile line H13A.
1.2 implementation method
Autumn in 1994; Above-mentioned trailing plants-indigo plant (94-1) and Ogu CMS leaf look minus green male sterile line (H13A) are sowed (last ten-days period in September) same period; In spring next year (late Febuary), when the about 30cm of plant tongue height (referring to Fig. 4), cut by about 40 ° inclined-plane is reciprocity in tongue stem middle part with scalpel; With cutting part as scion; Cut plant as stock reciprocal grafting (inclined-plane pastes back, film wrapped mutually and ties up with fine rule), scion becomes to live when blooming (referring to Fig. 5), by scion sterile flower (H13A) * stock fertile flower (94-1), stock sterile flower (H13A) * scion fertile flower (94-1) dual mode pollination hybridization; (H13A * 94-1) as contrast, the maturing stage is investigated the cross-fertile effect of different disposal mode with non-graft hybridization processing.
1.3 investigation result
With scion sterile flower (H13A) * stock fertile flower (94-1), the average result rate is 15.6%, and average ripening rate is 17.4%, is respectively 14.1 times and 15.8 times of non-grafting contrast; With stock sterile flower (H13A) * scion fertile flower (94-1), the average result rate is 0.6% with average ripening rate, descends 0.5 percentage point than non-grafting contrast.Therefore; The scion sterile flower is the hybridization mode of pollen donor as pollen acceptor, stock fertile flower; Its cross-fertile rate is significantly higher than reverse hybridized and contrast hybridization; This shows also grafting mainly shows with the stock to be that pollen donor, scion are on the hybridization mode of pollen acceptor to the effect that improves the cross-fertile rate, and this graft hybridization mode can not only improve the distant hybridization ripening rate significantly, and the seed normal development of distant hybrids is also had tangible effect.
1.4 observe the restorative performance of filial generation
Next year plantation scion sterile flower (H13A) * stock fertile flower (94-1) first familiar generation is observed the restorative performance of filial generation.Utilize in the hybridization mode of scion sterile flower (H13A) * stock fertile flower (94-1); Obtain 32 cabbage type rape Ogu CMS and trailing plants-blue distant hybrids altogether; Wherein 20 is big many ribs of grain kind of trailing plants-blue type, and 12 is circular kind of granule of cabbage type rape type; The distant hybrids that obtains is seeded in the land for growing field crops; Amount to into seedling 21 strains; Emerge into individual plant alive show all that leafing look shallow minus green (maternal H13A is dark minus green), leaf are given birth to seta, opened pale yellow flower, plant type is loose etc. and plant between the proterties characteristic, the crossbreed (containing big grain many ribs kind and the circular kind of granule) that is obtained by grafting is true hybrid.Carry out fertility investigation to plant flowering stage, and wherein 2 strains have the part recovery capability, and its stamen contains a small amount of pollen a little less than column cap, flower pesticide elongated (half-breadth of about normal anther).Above result shows; Present embodiment supplies the examination material Ogu CMS that trailing plants-indigo plant (94-1) carries to recover gene; Import among the cabbage type rape Ogu CMS through sexual hybridization, obtain the cabbage type rape distant hybrid progeny that importing has radish chromosome (carry on the chromosome and recover gene).
2. the recovery strain of cabbage type rape proterties screening
2.1 material
The cabbage type rape distant hybrid progeny that obtains in the above-mentioned steps 1; Be used for the former minus green male sterile line H13A that test cross uses, provide by Hu'nan Prov. Crops Research Inst..
2.2 implementation method
Adopt the radish sterile cytoplasm directly to identify the restorative performance (homogeneity recovery material selective breeding) of selecting material; 1995～2004 years serves as to select background with Ogu CMS leaf look minus green male sterile line (H13A); Mainly utilize leaf look minus green character screening cabbage type rape proterties; The educated strain that per generation is chosen obvious minus green constitutes new round selection colony, and the Ogu CMS leaf look minus green that shows as the cabbage type rape properties and characteristics until acquisition normally recovers strain (normally can educate).In the selection course, Ogu CMS minus green recovers the general self-fruitfulness extreme difference of strain, mainly replaces selfed seed with natural kind, each strain from generation to generation be colony's number between 20～30 strains, the total group number is between 50～100 strains.In kytoplasm (H13A) background of Ogu CMS, leaf look minus green proterties and the plant forms of filial generation demonstrate tangible dependency relation, and general leaf look minus green is obvious more, and the cabbage type rape characteristic is remarkable more.Screen recovering strain as selected marker with leaf look minus green continuously, plant forms is progressively turned to by type between planting and is the cabbage type rape type, and the fertility performance is carried out the transition to and spent in vain mosaic type and can educate (referring to Fig. 7) by early stage spend in vain half sterile (referring to Fig. 6); Can educate (referring to Fig. 8) to spending in vain normally again, Ogu CMS minus green recovers strain and has showed the directed selection to the cabbage type rape proterties.
2.3 main result
Through adopting the implementation method of above-mentioned steps 2.2; The Ogu CMS leaf look minus green that altogether acquisition shows as the cabbage type rape proterties after 9 generations (natural solid, generation test cross transformation, generation self-fertility comprising 7 generations) normally recovers strain; This Ogu CMS leaf look minus green normally recovers strain and on proterties, shows as leaf look minus green, opens white flowers, its flower pesticide tubbiness, full; Pollen amount is sufficient, has reached normally on the floral organ exophenotype and can educate standard.Above result shows, Ogu CMS leaf look minus green is normally recovered the cabbage type rape proterties of strain and selects to reach basic demand, does not obviously change yet but Ogu CMS leaf look minus green normally recovers unfavorable proterties such as the total fecundity extreme difference of strain, restorative instability.
3. the cabbage type rape Ogu CMS recovery material screening that isozygotys
3.1 material
Comprise mainly in the above-mentioned steps 2 that the Ogu CMS leaf look minus green that shows as cabbage type rape proterties (leaf look minus green, open and spend in vain) that obtains normally recovers strain; Leaf look normal Ogu-INRA CMS (H28A) is by Hu'nan Prov. Crops Research Inst.'s transformation from the Ogu-INRA CMS crossbreed offspring separated strain of external introduction; 20 parts of the breeding for double low content in rapeseed materials that is used to backcross are provided by Hu'nan Prov. Crops Research Inst..
3.2 implementation method
2005～2011 years; With leaf look normal Ogu-INRA CMS (H28A) serves as to select background; Main through strengthening colony's recovery rate that screening improves selected plant system; The educated strain (containing the inbred line through backcrossing) that per generation is chosen the higher strain of recovery rate system gets into the next generation, and selfing can be educated entirely until obtaining, the full recovery material that isozygotys that recovers of test cross.It is solid normal that Ogu-INRA CMS recovers strain, mainly be with the selfing main, to backcross be auxilliary the screening, each strain of selfing from generation to generation coefficient is between 30～50, the combination strain of backcrossing ties up to 10～20, the total group number is between 1000～6000 strains.
3.3 main result
In Ogu-INRA CMS leaf look normal male sterile line (H28A) kytoplasm background, marked change has taken place in proterties such as recovery strain ripening rate.Ogu-INRA CMS leaf look normal male sterile line (H28A) is spent Ogu CMS leaf look minus green in vain with minus green and is normally recovered strain hybridization, at hybridization F
1In the 24 group of hill bodies in generation; 2 strains separate to recover strain leaf look transfer to positive evergreen leaf, pattern transfer to yellow flower, flower pesticide form fully and cabbage type rape be as good as; It is solid from giving different friendship all can reach normal pollination, and the average fruit of angle fruit of being tied is about about 18 of 5cm, every fruit grain number.But the system of the most of selfing strain of each segregative generation thereafter recovers the strain rate and remains between 40%～60%, and the strain of backcrossing system recovers the strain rate and remains between 20%～40%.Altogether 201 selfing strain systems are screened in the period of 2007～2011 years five; The strain system that meets or exceeds 3:1 fertility separation ratio is 12; Only account for and select 6% of total strain coefficient, show that the linkage relationship that recovers gene and other radish gene is difficult to through conventional breaking easily from (returning) friendship method.
Adopting microcommunity, many strains is parallel system of selection, the selfing strain be average recovery rate by 2007 36% bring up to 2011 63%, fertility greater than the separated strain system of 3:1 by 6 of bringing up to 2011 of 2 in 2009.The selfing strain system that was numbered CLR650 in 2011 at first reaches homozygotic state; The recovery strain rate of this strain system 2009 was 76% (F:S=16:5); 2010 is 94% (F: S=34: 2); 2011 be that 100% (F: S=63: 0), 63 strain inbreeding populations all show as normally can educate (referring to Fig. 9, Figure 10, Figure 11).Randomly draw 10 individual plants and carry out selfing; And respectively with the corresponding test cross of male sterile line (H28A); 10 selfing strain systems all show as 100% and can educate, and 10 paired tester strains all show as 100% and recover, thereby are giving two links of test cross to verify the recovery capability that CLR650 had certainly.
4. cabbage type rape Ogu CMS recovers the Preliminary Applications of material
4.1 material
Comprise mainly in the above-mentioned steps 3 and to isozygoty the Ogu CMS that obtains to recover be CLR650 that male sterile line H20A, H31A, H33A by separating sterile strain transformation in the Ogu-INRA CMS filial generation are provided by Hu'nan Prov. Crops Research Inst..
4.2 implementation method
2010～2011 years; With H28A, H31A, H33A is female parent, and the recovery of isozygotying is that CLR650 is that male parent is joined hybrid combination in the pilot scale of gauze isolation environment, and production of hybrid seeds father, maternal row compare 1:3; Florescence is placed honeybee pollination, 40 square metres of the production of hybrid seeds sub-district areas of each combination (rolling over 0.06 mu).Behind the results seed, branch sub-district three repetition randomized arrangement are carried out the output comparative test, and 20 square metres of every sub-district areas (rolling over 0.03 mu) are contrast No. 4 with the assorted oil in POL CMS Hybrid Hunan, and carry out the proterties investigation by PR than test method and produce with survey.
4.3 main result
Utilize to recover be CLR650 produce Ogu CMS crossbreed obtained good yield result with the purity result, during the production of hybrid seeds of three hybrid combinations of this test is produced, combination H28A * CLR650, real 7.43 kilograms of the cell productions of receiving, 123.83 kilograms of folding per mu yields; Combination H31A * CLR650, real 8.10 kilograms of the cell productions of receiving, 135.00 kilograms of folding per mu yields; Combination H33A * CLR650, real 7.78 kilograms of the cell productions of receiving, 129.66 kilograms of folding per mu yields.Purity detecting is carried out in three hybrid combinations to results, and its purity of hybrid is respectively 100%, 99% and 99%, is significantly higher than the primary standard (90%) of Chinese cabbage type rape purity of hybrid quality.
Be that the crossbreed that CLR650 produces carries out in the output comparative test to recover, combination H28A * CLR650,15.8 kilograms of three sub-district effective yields, 175.5 kilograms of folding per mu yields; Combination H31A * CLR650,192.2 kilograms of three sub-district effective yields, folding per mu yield kilogram, combination H33A * CLR650; 16.4 kilograms of three sub-district effective yields, 182.2 kilograms of folding per mu yields, comparison is according to No. 4 (170.4 kilograms of folding per mu yields) raising the output 3.0% of the assorted oil in Hunan respectively; 12.8%, 6.9%, showed good yield potential.
It is that CLR650 is different with the Ogu CMS recovery material that adopts the cytoplasm integration technology to breed in the world that the cabbage type rape Ogu CMS that the inventive method obtains recovers; The recovery material system of the present invention's initiative directly utilizes sexual hybridization from recover genetic donor parent (the for example trailing plants of present embodiment-blue 94-1), to obtain to recover gene, and the original parent of employing and transfer method, transformation process all are different from existing Ogu CMS and recover the breeding technique of system.
The cabbage type rape Ogu CMS recovery that present embodiment is obtained is that CLR650 carries out the molecular labeling Preliminary Identification; Receive the patent protection contrast to recover material owing to lack with comparing; The method that adopts is with reference to the open primer of Hu (2008) with Primard-Brisset (2005); Choosing 1 specific mark (BnRFO-AS2F/BnRFO-AS2F) that recovers gene (Rf0) design based on Ogu CMS detects with the radish mark (ScH03, ScA14, SG34, BoIJon, PGIint) that recovers gene linkage with 5; And recovery system (RRH1, R113, R2000) molecular labeling linkage map of directly testing result and Primard-Brisset (2005) being enumerated in the text compares, and the molecular labeling primer sequence is seen table 1.
The specific mark of table 1:RF0 and the primer sequence of linked marker
Preliminary detection result shows; It is that CLR650 is to recovering the specific molecular marker (BnRFO-AS2F/BnRFO-AS2F) of gene (Rf0) design based on Ogu CMS that the Ogu CMS of transformation of the present invention recovers; Its selfing individual plant and test cross individual plant all are positive; Contrast the sterile strain reaction that then is negative, show that CLR650 has the total gene (Rf0) that recovers of Ogura CMS.In the detection of 5 chain radish molecular labelings, ScA14, SG34, three marks of BoIJon are positive, the reaction that is negative of ScH03, two marks of PGIint.Three linked marker figure that recover system of RRH1, R113, R2000 that this result and Primard-Brisset (2005) are enumerated compare (as shown in Figure 1); CLR650 is that RRH1 lacks two marks (ScH03, PGIint) than recovering; Than recovering is that R113 lacks a mark (PGIint), identical with R2000 in existing detection range.Show that thus CLR650 is that RRH1, R113 exist notable difference on the gene order of radish fragment with external similar recovery at least.
Therefore; Recover material compared with the cabbage type rape Ogu CMS that breeds in the world; The recovery of the present invention's screening is that CLR650 has identical Ogu CMS restore funcitons; All the tool radish recovers gene (Rf0); But then exist notable difference at it on the molecular labeling of radish fragment, the present invention has not only created a kind of new cabbage type rape Ogu CMS and has recovered the transfer method of system, and thus obtained Ogu CMS to recover be that CLR650 exists and is different from other and breeds the marker characteristic that recovers system.
Claims (7)
1. a cabbage type rape Ogu CMS recovery system reaches by the recovery material of deriving that obtains after its transformation; The preserving number that said cabbage type rape Ogu CMS recovery ties up to China Committee for Culture Collection of Microorganisms common micro-organisms center is CGMCCNo.6003, and it is CLR650 that its plant name is called Ogu CMS recovery.
2. a cabbage type rape Ogu CMS as claimed in claim 1 recovers the transfer method of system, may further comprise the steps:
(1) grafting imports the external source radish and recovers gene: with cabbage type rape Ogu CMS leaf look minus green male sterile line (Brassica Napus.L) and the Cruciferae resource that contains Ogu CMS external source recovery gene as the grafting parent; During grafting with the sterile strain of said Ogu CMS leaf look minus green male sterile line as scion; The educated strain of said Cruciferae resource is as stock; Scion become to be lived and when blooming; Get the hybridization of stock fertile flower powder and scion sterile flower, obtaining to import has the chromosomal cabbage type rape distant hybrid progeny of radish; Carry the recovery gene on the said radish chromosome;
(2) screening cabbage type rape proterties: obtain the cabbage type rape distant hybrid progeny from step (1); Select the educated strain of leaf look minus green to continue to cultivate; Screening through a plurality of generations; Radish is recovered the assortment of genes to cabbage type rape chromosome, and the Ogu CMS leaf look minus green that acquisition shows as the cabbage type rape proterties normally recovers strain;
(3) purifying Ogu CMS recovers system: with Ogu-INRA CMS leaf look normal male sterile line (Brassica Napus.L) as female parent; The Ogu CMS leaf look minus green of cultivating with above-mentioned steps (2) normally recovers strain and hybridizes as male parent; From filial generation, obtaining the leaf look is that separation green, that pattern is yellow recovers strain; Again through a plurality of generations give the screening of backcrossing certainly, obtaining selfing, test cross, to be the Ogu CMS recovery that can educate entirely be CLR650.
3. cabbage type rape Ogu CMS according to claim 2 recovers the transfer method of system, it is characterized in that: in said step (3), said from giving the offspring who separates that backcrosses all to adopt the parallel back-and-forth method of microcommunity, many strains system to screen.
4. cabbage type rape Ogu CMS according to claim 2 recovers the transfer method of system; It is characterized in that: in above-mentioned steps (1); The Cruciferae resource that the said Ogu of containing CMS external source is recovered gene is trailing plants-indigo plant (Raphanobrassica), and this trailing plants-indigo plant is the polyploid plant that radish and cabbage type rape natural crossing form.
5. cabbage type rape Ogu CMS according to claim 2 recovers the transfer method of system; It is characterized in that: in the said step (1); Carry out cabbage type rape and trailing plants-blue distant hybridization with grafting, it is on average tied the angle rate and reaches 15%～17%, and average ripening rate reaches 16%～18%.
6. cabbage type rape Ogu CMS according to claim 2 recovers the transfer method of system; It is characterized in that: in the said step (2); The Ogu CMS leaf look minus green that acquisition shows as the cabbage type rape proterties normally recovers strain and is meant that the mosaic type that can obtain to show as the cabbage type rape proterties through 5～6 generations recovers strain, can obtain the Ogu of Wild cabbage type proterties through 8～9 generations
CMS leaf look minus green normally recovers strain.
7. a cabbage type rape Ogu CMS recovery as claimed in claim 1 is the application of recovery material in cabbage type rape Ogu CMS pollination system of deriving that reaches by obtaining after its transformation.
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102986523A (en) * | 2012-11-30 | 2013-03-27 | 江苏太湖地区农业科学研究所 | Breeding method of novel germplasm resource of green tender flower stalk |
| CN103210833A (en) * | 2012-12-26 | 2013-07-24 | 吴文武 | Grafting hybridization improvement method |
| CN105145337A (en) * | 2015-08-21 | 2015-12-16 | 陕西省杂交油菜研究中心 | Culture method of excellent germplasm resource of rape |
| CN105961187A (en) * | 2016-05-12 | 2016-09-28 | 安徽农业大学 | Breeding method of main inflorescence polyhedron brassica napus cytoplasm male sterility restorer line |
| CN106613920A (en) * | 2017-02-20 | 2017-05-10 | 陕西省杂交油菜研究中心 | Method for breeding cabbage type rape radish cytoplasmic male sterility three-line |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1997002737A1 (en) * | 1995-07-07 | 1997-01-30 | Zenco (No.4) Limited | Cytoplasmic male sterility system producing canola hybrids |
| WO2004039988A1 (en) * | 2002-10-29 | 2004-05-13 | Genoplante-Valor | Ppr peptide sequences capable of restoring male fertility of plants bearing a male sterility-inducing cytoplasm |
| WO2005074671A1 (en) * | 2004-01-30 | 2005-08-18 | Syngenta Participations Ag | Improved fertility restoration for ogura cytoplasmic male sterile brassica and method |
| WO2009100178A2 (en) * | 2008-02-06 | 2009-08-13 | Pioneer Hi-Bred International, Inc. | New brassica ogura restorer lines with shortened raphanus fragment (srf) |
| CN101999316A (en) * | 2009-09-01 | 2011-04-06 | 沈昌健 | Breeding method of cytoplasmic male sterile line of brassica napus |
-
2012
- 2012-05-04 CN CN2012101370929A patent/CN102640700A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1997002737A1 (en) * | 1995-07-07 | 1997-01-30 | Zenco (No.4) Limited | Cytoplasmic male sterility system producing canola hybrids |
| WO2004039988A1 (en) * | 2002-10-29 | 2004-05-13 | Genoplante-Valor | Ppr peptide sequences capable of restoring male fertility of plants bearing a male sterility-inducing cytoplasm |
| WO2005074671A1 (en) * | 2004-01-30 | 2005-08-18 | Syngenta Participations Ag | Improved fertility restoration for ogura cytoplasmic male sterile brassica and method |
| WO2009100178A2 (en) * | 2008-02-06 | 2009-08-13 | Pioneer Hi-Bred International, Inc. | New brassica ogura restorer lines with shortened raphanus fragment (srf) |
| WO2009100178A3 (en) * | 2008-02-06 | 2009-11-12 | Pioneer Hi-Bred International, Inc. | New brassica ogura restorer lines with shortened raphanus fragment (srf) |
| CN101999316A (en) * | 2009-09-01 | 2011-04-06 | 沈昌健 | Breeding method of cytoplasmic male sterile line of brassica napus |
Non-Patent Citations (1)
| Title |
|---|
| 陈卫江等: "甘蓝型油菜萝卜细胞质雄性不育恢复材料的创制", 《中国农业科学》 * |
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| CN102986523B (en) * | 2012-11-30 | 2014-01-15 | 江苏太湖地区农业科学研究所 | Breeding method of novel germplasm resource of green tender flower stalk |
| CN103210833A (en) * | 2012-12-26 | 2013-07-24 | 吴文武 | Grafting hybridization improvement method |
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| CN111528100A (en) * | 2020-06-16 | 2020-08-14 | 上海市农业科学院 | Method for obtaining distant hybridization progeny of broccoli and brassica napus |
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