CN102771379B - Culture method of millet male sterile line of different-cytoplasmic-type anti-imazethapyr weedicide - Google Patents
Culture method of millet male sterile line of different-cytoplasmic-type anti-imazethapyr weedicide Download PDFInfo
- Publication number
- CN102771379B CN102771379B CN201210203605.1A CN201210203605A CN102771379B CN 102771379 B CN102771379 B CN 102771379B CN 201210203605 A CN201210203605 A CN 201210203605A CN 102771379 B CN102771379 B CN 102771379B
- Authority
- CN
- China
- Prior art keywords
- millet
- imazethapyr
- type
- kytoplasm
- cytoplasm
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a culture method of a millet male sterile line of different-cytoplasmic-type anti-imazethapyr weedicide. The method comprises the following steps that (1) millet breeding material cytoplasmic types are divided; (2) bridge male parents are created; (3) three-hybrid first generation of different-cytoplasmic-type anti-imazethapyr weedicide is created; and (4) the millet male sterile line of different-cytoplasmic-type anti-imazethapyr weedicide is created. The millet male sterile line with different cytoplasmic types is cultured through the method, the cytoplasmic types of the existing sterile materials are enriched, the basis is laid for the culture of different-cytoplasmic-type hybrid, difficult problems of single cytoplasmic type and easy suffering from specific diseases in the prior art can be solved, and the huge damage to the agriculture production caused by the cytoplasmic specific diseases is avoided. In addition, the millet male sterile line cultured by the method also has the characteristic of anti-imazethapyr effect and solves the problem of difficult weeding in the sterile line seed multiplication process.
Description
Technical field
The present invention relates to a kind of breeding method of millet male sterile line, be specifically related to a kind of breeding method of different kytoplasm type millet male sterile lines, the breeding method that more specifically relates to a kind of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, belongs to male sterile and utilizes research field.
Background technology
Liu Zhengli etc. follow the trail of discovery by pedigree, the male sterile line cytoplasm type of application in millet heterosis utilization research at present, spring paddy male sterile line is almost all derivative by " long 10A ", cytoplasm derives from " garden hen mouth oozes " (Crop science, 2011,51 (4): 1655-1663); Summer paddy male sterile line is almost all derivative by " glutinous millet 1A ", cytoplasm derives from " rheum officinale paddy " (Liu Zhengli etc., North China agronomy report, 2006,21 (supplementary issue): 103-109), millet male sterile line cytoplasm type is too single, and cytoplasm is matrilinear inheritance, and therefore the single of male sterility cytoplasm type must cause the single of crossbreed cytoplasm type.Infect disease because cytoplasm has specialization, infect once meet specialization disease, will produce to millet and bring crushing loss.
The breeding method of traditional millet male sterile line is the principle according to proterties complementation, do female parent with male sterile line, selecting the conventional variety that can make up male sterile line shortcoming to do male parent hybridizes, the sterile material that utilizes filial generation to separate, carry out bagging selfing, through the selection of continuous multi-generation, therefrom seed selection male sterile line.This traditional sterile line breeding method has advantages of that seed selection speed is fast, offspring Yi Wending, but because female parent is original male sterile line, and cytoplasm is matrilinear inheritance, therefore, the cytoplasm type of the male sterile line of seed selection is still original type, and the male sterile line of seed selection is not formulated out the male sterile line of novel cytoplasm type by this method.Therefore, traditional sterile line breeding method not only can not solve the problem that male sterile line cytoplasm type is too single, on the contrary due to the focus utilization of people to key male sterile line, makes the kytoplasm single problem of originating day by day serious; Simultaneously, because the conventional material of selecting mostly is the filial generation of improved variety, only there is the common character that some can overcome male sterile line shortcoming, lack special useful proterties, not only kytoplasm type is single to cause the male sterile line of seed selection, and lacks millet and produce the special useful proterties such as the antiweed be badly in need of, and hereditary basis is also very narrow, more and more nearer with conventional variety affiliation, be difficult for the strong advantage crossbreed of preparation.Up to the present, on millet, have no and carry out the report that different kytoplasm type male sterile lines are cultivated, more have no the report of cultivating different kytoplasm type anti-imazethapyr male sterile lines.
Summary of the invention
The object of the invention is to overcome that above-mentioned millet male sterile line cytoplasm type is single, the technological deficiency of antiweed not, and a kind of breeding method of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines is provided.
Principle of the present invention is that Millet Breeding material cytoplasm source chadogram is combined with the derivative evolution figure of female parent of Millet Breeding material, chadogram is carried out, on the basis of specific genetic similarity of rip cutting, the cytoplasm type of existing Millet Breeding material is divided definite; The sterile gene of existing male sterile line is imported in the millet normal fertile strain that contains anti-imazethapyr gene, initiative Bridge parent, by Bridge parent by sterile gene and anti-imazethapyr gene genetic to offspring; Take Bridge parent as male parent, do female parent with different kytoplasm types, the serial conventional material with objective trait, by female parent replace formulated out by novel kytoplasm and sterile gene, anti-miaow gene pyramiding to together with three handed over for 1 generations, then kill on the basis except antiweed offspring not at herbicide spraying, to sterile offspring's continuous bag sleeving selfing 4-6 generation of antiweed, be aided with continuous artificial orientation and select, can cultivate the anti-imazethapyr weed killer herbicide millet male sterile line of different kytoplasm types.
The main inventive point of the present invention is the Millet Breeding material type in different cytoplasm source to divide, utilize bridge male parent that sterile gene and anti-imazethapyr weed killer herbicide gene genetic are handed over to 1 generation to three of offspring, the serial different kytoplasm types of the maternal replacement initiative of utilization, and then cultivates the millet male sterile line of different kytoplasm type antiweeds.
The breeding method that the invention provides a kind of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, is characterized in that comprising the following steps: the division of I. Millet Breeding material cytoplasm type; II. the initiative of bridge male parent; III. different kytoplasm type anti-imazethapyr weed killer herbicides three are handed over the initiative in 1 generation; IV. the initiative of the anti-imazethapyr millet male sterile line of different kytoplasm types.
The breeding method of described a kind of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, is characterized in that the division of step I Millet Breeding material cytoplasm type comprises the steps: the structure of (1) cytoplasm source chadogram; (2) structure of maternal derivative evolution figure; (3) the molecule cluster in the definite and cytoplasm source of specific genetic similarity; (4) determining of Millet Breeding material cytoplasm type.
The breeding method of described a kind of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, is characterized in that the structure of the described cytoplasm source chadogram of step (1) in the division of step I Millet Breeding material cytoplasm type comprises the steps:
1. the cultivation of leaf tissue with obtain: the seed of Millet Breeding material is obtained to millet seedling in seedling-cultivating tray, in the respectively leaf tissue of clip breeding materials of 4 leaf phases;
2. the extraction of complete genome DNA: adopt the 1. complete genome DNA of Leaf tissue of CTAB method extraction step;
3. pcr amplification: adopt 20 μ l reaction systems, utilize the genomic DNA that 2. mitochondria special primer extracts step to carry out 37 reaction cycle pcr amplifications;
4. polymorphic detection: utilize gel electrophoresis to step 3. the pcr amplification product of Mitochondria carry out polymorphic detection;
5. the generation of cytoplasm source chadogram: to step 4. on electrophoresis pattern band clear and that can repeat be designated as " 1 ", the band that same position does not occur is designated as " 0 ", generate the original matrix being formed by " 1 " and " 0 ", carry out data analysis with NTSYS-pc2.1 software, carry out cluster analysis by the UPGMA method in SAHN program, the chadogram in cellulation matter source.
The breeding method of described a kind of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, it is characterized in that being configured to of the described derivative evolution figure of female parent of step (2) in the division of step I Millet Breeding material cytoplasm type: utilize pedigree back tracking method that Millet Breeding material is carried out to longitudinal arrangement by putting in order of chadogram in step (1), by upwards follow the trail of the female parent of breeding material by generation, until track farm variety, according to pedigree, breeding material derivative same female parent is progressively coupled together according to secondary relationship, form the maternal derivative figure of evolution.
The breeding method of described a kind of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, it is characterized in that the described specific genetic similarity of step (3) in the division of step I Millet Breeding material cytoplasm type determine and the molecule cluster in cytoplasm source is: the derivative figure of evolution of female parent and the kytoplasm of the Millet Breeding material building in step (1) of the Millet Breeding material that step (2) the is built chadogram of originating combines, and carry out comparative analysis, determine that substantially same maternal flavor can be gathered is the specific genetic similarity of a branch, kytoplasm source chadogram to the Millet Breeding material building in step (1) under this genetic similarity carries out rip cutting, Millet Breeding material is carried out to the molecule cluster in kytoplasm source, in phylogenetic tree, each branch represents a kytoplasm monoid.
The breeding method of described a kind of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, it is characterized in that described the definite of Millet Breeding material cytoplasm type of step (4) in the division of step I Millet Breeding material cytoplasm type is the phylogenetic tree in step (3) to be derived to evolution figure with the female parent building in step (2) be combined, each branch that kytoplasm is originated in phylogenetic tree contrasts one by one with corresponding maternal flavor, determine the cytoplasm type of each monoid material, complete the division to Millet Breeding material cytoplasm type.
The breeding method of described a kind of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, is characterized in that in the structure of the described cytoplasm source chadogram of step (1) in the division of step I Millet Breeding material cytoplasm type that the Millet Breeding material described in step is 1.: prolong paddy 12, 117A grows tall, Datong District's 29 purples, Ji paddy 26, Ji paddy 30, dam paddy 214, peace 9217, protect 182, Ji paddy 12, stone 06-766, prolong paddy 13, Ji paddy 20, Ji paddy 22, paddy 65, Ji paddy 21, Datong District 27, red paddy 10, hold paddy 12, Datong District 30, Henan paddy 6, Datong District 14, Zheng 9188, Datong District 28, iron 8240, brocade paddy 12, brocade paddy 14, towards 438, public paddy 68, iron 487, iron paddy 6, multiple 532, Fen, Shanxi 1, deep blue 156, Ji paddy 13, 229A grows tall, S80, Ji paddy 29, public paddy 70, stone 206065, Datong District 29 is green, peace 2491, emerging paddy 88, Shanxi 15, Fen, Shanxi 3, deep blue 344, Y61, stone 02399, C138, C208, towards paddy 12, K523, L70, the large grain of iron is yellow, K359, towards paddy 13, paddy 83, weighing apparatus 968, Ji 9403, Ji paddy 27, Ji 9409, Shandong paddy 10, Ji paddy 12, Ji paddy 11, paddy 6, stone 02521, stone 98700, stone 207382, stone 207393, stone 02530, stone 207286, peace 4852, paddy 3, Ji paddy 25, stone 207191, 146A grows tall, stone 207226, stone 206058, public paddy 65, C164, 203184 early, peace 2367, stone 202242, paddy 57, Ji paddy 24, Gufeng 2, stone 06-439, paddy 38, little scented rice, Ji paddy 19, K1130, dam paddy 214-2, paddy 11, K1011, long agriculture 36, Ji paddy 28, long agriculture 35, Shanxi paddy 35, select very much No. 5, select very much No. 2, Shanxi paddy 21, paddy 95, Shanxi paddy 29, towards paddy 14, K660, U.S.'s major part, paddy 10, K546, towards green-1, Shanxi paddy 16, totally 111 parts, stone 98622 and stone 97672.
The breeding method of described a kind of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, it is characterized in that in the structure of the described derivative evolution figure of female parent of step (2) in the division of step I Millet Breeding material cytoplasm type that in 111 parts of Millet Breeding materials, 12 parts derive from same maternal material Qinyuan hen mouth, 13 parts of material sources are in the yellow soft paddy of same maternal material, 63 parts of material sources were in same maternal material Japan 60 days, 6 parts of material sources are in the hard fringe paddy of same maternal material, 5 parts of material sources are in the black branch of same maternal material paddy, 12 parts of materials do not track its female parent, be convenient to study called after pedigree and lose I type and pedigree loss II type.
The breeding method of described a kind of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, it is characterized in that the described specific genetic similarity of step (3) in the division of step I Millet Breeding material cytoplasm type determine and the molecule cluster in cytoplasm source in specific genetic similarity be 0.74, under this genetic similarity, constructed chadogram in the structure of the cytoplasm source chadogram described in step (1) is carried out to rip cutting, be 7 branches by its rip cutting, each branch represents the molecule monoid in 1 kytoplasm source, the chadogram rip cutting of like this kytoplasm being originated is 7 monoids, rip cutting result reaches 98.2% with maternal derivative evolution figure uniformity.
The breeding method of described a kind of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, it is characterized in that in the determining of the described Millet Breeding material cytoplasm type of step (4) in the division of step I Millet Breeding material cytoplasm type by the specific genetic similarity described in step (3) determine and the molecule cluster in cytoplasm source in the structure of the phylogenetic tree that builds and the derivative evolution figure of the female parent described in step (2) constructed female parent derive evolution figure and combine, contrast one by one, determine the kytoplasm type of each monoid material, be respectively the Q type that derives from " garden hen mouth oozes ", derive from the R type of " Japan 60 days ", derive from the H type of " yellow soft paddy ", derive from the Y type of " hard fringe paddy ", derive from the S type of " Hei Zhigu ", also have two kinds because pedigree is lost, follow the trail of and originate less than female parent, name respectively as M I type and M II type.
The breeding method of described a kind of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, the initiative that it is characterized in that the bridge male parent described in Step II is: do female parent with millet height male sterility line, do male parent configuration hybrid combination with millet anti-imazethapyr material, ripe rear results F 1 seed; Plantation F 1 seed, in the 3-5 leaf phase, sprays imazethapyr weed killer herbicide and kills pseudostationary, and the plant normally surviving is the true hybrid that contains sterile gene and anti-imazethapyr gene, i.e. bridge male parent.
The breeding method of described a kind of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, it is characterized in that the millet height male nuclear sterile based material described in the initiative of Step II bridge male parent for the 229A that grows tall, the 146A that grows tall, the 117A that grows tall etc. one of them.
The breeding method of described a kind of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, it is characterized in that the millet anti-imazethapyr material described in the initiative of Step II bridge male parent be M200, M117, M302, M195, M150 etc. one of them.
The breeding method of described a kind of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, it is characterized in that the different kytoplasm type of the millet anti-imazethapyr weed killer herbicide three described in Step II I hands over the initiative in 1 generation to be: take the Millet Breeding material of the different kytoplasm types of dividing in step I as maternal, prepare different kytoplasms three take the bridge male parent of Step II initiative as male parent and hand over combination, obtain different kytoplasm type anti-imazethapyrs three hand over 1 generation seed.
The breeding method of described a kind of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, is characterized in that the initiative of the anti-imazethapyr male sterile line of the different kytoplasm types described in step IV comprises the steps:
1. three friendship 1 generation pseudostationary and the not removals of antiweed individual plant: the three friendship 1 generation seeds that obtain in plantation Step II I, spray imazethapyr weed killer herbicide in the 3-5 leaf phase, kill except pseudostationary and the filial generation of anti-imazethapyr not wherein, the heterozygosis fertile plant for anti-imazethapyr weed killer herbicide of surviving and the fertile plant of isozygotying, whole bagging selfings at heading stage, ripening stage, therefrom selects fine individual plant as three friendship 2 generation seeds;
2. offspring's selection: plantation step is the middle three friendship 2 generation seeds that obtain 1., spray imazethapyr weed killer herbicide in the 3-5 leaf phase, kill sterile strain and fertile plant except the not anti-imazethapyr weed killer herbicide wherein separating, what survive is two types of the sterile strain of anti-imazethapyr weed killer herbicide and the fertile plants of anti-imazethapyr weed killer herbicide, select sterile strain wherein heading stage, all baggings, ripening stage, is therefrom selected excellent sterile individual plant by head progeny row, continues to select for lower generation as three friendship 3 generation seeds; Through bagging selfing and the selection in 4-6 generation, can obtain the anti-imazethapyr millet male sterile line of stable multiple kytoplasm type so continuously.
The breeding method of described a kind of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, it is characterized in that growing tall 229A as maternal take millet male sterile line in the initiative of the bridge male parent described in Step II, take anti-imazethapyr millet weed killer herbicide intermediate materials M302 as male parent preparing hybrid, the initiative of Y type kytoplasm type anti-imazethapyr weed killer herbicide male sterile line is carried out in combination.
The breeding method of described a kind of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, it is characterized in that the multiple kytoplasm type of the millet anti-imazethapyr weed killer herbicide three described in Step II I hands over the initiative in 1 generation to be: with Y type kytoplasm Millet Breeding material Shanxi paddy 35 for maternal, hybridize with the bridge male parent of above-mentioned initiative, three of initiative Y type kytoplasm handed over for 1 generation.
The breeding method of described a kind of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, it is characterized in that growing tall 146A as maternal take millet male sterile line in the initiative of the bridge male parent described in Step II, take anti-imazethapyr millet weed killer herbicide intermediate materials M195 as male parent preparing hybrid, the initiative of S type kytoplasm type anti-imazethapyr weed killer herbicide male sterile line is carried out in combination.
The breeding method of described a kind of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, it is characterized in that the multiple kytoplasm type of the millet anti-imazethapyr weed killer herbicide three described in Step II I hands over the initiative in 1 generation to be: with S type kytoplasm Millet Breeding material multiple 532 for maternal, hybridize with the bridge male parent of above-mentioned initiative, three of initiative S type kytoplasm handed over for 1 generation.
Beneficial effect
Utilize the present invention can cultivate the anti-imazethapyr millet male sterile line of different kytoplasm types, enrich the kytoplasm type of millet male sterile line, for the crossbreed of cultivating different kytoplasm types lays the foundation, can solve because kytoplasm type is single, a difficult problem that infected by specialization disease, avoids the huge infringement bringing to agricultural production due to infecting of cytoplasm specialization disease.Simultaneously the present invention due to by sterile gene and anti-imazethapyr gene pyramiding in millet sterile material, by spraying imazethapyr weed killer herbicide, kill the weeds except male sterile line multiplication fields, can effectively solve the time-consuming difficult problem of male sterile line multiplication fields artificial weeding.
Accompanying drawing explanation
Fig. 1-1 and Fig. 1-2 are the mitochondrial pcr amplification polymorphic detection of Millet Breeding material collection of illustrative plates;
In figure, M is Marker, V1-V111 represents that respectively 111 parts of Millet Breeding materials prolong paddy 12, 117A grows tall, Datong District's 29 purples, Ji paddy 26, Ji paddy 30, dam paddy 214, peace 9217, protect 182, Ji paddy 12, stone 06-766, prolong paddy 13, Ji paddy 20, Ji paddy 22, paddy 65, Ji paddy 21, Datong District 27, red paddy 10, hold paddy 12, Datong District 30, Henan paddy 6, Datong District 14, Zheng 9188, Datong District 28, iron 8240, brocade paddy 12, brocade paddy 14, towards 438, public paddy 68, iron 487, iron paddy 6, multiple 532, Fen, Shanxi 1, deep blue 156, Ji paddy 13, 229A grows tall, S80, Ji paddy 29, public paddy 70, stone 206065, Datong District 29 is green, peace 2491, emerging paddy 88, Shanxi 15, Fen, Shanxi 3, deep blue 344, Y61, stone 02399, C138, C208, towards paddy 12, K523, L70, the large grain of iron is yellow, K359, towards paddy 13, paddy 83, weighing apparatus 968, Ji 9403, Ji paddy 27, Ji 9409, Shandong paddy 10, Ji paddy 12, Ji paddy 11, paddy 6, stone 02521, stone 98700, stone 207382, stone 207393, stone 02530, stone 207286, peace 4852, paddy 3, Ji paddy 25, stone 207191, 146A grows tall, stone 207226, stone 206058, public paddy 65, C164, 203184 early, peace 2367, stone 202242, paddy 57, Ji paddy 24, Gufeng 2, stone 06-439, paddy 38, little scented rice, Ji paddy 19, K1130, dam paddy 214-2, paddy 11, K1011, long agriculture 36, Ji paddy 28, long agriculture 35, Shanxi paddy 35, select very much No. 5, select very much No. 2, Shanxi paddy 21, paddy 95, Shanxi paddy 29, towards paddy 14, K660, U.S.'s major part, paddy 10, K546, towards green-1, Shanxi paddy 16, stone 98622, the DNA of stone 97672, A is standard band, and the band that the color of " band " was deeper than or was same as " A " is designated as " 1 ", and the band that is shallower than " A " is designated as " 0 ".
Fig. 2-1,2-2 and 2-3 are three parts of the cytoplasm source chadogram of 111 parts of Millet Breeding materials, and three is stitched together in order and obtains chadogram full figure;
In figure, 0.55,0.66,0.78,0.89,1.00 is genetic similarity
Fig. 3-1,3-2 and 3-3 are three parts of the derivative evolution figure of female parent of 111 parts of Millet Breeding materials, and three is stitched together in order and obtains maternal derivative evolution figure full figure;
In figure, what connect with solid line is the clear and definite material in maternal source; What connect with dotted line is that maternal source is indefinite, but molecule cluster they are gathered with corresponding solid line part is the material of a monoid; Qinyuan hen mouth, yellow soft paddy, Japan 60 days, hard fringe paddy, Hei Zhigu, pedigree are lost I type and are represented the different maternal sources of breeding material with pedigree loss II type.
Fig. 4-1,4-2 and 4-3 are three parts of the phylogenetic tree in the cytoplasm source of 111 parts of Millet Breeding materials, and three is stitched together in order and obtains phylogenetic tree full figure;
In figure, 0.55,0.66,0.78,0.89,1.00 is genetic similarity; L represents the specific genetic similarity 0.74 that chadogram is carried out to rip cutting; B1, B2, B3, B4, B5, B6, B7 are illustrated in specific genetic similarity 0.74 time to form after chadogram rip cutting 7 molecule monoids.
Fig. 5 is Millet Breeding material cytoplasm type division figure
In figure, the maternal flavor correspondence of B1 molecule monoid and Qinyuan hen mouth, location Q type; B2 molecule monoid and the maternal flavor correspondence of yellow soft paddy, location H type; B3 molecule monoid and Japan maternal flavor correspondence on the 60th, location R type; B4 molecule monoid and the maternal flavor correspondence of hard fringe paddy, location Y type; The maternal flavor pedigree that B5 molecule monoid is corresponding is lost, location M I type; B6 molecule monoid and the maternal flavor correspondence of black branch paddy, location S type; The maternal flavor pedigree that B7 molecule monoid is corresponding is lost, location M II type; A corresponding diagram 4-1, b corresponding diagram 4-2, c corresponding diagram 4-3; D corresponding diagram 3-1, e corresponding diagram 3-2, f corresponding diagram 3-3.
Below in conjunction with specific embodiment, the present invention is further elaborated, but the scope not limiting the present invention in any way.
Embodiment
In embodiment, agents useful for same is if no special instructions all purchased from raw work biotechnology Shanghai Co., Ltd, mitochondria special primer used is synthetic by giving birth to work biotechnology Shanghai Co., Ltd, NTSYS-pc2.1 software used is by State University of New York, Stony Brook development, the lucerne grass clean (former medicine contains 5% imazethapyr active ingredient) that imazethapyr weed killer herbicide used is produced purchased from Shandong Cynda Chemical Co., Ltd.111 parts of Millet Breeding materials used are from Xia Gu ecotope, China North China, Chun Gu ecotope, northwest, the Hebei of Chun Gu ecotope, northeast, Henan, Shandong, Liaoning, Jilin, Shanxi, Inner Mongol, 8 provinces and regions, Shaanxi (referring to table 1).Grow tall 229A, the 146A that grows tall, the 117A that grows tall of millet used height male nuclear sterile based material derives from Millet Research Inst., Shanxi Prov. Academy of Agricultural Science, and millet anti-imazethapyr material M200, M117, M302, M195, M150 are this institute resource material.
111 parts of lists for examination material of table 1
Embodiment 1: the cultivation of millet H type kytoplasm type anti-imazethapyr weed killer herbicide male sterile line
1, the division of Millet Breeding material kytoplasm type
(1) structure of cytoplasm source chadogram
1. the cultivation of leaf tissue with obtain: by prolonging paddy 12, the seed of the 111 parts of Millet Breeding materials (in table 1) such as 117A that grow tall plants respectively and obtained millet seedling in seedling-cultivating tray, in the 4 leaf phases leaf tissue of clip breeding materials respectively;
2. the extraction of complete genome DNA: adopt 1. complete genome DNA in Leaf tissue of CTAB method extraction step;
3. pcr amplification: adopt 20 μ l reaction systems, utilize 23 pairs of mitochondria special primers in the synthetic table 2 of raw work biotechnology Shanghai Co., Ltd to pass through 37 reaction cycle, the genomic DNA that 2. step is extracted carries out pcr amplification.20 μ l reaction systems comprise Taq enzyme (5U/ μ L) 0.2 μ L, 10 × buffer, 2.0 μ L, dNTP (2.0mM) 2.0 μ L, FPrime 1.0 μ L, RPrimer 1.0ML, DNA 2.0 μ L, ddH2O11.8 μ L.Pcr amplification carries out 37 circulations altogether.First circulates at 94 ℃ denaturation 4 minutes; Then, 94 ℃ of sex change 45 seconds, Tm ℃ of annealing 45 seconds, 72 ℃ are extended 1-4 minute, annealing temperature Tm ℃ with the extension time in table 2, circulate 35 times; Finally, 72 ℃ are extended 8 minutes;
Table 2 mitochondria special primer sequence and amplification condition thereof
4. polymorphic detection: utilize gel electrophoresis by step the pcr amplification product in 3. dyed blended by 10: 1 with ethidium bromide respectively after, get 10 μ L with liquid-transfering gun, in tbe buffer solution, carry out electrophoresis with 1.5% Ago-Gel, detect polymorphism; Add 5 μ LDL2000 Marker electrophoresis after 1 hour under 120V voltage, 400mA current condition, with EB dyeing 5 minutes, record (seeing Fig. 1) with gel imaging instrument observed result automated imaging;
5. the generation of cytoplasm source chadogram: to step 4. on electrophoresis pattern band clear and that can repeat be designated as " 1 ", the band that same position does not occur is designated as " 0 ", generate the original matrix (in table 3) being formed by " 1 " and " 0 ", carry out data analysis with NTSYS-pc2.1 software, carry out cluster analysis by the UPGMA method in SAHN program, generate kytoplasm source chadogram (seeing Fig. 2).
Table 3 Mitochondrial polymorphisms data original matrix
(2) structure of maternal derivative evolution figure: utilize pedigree back tracking method that 111 parts of Millet Breeding materials are carried out to longitudinal arrangement by putting in order in chadogram in step (1), by upwards following the trail of the female parent of breeding material by generation, until track farm variety.Through following the trail of, in 111 parts of materials 12 parts derive from same maternal material Qinyuan hen mouth, 13 parts of material sources in the yellow soft paddy of same maternal material, 63 parts of material sources in 60 days, 6 parts material sources of same maternal material Japan in the hard fringe paddy of same maternal material, 5 parts of material sources are in the black branch of same maternal material paddy, 12 parts of materials do not track its female parent, be convenient to study called after pedigree and lose I type and pedigree loss II type, according to pedigree, breeding material derivative same female parent is progressively coupled together according to secondary relationship, formed maternal derivative evolution figure (seeing Fig. 3).
(3) determining and the molecule cluster in cytoplasm source of specific genetic similarity: the kytoplasm source chadogram of the Millet Breeding material building in derivative the female parent of the Millet Breeding material building in step (2) figure of evolution and step (1) is carried out to binding analysis, in the time that genetic similarity is 0.74, can be a branch by deriving from 109 parts of breeding materials that same maternal flavor gathers, 0.74 is the specific genetic similarity that the cytoplasm source chadogram for examination material is carried out to rip cutting, kytoplasm source chadogram to Millet Breeding material under this genetic similarity carries out rip cutting, be 7 branches by its rip cutting, each branch represents the molecule monoid in 1 kytoplasm source, the chadogram rip cutting of like this kytoplasm being originated is 7 monoids (seeing Fig. 4).This rip cutting result reaches 98.2% with maternal derivative evolution figure uniformity.
(4) determining of cytoplasm type: the phylogenetic tree in step (3) is combined with the derivative evolution figure of the female parent building in step (2), 7 molecule monoids in kytoplasm source are contrasted one by one with the corresponding derivative system of female parent respectively, determine the kytoplasm type of each monoid material, be respectively the Q type that derives from " garden hen mouth oozes ", derive from the R type of " Japan 60 days ", derive from the H type of " yellow soft paddy ", derive from the Y type of " hard fringe paddy ", derive from the S type of " Hei Zhigu ", also have two kinds because pedigree is lost, follow the trail of and originate less than female parent, name respectively as MI type and M II type (seeing Fig. 5).
2, the initiative of bridge male parent: grow tall 146A as maternal take millet male sterile line, the combination take anti-imazethapyr millet weed killer herbicide intermediate materials M200 as male parent preparing hybrid, it is F 1, in these 4 leaf phases of F 1, spray 40 kilograms of former medicines of imazethapyr weed killer herbicide according to every mu: water=1: 200 imazethapyr weed killer herbicide, kill except pseudostationary, the plant normally surviving is true hybrid, true hybrid is the normal fertile strain that contains sterile gene and anti-imazethapyr weed killer herbicide gene, is F 1-bridge male parent of anti-imazethapyr weed killer herbicide of the present invention.
3, millet H type cytoplasm anti-imazethapyr weed killer herbicide three is handed over the initiative in 1 generation: the feature that is matrilinear inheritance according to cytoplasm, be maternal with the H type kytoplasm Millet Breeding material Datong District 29 in step 1, with the F 1 of " 146A × M200 grows tall " of initiative in step 2 be that male parent is hybridized, three of initiative H type kytoplasm handed over for 1 generations.
4, the initiative of the anti-imazethapyr male sterile line of millet H type kytoplasm: (1) three friendship 1 generation pseudostationary and the not removal of antiweed individual plant: the three friendship 1 generation seeds that obtain in plantation step 3, spray 40 kilograms of former medicines of imazethapyr weed killer herbicide in 4 leaf phases according to every mu: water=1: 200 imazethapyr weed killer herbicide, kill except pseudostationary and the filial generation of anti-imazethapyr not wherein, the heterozygosis fertile plant for anti-imazethapyr weed killer herbicide of surviving and the fertile plant of isozygotying, whole bagging selfings at heading stage, ripening stage, therefrom selects fine individual plant as three friendship 2 generation seeds, (2) offspring's selection: the three friendship 2 generation seeds that obtain in plantation step (1), spray 40 kilograms of former medicines of imazethapyr weed killer herbicide in 4 leaf phases according to every mu: water=1: 200 imazethapyr weed killer herbicide, kill sterile strain and fertile plant except the not anti-imazethapyr weed killer herbicide wherein separating, what survive is two types of the fertile plants of the sterile strain of anti-imazethapyr weed killer herbicide and anti-imazethapyr weed killer herbicide, select sterile strain wherein heading stage, all baggings, ripening stage, is therefrom selected excellent sterile individual plant by head progeny row, continue to select for lower generation as three friendship 3 generation seeds, through the selection in 4 generations, obtain the stable anti-miaow male sterile line " DZ2010N169A " with H type kytoplasm so continuously.
Embodiment 2: the cultivation of millet R type kytoplasm type anti-imazethapyr weed killer herbicide male sterile line
In embodiment 1 step 2, grow tall 146A as maternal take millet male sterile line, take anti-imazethapyr millet weed killer herbicide intermediate materials M117 as male parent preparing hybrid, the initiative of bridge male parent is carried out in combination; In embodiment 1 step 3, be maternal with the R type kytoplasm Millet Breeding material Ji paddy 12 in step 1, hybridize with the F 1 of the bridge male parent " 146A × M117 grows tall " of initiative in step 2, three of initiative R type kytoplasm handed over for 1 generation; In step 4 (2) offspring's of embodiment 1 selection, the selection through 5 generations so continuously, obtains the stable anti-miaow male sterile line " DZ20110634A " with R type kytoplasm.All the other steps are with embodiment 1.
Embodiment 3: the cultivation of millet Y type kytoplasm type anti-imazethapyr weed killer herbicide male sterile line
In embodiment 1 step 2, grow tall 229A as maternal take millet male sterile line, take anti-imazethapyr millet weed killer herbicide intermediate materials M302 as male parent preparing hybrid, the initiative of bridge male parent is carried out in combination; In embodiment 1 step 3, be maternal with the Y type kytoplasm Millet Breeding material Shanxi paddy 35 in step 1, hybridize with the F 1 of the bridge male parent " 229A × M302 grows tall " of initiative in step 2, three of initiative Y type kytoplasm handed over for 1 generation; In step 4 (2) offspring's of embodiment 1 selection, the selection through 5 generations so continuously, obtains the stable anti-miaow male sterile line " DZ20111260A " with Y type kytoplasm.All the other steps are with embodiment 1.
Embodiment 4: the cultivation of millet S type kytoplasm type anti-imazethapyr weed killer herbicide male sterile line
In embodiment 1 step 2, grow tall 146A as maternal take millet male sterile line, take anti-imazethapyr millet weed killer herbicide intermediate materials M195 as male parent preparing hybrid, the initiative of bridge male parent is carried out in combination; In embodiment 1 step 3, be maternal with the S type kytoplasm Millet Breeding material in step 1 multiple 532, hybridize with the F 1 of the bridge male parent " 146A × M195 grows tall " of initiative in step 2, three of initiative S type kytoplasm handed over for 1 generation; In step 4 (2) offspring's of embodiment 1 selection, the selection through 5 generations so continuously, obtains the stable anti-miaow male sterile line " DZ20112208A " with S type kytoplasm.All the other steps are with embodiment 1.
Embodiment 5: millet M
ithe cultivation of type kytoplasm type anti-imazethapyr weed killer herbicide male sterile line
In embodiment 1 step 2, grow tall 229A as maternal take millet male sterile line, take anti-imazethapyr millet weed killer herbicide intermediate materials M195 as male parent preparing hybrid, the initiative of bridge male parent is carried out in combination; In embodiment 1 step 3, use the M in step 1
itype kytoplasm Millet Breeding material Ji paddy 21 is maternal, hybridizes initiative M with the F 1 of the bridge male parent " 229A × M195 grows tall " of initiative in step 2
ithree of type kytoplasm handed over for 1 generation; In step 4 (2) offspring's of embodiment 1 selection, the selection through 6 generations so continuously, obtains the stable M that has
ithe anti-miaow male sterile line " DZ20110718A " of type kytoplasm.All the other steps are with embodiment 1.
Embodiment 6: millet M
iIthe cultivation of type kytoplasm type anti-imazethapyr weed killer herbicide male sterile line
In embodiment 1 step 2, grow tall 229A as maternal take millet male sterile line, take anti-imazethapyr millet weed killer herbicide intermediate materials M195 as male parent preparing hybrid, the initiative of bridge male parent is carried out in combination; In embodiment 1 step 3, use the M in step 1
iIthe large grain of type kytoplasm Millet Breeding material iron Huang is maternal, hybridizes initiative M with the F 1 of the bridge male parent " 229A × M195 grows tall " of initiative in step 2
iIthree of type kytoplasm handed over for 1 generation; In step 4 (2) offspring's of embodiment 1 selection, the selection through 6 generations so continuously, obtains the stable M that has
iIthe anti-miaow male sterile line " DZ20111369A of type kytoplasm.All the other steps are with embodiment 1.
Embodiment 7: the cultivation of millet Q type kytoplasm type anti-imazethapyr weed killer herbicide male sterile line
In embodiment 1 step 2, grow tall 117A as maternal take millet male sterile line, take anti-imazethapyr millet weed killer herbicide intermediate materials M150 as male parent preparing hybrid, the initiative of bridge male parent is carried out in combination; In embodiment 1 step 3, be maternal with the 117A that grows tall of the Q type kytoplasm Millet Breeding material in step 1, hybridize with the F 1 of the bridge male parent " 117A × M150 grows tall " of initiative in step 2, three of initiative Q type kytoplasm handed over for 1 generations; All the other steps are identical with embodiment 1, obtain the stable anti-miaow male sterile line " KMA91 " with Q type kytoplasm.
The embodiment that the present invention enumerates is intended to set forth further a kind of breeding method of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, and scope of the present invention is not formed to any restriction.
Claims (2)
1. a breeding method for different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines, is characterized in that comprising the following steps: the division of I. Millet Breeding material cytoplasm type; II. the initiative of bridge male parent; III. different kytoplasm type anti-imazethapyr weed killer herbicides three are handed over the initiative in 1 generation; IV. the initiative of the anti-imazethapyr millet male sterile line of different kytoplasm types;
The division of step I Millet Breeding material cytoplasm type comprises the steps: the structure of (1) cytoplasm source chadogram; (2) structure of maternal derivative evolution figure; (3) the molecule cluster in the definite and cytoplasm source of specific genetic similarity; (4) determining of Millet Breeding material cytoplasm type;
The structure of cytoplasm source chadogram described in step (1) in the division of step I Millet Breeding material cytoplasm type comprises the steps:
1. the cultivation of leaf tissue with obtain: the seed of Millet Breeding material is obtained to millet seedling in seedling-cultivating tray, in the respectively leaf tissue of clip breeding materials of 4 leaf phases;
2. the extraction of complete genome DNA: adopt the 1. complete genome DNA of Leaf tissue of CTAB method extraction step;
3. pcr amplification: adopt 20 μ l reaction systems, utilize the genomic DNA that 2. mitochondria special primer extracts step to carry out 37 reaction cycle pcr amplifications;
4. polymorphic detection: utilize gel electrophoresis to step 3. the pcr amplification product of Mitochondria carry out polymorphic detection;
5. the generation of cytoplasm source chadogram: to step 4. on electrophoresis pattern band clear and that can repeat be designated as " 1 ", the band that same position does not occur is designated as " 0 ", generate the original matrix being formed by " 1 " and " 0 ", carry out data analysis with NTSYS-pc2.1 software, carry out cluster analysis by the UPGMA method in SAHN program, the chadogram in cellulation matter source;
The described derivative evolution figure of female parent of step (2) in the division of step I Millet Breeding material cytoplasm type is configured to: utilize pedigree back tracking method that Millet Breeding material is carried out to longitudinal arrangement by putting in order of constructed chadogram in step I-step (1) above, by upwards follow the trail of the female parent of breeding material by generation, until track farm variety, according to pedigree, breeding material derivative same female parent is progressively coupled together according to secondary relationship, form the maternal derivative figure of evolution;
The described specific genetic similarity of step (3) in the division of step I Millet Breeding material cytoplasm type determine and the molecule cluster in cytoplasm source is: the derivative figure of evolution of female parent of the Millet Breeding material above building in step I-step (2) and the kytoplasm of the Millet Breeding material of structure in step I-step (1) the above chadogram of originating combines, and carry out comparative analysis, determine that substantially same maternal flavor can be gathered is the specific genetic similarity of a branch, kytoplasm source chadogram to the Millet Breeding material building in step I-step (1) above under this genetic similarity carries out rip cutting, Millet Breeding material is carried out to the molecule cluster in kytoplasm source, in molecule cluster, each branch represents a kytoplasm monoid,
In the molecule cluster in the definite and cytoplasm source of the specific genetic similarity described in the step (3) in the division of step I Millet Breeding material cytoplasm type, specific genetic similarity is 0.74, under this genetic similarity, constructed chadogram in the structure of the source of the cytoplasm described in step I-step (1) above chadogram is carried out to rip cutting, be 7 branches by its rip cutting, each branch represents the molecule monoid in 1 kytoplasm source, the chadogram rip cutting of like this kytoplasm being originated is 7 monoids, rip cutting result reaches 98.2% with maternal derivative evolution figure uniformity,
The described Millet Breeding material cytoplasm type of step (4) in the division of step I Millet Breeding material cytoplasm type definite derives evolution figure by the phylogenetic tree in step I-step (3) above with the female parent building in step I-step (2) to be above combined, each branch that kytoplasm is originated in phylogenetic tree contrasts one by one with corresponding maternal flavor, determine the cytoplasm type of each monoid material, complete Millet Breeding material cytoplasm type is divided;
The initiative of the bridge male parent described in Step II is: do female parent with millet height male nuclear sterile based material, do male parent configuration hybrid combination with millet anti-imazethapyr material, ripe rear results F 1 seed; Plantation F 1 seed, in the 3-5 leaf phase, sprays imazethapyr weed killer herbicide and kills pseudostationary, and the plant normally surviving is the true hybrid that contains sterile gene and anti-imazethapyr gene, i.e. bridge male parent;
Different kytoplasm type anti-imazethapyr weed killer herbicides three described in Step II I hand over the initiative in 1 generation to be: take the Millet Breeding material of the different kytoplasm types of dividing in above-mentioned steps I-step (4) for female parent, prepare different kytoplasms three take the bridge male parent of formulating in above-mentioned Step II as male parent and hand over combination, obtain different kytoplasm type anti-imazethapyrs three hand over 1 generation seed;
The initiative of the anti-imazethapyr millet male sterile line of the different kytoplasm types described in step IV comprises the steps:
1. three friendship 1 generation pseudostationary and the not removals of antiweed individual plant: the three friendship 1 generation seeds that obtain in plantation above-mentioned steps III, spray imazethapyr weed killer herbicide in the 3-5 leaf phase, kill except pseudostationary and the filial generation of anti-imazethapyr not wherein, the heterozygosis fertile plant for anti-imazethapyr weed killer herbicide of surviving and the fertile plant of isozygotying, whole bagging selfings at heading stage, ripening stage, therefrom selects fine individual plant as three friendship 2 generation seeds;
2. offspring's selection: plantation step is the middle three friendship 2 generation seeds that obtain 1., spray imazethapyr weed killer herbicide in the 3-5 leaf phase, kill sterile strain and fertile plant except the not anti-imazethapyr weed killer herbicide wherein separating, what survive is two types of the sterile strain of anti-imazethapyr weed killer herbicide and the fertile plants of anti-imazethapyr weed killer herbicide, select sterile strain wherein heading stage, all baggings, ripening stage, is therefrom selected excellent sterile individual plant by head progeny row, continues to select for lower generation as three friendship 3 generation seeds; Through bagging selfing and the selection in 4-6 generation, can obtain the anti-imazethapyr millet male sterile line of stable different kytoplasm types so continuously.
2. the breeding method of a kind of different kytoplasm type anti-imazethapyr weed killer herbicide millet male sterile lines according to claim 1, it is characterized in that: the different kytoplasm type anti-imazethapyr weed killer herbicides three described in Step II I hand over the initiative in 1 generation to be: use the Y type kytoplasm Millet Breeding material Shanxi paddy 35 that derives from " hard fringe paddy " for maternal, hybridize with the bridge male parent of initiative in claim 1, three of initiative Y type kytoplasm handed over for 1 generation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210203605.1A CN102771379B (en) | 2012-06-20 | 2012-06-20 | Culture method of millet male sterile line of different-cytoplasmic-type anti-imazethapyr weedicide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210203605.1A CN102771379B (en) | 2012-06-20 | 2012-06-20 | Culture method of millet male sterile line of different-cytoplasmic-type anti-imazethapyr weedicide |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102771379A CN102771379A (en) | 2012-11-14 |
| CN102771379B true CN102771379B (en) | 2014-05-14 |
Family
ID=47116696
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210203605.1A Expired - Fee Related CN102771379B (en) | 2012-06-20 | 2012-06-20 | Culture method of millet male sterile line of different-cytoplasmic-type anti-imazethapyr weedicide |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102771379B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113748982A (en) * | 2021-10-12 | 2021-12-07 | 张家口市农业科学院(河北省高寒作物研究所) | Method for creating male sterile line of millet containing different cytoplasm |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1198882A (en) * | 1998-06-15 | 1998-11-18 | 河北省农林科学院 | Creation and utilization of anti-phytocide germplasm of millet |
-
2012
- 2012-06-20 CN CN201210203605.1A patent/CN102771379B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1198882A (en) * | 1998-06-15 | 1998-11-18 | 河北省农林科学院 | Creation and utilization of anti-phytocide germplasm of millet |
Non-Patent Citations (6)
| Title |
|---|
| 刘正理等.谷子杂种优势群的构建方法及研究进展.《河北农业科学》.2010,第14卷(第11期),第102-104页. |
| 师志刚等.谷子抗咪唑乙烟酸新种质的初步研究.《河北农业科学》.2010,第14卷(第11期),第133-134、136页. |
| 李会霞等.谷子雄性不育系杂种优势利用研究.《甘肃农业科技》.2002,(第12 期),第7-9页. |
| 谷子抗咪唑乙烟酸新种质的初步研究;师志刚等;《河北农业科学》;20101231;第14卷(第11期);第133-134、136页 * |
| 谷子杂种优势群的构建方法及研究进展;刘正理等;《河北农业科学》;20101231;第14卷(第11期);第102-104页 * |
| 谷子雄性不育系杂种优势利用研究;李会霞等;《甘肃农业科技》;20021231(第12 期);第7-9页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102771379A (en) | 2012-11-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106386465B (en) | Method for breeding polyploid two-line hybrid rice | |
| CN104584890B (en) | A kind of scale inoculation identification method of MRDV resistance | |
| US20190191645A1 (en) | Method for cultivating perennial rice using asexual propagation characteristic of oryza longistaminata | |
| CN104255443B (en) | A kind of polymerization is containing the seed selection method for rice of anti-three kinds of disease genes | |
| CN105112517A (en) | Method for identifying corn haploid embryos and application of method | |
| CN106665332A (en) | Method for carrying out recurrent selection breeding on water-saving and drought-resisting rice by utilizing dominant nuclear male sterile material | |
| CN108496790B (en) | Method for cultivating rice blast-resistant two-line sterile line | |
| Wai et al. | Sources of resistance to bacterial wilt and restorer-of-fertility genotype for cytoplasmic male sterility in Capsicum pepper | |
| CN104303989B (en) | A kind of hybrid rice seed breeding process | |
| CN110122315A (en) | Utilize the method for the permanent sterility line breeding blast resisting multiple cross kind of rice temp-sensing | |
| CN102771379B (en) | Culture method of millet male sterile line of different-cytoplasmic-type anti-imazethapyr weedicide | |
| CN102972284A (en) | Novel method for obtaining artificially synthesized hexaploid wheat | |
| CN105104181A (en) | Haploid genome doubling method for maize | |
| CN114277175B (en) | Rapid and efficient wheat scab-resistant molecular design breeding method | |
| CN105010130A (en) | Breeding method of few-seed watermelons | |
| Yan et al. | Development and molecular cytogenetic characterization of cold-hardy perennial wheatgrass adapted to northeastern China | |
| CN109006456A (en) | A kind of selection of pimento genic male sterility dual purpose | |
| Nakazawa et al. | Genomic in situ hybridization (GISH) analysis of intergeneric hybrids in Colchicaceae | |
| CN101427651A (en) | Cultivation method for dual-purpose hybrid rice for flood and drought | |
| CN104429916B (en) | A kind of method that gamete breeding by crossing using multiplication chromosome cultivates triploid Dendrobidium huoshanness | |
| CN102750458B (en) | Cytoplasm type dividing method of millet breeding materials | |
| CN101982063A (en) | Method for creating 100% of self-fruitfulness germplasm of pear | |
| CN101810138A (en) | Breeding method of novel variety of dwarf bermudagrass | |
| CN104160945A (en) | Method for breeding novel species by hybridization of broad-leaf radish and petal-leaf radish | |
| CN108719047B (en) | Molecular breeding method for improving heat resistance of rice young ear in meiosis stage by using single-segment replacement line |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20140514 Termination date: 20170620 |