CN103190335A - Simple method for artificially synthesizing cabbage type rape - Google Patents
Simple method for artificially synthesizing cabbage type rape Download PDFInfo
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- CN103190335A CN103190335A CN2013101175530A CN201310117553A CN103190335A CN 103190335 A CN103190335 A CN 103190335A CN 2013101175530 A CN2013101175530 A CN 2013101175530A CN 201310117553 A CN201310117553 A CN 201310117553A CN 103190335 A CN103190335 A CN 103190335A
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Abstract
The invention discloses a simple method for artificially synthesizing a cabbage type rape. The simple method comprises the following steps of: when wild cabbage and Chinese cabbage are hybridized, carrying out fertilization on the stigma of a female parent; after 16-144 hours, directly spraying a colchicine aqueous solution with percent concentration of 0.05%-0.15% on young ovary; and after the ovary grows to naturally-matured silique, harvesting full seeds, i.e., obtaining artificially-synthesized wild cabbage type rape seeds. Therefore, a new means with convenience, fastness and high efficiency is provided for obtaining the new germplasm of the cabbage type rape.
Description
Technical field
The present invention relates to field of crop genetic breeding, particularly a kind of simple and quick method of artificially synthesized Brassica type rape.
Background technology
Artificially synthesized Brassica type rape is the effective way of widening rape genetic breeding germ plasm resource, also is research rape species origin and one of the important means that develop.In the process of synthetic cabbage type rape, because wild cabbage (CC=20) is inequality with incompatibility and the chromosome number of Chinese cabbage (AA=18) interspecific cross, heterozygote, heterozygote embryo are difficult to develop into normal embryo; Even obtained hybrid F1(AC=19) seed also can not educate, need could obtain the cabbage type rape (AACC=38) of the allotetraploid that can educate by chromosome doubling.At present, in order to obtain artificially synthesized Brassica type rape, often take following method: 1) directly extensive hybrid method, in wild cabbage and Chinese cabbage, make male parent with one, another of castration done maternal, once hybridizes 1000 and spends above, in the hope of obtaining the species hybrid cabbage type rape, the probability that this method obtains species hybrid is very low; 2) repeating the pollination method, or claim mentor method, is male parent with Chinese cabbage, wild cabbage is the maternal example that is, successively pollinates twice, awards pollen with self at the chapiter of wild cabbage earlier, the pollen of awarding again later with Chinese cabbage every five hours once must be species hybrid in this way approximately; 3) the rescue method of ovary or embryo culture, pollinated back about 15 days, and got ovary or embryo is cultivated, again monoploid hybrid seedling pre-cultivation in containing the MS medium of 0.01% colchicin handled about 10 days, carry out chromosome doubling, the cabbage type rape that is finally manually synthesized.
By these methods, although obtained some manually synthetic cabbage type rapes, all there are the following problems for the method for acquisition artificially synthesized Brassica type rape at present: workload is big, length consuming time, and complexity, efficient is low.The hereditary basis of the main oil crop cabbage type rape in the horn of plenty world, lay good basis for obtaining higher cabbage type rape heterosis utilization, it is simple to operate to press for a kind of energy of exploitation, and workload is little, the new method of the acquisition artificially synthesized Brassica type rape that efficient is high.
Summary of the invention
Technical problem to be solved by this invention is: incompatibility and workload at wild cabbage in the prior art and Chinese cabbage interspecific cross are big, length consuming time, complexity, inefficient problem, provide a kind of simple to operate, workload is little, the simple and quick method of the artificially synthesized Brassica type rape that efficient is high.
For solving the problems of the technologies described above, the technical solution adopted in the present invention is: a kind of simple and quick method of artificially synthesized Brassica type rape, it is with wild cabbage and Chinese cabbage hybridization the time, pollinate between back 16 hours-144 hours in the column cap of female parent, directly the tender ovary of children is sprayed the colchicine solution that mass percent concentration is 0.05%-0.15%, after treating that ovary development becomes the angle fruit of physical maturity, full seed in the fruit of results angle, namely.
One preferred embodiment of such scheme is that directly the tender ovary of children being sprayed mass percent concentration in back 16 hours in pollination is 0.15% colchicine solution 1 time.
One preferred embodiment of such scheme is that directly the tender ovary of children being sprayed mass percent concentration in back 24 hours in pollination is 0.1% colchicine solution 1 time, sprays 1 time every 24 hours afterwards, sprays altogether 6 times.
One preferred embodiment of such scheme is, directly the tender ovary of children being sprayed mass percent concentration respectively at pollination 24 hours and 48 hours two time points is 0.05% and 0.1% colchicine solution.
The described female parent of such scheme is wild cabbage or Chinese cabbage.
The Chinese cabbage of mentioning herein is turnip type rape.
Compared with prior art, the beneficial effect that the present invention has is: in the appropriate time of the present invention behind wild cabbage and Chinese cabbage hybridization pollination the young tender ovary after the pollination is directly sprayed colchicine solution, the incompatibility and the workload that have so not only overcome the wild cabbage that exists in the prior art and Chinese cabbage interspecific cross are big, length consuming time, complexity, inefficient problem, and the present invention is used for artificially synthesized Brassica type rape, simple to operate, workload is little, the efficient height.
Description of drawings
Fig. 1 is that the PCR of the artificially synthesized Brassica type rape of wild cabbage G3 * Chinese cabbage B1 acquisition detects figure.
Fig. 2 is that the PCR of the artificially synthesized Brassica type rape of wild cabbage G2 * Chinese cabbage B2-2 acquisition detects figure.
Fig. 3 is that the PCR of the artificially synthesized Brassica type rape of wild cabbage G4 * Chinese cabbage B1 acquisition detects figure.
Embodiment
Embodiment 1: utilize wild cabbage G3 strain and Chinese cabbage B1 strain artificially synthesized Brassica type rape
1. material plantation: on October 10th, 2010 wild cabbage G3 strain and Chinese cabbage B1 strain are planted in the rape experimental field, on March 10th, 2011, Chinese cabbage B1 enters flowering stage, and March 23, rigan blue G 3 entered flowering stage.
2. hybridization: be male parent with the Chinese cabbage B1 of blooming early, the wild cabbage G3 of blooming later relatively is for maternal.Between the morning 9-10 point, on the inflorescence of wild cabbage G3, choose that petal will launch but 11 in the bud do not opened, other bud of opening and not opening all removes; Thoroughly remove the stamen of 11 flowers then, the pollen of Chinese cabbage B1 is awarded on the column cap of wild cabbage G3, with sulfuric acid paper bag sack bagging, in case the pollen in other source is fallen on the column cap.
3. colchicine is handled: after pollinating 16 hours, remove sack, it is 0.15% colchicine solution that the young tender ovary of 11 flowers is sprayed mass percent concentration.Wait ovary development to become the fruit back, angle of physical maturity to receive seed afterwards, shrivelled seed and full seed are wherein arranged.
4. the detection of artificially synthesized Brassica type rape: 1 full planting seed that will obtain is in the nutritive cube of band soil, after seed germination has grown into three true leaves, extract the total DNA in the blade, utilization can be distinguished wild cabbage, the PCR method of Chinese cabbage and cabbage type rape is identified, the pcr amplification result shows, only amplify a less band from wild cabbage G3, only amplify a bigger band from Chinese cabbage B1, filial generation size 2 bands all increase out (Fig. 1), illustrate that what obtain after the hybridization is real artificial synthetic cabbage type rape, consequently obtain manually synthetic cabbage type rape GB-0 of 1 strain.
Embodiment 2: utilize wild cabbage G2 strain and Chinese cabbage B2-2 strain artificially synthesized Brassica type rape
The material plantation: on October 10th, 2011 wild cabbage G2 strain and Chinese cabbage B2-2 strain are planted in the rape experimental field, on March 10th, 2012, Chinese cabbage B2-2 enters flowering stage, and March 23, G2 entered flowering stage.
Hybridization: be male parent with the Chinese cabbage B2-2 of blooming early, the wild cabbage G2 of blooming later relatively is for maternal.Between the morning 9-10 point, on the inflorescence of wild cabbage G2, choose that petal will launch but 18 in the bud do not opened, other bud of opening and not opening all removes; Thoroughly remove the stamen of 18 flowers then, the pollen of Chinese cabbage B2-2 is awarded on the column cap of wild cabbage G2, with sulfuric acid paper bag sack bagging, in case the pollen in other source is fallen on the column cap.
Colchicine is handled: after pollinating 24 hours, remove sack, it is 0.1% colchicine solution that the young tender ovary of 18 flowers is sprayed mass percent concentration.Sprayed once every 24 hours afterwards, spray altogether 6 times, wait ovary development to become the fruit back, angle of physical maturity to receive seed, shrivelled seed and full seed are wherein arranged.
The detection of artificially synthesized Brassica type rape: 2 full planting seeds that will obtain are in the nutritive cube of band soil, after seed germination has grown into three true leaves, extract the total DNA in the blade, utilization can be distinguished wild cabbage, the PCR method of Chinese cabbage and cabbage type rape is identified, the pcr amplification result shows, only amplify a less band from wild cabbage G2, only amplify a bigger band from Chinese cabbage B2-2, filial generation size 2 bands all increase out (Fig. 2), illustrate that what obtain after the hybridization is real artificial synthetic cabbage type rape, its result obtains manually synthetic cabbage type rape GB-1 and GB-2 of 2 strains.
Embodiment 3: utilize wild cabbage G4 strain and Chinese cabbage B1 strain artificially synthesized Brassica type rape
The material plantation: on October 10th, 2011 wild cabbage G4 strain and Chinese cabbage B1 strain are planted in the rape experimental field, on March 10th, 2012, Chinese cabbage B1 enters flowering stage, and March 23, G4 entered flowering stage.
Hybridization: be male parent with the Chinese cabbage B1 of blooming early, the wild cabbage G4 of blooming later relatively is for maternal.Between the morning 9-10 point, on the inflorescence of wild cabbage G4, choose that petal will launch but 23 in the bud do not opened, other bud of opening and not opening all removes; Thoroughly remove the stamen of 23 flowers then, the pollen of Chinese cabbage B1 is awarded on the column cap of wild cabbage G4, with sulfuric acid paper bag sack bagging, in case the pollen in other source is fallen on the column cap.
Colchicine is handled: after pollinating 24 hours, remove sack, it is 0.05% colchicine solution that the young tender ovary of 23 flowers is sprayed mass percent concentration, and fill spray again after the 48 hours mass percent concentration of pollinating is 0.1% colchicine solution.Allow ovary development become the fruit back, angle of physical maturity to receive seed afterwards, shrivelled seed and full seed are wherein arranged.
The detection of artificially synthesized Brassica type rape: 3 full planting seeds that will obtain are in the nutritive cube of band soil, after seed germination has grown into three true leaves, extract the total DNA in the blade, utilization can be distinguished wild cabbage, the PCR method of Chinese cabbage and cabbage type rape is identified, the pcr amplification result shows, only amplify a less band from wild cabbage G4, only amplify a bigger band from Chinese cabbage B1, filial generation size 2 bands all increase out (as shown in Figure 3), illustrate that what obtain after the hybridization is real artificial synthetic cabbage type rape, its result obtains manually synthetic cabbage type rape GB4 of 3 strains, GB5 and GB6.
Claims (5)
1. the simple and quick method of an artificially synthesized Brassica type rape, it is characterized in that: this method is with wild cabbage and Chinese cabbage hybridization the time, pollinated back 16 hours-144 hours in the column cap of female parent, directly the tender ovary of children is sprayed the colchicine solution that mass percent concentration is 0.05%-0.15%, after treating that ovary development becomes the angle fruit of physical maturity, full seed in the fruit of results angle, namely.
2. the simple and quick method of artificially synthesized Brassica type rape as claimed in claim 1 is characterized in that: directly the tender ovary of children being sprayed mass percent concentration in back 16 hours in pollination is 0.15% colchicine solution 1 time.
3. the simple and quick method of artificially synthesized Brassica type rape as claimed in claim 1, it is characterized in that: directly the tender ovary of children being sprayed mass percent concentration in back 24 hours in pollination is 0.1% colchicine solution 1 time, sprayed 1 time every 24 hours afterwards, spray altogether 6 times.
4. the simple and quick method of artificially synthesized Brassica type rape as claimed in claim 1 is characterized in that: directly the tender ovary of children being sprayed mass percent concentration respectively at pollination 24 hours and 48 hours two time points is 0.05% and 0.1% colchicine solution.
5. as the simple and quick method of claim 1 or 2 or 3 or 4 described artificially synthesized Brassica type rapes, it is characterized in that: described female parent is wild cabbage or Chinese cabbage.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106718824A (en) * | 2016-11-16 | 2017-05-31 | 西南大学 | The method that high erucic acid cabbage type rape is prepared using artificial synthesis Brassica napus |
| CN106718823A (en) * | 2016-11-16 | 2017-05-31 | 西南大学 | A kind of method that utilization artificial synthesis Brassica napus prepare high oleic acid rape material |
| CN109042291A (en) * | 2018-07-28 | 2018-12-21 | 湖南科技大学 | A method of novel vegetable is created using Chinese cabbage and wild cabbage distant hybridization |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102301946A (en) * | 2011-07-26 | 2012-01-04 | 江苏省农业科学院 | Method for creating yellow-seeded brassica napus germplasm |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102301946A (en) * | 2011-07-26 | 2012-01-04 | 江苏省农业科学院 | Method for creating yellow-seeded brassica napus germplasm |
Non-Patent Citations (2)
| Title |
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| 《扬州大学硕士学位论文》 20081216 陆莉 一种快速生长的新型甘蓝型油菜的合成及其小孢子培养的研究 , * |
| 陆莉: "《扬州大学硕士学位论文》", 16 December 2008 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106718824A (en) * | 2016-11-16 | 2017-05-31 | 西南大学 | The method that high erucic acid cabbage type rape is prepared using artificial synthesis Brassica napus |
| CN106718823A (en) * | 2016-11-16 | 2017-05-31 | 西南大学 | A kind of method that utilization artificial synthesis Brassica napus prepare high oleic acid rape material |
| CN106718823B (en) * | 2016-11-16 | 2019-04-23 | 西南大学 | A method of high oleic acid rape material is prepared using artificial synthesis Brassica napus |
| CN109042291A (en) * | 2018-07-28 | 2018-12-21 | 湖南科技大学 | A method of novel vegetable is created using Chinese cabbage and wild cabbage distant hybridization |
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