CN1076161C - Technology for germ plasma preservation and quick breeding by group culture of pyrethrum - Google Patents
Technology for germ plasma preservation and quick breeding by group culture of pyrethrum Download PDFInfo
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- CN1076161C CN1076161C CN97119736A CN97119736A CN1076161C CN 1076161 C CN1076161 C CN 1076161C CN 97119736 A CN97119736 A CN 97119736A CN 97119736 A CN97119736 A CN 97119736A CN 1076161 C CN1076161 C CN 1076161C
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Abstract
The present invention relates to a technology for the rapid propagation of qualified pyrethrum seeds and the germplasm conservation of pyrethrum, which comprises: good clonal buds are adopted as explants; the buds are directly induced to germinate on an MS culture medium with additional hormones of different kinds and different concentrations; then, the combination of indoor cultivation, comprising proliferation, strengthening, rootage and the transition by taking test-tube plantlets out of a bottle, with outdoor cultivation, comprising field planting and cultivation, is carried out; qualified seed germplasm conservation is formed by indoor continuous cultivation by a group cultivation technology. The technology is stable and reliable, and has strong reproducibility. Through continuous cultivation of pyrethrum clonal varieties of scores of types, group cultivated seedlings grow strongly with strong resistance, the growth potential is better than that of seedlings cultivated by common seeds, and a high yield can be obtained.
Description
The present invention relates to plant tissue culture technique, particularly, relate to quick breeding by group culture of pyrethrum and quality saving technology.
Dalmatian chrysanthemum (Pyrethrum cinerariifolium Trev.) is a kind of herbaceos perennial, contains several effectively compositions---pyrethrins, cinerin and jasmone in its dried flower.Because of advantages such as it have efficiently, safety, residual life are short, it is a kind of up-and-coming natural pesticide plant, the breeding of Dalmatian chrysanthemum is mainly by two kinds of approach: the one, and sexual propagation, promptly multiply seedling of future generation by seed, if original seed is best in quality, can in one period, keep high yield.Yet Dalmatian chrysanthemum is self sterile, utilizes the seed of single variety for a long time, will cause deterioration of variety; If a plurality of kinds plant simultaneously, the purity of so original kind is not right to be kept, and will lose the advantage of original kind, for example Jiangsu Province, China plantation Dalmatian chrysanthemum that's how things stand.Another kind of breeding approach is vegetative propagation, promptly pass through the method for root division or cuttage, this class propagation method is subjected to the restriction of seasonality and natural conditions, in case maternal plant has infected root rot simultaneously, nematode etc., cause germ to spread soon by vegetative propagation, for example vegetative propagation is carried out in East Africa for a long time, now caused nematode seriously to spread, deterioration of variety, output of Dalmatian chrysanthemum and quality all are subjected to very big influence, as J.E.Parlevliet and J.G.Brewer at the summary in April, 1971 and C.W.Warai and all at Pyrethrum Post1991,18 (3): reported in 104.Therefore, necessary quick breeding and the quality saving problem of utilizing tissue culture technique to solve the Dalmatian chrysanthemum breeding.But the report that quick breeding of Dalmatian chrysanthemum breeding and the success of quality saving technology are not arranged so far.
The objective of the invention is at the above-mentioned problems in the prior art, a kind of Dalmatian chrysanthemum breeding breeding and quality saving technology fast are provided, this technology is not subjected to the restriction of natural conditions or regional disparity, reproduction coefficient is fast at a high speed, the rejuvenation, the purifying that help kind help the long preservation of germplasm.
In order to realize above-mentioned purpose of the present invention, the invention provides following technical scheme:
Quick breeding by group culture of pyrethrum and quality saving method, comprise that selecting clonal budling for use is explant, on minimal medium, directly induce and sprout, bud through propagation, increase strong, take root, the indoor cultivation of test-tube plantlet bottle outlet transition is to the outdoor cultivation of field planting, cultivation, form quality saving in indoor continuous culture, it is the basis that minimal medium is selected MS for use, sucrose is with 3% of volume, agar is with 0.8-1.0% of volume, and the bud inducing culture is selected 6-BA1.0mg/l or 6-BA1.0mg/l+NAA0.4mg/l for use; Proliferated culture medium is selected MS+6-BA1.0mg/l+NAA0.25-0.40mg/l or MS+6-BA0.1-2.0mg/l+NAA0.25mg/l for use; Increase strong medium and select MS or MS+6-BA1.0mg/l+NAA0.25mg/l or MS+6-BA0.5mg/l+NAA0.25mg/l root media MS or MS+NAA0.2-2.0mg/l, PH5.4-5.6, conventional autoclaving for use; The test-tube plantlet condition of culture selects 20-28 ℃; Illumination 12h/d, intensity of illumination 1500-1800Lux.
It is explant that said method can be selected the budling of the later stage plant that nourishes and grows for use.
Can select the season that the later stage enters the reproductive growth initial stage, plant begins to take out spray of nourishing and growing the period of drawing materials of said method budling.
The said method explant can preferentially be selected the budling of 0.5-1.5 centimetres of length.
The single medium that changes over to of bud that during the said method quality saving process initial culture is induced, it is the basis that medium is selected MS for use, sucrose is with 3% of volume, agar is with 0.8-1.0% of volume, preferably with 6-BA1.0mg/l or 6-BA1.0mg/l+NAA0.40mg/l, proliferated culture medium is selected MS+6BA1.0mg/l+NAA0.25mg/l+NAA0.25-0.40mg/l or MS+6-BA0.1-2.0mg/l+NAA0.25mg/l for use to the bud inducing culture.
The transition of said method test-tube plantlet bottle outlet was chosen in culture of rootage after 9-12 days, when the seedling cauline leaf is grown 4-6 centimetres, opened bottle cap in preceding 2-3 days emerging, treat that the leaf look deepens, be transplanted into and 2.5-3.0 centimetres aseptic perlitic is housed has lid to mould in the box, water spray, move into the greenhouse, regularly spray 0.1% bactericide, begin to make in a week humidity to remain on more than 90%, gradually open the lid, plant after 10-15 days in the earth bag thereafter; Growing moved on in the cool canopy after 7-10 days, increased illumination gradually, moved to open country through 14-20 days, again through field planting field in January.
In the technology of the present invention measure crucial principle based on:
Utilize tissue culture technique that breeding is carried out fast breeding institute time-consuming weak point, the reproduction coefficient height, the propagation of effective bud in each clone per generation is counted 3.6 times of average out to, each clone can breed for 8 generations at least in 1 year, whole transition stage survival rate average out to 94%, the effective breeding that can extrapolate in the bud 1 year is 3.6
8* 0.94=26518 strain.
The present invention shows by actual tests, tissue cultivating seedling (filial generation) has good genetic stability, compare by the investigation group training filial generation seedling (annual plant) of series and the multinomial genetic character of parental generation, the result has confirmed that group training filial generation seedling has shown with parental generation and has kept great uniformity, the statistical value of the multinomial biological character that filial generation showed and parental generation are very approaching, prove absolutely that tissue culture technology is a kind of technical measures desirable to quality saving.
The technical measures of key of the present invention are:
1, draw materials the time: Dalmatian chrysanthemum is a perennial herb, all can draw materials from early April August to next year, and area, routine Kunming does not have severe cold winter, and the advantage that Dalmatian chrysanthemum in the winter time can continued growth can be drawn materials for up to 9 months in 1 year.
2, the selection of culture medium prescription: the MS of medium is the basis, and characteristics and the needs according to training objective and variant cultivation stage add variety classeses such as 6-BA and NAA simultaneously;
3, incubation time: from single clone, the cultivation cycle average out in each generation (from the test-tube plantlet bottle outlet of drawing materials) 45 days;
4, incubation: A, indoor cultivation: each for the test-tube plantlet stage must through drawing materials, bud is induced, breed (rejuvenation), take root to bottle outlet average out to 45 days;
B, outdoor cultivation: need by bottle outlet transition, open country transition, land for growing field crops field planting stage, average out to 60-75 days.
5, the field planting of tissue cultivating seedling: transplant planting require to be selected clay with non-heavy magnesium carbonate or is given birth to common brick soil, non-low-lying land or rainy season chaor, require that the fertile state in soil is medium certain irrigation conditions;
6, the field management measure after the method for tissue cultivating seedling field planting and the field planting is identical with another patent application " high-yield cultivation technique for pyrethrum " of the present patent application person;
7, the technology of the present invention is reliable and stable, and is repeatable strong, through continuously tens kinds of Dalmatian chrysanthemum clone kinds having been cultivated the tissue cultivating seedling of strain more than 6000, and these tissue cultivating seedling robust growth, resistance, growth potential are better than the seedling of general cultivating seeds, obtain high yield.
Compared with prior art, beneficial effect of the present invention is:
1, be not subjected to the restriction of natural conditions or regional disparity: conventional propagation method is subjected to the weather of location and the restriction of geographical conditions, as cultivating seedling in the area, Kunming, sowing time should be before April-by the end of June, too early or mistake late sowing kind will directly influence the field planting of surviving of seedling or seedling, and the present invention is the seedling of breeding by tissue culture technique, be not subjected to the restriction of weather conditions, simultaneously tissue cultivating seedling can be by switching long preservation, in case can enter the cultivation of a new round when needing once more.
2, reproduction coefficient height, speed is fast, particularly very favourable to the procreation of new introduced variety or rare kind, for example from external introduction one new varieties, with prior art breeding, survival rate only is 5%, and through 150 days tissue culture, develop into the tissue cultivating seedling of strain more than 300 from original 1 strain, this is that prior art can't be accomplished in a short time.
3, help rejuvenation, the purifying of kind, be of value to the preservation of germplasm: conventional prior art such as asexual reproduction method can only be bred, and can't reach the long preservation of rejuvenation, purifying and germplasm; Utilize the technology of the present invention, can be weak, severe infections is viral, the kind of disease is carried out detoxification and cultivated on the medium of configuration, makes clone rejuvenation, purifying to growing, reach the purpose that the normal phase of germplasm preserves.
4, the present invention is to preventing deterioration of variety, and carrying out long-term breeding germ plasm resource preservation is one of highly effective technology.
Further specify essentiality content of the present invention with embodiment below, but content of the present invention is not limited thereto.
Embodiment one:
The budling of selecting good Dalmatian chrysanthemum choiceness for use is an explant, is selected in the later stage of nourishing and growing the period of drawing materials of budling to carry out the season that reproductive stage, plant begin to take out spray, the budling that the explant preferred length is 0.5-1.5 centimetres.On the MS medium of additional variety classes and concentration hormone, directly induce and sprout, then by propagation, increase strong, take root, the transition of test-tube plantlet bottle outlet, field planting, cultivation form quick breeding;
It is the basis that the medium of above-mentioned breeding is selected MS for use, sucrose 3%, and agar 0.8-1.0%, the bud inducing culture is preferably used 6-BA
1.0Or 6-BA
1.0+ NAA
0.40Proliferated culture medium is selected MS+6-BA for use
1.0+ NAA
0.25-0.40mg/l or MS+6-BA
0.1-2.0+ NAA
0.25Proliferated culture medium is selected MS+6-BA for use
0.1Mg/l+NAA
0.25Or MS+6-BA
0.5+ NAA
0.25Root media MS or MS+NAA
0.2-2.0, PH5.4-5.6, conventional autoclaving; 22-28 ℃ of test-tube plantlet condition of culture daytimes; 20-25 ℃ of nights; Illumination 12h/d, intensity of illumination 1500-1800Lux.
The transition of test-tube plantlet bottle outlet was chosen in culture of rootage after 9-12 days, when the seedling cauline leaf is grown 4-6 centimetres, open bottle cap in the choosing in preceding 2-3 days of emerging, treat the intensification of leaf look, be transplanted into aseptic perlitic the lid in the plastic casing of being equipped with 2.5-3.0 centimetres, water spray, move into the greenhouse, regularly spray 0.1% carbendazim, begin to make in a week humidity to remain on more than 90%, gradually open the lid, be transplanted to after 10-15 days in the earth bag thereafter; Growth moved on in the outdoor cool canopy after 14-20 days in the greenhouse, increased illumination gradually, just can descend ground through 7-19 days again, can the field planting field about January.
Carry out breeding of the first round, by bud induce, propagation, rejuvenation, cultivating process such as take root, cultivate nearly 6000 strains of tissue cultivating seedling, the bottle outlet survival rate reaches more than 95%, smooth transition behind the bottle outlet, late August moves to outdoor cropping.Tissue cultivating seedling robust growth after the field planting is neat, and by excellent investigation of a series of biology and statistics, group training seedling shows and the great uniformity of previous generation maternal plant, and the sub-seedling growth potential of same kind is neat, still keeps the obvious characteristic of original strain between each clone.
Embodiment two:
Drawn material: embodiment one breeds the tissue cultivating seedling that preserve the back, this batch seedling still keeps good vitality after preserving by more than 400 days, cultivate by series such as switching, propagation, bottle outlet passes through the transitional period smoothly, simultaneously can confirm fully that the present invention shows tangible excellent beneficial effect to the preservation of Dalmatian chrysanthemum breeding germplasm.
Claims (6)
1, Dalmatian chrysanthemum tissue culture propagation and quality saving method, comprise that selecting clonal budling for use is explant, on minimal medium, directly induce and sprout, it is characterized in that bud through propagation, increase strong, take root, the indoor cultivation of test-tube plantlet bottle outlet transition is to the outdoor cultivation of field planting, cultivation, form quality saving in indoor continuous culture, it is the basis that minimal medium is selected MS for use, sucrose is with 3% of volume, agar is with 0.8-1.0% of volume, and the bud inducing culture is selected 6-BA1.0mg/l or 6-BA1.0mg/l+NAA0.4mg/l for use; Proliferated culture medium is selected MS+6-BA1.0mg/l+NAA0.25-0.40mg/l or MS+6-BA0.1-2.0mg/l+NAA0.2mg/l for use; Increase strong medium and select MS or MS+6-BA0.1mg/l+NAA0.25mg/l or MS+6-BA0.5mg/l+NAA0.25mg/l for use; Root media MS or MS+NAA0.2-2.0mg/l, PH5.4-5.6, conventional autoclaving; The test-tube plantlet condition of culture selects 20-28 ℃; Illumination 12h/d, intensity of illumination 1500-1800Lux.
2, method according to claim 1, it is characterized in that selecting for use the nourish and grow budling of later stage plant is explant.
3, method according to claim 1 and 2 is characterized in that select to nourish and grow the season that the later stage enters the reproductive growth initial stage, plant begins to take out spray in period of drawing materials of budling.
4, method according to claim 1 and 2 is characterized in that explant selects the budling of 0.5-1.5 centimetres of length.
5, method according to claim 1, the single medium that changes over to of bud that when it is characterized in that quality saving the process initial culture is induced, it is the basis that medium is selected MS for use, sucrose is with 3% of volume, agar is with 0.8-1.0% of volume, preferably with 6-BA1.0mg/l or 6-BA1.0mg/l+NAA0.40mg/l, proliferated culture medium is selected MS+6-BA1.0mg/l+NAA0.25mg/l+NAA0.25-0.40mg/l or MS+6-BA0.1-2.0mg/l+NAA0.25mg/l for use to the bud inducing culture.
6, method according to claim 1, it is characterized in that the transition of test-tube plantlet bottle outlet was chosen in culture of rootage after 9-12 days, during long 4-6 centimetres of seedling cauline leaf, opened bottle cap in preceding 2-3 days emerging, treat the intensification of leaf look, be transplanted into and 2.5-3.0 centimetres aseptic perlitic is housed has lid to mould in the box, water spray moves into the greenhouse, regularly spray 0.1% bactericide, begin to make in a week humidity to remain on more than 90%, open the lid gradually thereafter, plant after 10-15 days in the earth bag; Growing moved on in the cool canopy after 7-10 days, increased illumination gradually, moved to open country through 14-20 days, again through field planting field in January.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN97119736A CN1076161C (en) | 1997-10-10 | 1997-10-10 | Technology for germ plasma preservation and quick breeding by group culture of pyrethrum |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN97119736A CN1076161C (en) | 1997-10-10 | 1997-10-10 | Technology for germ plasma preservation and quick breeding by group culture of pyrethrum |
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| Publication Number | Publication Date |
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| CN1178067A CN1178067A (en) | 1998-04-08 |
| CN1076161C true CN1076161C (en) | 2001-12-19 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN97119736A Expired - Fee Related CN1076161C (en) | 1997-10-10 | 1997-10-10 | Technology for germ plasma preservation and quick breeding by group culture of pyrethrum |
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Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100420369C (en) * | 2006-04-30 | 2008-09-24 | 江汉大学 | Method for obtaining hormone-free tissue culture detoxified seedling of chrysanthemum for tea use |
| CN102783418B (en) * | 2012-08-21 | 2013-09-04 | 云南南宝生物科技有限责任公司 | Tissue culture method for pyrethrum cinerariifolium |
| CN103141384A (en) * | 2013-02-27 | 2013-06-12 | 上海交通大学 | Rapid tissue culture propagation method of pot chrysanthemum cultivars |
| CN104170743A (en) * | 2014-09-09 | 2014-12-03 | 安徽科技学院 | Culture medium for preserving tissue culture seedlings of stevia rebaudiana |
| CN104304023B (en) * | 2014-10-24 | 2016-05-25 | 柳州市天姿园艺有限公司 | Ground is by chrysanthemum special culture media |
| CN105475144B (en) * | 2016-02-26 | 2017-09-29 | 邓珂 | Cherish Cornmarigold tissue culture medium (TCM) |
| CN108967160A (en) * | 2018-09-12 | 2018-12-11 | 广州田园牧歌农林有限公司 | A kind of Zengcheng honey chrysanthemum test tube seedling transplantation method |
| CN109287489B (en) * | 2018-11-28 | 2021-08-20 | 云南省农业科学院花卉研究所 | Standardized in-vitro culture method of pyrethrum seedlings |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH02238829A (en) * | 1989-03-10 | 1990-09-21 | Noyaku Bio Technol Kaihatsu Gijutsu Kenkyu Kumiai | Raising seedling of dalmatian pyrethrum |
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1997
- 1997-10-10 CN CN97119736A patent/CN1076161C/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH02238829A (en) * | 1989-03-10 | 1990-09-21 | Noyaku Bio Technol Kaihatsu Gijutsu Kenkyu Kumiai | Raising seedling of dalmatian pyrethrum |
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| CN1178067A (en) | 1998-04-08 |
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