CN103141388A - Tissue culture method for ornithogalum caudatum - Google Patents
Tissue culture method for ornithogalum caudatum Download PDFInfo
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Abstract
The invention relates to a tissue culture method for ornithogalum caudatum. The method comprises the following steps of: preparing culture media, selecting and sterilizing explants, primarily cultivating, inducing adventitious buds, growing of the adventitious buds, performing root induction on the adventitious buds, and domesticating and transplanting tissue culture seedlings. The basic culture medium selects an MS culture medium; the dosage of saccharose is 20-40 g/L; a coagulator is agar powder, and the dosage of the coagulator is 8-9 g/L; the pH of the culture medium is adjusted to 5.7-5.8 before being sub-charged; the initial culture medium comprises 0.1-0.5 mg/L of MS+6-BA and 0.05-0.2 mg/L of NAA; the adventitious bud induction culture medium comprises 2.0-4.0 mg/L of MS+6-BA and 0.05-0.2 mg/L of NAA; and the adventitious bud increment culture medium comprises 0.5-3.0 mg/L of MS+6-BA and 0.05-0.2 mg/L of NAA; and the rooting medium comprises 0-0.5mg/L of MS+IBA, or 0-0.5mg/L of 1/2MS+IBA. The method has the advantages that a built tissue culture and rapid propagation technique system for ornithogalum caudatum can reach a growth coefficient of 3-5, a rooting rate of 100% and a transplanting survival rate of 100%, and the tissue culture seedlings are healthy and even. Therefore, the method is suitable for detoxification and large-scale production of excellent ornithogalum caudatum seedlings.
Description
Technical field
The present invention relates to plant tissue culture technique, plant stem apex culture technique, is mainly a kind of method that Urginea maritima tissue is cultivated.
Background technology
Urginea maritima (Ornithogalum caudatum), the another name Ornithogalum caudatum, be commonly called as the glass globe daisy, cucurbit orchid etc., for Liliaceae, Ornithogalum caudatum's platymiscium, perennial napiform root herbage flower, originate in the south, Africa, happiness sunlight, avoid direct sunlight summer, also partly cloudy cold-resistant, good moist environment, the rear leafage of heavy frost in winter still keeps green.When Urginea maritima blooms, plant height is not about 30～40cm, while blooming scape can to reach 50～60cm high, bloom the general annual the first tenday period of a month in mid-April to May, the taper inflorescence, flower adularescent, orange and polyphyll kind, quiet and tastefully laid out, simple, the florescence is long, be to arrange natural garden style and rock garden and excellent material, also be applicable to cut-flower and potted plant viewing and admiring.Simultaneously, Urginea maritima also has certain will its leaf and following bulb section or piece being steep in wine with being worth, and can relax the muscles and stimulate the blood circulation, for Orally taken and externally, for osteopatia sprain, there is a great strain on the muscles that good restitution is arranged.
At present, the breeding of Urginea maritima is mainly undertaken by seed and bulb separation modes of reproduction, but planting ball can be used for breeding as kind of a ball general annual 8～September, and seedling need to be cultivated and just can bloom in 3～4 years, kind ball, seed production are subject to season, time limit restriction seriously, annual breeding limited amount, do not utilize the popularization of Urginea maritima simultaneously.Simultaneously, due to Urginea maritima kind ball, for a long time in underground, easily be subject to the infection of various viruses, if utilize kind of a ball to carry out vegetative propagation, the poison once the kind feel of the ball is caught an illness, just be difficult to eradicate by chemicals, more seriously reduce the ornamental value of Urginea maritima.
Plant tissue culture technique is considered to the most efficient plant seedling fast breeding technique means at present, at present, there is not yet both at home and abroad and utilizes the plant tissue culture technology Urginea maritima to be carried out to the bibliographical information of Fast-propagation.
Summary of the invention
The object of the invention is to overcome the deficiency that prior art exists, and a kind of method that provides green onion tissue to cultivate solves the good Urginea maritima tissue culture regeneration system of setting up, for the production of Urginea maritima seedling, new varieties initiative and quality genetic improvement etc. are carried out technical support.
The objective of the invention is to complete by following technical solution.The method that this Urginea maritima tissue is cultivated, the method comprises following step:
1), the choosing and processing of explant: choose the underground bulb of Urginea maritima and still unopened colored fringe as starting material, bulb is peelled off to outside 1～2 layer of scale, Hua Sui is cut into 1cm length, as group training explant;
2), first culture: on superclean bench, will be through the explant of surface sterilization in being transferred to the aseptic inoculation dish, suck the explant surface moisture with aseptic filter paper, be inoculated in just and carry out just culture in culture base MS+6-BA0.1～0.5mg/L+NAA0.05～0.2mg/L, 25 ± 2 ℃ of cultivation temperature, light application time is 12～14h/d, and intensity of illumination is 45～60 μ molm
-2s
-1;
3) adventitious bud inducing: that first culture obtains is clean, survive material is inoculated in adventitious bud induction culture base MS+6-BA2.0～4.0mg/L+NAA0.05～0.2mg/L on aseptic working platform, carry out inducing of indefinite bud, 25 ± 2 ℃ of cultivation temperature, light application time is 12～14h/d, and intensity of illumination is 45～60 μ molm
-2s
-1;
4) adventitious bud proliferation: adventitious bud proliferation: on aseptic working platform, 2～3 one clump of the high indefinite bud of 2～3cm that adventitious bud induction culture is obtained, utilize aseptic scalpel that the indefinite bud base portion is caused to some wounds gently, be inoculated in the adventitious bud proliferation medium and breed cultivation, the adventitious bud proliferation medium is MS+6-BA0.5～3.0mg/L+NAA0.05～0.2mg/L, 25 ± 2 ℃ of cultivation temperature, light application time is 12～14h/d, intensity of illumination is 45～60 μ molm
-2s
-1;
5), adventitious bud rooting is induced: the indefinite bud of growing thickly that height is grown into to 3～5cm from base portion is separated into single indefinite bud, carry out inducing of adventive root in the access root media, root media is MS+IBA0～0.5mg/L or 1/2MS+IBA0～0.5mg/L, 25 ± 2 ℃ of cultivation temperature, light application time is 12～14h/d, and intensity of illumination is 45～60 μ molm
-2s
-1;
6), the domestication of group training seedling and transplanting: after the indefinite bud base portion grows the adventive root of 3～5 1～2cm, to organize training and move to greenhouse, scattered light is opened bottle cap after cultivating 5d, cultivate again 1～2d, complete the front domestication of group training transplantation of seedlings, then will organize the training seedling and take out from blake bottle, clean group training seedling surface agar, plant in matrix, 15d is cultivated in moisturizing more than 90%, then in greenhouse, normally cultivates.
Just culture, adventitious bud inducing and propagation, adventitious bud rooting induce minimal medium to be the MS medium, and dosage of sucrose is 20～40g/L, and coagulating agent is agar powder, and consumption is 8～9g/L, before the medium pH packing, is adjusted into 5.7～5.8.
The effect that the present invention is useful is:
1, carry out the Urginea maritima quick reproduction technique by tissue culture technique, produce the restriction that is not subject to area, season, weather and maternal plant growth year, be convenient to factorial seedling growth, can be produced according to order, production time and scale are controlled, for applying of Urginea maritima provides sufficient high quality seedling guarantee.
2, reproduction coefficient reaches 3～5, rooting rate 100%, and transplanting survival rate 100%, reach the requirement that the flower seed plantlet factorial seedling growth is produced, and has not yet to see the bibliographical information of cultivating about the Urginea maritima tissue.
3, the Urginea maritima adventitious bud inducing adopts direct organ to occur, and without more organizing wound to induce and callus induction link, reduces the generation of offspring's variation in the plant tissue culture process, and offspring's seedling genetic background is consistent, keeps to greatest extent maternal merit.
4, be conducive to that ripening rate is low, the popularization of even acarpous good Urginea maritima kind.
5, set up good tissue-culturing rapid propagation system, lay the foundation for utilizing from now on Plant Biotechnology Urginea maritima to be carried out to the research work such as rearing new variety, genetic improvement.
Embodiment
Below by embodiment, the present invention is further elaborated, and embodiment will help to understand better the present invention, but the present invention is not limited only to following embodiment.
The method that a kind of green onion tissue of the present invention is cultivated, step is as follows:
1, medium and condition of culture: first culture, adventitious bud inducing and propagation, adventitious bud rooting induce minimal medium to be the MS medium, dosage of sucrose is 20～40g/L, coagulating agent is agar powder, and consumption is 8～9g/L, before the medium pH packing, is adjusted into 5.7～5.8; Just the culture base is MS+6-BA0.1～0.5mg/L+NAA0.05～0.2mg/L; The adventitious bud induction culture base is MS+6-BA2.0～4.0mg/L+NAA0.05～0.2mg/L; The adventitious bud proliferation medium is MS+6-BA0.5～3.0mg/L+NAA0.05～0.2mg/L; Root media is MS+IBA0～0.5mg/L or 1/2MS+IBA0～0.5mg/L; Just culture, adventitious bud inducing and propagation, adventitious bud rooting condition of culture are, 25 ± 2 ℃ of cultivation temperature, and light application time is 12～14h/d, intensity of illumination is 45～60 μ molm
-2s
-1.
2, the processing of explant and sterilization: choose the underground bulb of Urginea maritima and still unopened colored fringe as starting material, bulb is peelled off to outside 1～2 layer of scale, Hua Sui is cut into 1cm left and right length, the explant of handling well cleans 20min with liquid detergent solution, constantly stir gently during this time, then flowing water rinses the explant material 20～30min cleaned through liquid detergent, during the explant that will clean through flowing water on the superclean bench of process sterilization is transferred to aseptic conical flask, with 75% alcohol-pickled 60s, constantly rock gently during this time conical flask, removal is attached to the explant blibbing, pour out 75% alcohol, use again 0.1%HgCl
2solution soak-out material 8～10min, or be 1%NaClO solution soak-out material 10～15min with effective chlorine density, constantly rock gently conical flask between soak period, remove and be attached to the explant blibbing, pour out 0.1%HgCl
2solution or 1%NaClO solution, the explant of disinfecting by the sterile water wash process 4～5 times, with sterile water immersion, standby.
3, first culture: on superclean bench, will be through the explant of surface sterilization in being transferred to the aseptic inoculation dish, suck the explant surface moisture with aseptic filter paper, be inoculated in just and carry out just culture in culture base MS+6-BA0.1～0.5mg/L+NAA0.05～0.2mg/L, 25 ± 2 ℃ of cultivation temperature, light application time is 12～14h/d, and intensity of illumination is 45～60 μ molm
-2s
-1.
4, adventitious bud inducing: that first culture obtains is clean, survive material is inoculated in adventitious bud induction culture base MS+6-BA2.0～4.0mg/L+NAA0.05～0.2mg/L on aseptic working platform, carry out inducing of indefinite bud, 25 ± 2 ℃ of cultivation temperature, light application time is 12～14h/d, and intensity of illumination is 45～60 μ molm
-2s
-1.
5, adventitious bud proliferation: on aseptic working platform, 2～3 one clump of the high indefinite bud of 2～3cm that adventitious bud induction culture is obtained, utilize aseptic scalpel that the indefinite bud base portion is caused to some wounds gently, be inoculated in the adventitious bud proliferation medium and breed cultivation, the adventitious bud proliferation medium is MS+6-BA0.5～3.0mg/L+NAA0.05～0.2mg/L, 25 ± 2 ℃ of cultivation temperature, light application time is 12～14h/d, intensity of illumination is 45～60 μ molm
-2s
-1.
6, adventitious bud rooting is induced: the indefinite bud of growing thickly that height is grown into to 3～5cm from base portion is separated into single indefinite bud, carry out inducing of adventive root in the access root media, root media is MS+IBA0～0.5mg/L or 1/2MS+IBA0～0.5mg/L, 25 ± 2 ℃ of cultivation temperature, light application time is 12～14h/d, and intensity of illumination is 45～60 μ molm
-2s
-1, rooting rate 100%.
7, the domestication of group training seedling and transplanting: after the indefinite bud base portion grows the adventive root of 3～5 1～2cm, to organize training and move to greenhouse, scattered light is opened bottle cap after cultivating 5d, then cultivates 1～2d, completes the front domestication of group training transplantation of seedlings, then will organize the training seedling takes out from blake bottle, clean group training seedling surface agar, plant in matrix, 15d is cultivated in moisturizing more than 90%, then normally cultivate transplanting survival rate 100% in greenhouse.
Finally, it should be pointed out that above example is only the more representational example of the present invention.Obviously, technical scheme of the present invention is not limited to above-mentioned example, and many distortion can also be arranged, and all distortion that those of ordinary skill in the art can directly derive or associate from content disclosed by the invention, all should think protection scope of the present invention.
Claims (2)
1. the method that a Urginea maritima tissue is cultivated, it is characterized in that: the method comprises following step:
1), the choosing and processing of explant: choose the underground bulb of Urginea maritima and still unopened colored fringe as starting material, bulb is peelled off to outside 1～2 layer of scale, Hua Sui is cut into 1cm length, as group training explant;
2), first culture: on superclean bench, will be through the explant of surface sterilization in being transferred to the aseptic inoculation dish, suck the explant surface moisture with aseptic filter paper, be inoculated in just and carry out just culture in culture base MS+6-BA0.1～0.5mg/L+NAA0.05～0.2mg/L, 25 ± 2 ℃ of cultivation temperature, light application time is 12～14h/d, and intensity of illumination is 45～60 μ molm
-2s
-1;
3) adventitious bud inducing: that first culture obtains is clean, survive material is inoculated in adventitious bud induction culture base MS+6-BA2.0～4.0mg/L+NAA0.05～0.2mg/L on aseptic working platform, carry out inducing of indefinite bud, 25 ± 2 ℃ of cultivation temperature, light application time is 12～14h/d, and intensity of illumination is 45～60 μ molm
-2s
-1;
4) adventitious bud proliferation: adventitious bud proliferation: on aseptic working platform, 2～3 one clump of the high indefinite bud of 2～3cm that adventitious bud induction culture is obtained, utilize aseptic scalpel that the indefinite bud base portion is caused to some wounds gently, be inoculated in the adventitious bud proliferation medium and breed cultivation, the adventitious bud proliferation medium is MS+6-BA0.5～3.0mg/L+NAA0.05～0.2mg/L, 25 ± 2 ℃ of cultivation temperature, light application time is 12～14h/d, intensity of illumination is 45～60 μ molm
-2s
-1;
5), adventitious bud rooting is induced: the indefinite bud of growing thickly that height is grown into to 3～5cm from base portion is separated into single indefinite bud, carry out inducing of adventive root in the access root media, root media is MS+IBA0～0.5mg/L or 1/2MS+IBA0～0.5mg/L, 25 ± 2 ℃ of cultivation temperature, light application time is 12～14h/d, and intensity of illumination is 45～60 μ molm
-2s
-1;
6), the domestication of group training seedling and transplanting: after the indefinite bud base portion grows the adventive root of 3～5 1～2cm, to organize training and move to greenhouse, scattered light is opened bottle cap after cultivating 5d, cultivate again 1～2d, complete the front domestication of group training transplantation of seedlings, then will organize the training seedling and take out from blake bottle, clean group training seedling surface agar, plant in matrix, 15d is cultivated in moisturizing more than 90%, then in greenhouse, normally cultivates.
2. the method that Urginea maritima tissue according to claim 1 is cultivated, it is characterized in that: first culture, adventitious bud inducing and propagation, adventitious bud rooting induce minimal medium to be the MS medium, dosage of sucrose is 20～40g/L, coagulating agent is agar powder, consumption is 8～9g/L, before the medium pH packing, is adjusted into 5.7～5.8.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104521754A (en) * | 2014-12-10 | 2015-04-22 | 湖北省农业科学院经济作物研究所 | Rapid propagation method of Allium ovalifolium on high mountain |
| CN105815214A (en) * | 2016-03-21 | 2016-08-03 | 云南省农业科学院农业环境资源研究所 | Leaf seedling rapid breeding method of ornithogalum caudatum |
| CN106035080A (en) * | 2016-05-30 | 2016-10-26 | 甘肃省治沙研究所 | Allium mongolicum regenerated seedling and allium mongolicum callus acquisition method |
| CN108617510A (en) * | 2018-05-10 | 2018-10-09 | 漳州市农业科学研究所 | A method of improving wide leaf spring grass tissue-cultured seedling transplanting survival rate |
| CN109430059A (en) * | 2018-12-25 | 2019-03-08 | 云南省农业科学院花卉研究所 | A kind of cut-flower Ornithogalum caudatum in vitro culture quick-breeding method |
| CN115843692A (en) * | 2022-12-29 | 2023-03-28 | 浙江省农业科学院 | Allium fistulosum tissue culture method |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104521754A (en) * | 2014-12-10 | 2015-04-22 | 湖北省农业科学院经济作物研究所 | Rapid propagation method of Allium ovalifolium on high mountain |
| CN104521754B (en) * | 2014-12-10 | 2016-06-15 | 湖北省农业科学院经济作物研究所 | High mountain ovum leaf fragrant-flowered garlic method for quickly breeding |
| CN105815214A (en) * | 2016-03-21 | 2016-08-03 | 云南省农业科学院农业环境资源研究所 | Leaf seedling rapid breeding method of ornithogalum caudatum |
| CN105815214B (en) * | 2016-03-21 | 2018-01-02 | 云南省农业科学院农业环境资源研究所 | A kind of blade seedling rapid propagation method of Ornithogalum caudatum |
| CN106035080A (en) * | 2016-05-30 | 2016-10-26 | 甘肃省治沙研究所 | Allium mongolicum regenerated seedling and allium mongolicum callus acquisition method |
| CN108617510A (en) * | 2018-05-10 | 2018-10-09 | 漳州市农业科学研究所 | A method of improving wide leaf spring grass tissue-cultured seedling transplanting survival rate |
| CN108617510B (en) * | 2018-05-10 | 2021-06-29 | 漳州市农业科学研究所 | Method for improving transplanting survival rate of tissue culture seedlings of broad-leaf spring grass |
| CN109430059A (en) * | 2018-12-25 | 2019-03-08 | 云南省农业科学院花卉研究所 | A kind of cut-flower Ornithogalum caudatum in vitro culture quick-breeding method |
| CN115843692A (en) * | 2022-12-29 | 2023-03-28 | 浙江省农业科学院 | Allium fistulosum tissue culture method |
| CN115843692B (en) * | 2022-12-29 | 2023-09-29 | 浙江省农业科学院 | Tissue culture method for allium fistulosum |
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Application publication date: 20130612 |