CN103141388A - Tissue culture method for ornithogalum caudatum - Google Patents

Tissue culture method for ornithogalum caudatum Download PDF

Info

Publication number
CN103141388A
CN103141388A CN2013100740529A CN201310074052A CN103141388A CN 103141388 A CN103141388 A CN 103141388A CN 2013100740529 A CN2013100740529 A CN 2013100740529A CN 201310074052 A CN201310074052 A CN 201310074052A CN 103141388 A CN103141388 A CN 103141388A
Authority
CN
China
Prior art keywords
culture
adventitious bud
bud
adventitious
explant
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.)
Pending
Application number
CN2013100740529A
Other languages
Chinese (zh)
Inventor
汪一婷
陈志�
牟豪杰
吕永平
周迪江
陈剑平
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Zhejiang Academy of Agricultural Sciences
Original Assignee
Zhejiang Academy of Agricultural Sciences
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Zhejiang Academy of Agricultural Sciences filed Critical Zhejiang Academy of Agricultural Sciences
Priority to CN2013100740529A priority Critical patent/CN103141388A/en
Publication of CN103141388A publication Critical patent/CN103141388A/en
Pending legal-status Critical Current

Links

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

A kind of method that Urginea maritima tissue is cultivated
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.
CN2013100740529A 2013-03-08 2013-03-08 Tissue culture method for ornithogalum caudatum Pending CN103141388A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN2013100740529A CN103141388A (en) 2013-03-08 2013-03-08 Tissue culture method for ornithogalum caudatum

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN2013100740529A CN103141388A (en) 2013-03-08 2013-03-08 Tissue culture method for ornithogalum caudatum

Publications (1)

Publication Number Publication Date
CN103141388A true CN103141388A (en) 2013-06-12

Family

ID=48539950

Family Applications (1)

Application Number Title Priority Date Filing Date
CN2013100740529A Pending CN103141388A (en) 2013-03-08 2013-03-08 Tissue culture method for ornithogalum caudatum

Country Status (1)

Country Link
CN (1) CN103141388A (en)

Cited By (6)

* Cited by examiner, † Cited by third party
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

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1507770A (en) * 2002-12-18 2004-06-30 薛留芳 Cultivation method of Rohdea japonica with special modelling
JP2008048655A (en) * 2006-08-24 2008-03-06 Akita Prefectural Univ Method for producing ornithogalum thyrsoides bulb
US20100138969P1 (en) * 2008-12-01 2010-06-03 Rodolfo Valdoz Bautista Ornithogalum plant named 'Bethlehem'

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1507770A (en) * 2002-12-18 2004-06-30 薛留芳 Cultivation method of Rohdea japonica with special modelling
JP2008048655A (en) * 2006-08-24 2008-03-06 Akita Prefectural Univ Method for producing ornithogalum thyrsoides bulb
US20100138969P1 (en) * 2008-12-01 2010-06-03 Rodolfo Valdoz Bautista Ornithogalum plant named 'Bethlehem'

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
杨国会等: "不同激素水平对虎眼万年青组织培养的影响", 《湖北农业科学》, vol. 50, no. 1, 31 January 2011 (2011-01-31) *
韦三立: "《花卉组织培养》", 31 January 2001, article "虎眼万年青的组织培养", pages: 52 - 19 *

Cited By (10)

* Cited by examiner, † Cited by third party
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
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

Similar Documents

Publication Publication Date Title
CN103141388A (en) Tissue culture method for ornithogalum caudatum
CN106718883A (en) A kind of Yunnan Chinese catalpa and the miniature engrafting method of test tube seedling of Chinese catalpa
CN103141387A (en) Method for cultivating haworthia maughanii tissue
CN106417011A (en) Wild bletilla striata tissue culture rapid propagation method
CN102440192B (en) Culture media for peony high frequency embryogenic callus differentiation, and culture method thereof
CN104585036B (en) The tissue culture and rapid propagation method of the fiery axillalry bud on Malus spectabilis plateau, a kind of North America
CN106489738B (en) A kind of production method of spindle tree leaf regeneration plant
CN102960243A (en) Method for tissue culture and rapid propagation by basal disc-free scales of Lycoris chinensis
CN107197746A (en) A kind of mating system of China fir field excellent resources
CN102217538B (en) Method for treating walnut tissue culture stem segments inside test tubes and rooting outside test tubes
CN103270947B (en) Duvalia angustiloba tissue culturing method
CN106665367B (en) A kind of Golden Bell Tree quick breeding method for tissue culture
CN103548695B (en) A kind of meadowrueleaf corydalis root quick breeding method for tissue culture
CN102613087A (en) Method for culturing and breeding Correa carmen by using biological tissue
CN104488722A (en) Quick propagation method for tissue culture of staurogyne sp
CN104604689A (en) Method for rapidly obtaining chicory explants and improving callus rate of chicory explants
CN104012406A (en) Regeneration in-vitro method for sweet cherry variety wanhongzhu
CN107278893A (en) A kind of apple rootstock SH40 rapid propagation methods
CN105284622B (en) A kind of method that quick acquisition Rhizoma Iridis Tectori hybridizes superior clone
CN102232359B (en) In-vitro rapid propagation method of double-petal Jasminum sambac
CN107223566A (en) A kind of Wulian poplar method for tissue culture
CN109247147A (en) A kind of method that tea tree rapid cuttage is taken root
CN104604735A (en) Tissue culture and rapid propagation method for American lagerstroemia indica pink velour
CN104488709A (en) Method for culturing bulb tissues of tulbaghia violacea floral leaf
CN104488710B (en) A kind of method of Calotropis gigantea tissue cultures

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C12 Rejection of a patent application after its publication
RJ01 Rejection of invention patent application after publication

Application publication date: 20130612