CN102232359B - In-vitro rapid propagation method of double-petal Jasminum sambac - Google Patents
In-vitro rapid propagation method of double-petal Jasminum sambac Download PDFInfo
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
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- 239000002363 auxin Substances 0.000 claims description 11
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- JTEDVYBZBROSJT-UHFFFAOYSA-N indole-3-butyric acid Chemical compound C1=CC=C2C(CCCC(=O)O)=CNC2=C1 JTEDVYBZBROSJT-UHFFFAOYSA-N 0.000 claims description 10
- 238000004659 sterilization and disinfection Methods 0.000 claims description 10
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 claims description 9
- 238000005406 washing Methods 0.000 claims description 8
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- UZKQTCBAMSWPJD-UQCOIBPSSA-N trans-Zeatin Natural products OCC(/C)=C\CNC1=NC=NC2=C1N=CN2 UZKQTCBAMSWPJD-UQCOIBPSSA-N 0.000 claims description 6
- UZKQTCBAMSWPJD-FARCUNLSSA-N trans-zeatin Chemical compound OCC(/C)=C/CNC1=NC=NC2=C1N=CN2 UZKQTCBAMSWPJD-FARCUNLSSA-N 0.000 claims description 6
- 229940023877 zeatin Drugs 0.000 claims description 6
- 238000004140 cleaning Methods 0.000 claims description 5
- 229960002523 mercuric chloride Drugs 0.000 claims description 5
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 claims description 5
- 239000007921 spray Substances 0.000 claims description 5
- 239000012153 distilled water Substances 0.000 claims description 4
- 229930024421 Adenine Natural products 0.000 claims description 3
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 claims description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 3
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- 229960000643 adenine Drugs 0.000 claims description 3
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- 238000011010 flushing procedure Methods 0.000 claims description 3
- IXORZMNAPKEEDV-UHFFFAOYSA-N gibberellic acid GA3 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(C=CC3O)C1C3(C)C(=O)O2 IXORZMNAPKEEDV-UHFFFAOYSA-N 0.000 claims description 3
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Abstract
The invention provides a rapid propagation method of double-petal Jasminum sambac, which is used for large-scale production and propagation to meet the market needs. The tissue culture of double-petal Jasminum sambac comprises the following steps: A, disinfecting explants; B, obtaining sterile seedlings; C, growing and cultivating axillary buds; D, inducing cluster buds; E, rooting and cultivating; and F, exercising and transplanting seedlings. As the double-petal Jasminum sambac is propagated by the plant tissue culture method, the propagation is not influenced by external conditions and can be performed in all seasons while the occupied land of the seedlings is saved and the production cost is reduced; and moreover, all good characters of the parent can be maintained, and the hereditary character is stable. By using the method, a great number of good test tube seedlings can be formed in short time for large-scale and factory production, thereby providing abundant raw materials for the industries such as ornamental horticulture, spices, pharmacy and the like.
Description
Technical field
The present invention relates to bivalve jasmine method for quickly breeding, belong to artificial propagation and the cultivation method technical field of plant.
Background technology
Jasmine (Jasminum sambac L.) is perennial evergreen undershrub of Oleaceae (Oleaceae) gelsemium or the undershrub that slightly becomes the liana shape; Property happiness warm and moist; Originate in an India and an Arabic band; After introducing China,, adapted to China's south most of areas weather through long cultivation domestication.As a kind of important fragrant plant, Jasmine is the important source material of refining essential oil and basement system jasmine tea.At home, since the root of Jasmine or complete stool have treatment diarrhoea, alleviate medical value such as abdominal pain, anti-inflammatory and cultivated application as a kind of central bay herbal medicine.Also have the researcher to find that the jasmine tea fragrance of low concentration can effectively be releived because the pain that vegetative nerve is movable and mood is brought.Except medical value.Jasmine still is a kind of famous ornamental plants, is not only Philippine's national flower, also is chosen as Jiangsu Province in 2007 simultaneously and spends.Therefore, jasmine receives much concern owing to its important use is worth.
Because the ripening rate of jasmine is very low, in production application, all breeds through cuttage and seedling culture.But long-term cottage propagation is to cause weakening and viruliferous major reason of plant vigor, and has a strong impact on resistivity and fragrance of a flower quality and the quality of jasmine to adverse circumstance, and has restricted the breeding process of jasmine.
Summary of the invention
The object of the present invention is to provide a kind of bivalve jasmine method for quickly breeding, carry out large-scale production and train the needs of the research work of other subjects that are the basis with bivalve jasmine group.
Technical scheme of the present invention is following, and the main feature of this kind bivalve jasmine method for quickly breeding is:
A explant sterilization: choose the full and spray that do not sprout as yet of axillalry bud, pluck blade, then branch is cut the segment into about 2cm; Be with a pair of axillalry bud for every section, the Amway cleaning solution with 0.1% shakes washing 15～20min on shaking table, behind flowing water flushing 30～60min; Again on superclean bench with 75% alcohol rinsing 30s, after the sterile water wash 4 times, with containing 0.1% mercuric chloride sterilization 50-60min; Aseptic water washing 4-5 time, blot surface moisture with aseptic filter paper at last;
The B axillalry bud is induced: the bivalve jasmine stem section that disinfects is cut into the band bud sections of 1.0-1.5cm, is inoculated in the following medium: WPM minimal medium, sucrose (table sugar) 20～40g/L, proper pH value; 28 ± 1 ℃ of cultivation temperature, intensity of illumination 1000～3000lx, light application time 10～16h carries out illumination cultivation;
The C axillalry bud is cultivated: the aseptic seedling stem apex of cultivating about 20 days behind the axillary bud sprouting is cut with scissors, be inoculated in the following medium: WPM minimal medium, sucrose (table sugar) 20～40g/L, auxin 0.2～2.0mg/L, proper pH value; 28 ± 1 ℃ of cultivation temperature,
The D inducing clumping bud: the bud seedling that will grow to about 8cm downcuts, and is divided into the segment that has a pair of axillalry bud: the WPM minimal medium, and auxin 0.5～1.0mg/L carried out illumination cultivation 30～40 days;
E culture of rootage: choose healthy and strong test-tube plantlet; After pruning; After containing auxin 400～500mg/L immersion 10-15min, be inoculated in the following medium: 1/2WPM or WPM minimal medium, sucrose (table sugar) 20～40g/L carried out illumination cultivation 50～60 days.
F acclimatization and transplants: the tissue culture bottle lid is opened, and added the distilled water of 5～10ml, carry out nonsterile conditions domestication 2-3d; With tweezers tissue cultivating seedling is taken out from bottle, is transplanted in the cultivation matrix of having sterilized, shelter from heat or light the domestication 3 weeks, during the irregular 1/2WPM nutrient solution that sprays.
The elongation technology scheme of described bivalve jasmine method for quickly breeding comprises: the auxin that the bivalve jasmine relates in the described medium of breeding fast among pH value scope among sterilization reagent in the steps A and time, the step B and step B, C, D, the E, comprise 6-Bian Ji adenine (6-BA), indolebutyric acid (IBA), methyl (NAA), gibberellin (GA3), zeatin (ZT) wherein one or more.
The elongation technology scheme of described bivalve jasmine method for quickly breeding comprises: the seed disinfection in the steps A is shaken washing 15～20min and is contained 0.1% mercuric chloride sterilization, 50～60min with the Amway cleaning solution of reagent 0.1% on shaking table.
The elongation technology scheme of described bivalve jasmine method for quickly breeding comprises; PH value scope is 5.8～7.0 among the step B.
The elongation technology scheme of described bivalve jasmine method for quickly breeding comprises: it is characterized in that the involved auxin of step C comprises indolebutyric acid (IBA) 1.0～2.0mg/L, 6-Bian Ji adenine (6-BA) 0.1～0.5mg/L, gibberellin (GA
3) 0.5～1.0mg/L.
The elongation technology scheme of described bivalve jasmine method for quickly breeding comprises: involved auxin zeatin (ZT) 0.5～1.0mg/L among the step D, indolebutyric acid (IBA) 0.1～0.5mg/L.
The elongation technology scheme of described bivalve jasmine method for quickly breeding comprises: involved auxin methyl (NAA) 400～500mg/L in the step e, soak time is 10-15min.
The elongation technology scheme of described bivalve jasmine method for quickly breeding comprises: add distilled water 5～10ml in the step F, bacterium domestication 2-3 days are arranged.
Adopt the method breeding bivalve jasmine of Plant Tissue Breeding, not only can break away from the influence of external condition, the four seasons can both produce, and can practice thrift the occupation of land of growing seedlings, and reduce production costs.Plant tissue culture technique is a totipotency of utilizing cell, takes one block organization of cell clique on the plant corpus, through the control of artificial condition, make these tissues form millions upon millions of plant, and preserved whole merits of parent, and stabilization characteristics of genetics.This method has kept the plant activity of bivalve jasmine, can form a large amount of good test-tube plantlets in a short time, carries out scale, batch production production.
Following constipation closes embodiment and accompanying drawing, and specific embodiments of the invention is done further to detail.
Description of drawings
Fig. 1 is the bivalve jasmine plant figure of usefulness of drawing materials;
Fig. 2 is the axillalry bud figure (condition of culture: WPM minimal medium, table sugar 30g/L, 16h/d illumination cultivation) that induces 2-3 to sprout after week;
Fig. 3 cultivates axillalry bud seedling figure (condition of culture: WPM+IBA2mg/L+6-BA0.5mg/L+GA30.7mg/L+ table sugar 30g/L, 16h/d illumination cultivation) in the optimum growh medium;
Fig. 4 is inducing clumping bud figure (condition of culture: WPM+ZT1mg/L+IBA0.5mg/L+ table sugar 30g/L, 16h/d illumination cultivation);
Fig. 5 is the situation map of taking root after tissue cultivating seedling is handled in step e;
Fig. 6 is that the group training obtains complete plant figure;
Fig. 7 is the transplanting tissue cultivating seedling figure through the domestication of refining seedling.
Embodiment
Instance below in conjunction with the bivalve jasmine breeds is fast explained embodiment of the present invention.
The inducing and cultivating of embodiment 1 axillalry bud
1, get the tender stem segments of bivalve jasmine, the Amway cleaning solution with 0.1% shakes washing 15～20min on shaking table, behind the flowing water flushing 60min, again on superclean bench with 75% alcohol rinsing 30s, after the sterile water wash 4 times, with containing 0.1%HgCL
2Sterilization 50～60min (in mercuric chloride, adding several Tween 80s), aseptic water washing 4-5 time blots the surface of the seed moisture with aseptic filter paper at last.
2, the bivalve jasmine stem section that disinfects directly is inoculated in the axillalry bud inducing culture: add sucrose (table sugar) 30g/L, proper pH value in the WPM minimal medium; 28 ± 1 ℃ of cultivation temperature, intensity of illumination 1000～3000lx, light application time 10～16h carries out illumination cultivation.
3, cut with scissors cultivating 20 days left and right sides aseptic seedling stem apexs behind the axillary bud sprouting, be inoculated in the axillary bud growth medium of the present invention, concrete prescription is for adding plant hormone IBA2.0mg/L, BA0.5mg/L and GA in the WPM minimal medium
30.7mg/L sucrose 30g/L carries out illumination cultivation.
Embodiment 2 grow thickly inducing of bud and taking root, transplanting of test-tube plantlet
1, cultivates after 30 days, treat that the bud seedling grows to 6-8cm, the renewed vaccination of bud seedling in inducing clumping bud medium of the present invention, is carried out enrichment culture.The inducing clumping bud medium is to be minimal medium with the WPM medium, and adds auxin ZT1mg/L and IBA0.5mg/L, and sucrose (table sugar) 30g/L carries out enrichment culture.Cultivate more than 30 days, inductivity reaches 98%, and most of bud seedling can induce the bud number of growing thickly and reach 3～5.
2, choose the test-tube plantlet of robust growth, it is immersed in 10min in the solution that contains NAA450mg/L of the bacterium of going out, be inoculated into then on the 1/2MS minimal medium, sucrose (table sugar) 15g/L carries out root induction.Cultivate after 40 days, rooting rate reaches 86%, and the average root number reaches 18.5.
3, after bivalve jasmine tissue cultivating seedling culture of rootage 1.5-2 month, begin to carry out acclimatization and transplants; At first tissue culture bottle is moved into illumination box, carry out alternating temperature and become light acclimation 5-7 days, again the tissue culture bottle lid is opened; And add 10ml distilled water, carry out nonsterile conditions domestication 2-3 days.Then with tissue cultivating seedling be transplanted to the cultivation matrix of having sterilized (mire: perlite: garden mould=1: 1: 5), place in the greenhouse that does not have the light of heating, shelter from heat or light 3 weeks of domestication, during the irregular 1/2WPM nutrient solution that sprays, transplant the back survival rate and reach 78%.
The bivalve jasmine stem with bud tissue-culturing rapid propagation situation of above instance is seen accompanying drawing.
Claims (3)
1. the method for in-vitro rapid propagation of bivalve jasmine is characterized in that:
A explant sterilization: choose the full and spray that do not sprout as yet of axillalry bud, pluck blade, then branch is cut the segment into about 2cm; Be with a pair of axillalry bud for every section, the Amway cleaning solution with 0.1% shakes washing 15～20min on shaking table, behind flowing water flushing 30～60min; Again on superclean bench with 75% alcohol rinsing 30s, after the sterile water wash 4 times, with containing 0.1% mercuric chloride sterilization 50-60min; Aseptic water washing 4-5 time, blot surface moisture with aseptic filter paper at last;
The B axillalry bud is induced: the bivalve jasmine stem section that disinfects is cut into the band bud sections of 1.0-1.5cm, is inoculated in the following medium: WPM minimal medium, sucrose 20～40g/L, proper pH value; 28 ± 1 ℃ of cultivation temperature, intensity of illumination 1000～3000lx, light application time 10～16h carries out illumination cultivation;
The C axillalry bud is cultivated: the aseptic seedling stem apex of cultivating about 20 days behind the axillary bud sprouting is cut with scissors; Be inoculated in the following medium: the WPM minimal medium; Sucrose 20～40g/L; Auxin indolebutyric acid (IBA) 1.0～2.0mg/L, 6-Bian Ji adenine (6-BA) 0.1～0.5mg/L, gibberellin (GA
3) 0.5～1.0mg/L, proper pH value; 28 ± 1 ℃ of cultivation temperature;
The D inducing clumping bud: the bud seedling that will grow to about 8cm downcuts; Be divided into the segment that has a pair of axillalry bud; Be inoculated in the following medium: WPM minimal medium, sucrose 30g/L, auxin zeatin (ZT) 0.5～1.0mg/L; Indolebutyric acid (IBA) 0.1～0.5mg/L carried out illumination cultivation 30～40 days;
E culture of rootage: choose healthy and strong test-tube plantlet; After pruning; Be inoculated in the following medium after in the solution that contains auxin methyl (NAA) 400～500mg/L, soaking 10-15min: 1/2WPM or WPM minimal medium, sucrose 20～40g/L carried out illumination cultivation 50～60 days;
F acclimatization and transplants: the tissue culture bottle lid is opened, and added the distilled water of 5～10ml, carry out nonsterile conditions domestication 2-3d; With tweezers tissue cultivating seedling is taken out from bottle, is transplanted in the cultivation matrix of having sterilized, shelter from heat or light the domestication 3 weeks, during the irregular 1/2WPM nutrient solution that sprays.
2. according to the method for in-vitro rapid propagation of the said bivalve jasmine of claim 1, it is characterized in that the seed disinfection in the steps A is shaken washing 15～20min and 0.1% mercuric chloride sterilization, 50～60min with the Amway cleaning solution of reagent 0.1% on shaking table.
3. the method for in-vitro rapid propagation of bivalve jasmine according to claim 1 is characterized in that pH value scope involved among the step B is 5.8～7.0.
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| CN102499043A (en) * | 2011-11-21 | 2012-06-20 | 江苏省农业科学院 | Cutting water culture rooting method for jasminum sambac |
| CN103238518B (en) * | 2013-04-28 | 2015-03-25 | 福建省农业科学院农业工程技术研究所 | Method for induction and subculture maintenance on jasmine flower calluses |
| CN109380123B (en) * | 2018-12-25 | 2021-09-21 | 福建农林大学 | Formula and application of subculture medium for tissue culture seedlings of pearl color osmanthus fragrans |
| CN110663555B (en) * | 2019-11-11 | 2022-08-26 | 绵阳师范学院 | Rapid propagation method of odorous jasminum grandiflorum |
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| JPH0257130A (en) * | 1988-08-19 | 1990-02-26 | Fuji Spinning Co Ltd | Production of adventitious embryo and/or adventitious bud of jasmine plant |
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Non-Patent Citations (3)
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| JP平2-57130A 1990.02.26 |
| 孙艳妮等."茉莉离体快繁体系的建立".《浙江农业学报》.2009,第21卷(第4期),第390-394页. |
| 蔡汉等."茉莉离体微繁及无糖生根技术".《江苏农业学报》.2007,第23卷(第5期),第464-468页. |
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