CN104255517A - Rapid propagation method of suspension cells of euphorbia lathyris - Google Patents
Rapid propagation method of suspension cells of euphorbia lathyris Download PDFInfo
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- CN104255517A CN104255517A CN201410540511.2A CN201410540511A CN104255517A CN 104255517 A CN104255517 A CN 104255517A CN 201410540511 A CN201410540511 A CN 201410540511A CN 104255517 A CN104255517 A CN 104255517A
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- euphorbia lathyris
- suspension cell
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- lathyris
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Abstract
The invention researches a rapid propagation method of suspension cells of euphorbia lathyris. The method comprises steps as follows: obtaining of sterile leaves of the euphorbia lathyris, induction of calluses, culture of the suspension cells, mutation of the suspension cells and the like. The euphorbia lathyris cells produced with the method are high in viability, short in growth cycle and uniform and stable in shape, and a foundation is laid for industrial production of medicinal plant cells.
Description
Technical field
The present invention relates to the quick-breeding method that the suspension of a kind of Euphorbia lathyris is cultivated, belong to biological technical field.
Background technology
Euphorbia lathyris, also known as moleplant seed, Euphorbiaceae,
euphorbia lathylris L, biennial herb, complete stool is without hair.Root column, stem is upright.Leaf alternately to life, wire lanceolar, Quan Yuan.Inflorescence Dan Sheng, nearly mitriform.Male flower is most, female flower 1 piece.Capsule three prismatic is spherical, Glabrous.Seed column to ossphere shape, brown or taupe.The florescence 4-7 month, the fruit phase 6-9 month.Produce most of China provinces and regions, cultivation or ease are for wild extensive distribution or cultivate in Europe, north African, the Central Asia, East Asia and South and North America.Herb is poisonous.Tool medical value.Euphorbia lathyris seed oil content generally reaches about 45%, and high reaches more than 48%.The fatty acid composition of Euphorbia lathyris oil is similar with the molecular composition of substitute of diesel fuel, and its main component is sesquiterpene terpene, is rich in ingenol 3 one hexadecane ester, plant milk is rich in a large amount of olefines hydrocarbon, Euphorbia lathyris happiness is warm, illumination and middle raw environment, and resistance is stronger, easy cultivation, should be moistening, water funk flood, requires not tight to soil, sandy loam, loess, in vain meals are native, wheatland is native, but best with sandy loam humus soil.Be born in paddy field, low humidity dry land and rand.In the seed of Euphorbia lathyris, stem, leaf and stem white milk all can be used as medicine, detumescence of relieving oedema or abdominal distension through diuresis or purgation, broken disease desinsection, catharsis, calmness, analgesia, anti-inflammatory, the effect such as antibacterial, antitumor.Clinical report, advanced schistosomiasis ascites, venomous snake bite, women can be treated through the disease such as closing.Medicinal material is the seed of euphorbia plant Euphorbia lathyris.Between 8 ~ September, after seed maturity, extract herb, dry, lay seed, remove impurity.Nature and flavor are pungent, temperature, poisonous.Mainly seminal propagation.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of method for quickly breeding of Euphorbia lathyris suspension cell, and the Euphorbia lathyris cytoactive prepared by the method is high, and growth cycle is short, cell shape stable homogeneous, for Medicinal Plant Cell suitability for industrialized production lays the foundation.
Technical problem to be solved by this invention is realized by following scheme:
Get the blade that Euphorbia lathyris children is tender, static 15min in bleaching powder, running water surface smut, on superclean bench 0.2% mercuric chloride sterilization 10min, aseptic water washing 5 times, blot stand-by, callus induction is carried out in the blade access Miller+ZT0.5mg/L+KT1.5mg/L+0.4mg/ml penicillin medium that the Euphorbia lathyris children disinfected is tender, additional saccharose 30g/L, agar 6.5g/L, pH6.0, intensity of illumination 1000lx, light application time 6h/d, temperature 22 DEG C, the callus derived is put into liquid nutrient medium Miller+0.3mg/LIAA+1.0-1.5mg/L2ip+100mg/L lactoalbumin hydrolysate and is carried out liquid suspension cell chulture, constant temperature oscillator 100r/min, inoculum concentration is 2.5g, cultivation cycle is 40d, additional saccharose 30g/L, pH5.8, illumination 2500lx, light application time 15h/d, temperature 24 DEG C, cultivate the suspension cell after month put into filtration sterilization after ethylmethane sulfonate solution, concentration is 0.3%-1%, processing time is 1-5h, add after process in fresh medium and cultivate.
The suspension cell activity adopting the present invention to prepare is high, and the cycle is short, and output is large, pollutes little, is beneficial to implant mass.
Below in conjunction with embodiment, the present invention is further elaborated, but the scope of protection of present invention is not limited to following embodiments.
Embodiment
Embodiment 1
Get the blade that Euphorbia lathyris children is tender, static 15min in bleaching powder, running water surface smut, on superclean bench 0.2% mercuric chloride sterilization 10min, aseptic water washing 5 times, blot stand-by, callus induction is carried out in the blade access Miller+ZT0.5mg/L+KT1.5mg/L+0.4mg/ml penicillin medium that the Euphorbia lathyris children disinfected is tender, additional saccharose 30g/L, agar 6.5g/L, pH6.0, intensity of illumination 1000lx, light application time 6h/d, temperature 22 DEG C, the callus derived is put into liquid nutrient medium Miller+0.3mg/LIAA+1.0mg/L2ip+100mg/L lactoalbumin hydrolysate and is carried out liquid suspension cell chulture, constant temperature oscillator 100r/min, inoculum concentration is 2.5g, cultivation cycle is 40d, additional saccharose 30g/L, pH5.8, illumination 2500lx, light application time 15h/d, temperature 24 DEG C, cultivate the suspension cell after month put into filtration sterilization after ethylmethane sulfonate solution, concentration is 0.3%, processing time is 1h, add after process in fresh medium and cultivate, growth rate increases by 70%.
Embodiment 2
Get the blade that Euphorbia lathyris children is tender, static 15min in bleaching powder, running water surface smut, on superclean bench 0.2% mercuric chloride sterilization 10min, aseptic water washing 5 times, blot stand-by, callus induction is carried out in the blade access Miller+ZT0.5mg/L+KT1.5mg/L+0.4mg/ml penicillin medium that the Euphorbia lathyris children disinfected is tender, additional saccharose 30g/L, agar 6.5g/L, pH6.0, intensity of illumination 1000lx, light application time 6h/d, temperature 22 DEG C, the callus derived is put into liquid nutrient medium Miller+0.3mg/LIAA+1.5mg/L2ip+100mg/L lactoalbumin hydrolysate and is carried out liquid suspension cell chulture, constant temperature oscillator 100r/min, inoculum concentration is 2.5g, cultivation cycle is 40d, additional saccharose 30g/L, pH5.8, illumination 2500lx, light application time 15h/d, temperature 24 DEG C, cultivate the suspension cell after month put into filtration sterilization after ethylmethane sulfonate solution, concentration is 0.5%, processing time is 3h, add after process in fresh medium and cultivate, growth rate increases by 68%.
Embodiment 3
Get the blade that Euphorbia lathyris children is tender, static 15min in bleaching powder, running water surface smut, on superclean bench 0.2% mercuric chloride sterilization 10min, aseptic water washing 5 times, blot stand-by, callus induction is carried out in the blade access Miller+ZT0.5mg/L+KT1.5mg/L+0.4mg/ml penicillin medium that the Euphorbia lathyris children disinfected is tender, additional saccharose 30g/L, agar 6.5g/L, pH6.0, intensity of illumination 1000lx, light application time 6h/d, temperature 22 DEG C, the callus derived is put into liquid nutrient medium Miller+0.3mg/LIAA+1.5mg/L2ip+100mg/L lactoalbumin hydrolysate and is carried out liquid suspension cell chulture, constant temperature oscillator 100r/min, inoculum concentration is 2.5g, cultivation cycle is 40d, additional saccharose 30g/L, pH5.8, illumination 2500lx, light application time 15h/d, temperature 24 DEG C, cultivate the suspension cell after month put into filtration sterilization after ethylmethane sulfonate solution, concentration is 1%, processing time is 5h, add after process in fresh medium and cultivate, growth rate increases by 65%.
Claims (3)
1. a method for quickly breeding for Euphorbia lathyris suspension cell, comprise the acquisition of the aseptic blade of Euphorbia lathyris, the induction of callus, the cultivation of suspension cell, the mutagenesis etc. of suspension cell, its key step is as follows:
(1) blade that Euphorbia lathyris children is tender is got, disinfection;
(2) get in the tender blade access Miller+ZT0.5mg/L+KT1.5mg/L+0.4mg/ml penicillin medium of Euphorbia lathyris children that step (1) disinfected and carry out callus induction, additional saccharose 30g/L, agar 6.5g/L, pH6.0, intensity of illumination 1000lx, light application time 6h/d, temperature 22 DEG C;
(3) get callus that step (2) derives to put into liquid nutrient medium Miller+0.3mg/LIAA+1.0-1.5mg/L2ip+100mg/L lactoalbumin hydrolysate and carry out liquid suspension cell chulture, additional saccharose 30g/L, pH5.8, illumination 2500lx, light application time 15h/d, temperature 24 DEG C;
(4) get step (3) cultivate the suspension cell after month put into filtration sterilization after ethylmethane sulfonate solution, concentration is 0.3%-1%, and the processing time is 1-5h, adds in fresh medium and cultivate after process.
2. according to the method for quickly breeding of a kind of Euphorbia lathyris suspension cell according to claim 1, it is characterized in that: the acquisition of the aseptic blade of Euphorbia lathyris described in step (1) is for getting the tender blade of Euphorbia lathyris children, static 15min in bleaching powder, running water surface smut, on superclean bench 0.2% mercuric chloride sterilization 10min, aseptic water washing 5 times, blots stand-by.
3. according to the method for quickly breeding of a kind of Euphorbia lathyris suspension cell according to claim 1, it is characterized in that: in step (3), the condition of Euphorbia lathyris suspension cell culture is constant temperature oscillator 100r/min, and inoculum concentration is 2.5g, and cultivation cycle is 40d.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410540511.2A CN104255517A (en) | 2014-10-14 | 2014-10-14 | Rapid propagation method of suspension cells of euphorbia lathyris |
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| CN201410540511.2A CN104255517A (en) | 2014-10-14 | 2014-10-14 | Rapid propagation method of suspension cells of euphorbia lathyris |
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| CN104255517A true CN104255517A (en) | 2015-01-07 |
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| CN201410540511.2A Pending CN104255517A (en) | 2014-10-14 | 2014-10-14 | Rapid propagation method of suspension cells of euphorbia lathyris |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104782495A (en) * | 2015-05-02 | 2015-07-22 | 冯文杰 | Tissue culture and rapid propagation method for euphorbia lathyris |
| CN110551676A (en) * | 2019-09-07 | 2019-12-10 | 江苏省中国科学院植物研究所 | Euphorbia lathyris callus cell suspension culture method capable of extracting ingenol |
-
2014
- 2014-10-14 CN CN201410540511.2A patent/CN104255517A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104782495A (en) * | 2015-05-02 | 2015-07-22 | 冯文杰 | Tissue culture and rapid propagation method for euphorbia lathyris |
| CN110551676A (en) * | 2019-09-07 | 2019-12-10 | 江苏省中国科学院植物研究所 | Euphorbia lathyris callus cell suspension culture method capable of extracting ingenol |
| CN110551676B (en) * | 2019-09-07 | 2023-06-20 | 江苏省中国科学院植物研究所 | Suspension culture method for euphorbia lathyris callus cells capable of extracting and obtaining ingenol |
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Application publication date: 20150107 |