CN104255478A - Method for quickly reproducing Murraya exotica L. suspension cells - Google Patents
Method for quickly reproducing Murraya exotica L. suspension cells Download PDFInfo
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- CN104255478A CN104255478A CN201410463221.2A CN201410463221A CN104255478A CN 104255478 A CN104255478 A CN 104255478A CN 201410463221 A CN201410463221 A CN 201410463221A CN 104255478 A CN104255478 A CN 104255478A
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- kamuning
- suspension cell
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Abstract
The invention provides a method for quickly reproducing Murraya exotica L. suspension cells. The method comprises the following steps: disinfecting sterile leaves of Murraya exotica L.; inducing callus tissues; culturing liquid suspension cells; optimizing suspension cell culture, and the like. The Murraya exotica L. suspension cells prepared by the method have high growing speed; and the method can be used for preparing relatively high suspension cells and establishing a quickly growing suspension cell culture system to maintain the sustainable development and utilization of Murraya exotica L. suspension cells.
Description
Technical field
The present invention relates to the quick-breeding method that the suspension of a kind of kamuning is cultivated, belong to biological technical field.
Background technology
Kamuning, also known as: stone capsicum, nine autumns perfume, nine tree perfume etc., Rutaceae,
murraya exotica L., dungarunga, kamuning happiness is warm, and the temperature of optimum growth is 20-32 DEG C, does not resist cold.In being common in the shrub of the not far level land of bank off sea, gentle slope, hillock.Sandy soil, on the sunny side place are born in happiness.Kamuning originates in: the ground such as Yunnan, Guizhou, Hunan, Guangdong, Guangxi, Fujian, Hainan, TaiWan, China, and some other torrid zone, Asia and subtropical zone.Produce Taiwan, Fujian, Guangdong, Hainan, south, provinces and regions, Guangxi five.Murraya jasminorage leaf is containing multiple Coumarins compound, be separated from each local specialties such as Guangdong, Yunnan, Hainan, Taiwan and obtained: isomexoticin, murpanidin, murpanicin, come into leaves coumurrayin glycol, the kamuning that comes into leaves aldehyde, 5,7-dimethoxy-8-(3 '-methyl-2'-ketone group butyl) coumarin, Hainan coumurrayin etc., the promoting flow of qi and blood circulation; Blood stasis removing analgesic; Removing toxicity for detumescence.Main gastral cavity pain; Tumbling and swelling; Sore carbuncle; Snake bite and insect sting.Cure mainly treating swelling and pain by traumatic injury, treating rheumatic ostealgia, stomachache, toothache, tetanus, Japanese Type-B encephalitis, worm, snake bite, local anaesthesia.For stomachache, arthralgia due to wind-dampness; Control toothache outward, tumbling and swelling, worm snake bite.Implantation methods is seminal propagation, propagation by layering, cottage propagation.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of method for quickly breeding of kamuning suspension cell, kamuning suspension cell growth speed prepared by the method is fast, higher suspension cell can be obtained, set up a plant cell suspension cultures grown fast, maintain its sustainable exploitation and use.
Technical problem to be solved by this invention is realized by following scheme:
Get the blade that kamuning children is tender, running water 20min, on superclean bench 0.1% mercuric chloride process 8min, aseptic water washing 5 times, blotting paper blots, callus induction is carried out in the blade access MSB+IAA1.0mg/L+6-BA10mg/L+2mg/LKT medium that the kamuning children disinfected is tender, additional saccharose 30g/L, agar 7g/L, pH5.8, darkroom is cultivated, temperature 25 DEG C, relative moisture 75%, the callus derived puts into liquid nutrient medium MSB+4-5 μm ol/L2, 4-D+1-1.5 μm ol/L6-BA carries out liquid suspension cell chulture, additional saccharose 50g/L, pH5.8, illumination 4500lx, temperature 25 DEG C, inoculum density is 8%, 125ml medicine bottle dress 25ml medium, rotating speed 110r/min, the cell out of suspension cultivation after two weeks puts into the liquid nutrient medium that addition of sodium acetate 10mg/L+ benzenpropanoic acid sodium 10mg/L, the growth rate of suspension cell is measured after 30d.
The suspension cell growth rate 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 kamuning children is tender, running water 20min, on superclean bench 0.1% mercuric chloride process 8min, aseptic water washing 5 times, blotting paper blots, callus induction is carried out in the blade access MSB+IAA1.0mg/L+6-BA10mg/L+2mg/LKT medium that the kamuning children disinfected is tender, additional saccharose 30g/L, agar 7g/L, pH5.8, darkroom is cultivated, temperature 25 DEG C, relative moisture 75%, the callus derived puts into liquid nutrient medium MSB+4 μm ol/L2, 4-D+1 μm of ol/L6-BA carries out liquid suspension cell chulture, additional saccharose 50g/L, pH5.8, illumination 4500lx, temperature 25 DEG C, inoculum density is 8%, 125ml medicine bottle dress 25ml medium, rotating speed 110r/min, the cell out of suspension cultivation after two weeks puts into the liquid nutrient medium that addition of sodium acetate 10mg/L+ benzenpropanoic acid sodium 10mg/L, the growth rate of suspension cell is measured after 30d, growth rate improves 50%.
Embodiment 2
Get the blade that kamuning children is tender, running water 20min, on superclean bench 0.1% mercuric chloride process 8min, aseptic water washing 5 times, blotting paper blots, callus induction is carried out in the blade access MSB+IAA1.0mg/L+6-BA10mg/L+2mg/LKT medium that the kamuning children disinfected is tender, additional saccharose 30g/L, agar 7g/L, pH5.8, darkroom is cultivated, temperature 25 DEG C, relative moisture 75%, the callus derived puts into liquid nutrient medium MSB+5 μm ol/L2, 4-D+1.5 μm ol/L6-BA carries out liquid suspension cell chulture, additional saccharose 50g/L, pH5.8, illumination 4500lx, temperature 25 DEG C, inoculum density is 8%, 125ml medicine bottle dress 25ml medium, rotating speed 110r/min, the cell out of suspension cultivation after two weeks puts into the liquid nutrient medium that addition of sodium acetate 10mg/L+ benzenpropanoic acid sodium 10mg/L, the growth rate of suspension cell is measured after 30d, growth rate improves 45%.
Embodiment 3
Get the blade that kamuning children is tender, running water 20min, on superclean bench 0.1% mercuric chloride process 8min, aseptic water washing 5 times, blotting paper blots, callus induction is carried out in the blade access MSB+IAA1.0mg/L+6-BA10mg/L+2mg/LKT medium that the kamuning children disinfected is tender, additional saccharose 30g/L, agar 7g/L, pH5.8, darkroom is cultivated, temperature 25 DEG C, relative moisture 75%, the callus derived puts into liquid nutrient medium MSB+4 μm ol/L2, 4-D+1.5 μm ol/L6-BA carries out liquid suspension cell chulture, additional saccharose 50g/L, pH5.8, illumination 4500lx, temperature 25 DEG C, inoculum density is 8%, 125ml medicine bottle dress 25ml medium, rotating speed 110r/min, the cell out of suspension cultivation after two weeks puts into the liquid nutrient medium that addition of sodium acetate 10mg/L+ benzenpropanoic acid sodium 10mg/L, the growth rate of suspension cell is measured after 30d, growth rate improves 47%.
Claims (3)
1. a method for quickly breeding for kamuning suspension cell, comprise the sterilization of the aseptic blade of kamuning, the induction of callus, the cultivation of liquid suspension cell, the optimization of suspension cell culture, its key step is as follows:
(1) blade that kamuning children is tender is got, disinfection;
(2) get in the tender blade access MSB+IAA1.0mg/L+6-BA10mg/L+2mg/LKT medium of kamuning children that step (1) disinfected and carry out callus induction, additional saccharose 30g/L, agar 7g/L, pH5.8, darkroom is cultivated, temperature 25 DEG C, relative moisture 75%;
(3) get callus that step (2) derives to put into liquid nutrient medium MSB+4-5 μm ol/L2,4-D+1-1.5 μm ol/L6-BA and carry out liquid suspension cell chulture, additional saccharose 50g/L, pH5.8, illumination 4500lx, temperature 25 DEG C;
(4) get the cell out of step (3) suspension cultivation after two weeks and put into the liquid nutrient medium that addition of sodium acetate 10mg/L+ benzenpropanoic acid sodium 10mg/L.
2. according to the method for quickly breeding of a kind of kamuning suspension cell according to claim 1, it is characterized in that: the acquisition of the aseptic blade of kamuning described in step (1) is for getting the tender blade of kamuning children, running water 20min, on superclean bench 0.1% mercuric chloride process 8min, aseptic water washing 5 times, blotting paper blots.
3. according to the method for quickly breeding of a kind of kamuning suspension cell according to claim 1, it is characterized in that: in step (3), the condition of kamuning suspension cell culture is that after aseptic suction filtration, inoculum density is 8%, 125ml medicine bottle dress 25ml medium, rotating speed 110r/min, cultivation cycle is 30d.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410463221.2A CN104255478A (en) | 2014-09-12 | 2014-09-12 | Method for quickly reproducing Murraya exotica L. suspension cells |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201410463221.2A CN104255478A (en) | 2014-09-12 | 2014-09-12 | Method for quickly reproducing Murraya exotica L. suspension cells |
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| CN104255478A true CN104255478A (en) | 2015-01-07 |
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| CN201410463221.2A Pending CN104255478A (en) | 2014-09-12 | 2014-09-12 | Method for quickly reproducing Murraya exotica L. suspension cells |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106069759A (en) * | 2016-06-22 | 2016-11-09 | 南京泽朗生物科技有限公司 | A kind of suspension cell and the method for quickly breeding of regeneration plant |
| CN106106148A (en) * | 2016-06-22 | 2016-11-16 | 南京泽朗生物科技有限公司 | A kind of method for quickly breeding of mao of leaf Radix seu caulis chonemorphae valvatae suspension cell culture |
| CN106718890A (en) * | 2016-12-02 | 2017-05-31 | 河池学院 | A kind of method for tissue culture suitable for Chinese herbal medicine murraya paniculataJack seedling |
| CN107568067A (en) * | 2017-10-11 | 2018-01-12 | 陈金水 | A kind of method for building up of kamuning vitro Regeneration System |
-
2014
- 2014-09-12 CN CN201410463221.2A patent/CN104255478A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106069759A (en) * | 2016-06-22 | 2016-11-09 | 南京泽朗生物科技有限公司 | A kind of suspension cell and the method for quickly breeding of regeneration plant |
| CN106106148A (en) * | 2016-06-22 | 2016-11-16 | 南京泽朗生物科技有限公司 | A kind of method for quickly breeding of mao of leaf Radix seu caulis chonemorphae valvatae suspension cell culture |
| CN106718890A (en) * | 2016-12-02 | 2017-05-31 | 河池学院 | A kind of method for tissue culture suitable for Chinese herbal medicine murraya paniculataJack seedling |
| CN107568067A (en) * | 2017-10-11 | 2018-01-12 | 陈金水 | A kind of method for building up of kamuning vitro Regeneration System |
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| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20150107 |
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| WD01 | Invention patent application deemed withdrawn after publication |