CN101040602A - Quick propagating method of blumea riparia(Bl.)DC medicine material - Google Patents
Quick propagating method of blumea riparia(Bl.)DC medicine material Download PDFInfo
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- CN101040602A CN101040602A CN 200710034891 CN200710034891A CN101040602A CN 101040602 A CN101040602 A CN 101040602A CN 200710034891 CN200710034891 CN 200710034891 CN 200710034891 A CN200710034891 A CN 200710034891A CN 101040602 A CN101040602 A CN 101040602A
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Abstract
The invention provides a method for quickly cultivating Blumea riparia medicinal material, which comprises that collecting Blumea riparia explants in wild, which it contains stem sharp, young stem, old stem, blades, bulbs, and seeds, to be washed and disinfected, and using plant organism quick cultivating method and grafting cultivating method to quickly cultivate the Blumea riparia, cultivating seeds, while the obtained seed can be directly used in field. The invention has high cultivate speed, short period as 2-3 months, with high survival rate more than 90%, simple operation, simple device, low cost, and application in whole year. And the product cultivated by the invention has stable gene character, without variation.
Description
Technical field
The present invention relates to a kind of plant regeneration method, especially a kind of method that is exclusively used in quick breeding of blumea riparia and cultivation by tissue culture technique.
Background technology
Blumea riparia is composite family Blumea balsamifera platymiscium Bloumea megacephala (Randeria) Chang et Tseng through expert appraisal, have another name called magnificent Blumea balsamifera, pipe bud, it is the primary raw material of Guangxi Guixi Pharmaceutical Co., Ltd's main product blumea riparia particle, all herbal medicine, that product has is promoting blood circulation and removing blood stasis, the effect of blood-arresting catamenia-regulating, be used for the treatment of stagnation of blood stasis, menorrhalgia, menstrual period are long, gynecologic blood diseases such as post partum lochiorrhea treatment total effective rate reaches more than 96%, the product annual sales revenue surpasses 6,000 ten thousand yuan, is the high plant of a kind of economic worth.
The blumea riparia medicinal material requires harsh to growing environment, is suitable in the winter time 〉=-2 ℃, and summer≤35 ℃, warm, with heavy rainfall subtropical area growth, the cloudy happiness of happiness is wet, avoids intense light irradiation, and cultivation is difficulty quite, and among the people up to now do not have a cultivation history.Since blumea riparia particle medicament in 1994 was produced in batches, the blumea riparia medicinal raw material derived from wild herb resource, and annual requirement reaches more than 2000 tons.Through digging for many years, the resources of natural crude drugs gradually reduce, and the quality instability of medicinal material, can not satisfy the production demand that has enlarged, need carry out the source of large-area artificial cultivation as medicinal raw material.At present, wild blumea riparia is mainly by seminal propagation, because the seed embryo tissue development of this medicinal material is bad, and the double influence that is subjected to factors such as season, growing environment, the reproduction rate of seed is low, and germination rate is lower than 5%, can not satisfy the tame needs of large tracts of land; In addition, owing to be the sexual hybridization breeding, thereby make the offspring produce genetic variation, and cause the quality instability, be unfavorable for the quality control of drug production process.Carry out breeding by the seminal propagation method, reproduction speed is slow, and survival rate is low, is difficult to satisfy the requirement of implant mass, therefore is badly in need of the reproduction rate height, and fireballing propagation method is carried out seedling production.
By retrieval, up to now, domesticly do not see relevant for the report of relevant breeding of blumea riparia and cultivation research does not have the research record of being correlated with abroad yet.
Summary of the invention
The purpose of this invention is to provide a kind of effective ways of blumea riparia tissue cultivating seedling breeding, the seedling that enrichment culture forms can be directly used in field planting, produces the blumea riparia medicinal material, and speed is fast, and is effective, the survival rate height.
In the present invention, we have adopted plant tissue cell's breeding method and cottage propagation method to carry out artificial propagation, belong to vegetative propagation, the medicinal material heredity that obtains is relatively stable, cooperate culture technique unified, standard again, the quality of medicinal material of producing is stable, meets the requirement of pharmaceutical producing enterprise.After deliberation, this medicinal material is bred by the mode of cuttage, and after treatment, germination rate can reach 70～80%, and transplanting survival rate can reach 90%; Adopt plant cell tissue's cultured method, can make blumea riparia tissue cultivating seedling propagation at short notice rapidly, its reproduction coefficient can reach 4.0～6.0, and transplanting survival rate also can reach more than 90%.Grow seedlings by above two kinds of methods, not only can obtain a large amount of seedlings, and after planting transplantation of seedlings, growth is rapid, the survival rate height.
The propagation method of blumea riparia medicinal material provided by the present invention has 2 kinds, is respectively plant tissue cell and cultivates breeding method and cottage propagation method.
Will be from the blumea riparia explant of field acquisition, comprising the stem apex of blumea riparia, tender stem, old stem, blade, petal, seed, sterilization is cleaned at each position, by plant cell tissue's cultivation rapid breeding method and cottage propagation method blumea riparia is bred rapidly, cultivate seedling, the seedling that is obtained is directly used in the land for growing field crops and produces.
1, breeding method is cultivated by plant tissue cell
It is with the blumea riparia explant from field acquisition that rapid breeding method is cultivated by plant cell tissue, clean sterilization comprising the stem apex of blumea riparia, tender stem, old stem, blade, petal, each position of seed, clean sterilization, the explant that cuts suitable specification is inoculated on the medium that has added plant hormone and carbohydrate, carry out the inducing of the inducing of callus, test-tube plantlet, the fast breeding cultivation of test-tube plantlet, the strong seedling culture of test-tube plantlet, the culture of rootage of test-tube plantlet, carry out acclimatization and transplants at last, obtain the test tube seedling.
It is the MS medium that the used medium of rapid breeding method is cultivated by plant cell tissue, plant hormone is BA, NAA, Met, KT, 2,4-D one or more hormons wherein use, carbohydrate is a sucrose, agar content is 3～7g/L, pH value 4～7, and each stage condition of culture is: 25～28 ℃ of temperature, illumination 1000～1200Lux, light application time is 10～12h/d.
The concrete operations step is as follows:
(1) inducing of callus: the explant of fetching from the field is inoculated in MS+BA1.0～2.0mg/L+NAA0.1～0.5mg/L+ sucrose 2～3% (mass percent, as follows) through behind the sterilization, secretly cultivates after 15～25 days and induces callus;
(2) differentiation of calli: with the callus that induces change over to differential medium MS+KT1.0～3.0mg/L+BA1.0～2.0mg/L+NAA0.1～0.5mg/L+ sucrose after 2～3%, 20～30 days callus begin to differentiate budlet;
(3) shoot proliferation of test-tube plantlet: test-tube plantlet is changed on MS+BA0.5～2.0mg/L+NAA0.1～1.0mg/L+Met0.5～2.0mg/L+ sucrose 2～3% proliferated culture mediums, inoculate back 25～30 days from the young shoot base portion and sprout the bud of growing thickly, shoot proliferation gradually;
(4) strong sprout of test-tube plantlet: receive on MS+BA0.5～4.0mg/L+NAA0.5～4.0mg/L+Met0.5～2.0mg/L+ sucrose 2～3% strong seedling culture bases after the test-tube plantlet of enrichment culture cut branch, the inoculation back obtained healthy and strong test-tube plantlet in 25～30 days;
(5) taking root of test-tube plantlet: the aseptic seedling after the strong seedling culture is inoculated on MS+NAA0.1～0.5mg/L+ sucrose 2～3% root medias, and inoculation back 25～30 days sends out roots from the base portion of seedling, becomes whole plant.
(6) agar content is 5g/L in above-mentioned each medium, pH value 5.8, and each stage condition of culture is: except inducing of callus is dark cultivation, all the other each stages cultivate and are the light cultivation, cultivation temperature is 25～28 ℃, illumination 1000～1200Lux, and light application time is 10～12h/d.
2, cottage propagation method
Cuttage breeding method is with the blumea riparia vigorous branches from field acquisition, and (15～40cm), the immersion treatment of carrying out root induction is carried out cuttage then, obtains vegetative seedling to intercept suitable length.
The used plant hormone of cuttage breeding method is NAA, and IAA, IBA one or more hormons wherein use.
The concrete operations step is as follows:
(1) choosing of vigorous branches: the blumea riparia branch that will adopt back from the field, choose the branch that diameter is 0.25～0.5cm, be cut into 15～30cm segment.
(2) processing before the cuttage: the branch of intercepting is cleaned,, soaked in the solution that IAA, IBA are configured to 4～8 hours by NAA, NAA in the solution, IAA, IBA concentration respectively is 0.5～2.5mg/L.
(3) cuttage and seedling culture: the branch of handling well is inserted in Sha Chizhong, the sunshade cultivation of preserving moisture, the petiole base position at branch about 2 weeks grows tender shoots, continues to cultivate about 4 weeks to grow new root, and transplanting survival rate reaches more than 90%.
The present invention has following advantage:
1. the present invention mainly utilizes stem apex, stem section, blade, the petal of blumea riparia to carry out tissue culture as explant, systematically explores the best cultivating system that a cover is bred fast to blumea riparia relatively comprehensively.
2. reproduction speed is fast, and reproduction coefficient reaches 4.0～6.0.
3. the cycle is lacked about 2～3 months of one-period.
4. transplanting survival rate height, tissue cultivating seedling ground transplanting survival rate has all reached more than 90%.
5. method of operating is simple, only needs simple facility, and cost is low.Utilize the present invention, the test-tube plantlet of its transplanting can be directly used in the land for growing field crops and produce.All can operate the whole year.
6. by the seedling of above plant tissue culture technique acquisition, hereditary capacity is stable, can not morph, and is applicable to and commercially produces.
7. method for quickly breeding provided by the present invention and culture technique are applicable to all kinds blumea riparia kind.
Embodiment
Embodiment 1:
1. inducing of callus: the explant different parts after the sterilization treatment is inserted in the inducing culture, and the long approximately 2cm of the stem apex of explant clip, the long approximately 3.5cm of tender stem, the long approximately 3.5cm of old stem, blade size are 1.0 * 0.6cm
2, petal length is about 1.0cm.Explant inoculation quantity is decided on the different explants position, and general stem apex is 3/bottle, and tender stem is 4 sections/bottle, and old stem is 4 sections/bottle, and blade is 10 pieces/bottle, and petal is 10/bottle.The prescription of inducing culture is a MS+BA1.0mg/L+NAA0.1mg/L+ sucrose 2.0%+ agar powder 0.5%; (wherein the MS medium is general minimal medium; the present invention selects for use plant hormone market also on sale; 2.4-D; NAA is that China Medicine (Group) Shanghai Chemical Reagent Co., produces; 6-BA; KT; IBA is Sanland international Inc. production; IAA is that worker's biotechnology Co., Ltd is given birth in Shanghai; Met is that the safe foundation development in science and technology center that shakes, Beijing produces) condition of culture cultivates for dark; explant induction goes out callus after 15 days, and the inductivity of callus reaches as high as 93%.
2. differentiation of calli: as callus length to 1.0 * 0.5cm
2During size, the clip callus is inoculated in differential medium, differential medium is a MS+KT1.0mg/L+BA1.0mg/L+NAA0.1mg/L+ sucrose 2.0%+ agar powder 0.5%, condition of culture is, temperature is 25～28 ℃, pH value 5.8, illumination 1000～1200Lux, light application time is 10～12h/d, and callus differentiates budlet after 25 days.
3. the shoot proliferation of test-tube plantlet: when the test-tube plantlet of blumea riparia long during to 1.5cm, the clip sprout changes on the proliferated culture medium of MS+BA0.5mg/L+NAA0.1mg/L+Met0.5mg/L+ sucrose 2.0%+ agar powder 0.5%, condition of culture is the same, inoculate back 30 days and sprout the bud of growing thickly gradually from the young shoot base portion, average each young shoot can break up about 7～8 strains, again young shoot is cut and changed over to identical medium and repeat to cultivate a large amount of breedings.
4. strong sprout of test-tube plantlet: receive on the MS+BA4.0mg/L+NAA4.0mg/L+Met2.0mg/L+ sucrose 3.0%+ agar powder 0.5% strong seedling culture base after the test-tube plantlet of enrichment culture cut, condition of culture is the same, inoculates back 25 days and obtains healthy and strong test-tube plantlet;
5. taking root of test-tube plantlet: the aseptic seedling after the strong seedling culture is inoculated on MS+NAA0.5mg/L+ sucrose 3.0%+ agar powder 0.5% root media, condition of culture is the same, inoculates back 25 days, grows the adventive root of light green color or white from the base portion of seedling, long and many, rooting rate reaches 100%.
6. acclimatization and transplants: the tissue cultivating seedling that growth potential is more vigorous takes out from culturing room, is positioned in the seeding room, and indoor temperature remains on about 25 ℃, relative moisture remains on about 75%, avoids the sunlight direct projection, at room temperature place 5～7 days after, to cultivate bottle cap opens and carries out 2～3 days hardenings, take out seedling then, clean the medium that is bonded at root, be transplanted in the nutritive cube that sterilization matrix is housed, water sufficient water, the humidity that keeps matrix constantly is positioned under the awning and cultivates, and temperature remains on about 26 ℃.Water one time of nutrition liquid weekly, grow new root about 15 days, survival rate reaches more than 90%.
Embodiment 2:
1. inducing of aseptic seedling: will be inoculated on the inducing culture through the blumea riparia stem section of sterilization treatment, the inducing culture based formulas is a MS+BA1.0mg/L+NAA0.1mg/L+ sucrose 2.5%+ agar powder 0.5%, condition of culture is, temperature is 25～28 ℃, pH value 5.8, illumination 1000～1200Lux, light application time is 10～12h/d, the axillalry bud punishment from the stem section after 30 days dissolves budlet.
2. the shoot proliferation of test-tube plantlet: when the test-tube plantlet of blumea riparia long during to 1.5cm, the clip sprout changes on the proliferated culture medium of MS+BA1.0mg/L+NAA0.2mg/L+Met1.0mg/L+ sucrose 2.5%+ agar powder 0.5%, condition of culture is the same, inoculate back 20 days and sprout the bud of growing thickly gradually from the young shoot base portion, average each young shoot can break up about 7～8 strains, again young shoot is cut and changed over to identical medium and repeat to cultivate a large amount of breedings.
3. strong sprout of test-tube plantlet: receive on the MS+BA0.5mg/L+NAA0.3mg/L+Met1.5mg/L+ sucrose 2.5%+ agar powder 0.5% strong seedling culture base after the test-tube plantlet of enrichment culture cut, condition of culture is the same, inoculates back 20 days and obtains healthy and strong test-tube plantlet;
4. taking root of test-tube plantlet: the aseptic seedling after the strong seedling culture is inoculated on MS+NAA0.2mg/L+ sucrose 2.5% root media, condition of culture is the same, inoculates back 20 days, grows the adventive root of light green color or white from the base portion of seedling, long and many, rooting rate reaches 100%.
5. acclimatization and transplants: the tissue cultivating seedling that growth potential is more vigorous takes out from culturing room, is positioned in the seeding room, and indoor temperature remains on about 25 ℃, relative moisture remains on about 75%, avoids the sunlight direct projection, at room temperature place 5～7 days after, to cultivate bottle cap opens and carries out 2～3 days hardenings, take out seedling then, clean the medium that is bonded at root, be transplanted in the nutritive cube that sterilization matrix is housed, water sufficient water, the humidity that keeps matrix constantly is positioned under the awning and cultivates, and temperature remains on about 26 ℃.Water one time of nutrition liquid weekly, grow new root about 15 days, survival rate reaches more than 90%.
Embodiment 3:
1. choosing of vigorous branches: the blumea riparia branch that will adopt back from the field, choose the branch that diameter is 0.25～0.5cm, be cut into 15～40cm segment.
2. the processing before the cuttage: the branch of intercepting is cleaned,, soaked 5 hours in the solution that IAA, IBA are configured to by NAA, NAA in the solution, IAA, IBA concentration respectively is 0.5～2.5mg/L.
3. cuttage and seedling culture: the branch of handling well is inserted in Sha Chizhong, the sunshade cultivation of preserving moisture, the petiole base position at branch about 2 weeks grows tender shoots, continues to cultivate about 4 weeks to grow new root, and transplanting survival rate reaches more than 90%.
Claims (5)
1, a kind of method for quickly breeding of blumea riparia medicinal material is characterized in that:
Will be from the blumea riparia explant of field acquisition, comprising the stem apex of blumea riparia, tender stem, old stem, blade, petal, seed, sterilization is cleaned at each position, by plant cell tissue's cultivation rapid breeding method and cottage propagation method blumea riparia is bred rapidly, cultivate seedling, the seedling that is obtained is directly used in the land for growing field crops and produces.
2, the method for quickly breeding of blumea riparia medicinal material according to claim 1, it is characterized in that: it is with the blumea riparia explant from field acquisition that rapid breeding method is cultivated by described plant cell tissue, stem apex comprising blumea riparia, tender stem, old stem, blade, petal, sterilization is cleaned at each position of seed, clean sterilization, cutting explant is inoculated on the medium that has added plant hormone and carbohydrate, carry out inducing of callus, inducing of test-tube plantlet, the fast breeding of test-tube plantlet is cultivated, the strong seedling culture of test-tube plantlet, the culture of rootage of test-tube plantlet, carry out acclimatization and transplants at last, obtain the test tube seedling.
3, the method for quickly breeding of blumea riparia medicinal material according to claim 2, it is characterized in that: it is the MS medium that the used medium of rapid breeding method is cultivated by plant cell tissue, plant hormone is BA, NAA, Met, KT, 2,4-D one or more hormons wherein use, and carbohydrate is a sucrose, and agar content is 3～7g/L, pH value 4～7, each stage condition of culture is: 25～28 ℃ of temperature, and illumination 1000～1200Lux, light application time is 10～12h/d.
4, the method for quickly breeding of blumea riparia medicinal material according to claim 1, it is characterized in that: described cuttage breeding method is with the blumea riparia vigorous branches from field acquisition, the segment of intercepting 15～40cm, carry out the immersion treatment of root induction, carry out cuttage then, obtain vegetative seedling.
5, the method for quickly breeding of blumea riparia medicinal material according to claim 3 is characterized in that: the used plant hormone of cuttage breeding method is NAA, and IAA, IBA one or more hormons wherein use.
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102550421A (en) * | 2012-03-01 | 2012-07-11 | 广西壮族自治区药用植物园 | Fast breeding and cultivating method of Blumea aromatica seedlings |
| CN103250645A (en) * | 2013-05-23 | 2013-08-21 | 贵州一合生物技术有限公司 | Rapid propagation and transplantation method of blumea balsamifera |
| CN103385110A (en) * | 2013-08-21 | 2013-11-13 | 贵州省亚热带作物研究所 | Raising method for blumea balsamifera seedlings |
| CN103688856A (en) * | 2013-12-13 | 2014-04-02 | 广西桂西制药有限公司 | Rapid propagation method of Blumea riparia (Bl.) DC medicinal material |
| CN103688715A (en) * | 2013-12-10 | 2014-04-02 | 广西万寿堂药业有限公司 | Method for raising seedlings of Yunnan-Guangxi balsamiferou blumea herb leaves by cutting |
| CN104756706A (en) * | 2015-04-03 | 2015-07-08 | 贵州大学 | Transplanting method of blumea balsamifera seedlings |
| CN105309315A (en) * | 2015-11-26 | 2016-02-10 | 中国热带农业科学院热带作物品种资源研究所 | Embryoid-approach blumea balsamifera tissue culture method |
| CN105510330A (en) * | 2015-11-24 | 2016-04-20 | 中国热带农业科学院热带作物品种资源研究所 | Method for distinguishing Hainan and Guizhou province germplasm Blumea balsamifera |
| CN107549018A (en) * | 2017-10-30 | 2018-01-09 | 湖北理工学院 | Chinese mugwort tissue culture method |
| CN109156355A (en) * | 2018-10-10 | 2019-01-08 | 东北农业大学 | A kind of method that IBA induces two strain callus from stem segment |
| CN115152441A (en) * | 2022-06-13 | 2022-10-11 | 贵州中医药大学 | Cuttage propagation method for Miao medicine blumea balsamifera |
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2007
- 2007-04-25 CN CN 200710034891 patent/CN101040602A/en active Pending
Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102550421B (en) * | 2012-03-01 | 2013-04-24 | 广西壮族自治区药用植物园 | Fast breeding and cultivating method of Blumea aromatica seedlings |
| CN102550421A (en) * | 2012-03-01 | 2012-07-11 | 广西壮族自治区药用植物园 | Fast breeding and cultivating method of Blumea aromatica seedlings |
| CN103250645A (en) * | 2013-05-23 | 2013-08-21 | 贵州一合生物技术有限公司 | Rapid propagation and transplantation method of blumea balsamifera |
| CN103250645B (en) * | 2013-05-23 | 2014-07-23 | 贵州一合生物技术有限公司 | Rapid propagation and transplantation method of blumea balsamifera |
| CN103385110A (en) * | 2013-08-21 | 2013-11-13 | 贵州省亚热带作物研究所 | Raising method for blumea balsamifera seedlings |
| CN103688715A (en) * | 2013-12-10 | 2014-04-02 | 广西万寿堂药业有限公司 | Method for raising seedlings of Yunnan-Guangxi balsamiferou blumea herb leaves by cutting |
| CN103688856B (en) * | 2013-12-13 | 2016-05-18 | 广西桂西制药有限公司 | A kind of method for quickly breeding of blumea riparia medicinal material |
| CN103688856A (en) * | 2013-12-13 | 2014-04-02 | 广西桂西制药有限公司 | Rapid propagation method of Blumea riparia (Bl.) DC medicinal material |
| CN104756706A (en) * | 2015-04-03 | 2015-07-08 | 贵州大学 | Transplanting method of blumea balsamifera seedlings |
| CN105510330B (en) * | 2015-11-24 | 2018-07-17 | 中国热带农业科学院热带作物品种资源研究所 | A method of distinguishing Guizhou Province fine jade germplasm Blumea balsamifera |
| CN105510330A (en) * | 2015-11-24 | 2016-04-20 | 中国热带农业科学院热带作物品种资源研究所 | Method for distinguishing Hainan and Guizhou province germplasm Blumea balsamifera |
| CN105309315A (en) * | 2015-11-26 | 2016-02-10 | 中国热带农业科学院热带作物品种资源研究所 | Embryoid-approach blumea balsamifera tissue culture method |
| CN107549018A (en) * | 2017-10-30 | 2018-01-09 | 湖北理工学院 | Chinese mugwort tissue culture method |
| CN109156355A (en) * | 2018-10-10 | 2019-01-08 | 东北农业大学 | A kind of method that IBA induces two strain callus from stem segment |
| CN115152441A (en) * | 2022-06-13 | 2022-10-11 | 贵州中医药大学 | Cuttage propagation method for Miao medicine blumea balsamifera |
| CN115152441B (en) * | 2022-06-13 | 2023-12-08 | 贵州中医药大学 | Cutting propagation method of blumea balsamifera serving as seedling medicine |
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