CN101180949B - Stem tip tissue rapid cultivating and seedling method of algam dendrobium nobile - Google Patents
Stem tip tissue rapid cultivating and seedling method of algam dendrobium nobile Download PDFInfo
- Publication number
- CN101180949B CN101180949B CN2007100664801A CN200710066480A CN101180949B CN 101180949 B CN101180949 B CN 101180949B CN 2007100664801 A CN2007100664801 A CN 2007100664801A CN 200710066480 A CN200710066480 A CN 200710066480A CN 101180949 B CN101180949 B CN 101180949B
- Authority
- CN
- China
- Prior art keywords
- seedling
- culture
- illumination
- stem
- concentration
- 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.)
- Expired - Fee Related
Links
- 238000000034 method Methods 0.000 title claims description 8
- 240000004638 Dendrobium nobile Species 0.000 title description 5
- 230000001939 inductive effect Effects 0.000 claims abstract description 9
- 230000004069 differentiation Effects 0.000 claims abstract description 7
- 238000009395 breeding Methods 0.000 claims abstract description 6
- 241000026010 Dendrobium candidum Species 0.000 claims description 19
- 238000005286 illumination Methods 0.000 claims description 19
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims description 15
- 241000234295 Musa Species 0.000 claims description 12
- 235000021015 bananas Nutrition 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 12
- 238000005406 washing Methods 0.000 claims description 8
- 230000001488 breeding effect Effects 0.000 claims description 5
- 244000061456 Solanum tuberosum Species 0.000 claims description 4
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 4
- 239000006160 differential media Substances 0.000 claims description 4
- 239000001963 growth medium Substances 0.000 claims description 4
- 239000000463 material Substances 0.000 claims description 4
- 239000002609 medium Substances 0.000 claims description 4
- 229960002523 mercuric chloride Drugs 0.000 claims description 4
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 claims description 4
- 230000007613 environmental effect Effects 0.000 claims description 2
- 239000012467 final product Substances 0.000 claims description 2
- 150000004676 glycans Chemical class 0.000 abstract description 3
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 229920001282 polysaccharide Polymers 0.000 abstract description 3
- 239000005017 polysaccharide Substances 0.000 abstract description 3
- 229930013930 alkaloid Natural products 0.000 abstract description 2
- 150000003797 alkaloid derivatives Chemical class 0.000 abstract description 2
- 241001076416 Dendrobium tosaense Species 0.000 abstract 3
- 241001523681 Dendrobium Species 0.000 abstract 2
- 230000002068 genetic effect Effects 0.000 abstract 1
- 238000005728 strengthening Methods 0.000 abstract 1
- 238000005516 engineering process Methods 0.000 description 4
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 210000001124 body fluid Anatomy 0.000 description 2
- 239000010839 body fluid Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 244000025254 Cannabis sativa Species 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 208000015220 Febrile disease Diseases 0.000 description 1
- 206010020649 Hyperkeratosis Diseases 0.000 description 1
- 241001562709 Muhlenbergia torreyi Species 0.000 description 1
- 240000001519 Verbena officinalis Species 0.000 description 1
- 235000018718 Verbena officinalis Nutrition 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 206010036067 polydipsia Diseases 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001902 propagating effect Effects 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
Abstract
The invention provides a breeding method for fast propagation of tissue cultured a stem tip of dendrobium officinale K. Kimura et Migo, which follows the culture steps of protocorm inducing, protocorm reproducing, protocorm differentiation, seedling strengthening and rooting. After rooting, the seedlings can be transplanted after hardening. The invention shortens the cultivation period of the Dendrobium officinale K. Kimura et Migo, which is only 5 to 6 months from protocorm to rooted seedling. The invention greatly reduces production cost. And the dendrobium seedlings nurtured by the invention have stable genetic traits, and the content of polysaccharides and Dendrobium alkaloid is similar to a wild Dendrobium officinale K. Kimura et Migo.
Description
Technical field
The present invention relates to a kind of seedling-cultivating method of dendrobium candidum stem apex quick breeding by group culture, belong to technical field of bioengineering.
Background technology
The stem of noble dendrobium is China's tradition rare Chinese medicine, be divided into dozens of kinds such as iron sheet, yellow grass, Jin Chai, Huoshan, ring grass, horsewhip, wherein dendrobium candidum (Dendrobium candidum Wall.ex Lindl) is the superfine product in the stem of noble dendrobium, mainly is distributed in the south of the River and southwestern each province.That dendrobium candidum has is nourishing Yin and clearing heat, the beneficial stomach that promotes the production of body fluid, function such as moisten the lung and relieve the cough, and is used for consumption of body fluid caused by febrile disease, dry polydipsia, various disease conditions such as abnormal heat after being ill.Modern pharmacology studies show that, that the contained polysaccharide of dendrobium candidum has is antitumor, anti-ageing, strengthen effects such as body immunity and hemangiectasis.Dendrobium candidum is given birth on the overhanging cliff more; because minimum, the no endosperm of seed; sprouting under the natural conditions needs and some mycosymbiosis; the natural propagation rate is low; and China for many years to the collection capacity of dendrobium candidum much larger than its amount of growth, owing to excavate depletion of natural resources for a long time; be on the verge of disappearance, classified as the natural crude drugs kind of focused protection by country.For protecting this rare herbal species, domestic many scientific research personnel have carried out the research of quick propagating technology to dendrobium candidum, form callus, protocorm but study mostly to concentrate on by the wild seed sprouting, carry out differentiation culture then and obtain regrowth.There are shortcomings such as cultivation period length, cost height in these technology.Therefore, how to shorten the cultivation period of tissue cultivating seedling, become the key that dendrobium candidum is realized artificial large-scale planting.
Summary of the invention
For overcoming the above-mentioned defective that prior art exists, the invention provides a kind of seedling-cultivating method of dendrobium candidum stem apex quick breeding by group culture.
The present invention finishes by following technical scheme: a kind of seedling-cultivating method of dendrobium candidum stem apex quick breeding by group culture is characterized in that through following process steps:
A, ball stem inducing culture
Choose healthy and strong wild dendrobium candidum seedling, with its stem apex or bud point as explant, rinse well with flowing water earlier, sterilized 30～60 seconds with 70% ethanolic solution again, with aseptic water washing one time, material is put into bottle, be that 0.1% mercuric chloride soaks 6～10min with concentration, use aseptic water washing then 4～5 times, be inoculated in the following aseptic inducing culture: B
5+ KT1.5～2.0mg/L+NAA0.4～0.8mg/L+ concentration is 10～15% bananas juice, at temperature 25-28 ℃, intensity of illumination 1800～2000lx, light application time is under 12 hours the condition, to cultivate 55～65 days, can form the ball stem;
B, ball stem enrichment culture
Well-grown ball stem is transferred in the following proliferated culture medium: B
5+ NAA1.5～2.0mg/L+KT0.8～1.0mg/L+ concentration is 10～15% potato juice, carries out enrichment culture, and the environmental condition of cultivation is: temperature 25-28 ℃, and intensity of illumination 2000～2200lx, illumination 13 hours, cultivation cycle is 35-45 days;
C, ball stem differentiation culture
The ball stem is changed in the following differential medium: B
5+ NAA0.8～1.0mg/L+KT0.1～0.5mg/L+ concentration is 10～15% bananas juice, 25-28 ℃ of control temperature, and intensity of illumination 2000～2200lx, light application time 12 hours was cultivated 55～65 days, obtained breaking up seedling;
D, strong sprout and culture of rootage
To break up seedling changes in the following strong plantlets and rootage medium: B
5+ NAA1.0～1.5mg/L+ concentration is 10～15% bananas juice, 25-28 ℃ of control temperature, and intensity of illumination 1800～2000lx, light application time 12 hours was cultivated 35～45 days, obtained the seedling of taking root, and got final product bottle outlet hardening, transplanting.
The present invention has following advantage and good effect:
(1) the present invention has shortened the cultivation period of dendrobium candidum seedling, and the cultivation period from the ball stem to the seedling of taking root is 5～6 months only, greatly reduces production cost.
(2) the present invention adopts the stem of noble dendrobium seedling that the stem apex tissue culture technology is cultivated, stabilization characteristics of genetics, and its polysaccharide and stem of noble dendrobium alkaloid are close with wild dendrobium candidum.
Embodiment
Below in conjunction with specific embodiment, the present invention is described further.
Embodiment 1
Choose healthy and strong wild dendrobium candidum seedling, with its stem apex or bud point as explant, rinse well with flowing water earlier, sterilized 30 seconds with 70% ethanolic solution again, with aseptic water washing one time, material is put into bottle, soak 6min with 0.1% mercuric chloride then, use aseptic water washing afterwards 4 times, be inoculated in the following aseptic inducing culture: B
5+ KT2.0mg/L+NAA0.8mg/L+ concentration is 10% bananas juice, is that 25-28 ℃, intensity of illumination are that 1800lx, light application time are under 12 hours the condition in temperature, inducing culture 55 days, the ball stem; Well-grown ball stem is transferred in the following proliferated culture medium: B
5+ NAA2.0mg/L+KT1.0mg/L+ concentration is 10% potato, is 25-28 ℃ in temperature, intensity of illumination 2000lx, and light application time is under 13 hours the condition, enrichment culture 35 days, growth coefficient 8.5; The ball stem is changed in the following differential medium: B
5+ NAA1.0mg/L+KT0.1mg/L+ concentration is 10% bananas juice, 25-28 ℃ of temperature of control, and intensity of illumination 2000lx, light application time 12 hours, differentiation culture 60 days, obtaining highly is differentiation seedling about 1 centimetre; To break up seedling changes in the following strong plantlets and rootage medium: B
5+ NAA1.0mg/L+ concentration is 10% bananas juice, 25-28 ℃ of temperature of control, and intensity of illumination 1800lx, light application time 12 hours was cultivated 35 days, and obtaining highly is 3～5 centimetres the seedling of taking root, and wherein the seedling of taking root about 5 centimetres accounts for 65%.
Embodiment 2
Choose healthy and strong wild dendrobium candidum seedling, with its stem apex or bud point as explant, rinse well with flowing water earlier, sterilized 55 seconds with 70% ethanolic solution again, with aseptic water washing one time, material is put into bottle, soak 10min with 0.1% mercuric chloride then, use aseptic water washing afterwards 5 times, be inoculated in the following aseptic inducing culture: B
5+ KT1.5mg/L+NAA0.4mg/L+ concentration is 15% bananas juice, is that 25-28 ℃, intensity of illumination are that 2000lx, light application time are under 12 hours the condition in temperature, inducing culture 65 days, the ball stem; Well-grown ball stem is transferred in the following proliferated culture medium: B
5+ NAA1.5mg/L+KT0.8mg/L+ concentration is to carry out enrichment culture, condition of culture in 15% the potato juice: temperature 25-28 ℃, intensity of illumination 2200lx under 13 hours conditions of illumination, cultivated growth coefficient 9 45 days; The ball stem is changed in the following differential medium: B
5+ NAA0.8mg/L+KT0.5mg/L+ concentration is 15% bananas juice, 25-28 ℃ of temperature of control, and intensity of illumination 2200lx, light application time 12 hours was cultivated 65 days, and obtaining highly is differentiation seedling about 1 centimetre; To break up seedling changes in the following strong plantlets and rootage medium: B
5+ NAA1.5mg/L+ concentration is 15% bananas juice, 25-28 ℃ of temperature of control, and intensity of illumination 2000lx, light application time 12 hours was cultivated 45 days, and obtaining highly is the seedling of taking root about 3～5 centimetres, and wherein the seedling of taking root about 5 centimetres accounts for 68%.
Claims (1)
1. the seedling-cultivating method of a dendrobium candidum stem apex quick breeding by group culture is characterized in that through following process steps:
A, ball stem inducing culture
Choose healthy and strong wild dendrobium candidum seedling, with its stem apex or bud point as explant, rinse well with flowing water earlier, sterilized 30～60 seconds with 70% ethanolic solution again, with aseptic water washing one time, material is put into bottle, be that 0.1% mercuric chloride soaks 6-10min with concentration, use aseptic water washing then 4～5 times, be inoculated in the following aseptic inducing culture: B
5+ KT1.5～2.0mg/L+NAA0.4～0.8mg/L+ concentration is 10～15% bananas juice, at temperature 25-28 ℃, intensity of illumination 1800～2000lx, light application time is under 12 hours the condition, to cultivate 55-65 days, can form the ball stem;
B, ball stem enrichment culture
Well-grown ball stem is transferred in the following proliferated culture medium: B
5+ NAA1.5～2.0mg/L+KT0.8～1.0mg/L+ concentration is 10～15% potato juice, carries out enrichment culture, and the environmental condition of cultivation is: temperature 25-28 ℃, and intensity of illumination 2000～2200lx, illumination 13 hours, cultivation cycle is 35-45 days;
C, ball stem differentiation culture
The ball stem is changed in the following differential medium: B
5+ NAA0.8～1.0mg/L+KT0.1～0.5mg/L+ concentration is 10～15% bananas juice, 25-28 ℃ of control temperature, and intensity of illumination 2000～2200lx, light application time 12 hours was cultivated 55-65 days, obtained breaking up seedling;
D, strong sprout and culture of rootage
To break up seedling changes in the following strong plantlets and rootage medium: B
5+ NAA1.0～1.5mg/L+ concentration is 10～15% bananas juice, 25-28 ℃ of control temperature, and intensity of illumination 1800～2000lx, light application time 12 hours was cultivated 35-45 days, obtained the seedling of taking root, and got final product bottle outlet hardening, transplanting.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007100664801A CN101180949B (en) | 2007-12-20 | 2007-12-20 | Stem tip tissue rapid cultivating and seedling method of algam dendrobium nobile |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007100664801A CN101180949B (en) | 2007-12-20 | 2007-12-20 | Stem tip tissue rapid cultivating and seedling method of algam dendrobium nobile |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101180949A CN101180949A (en) | 2008-05-21 |
| CN101180949B true CN101180949B (en) | 2011-06-08 |
Family
ID=39446483
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2007100664801A Expired - Fee Related CN101180949B (en) | 2007-12-20 | 2007-12-20 | Stem tip tissue rapid cultivating and seedling method of algam dendrobium nobile |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101180949B (en) |
Families Citing this family (19)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101699984B (en) * | 2009-11-23 | 2012-05-30 | 中国农业大学 | Artificial light cultivating method of dendrobium candidum |
| CN102217551B (en) * | 2011-06-14 | 2013-02-06 | 广东省农业科学院花卉研究所 | Tissue culture quick reproduction method for dendrobium chrysotoxum lindl bud tips |
| CN102613086B (en) * | 2012-03-31 | 2013-12-11 | 南京农业大学 | Hormone-free tissue culture method for dendrobium candidum |
| CN103155871B (en) * | 2013-03-07 | 2014-06-04 | 华中科技大学 | Dendrobium officinale sprout rapid propagation method with high efficiency |
| CN103181316A (en) * | 2013-03-11 | 2013-07-03 | 安徽新津铁皮石斛开发有限公司 | Precursor culture medium for wild dendrobium candidum |
| CN105961199A (en) * | 2013-09-16 | 2016-09-28 | 周杰 | Tissue culture method of dendrobium officinale |
| CN103609445A (en) * | 2013-11-27 | 2014-03-05 | 苏州田园农业技术开发有限公司 | Efficient propagation method of Dendrobium officinale |
| CN103621404B (en) * | 2013-11-27 | 2016-07-13 | 铄洋生物科技（上海）股份有限公司 | A kind of inoculation method of Herba Dendrobii |
| CN103891617B (en) * | 2014-04-04 | 2016-03-09 | 广东国方医药科技有限公司 | A kind of Freezing inducement method of dendrobium candidum protocorm |
| CN104041410A (en) * | 2014-06-05 | 2014-09-17 | 卞佳林 | Strong strawberry seedling rooting culture method |
| CN104041411A (en) * | 2014-06-05 | 2014-09-17 | 卞佳林 | Seedling hardening and rooting culture medium for strawberry |
| CN104322374A (en) * | 2014-11-12 | 2015-02-04 | 柳州市天姿园艺有限公司 | Culture medium solution for tissue culture seedling of dendrobium officinale for propagating by using pedicel |
| CN104429974B (en) * | 2014-12-25 | 2016-03-30 | 安徽同创现代农业投资发展有限公司 | The root media that a kind of candidum tissue culturing seedling is cultivated |
| CN105663762A (en) * | 2016-01-11 | 2016-06-15 | 杭州太仁堂生物科技股份有限公司 | Formula of wild-like fresh Dendrobium officinale safflower Chinese angelica oral liquid and preparation method thereof |
| CN106550751A (en) * | 2016-11-10 | 2017-04-05 | 唐翔 | A kind of dendrobium candidum Emulational culture method |
| CN106561246A (en) * | 2016-11-10 | 2017-04-19 | 唐翔 | High-yield and high-efficiency dendrobium chrysanthum emulational culture method |
| CN106561244A (en) * | 2016-11-10 | 2017-04-19 | 唐翔 | Biomimetic tissue culturing planting method of fimbriate dendrobium |
| CN106561247A (en) * | 2016-11-10 | 2017-04-19 | 唐翔 | Cultivation method for imitated wild Dendrobium nobile Lindl |
| CN108308039A (en) * | 2018-05-16 | 2018-07-24 | 芜湖市三山区绿色食品产业协会 | Tissue culture medium (TCM) and its preparation method and application for dendrobium candidum tissue culture |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1433669A (en) * | 2002-01-25 | 2003-08-06 | 中国医学科学院药用植物研究所 | Application of mycorrhizal fungi in dendrobe cultivation |
-
2007
- 2007-12-20 CN CN2007100664801A patent/CN101180949B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1433669A (en) * | 2002-01-25 | 2003-08-06 | 中国医学科学院药用植物研究所 | Application of mycorrhizal fungi in dendrobe cultivation |
Non-Patent Citations (4)
| Title |
|---|
| 刘骅等.铁皮石斛试管苗壮苗培养基的研究.《中国中药杂志》.1998,第23卷(第11期),654-657. * |
| 周俊辉等.铁皮石斛的组织培养与快速繁殖研究.《仲恺农业技术学院学报》.2005,第18卷(第1期),23-26. * |
| 张泉锋等.铁皮石斛培养的产业化研究.《中草药》.2004,第35卷(第4期),438-440. * |
| 徐桂萍等.黄草石斛的组培和快速繁殖.《烟台教育学院学报》.2004,第10卷(第4期),59-63. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101180949A (en) | 2008-05-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101180949B (en) | Stem tip tissue rapid cultivating and seedling method of algam dendrobium nobile | |
| CN103416305B (en) | Hormone-free tissue culture and rapid propagation method of anoectochilus formosanus seedlings | |
| CN101564008B (en) | Hormone-free cultivation and rapid propagation method of dendrobium candidum axenic seedlings | |
| CN100998311B (en) | Method for artificially breeding edulis mono-garlic clove orchid | |
| CN102119655B (en) | Natural light rapid breeding method for dendrobium officinale | |
| CN104082138B (en) | A kind of tissue cultivation rapid breeding method of Radix Aristolochiae Tagalae | |
| CN103535281B (en) | Tissue culture medium of sealwort roots and stems and in-vitro regeneration method | |
| CN103749302A (en) | Induced acclimation and cultivation method for salt-tolerant bamboo reed seedlings | |
| CN102630562A (en) | Tissue culture and rapid propagation method of Jiuhua Polygonatum sibiricum | |
| CN102948367B (en) | Method for performing in-vitro culturing and rapid propagating on bergenia crassifolia | |
| CN102835318A (en) | Technology for rapid sugar-free micropropagation of dendrobium officinale kimura et migo under non-aseptic condition | |
| CN105123529A (en) | Rapid propagation and efficient cultivation method of Bletilla striata | |
| CN105104190A (en) | Anoectochilus roxburghii tissue culture seedling method | |
| CN104137779B (en) | A kind of method of Rhizoma Sapium japonicum rapidly inducing stem segments regeneration plant | |
| CN105309311A (en) | Method for breeding improved variety of scrophularia ningpoensis Hemsl. | |
| CN101983557B (en) | In vitro quick breeding method of seedling stem of santal seed embryo | |
| CN105494098B (en) | A kind of method of quick breeding tuniclike psammosilene root seedling | |
| CN103460971B (en) | Method for improving transplanting survival rate of trichosanthes kirilowii tissue culture seedlings | |
| CN101953300B (en) | Tissue culture method for Curcuma wenyujin No.1 | |
| CN100425126C (en) | Fast lavandulol regeneration | |
| CN105409771A (en) | Rapid propagation method for Gymnadenia conopsea (L.) R. Br. | |
| CN104488723A (en) | Tissue-culture and rapid-propagation method of epimedium koreanum nakai | |
| CN105145365A (en) | Green globular body tissue culture propagation method for Alsophila costularis | |
| CN102893872A (en) | Tissue culture method for domesticated seedlings of iris pallida | |
| CN102067817A (en) | Method for quickly cultivating medicinal anectochilus roxburhii |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20110608 Termination date: 20131220 |