CN102144566A - Method for culturing test tube plantlet of Xingren Anoectochilus - Google Patents
Method for culturing test tube plantlet of Xingren Anoectochilus Download PDFInfo
- Publication number
- CN102144566A CN102144566A CN 201110078382 CN201110078382A CN102144566A CN 102144566 A CN102144566 A CN 102144566A CN 201110078382 CN201110078382 CN 201110078382 CN 201110078382 A CN201110078382 A CN 201110078382A CN 102144566 A CN102144566 A CN 102144566A
- Authority
- CN
- China
- Prior art keywords
- tube plantlet
- xingren
- medium
- test tube
- test
- 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.)
- Pending
Links
Abstract
The invention discloses a method for culturing a test tube plantlet of Xingren Anoectochilus, which comprises the steps of: selecting a stem section with an axillary bud as an explant, carrying out collateral bud induction and enrichment culture, grafting the collateral bud into a strong seedling rooting culture medium for strong seedling rooting cultivation, and obtaining the test tube plantlet, wherein the culture medium used for collateral bud induction and enrichment culture is MS+(2-5)mg/l 6-BA (butyl acrylate)+(0.2-0.5)mg/l NAA (naphthyl acetic acid)+8g/l agar+30g/l sugar pH: 5.7-6.0; and the strong seedling rooting medium is (1/2MS-MS)+(0.01-0.05)mg/l 6-BA+(0.05-0.10)mg/l NAA+8g/l agar+(15-25)g/l sugar pH: 5.7-6.0. The Xingren Anoectochilus has the propagation coefficient of more than 10 times, so that the test tube plantlet is thick and strong and has the transplant survival rate of more than 95%.
Description
Technical field
The present invention relates to plant tissue and cultivate technical field, particularly relate to the breeding method of Xingren roxburgh anoectochilus terminal bud test-tube plantlet.
Background technology
The Xingren roxburgh anoectochilus terminal bud (
A.xingrenensisZ.H.Tsi et X.H.Jin) be the orchid family (Orchidaceae), open the lip Cymbidium (
Anoectochilus) herbaceos perennial, open general designation roxburgh anoectochilus terminal buds such as lip orchid, anoectochilus formosanus, Kaohsiung roxburgh anoectochilus terminal bud with floral leaf, among the people do medicinal, but anti-inflammatory, detoxifcation, hemostasis step-down, cardiac stimulant diuresis, oxyhepatitis, chronic hepatitis, acute and chronic nephritis, kidney failure, hypertension, diabetes, gynecological disease, cancer etc. there is very significantly curative effect, therefore at the title that " king of medicine, refreshing medicine, gold grass " arranged among the people.According to reports, amino acid composition, composition, content and activity of fighting against senium content of elements all are higher than American Ginseng and wild ginseng in the roxburgh anoectochilus terminal bud.Roxburgh anoectochilus terminal bud is as a kind of precious medicine, and price is very high always, and bright at home careless procurement price reached 400-600 unit/kg in 2010, and the hay price is more up to 1500-2000 unit/kg.Roxburgh anoectochilus terminal bud plant type in Xingren is small and exquisite in addition, and blade profile is attractive in appearance, has high ornamental value.
Xingren roxburgh anoectochilus terminal bud seed only could be sprouted under the situation of mycosymbiosis, and germination rate is low.At natural world, roxburgh anoectochilus terminal bud is the plant of happiness foods such as many worms, bird, mouse, snake.Because mankind's activity, ecotope is damaged, and orders about down in interests, suffers destructive collection again, now has been in critically endangered state.
Summary of the invention
Technical problem to be solved by this invention is to overcome the existing low and endangered defective of wild Xingren roxburgh anoectochilus terminal bud seed germination rate, the kind breeding method of a kind of Xingren roxburgh anoectochilus terminal bud test-tube plantlet is provided, this method is through the screening to medium, optimize and be fit to the medium that Xingren roxburgh anoectochilus terminal bud test-tube plantlet is cultivated, reproduction coefficient reaches more than 10 times, seedling is sturdy, and transplanting survival rate is more than 95%.
In order to solve the problems of the technologies described above, the present invention adopts following technical scheme:
The breeding method of Xingren of the present invention roxburgh anoectochilus terminal bud test-tube plantlet: the stem section of select tape axillalry bud is made explant, carries out inducing and enrichment culture from sprouting, and will be transferred to from sprouting then to carry out the strengthening seedling and rooting cultivation the strengthening seedling and rooting medium, promptly gets test-tube plantlet, wherein:
Induce the medium with enrichment culture to be from sprouting:
MS+(2～5) mg/l 6-BA+(0.2～0.5) mg/l NAA+8g/l agar+30g/l sugar pH:5.7～6.0;
The strengthening seedling and rooting medium is:
(mg/l 6-BA+(0.05～0.10) mg/l NAA+8g/l agar of 1/2MS～MS)+(0.01～0.05)+(15～25) g/l sugar pH:5.7～6.0.
In the breeding method of above-mentioned Xingren roxburgh anoectochilus terminal bud test-tube plantlet, when carrying out the strengthening seedling and rooting cultivation, its condition of culture is illumination 16h/d, intensity of illumination 500～1200lux, and 23～27 ℃ of temperature were carried out 1 time to culturing room and sterilized in per 2 months.
Compared with prior art, the screening of the medium that the present invention is adopted when test-tube plantlet is cultivated, optimized the medium that suitable Xingren roxburgh anoectochilus terminal bud test-tube plantlet is cultivated, reproduction coefficient is reached more than 10 times, and the gained test-tube plantlet is sturdy, and the transplanting survival rate of test-tube plantlet can reach more than 95%, in addition, what the present invention adopted is conventional test-tube plantlet breeding method, and this method is simple, ripe, is fit to very much apply.
Embodiment
Describe technical scheme of the present invention in detail below in conjunction with embodiment, the breeding method of Xingren roxburgh anoectochilus terminal bud test-tube plantlet:
(1) foundation of explant selection and sterile system: the plant of selecting robust growth, get the stem section of the long band axillalry bud of 1cm and make explant, the stem section is cleaned out, put into washing powder water rinsing 3～5 minutes earlier, then with flowing water flushing 30 minutes, in superclean bench, with 75% alcohol moistening 5～8 seconds, take out after putting into 0.1% HgCl solution and sterilizing 9～13 minutes, with aseptic water washing 5～6 times;
(2) induce and enrichment culture from sprouting: the explant after the aseptic process is inoculated in from the inducing culture of sprouting, carry out from the inducing culture of sprouting, general inoculation began to sprout in 7～10 days, when sprouting 0.5cm length from sprouting, it is cut open, be forwarded to and make further enrichment culture on the proliferated culture medium, the albefaction seedling that occurs when removing enrichment culture during switching;
Induce screening with the enrichment culture medium from sprouting:
Test is by with the improved culture medium of MS, 1/2MS, MS (macroelement reduces by half, trace element double), and the consumption of 6-BA is from 1～6mg/L, and the consumption of NAA passes through a large amount of screening tests such as orthogonal experiment from 0.1～1.5mg/L.Draw following comparatively ideal medium combination:
MS+(2～5) mg/l 6-BA+(0.2～0.5) mg/l NAA+8g/l agar+30g/l sugar pH:5.7～6.0;
(3) strengthening seedling and rooting is cultivated: from the process of growth of sprouting, while is president's adventive root also, when growing up to the high seedling of 1-2cm from sprouting, be forwarded to and carry out strong plantlets and rootage in the strong plantlets and rootage medium and cultivate, culturing room carried out 1 sterilization in per 2 months, if illumination is strong excessively, the albinism of test-tube plantlet is serious, and the Xingren roxburgh anoectochilus terminal bud is contaminated easily, so per 2 months carry out disinfection to culturing room, can adopt formaldehyde fumigation, culturing room also can adopt ozonization equipment to carry out sterilization;
The screening of strong plantlets and rootage medium:
The strong plantlets and rootage medium is from MS, 1/2MS, 1/4MS, and the consumption of sugar is from 15～30g, and the consumption of 6-BA is from 0.01～0.1, and the NAA consumption filters out following comparatively ideal medium combination from 0.01～0.5mg/L by test of many times such as orthogonal experiments:
(mg/l 6-BA+(0.05～0.10) mg/l NAA+8g/l agar of 1/2MS～MS)+(0.01～0.05)+(15～25) g/l sugar pH:5.7～6.0.
(4) successive transfer culture: in time carry out successive transfer culture behind the strengthening seedling and rooting, guarantee the nutrient supply of medium,, in time change if polluting appears in medium.
(5) acclimatization and transplants: place the normal temperature lower refining seedling after 1 week on blake bottle, take out seedling, clean medium, with plantation comparative trials such as the peat composed of rotten mosses, perlite, wood chip, leaves, wherein the peat composed of rotten mosses is planted roxburgh anoectochilus terminal bud, and survival rate is the highest, can reach more than 95%.
Condition of culture in the above-mentioned cultural method is illumination 16h/d, intensity of illumination 500～1200lux, 23～27 ℃ of temperature.
Claims (2)
1. the breeding method of an Xingren roxburgh anoectochilus terminal bud test-tube plantlet, it is characterized in that: the stem section of select tape axillalry bud is made explant, carries out inducing and enrichment culture from sprouting, and will be transferred to from sprouting then to carry out the strengthening seedling and rooting cultivation the strengthening seedling and rooting medium, promptly get test-tube plantlet, wherein:
Induce the medium with enrichment culture to be from sprouting:
MS+(2～5) mg/l 6-BA+(0.2～0.5) mg/l NAA+8g/l agar+30g/l sugar pH:5.7～6.0;
The strengthening seedling and rooting medium is:
(mg/l 6-BA+(0.05～0.10) mg/l NAA+8g/l agar of 1/2MS～MS)+(0.01～0.05)+(15～25) g/l sugar pH:5.7～6.0.
2. according to the breeding method of the described Xingren of claim 1 roxburgh anoectochilus terminal bud test-tube plantlet, it is characterized in that: the condition of culture that strengthening seedling and rooting is cultivated is illumination 16h/d, intensity of illumination 500～1200lux, and 23～27 ℃ of temperature were carried out 1 time to culturing room and sterilized in per 2 months.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110078382 CN102144566A (en) | 2011-03-30 | 2011-03-30 | Method for culturing test tube plantlet of Xingren Anoectochilus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110078382 CN102144566A (en) | 2011-03-30 | 2011-03-30 | Method for culturing test tube plantlet of Xingren Anoectochilus |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102144566A true CN102144566A (en) | 2011-08-10 |
Family
ID=44419204
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201110078382 Pending CN102144566A (en) | 2011-03-30 | 2011-03-30 | Method for culturing test tube plantlet of Xingren Anoectochilus |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102144566A (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102511388A (en) * | 2011-11-25 | 2012-06-27 | 贵州师范大学 | Method for breeding anti-reversible tissue culture seedlings of anoectochilus roxburghii |
| CN103120126A (en) * | 2013-02-04 | 2013-05-29 | 南京工业大学 | Method for carrying out tissue cultivation propagation of anoectochilus roxburghii by intermittent submerged bioreactor |
| CN103688854A (en) * | 2013-12-06 | 2014-04-02 | 四川省自然资源科学研究院 | Tissue culture rapid propagation method of Emei anoectochilus formosanus |
| CN104904600A (en) * | 2015-06-03 | 2015-09-16 | 吴华球 | Tissue-culturing and rapid-propagating method of anoectochilus formosanus |
| CN105475129A (en) * | 2015-05-29 | 2016-04-13 | 三明市农业科学研究院 | Tissue-culture rapid propagation method for arundina graminifolia |
| CN105660400A (en) * | 2016-01-25 | 2016-06-15 | 宁波易中禾生物技术有限公司 | Strengthening and weight increasing method for anoectochilus roxburghii tissue cultured seedlings |
| CN105766647A (en) * | 2016-04-05 | 2016-07-20 | 福建农林大学 | Tissue culture method for club moss |
| CN106386487A (en) * | 2016-09-08 | 2017-02-15 | 福建虎伯寮生物集团有限公司 | Anoectochilus roxburghii tissue culturing method |
| CN106613995A (en) * | 2017-01-18 | 2017-05-10 | 福建农林大学 | Tissue culture method for culturing anoectochilus roxburghii by bacteriostatic culture medium |
| CN109042341A (en) * | 2018-10-24 | 2018-12-21 | 云南善源生物科技发展有限公司 | A kind of roxburgh anoectochilus terminal bud culture medium and preparation method thereof |
-
2011
- 2011-03-30 CN CN 201110078382 patent/CN102144566A/en active Pending
Non-Patent Citations (3)
| Title |
|---|
| 《植物生理学通讯》 19981231 陈穗云 等 台湾金线莲的离体快速繁殖 443 1-2 第34卷, 第6期 * |
| 《现代园艺》 20081031 陈永快 等 福建金线莲和台湾金线莲的组培快繁技术 9-12 1-2 , 第10期 * |
| 《贵州科学》 20070630 张以忠 等 金线莲研究的现状与展望 81-84 1-2 第25卷, 第2期 * |
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102511388A (en) * | 2011-11-25 | 2012-06-27 | 贵州师范大学 | Method for breeding anti-reversible tissue culture seedlings of anoectochilus roxburghii |
| CN103120126A (en) * | 2013-02-04 | 2013-05-29 | 南京工业大学 | Method for carrying out tissue cultivation propagation of anoectochilus roxburghii by intermittent submerged bioreactor |
| CN103120126B (en) * | 2013-02-04 | 2014-05-07 | 南京工业大学 | Method for carrying out tissue cultivation propagation of anoectochilus roxburghii by intermittent submerged bioreactor |
| CN103688854B (en) * | 2013-12-06 | 2016-03-30 | 四川省自然资源科学研究院 | The tissue culture and rapid propagation method of high eyebrow roxburgh anoectochilus terminal bud |
| CN103688854A (en) * | 2013-12-06 | 2014-04-02 | 四川省自然资源科学研究院 | Tissue culture rapid propagation method of Emei anoectochilus formosanus |
| CN105475129A (en) * | 2015-05-29 | 2016-04-13 | 三明市农业科学研究院 | Tissue-culture rapid propagation method for arundina graminifolia |
| CN104904600A (en) * | 2015-06-03 | 2015-09-16 | 吴华球 | Tissue-culturing and rapid-propagating method of anoectochilus formosanus |
| CN105660400A (en) * | 2016-01-25 | 2016-06-15 | 宁波易中禾生物技术有限公司 | Strengthening and weight increasing method for anoectochilus roxburghii tissue cultured seedlings |
| CN105660400B (en) * | 2016-01-25 | 2018-02-13 | 宁波易中禾生物技术有限公司 | A kind of strong sprout weight increasing method of roxburgh anoectochilus terminal bud tissue-cultured seedling |
| CN105766647A (en) * | 2016-04-05 | 2016-07-20 | 福建农林大学 | Tissue culture method for club moss |
| CN105766647B (en) * | 2016-04-05 | 2018-01-12 | 福建农林大学 | A kind of hectolitre pine method for tissue culture |
| CN106386487A (en) * | 2016-09-08 | 2017-02-15 | 福建虎伯寮生物集团有限公司 | Anoectochilus roxburghii tissue culturing method |
| CN106613995A (en) * | 2017-01-18 | 2017-05-10 | 福建农林大学 | Tissue culture method for culturing anoectochilus roxburghii by bacteriostatic culture medium |
| CN109042341A (en) * | 2018-10-24 | 2018-12-21 | 云南善源生物科技发展有限公司 | A kind of roxburgh anoectochilus terminal bud culture medium and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102144566A (en) | Method for culturing test tube plantlet of Xingren Anoectochilus | |
| CN102246694B (en) | Tissue culture method of gynura divaricata | |
| CN102119660A (en) | Method for rooting culture,seedling adaptation and transplantation of alpine rose tissue-cultured seedling in greenhouse | |
| CN102499088B (en) | Method for quickly breeding seedlings of Guangxi anoectochilus roxburghii capsules by utilizing Guangxi anoectochilus roxburghii capsules | |
| CN103270952B (en) | Sinkiang drug mulberry tissue culture and rapid propagation culture medium | |
| CN102884982A (en) | Hormone-free rapid propagation organic production method of anoectochilus formosanus germchit | |
| CN109258460A (en) | Micro-stem tip culture combines the breeding method of heat treatment acquisition Zengcheng honey chrysanthemum detoxic seedling | |
| CN102845313A (en) | Method for quickly in-vitro actinidia kolomikta propagating | |
| CN104585027B (en) | A kind of hardening off method of Rhizoma Zingiberis Recens tissue cultured seedling | |
| CN102144555A (en) | Rapid propagation method for radix puerariae | |
| CN1985582A (en) | Tissue culture seedling growing process for trichosanthes | |
| CN101983557B (en) | In vitro quick breeding method of seedling stem of santal seed embryo | |
| CN103460971B (en) | Method for improving transplanting survival rate of trichosanthes kirilowii tissue culture seedlings | |
| CN106718877A (en) | A kind of flourishing torch root tuber of aromatic turmeric high quality seedling rapid propagation method | |
| CN100429972C (en) | Tissue culture propagation process of late spring michelia | |
| CN107821165A (en) | A kind of tissue culture propagation and purposes for ceasing the tuber of pinellia | |
| CN103004595A (en) | Twig cuttage breeding method for ginseng fruit | |
| CN108739380B (en) | Method for one-time seedling formation of rhizoma bletillae tissue culture seedlings | |
| CN102144569B (en) | Method for rapidly propagating root and rhizome of Divaricate Gynura | |
| CN106508661A (en) | Method for constructing anoectochilus formosanus polyploidy resource | |
| CN106258993A (en) | A kind of blueberry tissue culture method | |
| CN105993952A (en) | Rapid breeding method of Euryodendron excelsum cultivation seedlings | |
| CN112119913A (en) | Virus-free propagation method for sweet potatoes | |
| CN105432466B (en) | Method with plant regeneration occurs for a kind of pittosporum tobira somatic embryo | |
| CN108391591B (en) | Tissue culture and rapid propagation method for tabebuia flavedo |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20110810 |