CN101785432A - Adventitious bud inducing and rooting medium for radicle of Carya cathayensis Sarg and tissue culture method - Google Patents
Adventitious bud inducing and rooting medium for radicle of Carya cathayensis Sarg and tissue culture method Download PDFInfo
- Publication number
- CN101785432A CN101785432A CN201019146041A CN201019146041A CN101785432A CN 101785432 A CN101785432 A CN 101785432A CN 201019146041 A CN201019146041 A CN 201019146041A CN 201019146041 A CN201019146041 A CN 201019146041A CN 101785432 A CN101785432 A CN 101785432A
- Authority
- CN
- China
- Prior art keywords
- adventitious bud
- medium
- radicle
- culture
- days
- 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.)
- Granted
Links
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses an adventitious bud inducing and rooting medium for radicle of Carya cathayensis Sarg, which comprises four culture media for different induction periods: (1) a primary culture medium, (2) a radicle adventitious bud induction culture medium, (3) an adventitious bud enrichment culture medium, (4) an adventitious bud rooting medium. The four culture media all takes MS as a basic culture medium and are added with sucrose, agar A and other substances and have a pH value of 5.7. A tissue culture method for adventitious bud inducing and rooting of the radicle of the Carya cathayensis Sarg with the four culture media comprises the following five steps: 1, collecting and disinfecting explants; 2, performing primary culture; 3, inducing the adventitious bud of the radicle; 4, performing adventitious bud enrichment culture; and 5, performing rooting induction. The culture of the five steps lays a solid foundation for rapidly culturing saplings of the Carya cathayensis Sarg with good quality. When the medium is used for tissue culture, regenerated plantlet can maintain the excellent properties of parents; the propagation speed of the saplings is improved; the propagation number is expanded; seeds are effectively utilized; and the seedling period is shortened so as to contribute to meeting social requirements.
Description
Technical field
The present invention relates to a kind of is explant with the cathay hickory rataria, grows to form radicle, forms the medium and the tissue culture method of indefinite bud, root induction then.
Background technology
Cathay hickory (Carya cathayensis Sarg.) is the good economic tree of the fruit of Juglandaceae hickory, wooden dual-purpose, and under natural environment, cathay hickory is by seminal propagation.The cathay hickory dry fruit can be processed into various characteristic typical local food, health products, and the cathay hickory oil content is rich in unsaturated fatty acid up to 70%～74%, contains crude protein 7.8%～9.6% and contains necessary for human body mineral elements such as K, Ca, Mg, Na.Cathay hickory wooden hard, texture is beautiful, anticorrosive, of many uses in military project, boats and ships and building industry, be a kind of very good economic tree of China.But the distributed areas of hickory are narrow, are the endemic tree of China, and have only the ground, South Mountain, Anhui of mountain region, northwest, Zhejiang and boundary to distribute to some extent.What demand was prosperous, product is few is particularly thorny.For a long time, the breeding of cathay hickory mainly is to grow directly from seeds and propagation by grafiting, and the breeding of growing directly from seeds is difficult to keep the good characteristic of kind.Because of being rich in tannin, otch is oxidizing brown stain very easily again for cathay hickory, and is higher to the specification requirement of grafting, and grafting is subjected to the restriction of season and weather again, and reproduction speed is difficult to satisfy the demand of production development.
Forestry circle, numerous scholars have done many new explorations and trial to the breeding of cathay hickory, insert Mr.s such as propagation technique, Pei Dong as the root of Mr.'s Huang Youjun treelet the tender stem-root technology of test tube of walnut and the root of treelet are inserted propagation technique etc., each has made useful contribution to society.By retrieval, do not find that so far cultivating indefinite bud and breeding by the cathay hickory rataria through test tube takes root and breed the relevant report of plant.
Summary of the invention
Economic worth at the cathay hickory seeds more and more is familiar with by people, spontaneous the springing up of enthusiasm that development cathay hickory produces, rely on traditional grow directly from seeds breeding and propagation by grafiting, from quality and quantitatively all can't satisfy reality demand, for this reason, it is explant with the cathay hickory rataria that the technical problem to be solved in the present invention provides a kind of, induces the medium and the tissue culture method of radicle growth, indefinite bud formation and adventitious bud rooting.
Technical problem of the present invention solves by following technical solution:
1. this cathay hickory radicle adventitious bud inducing and root media comprise not isogeneous induction four kinds of medium in period, and the raw material and the additional amount thereof of various medium are respectively:
(1) initial culture base: with MS is minimal medium, and additional saccharose 20～30g/L, Agar type A are agar A 6.5～8.5g/L, and the pH value of medium is 5.7;
(2) radicle adventitious bud induction culture base: MS additional saccharose 20～30g/L, agar A 6.5～8.5g/L, Picloram are 4-amino-3,5, and 6-trichloropyridine carboxylic acid 0.01mg/L, 6-BA are 6-aminoadenine 1.0～3.0mg/L, pH value 5.7;
(3) adventitious bud proliferation medium: MS additional saccharose 20～30g/L, agar A6.5～8.5g/L, Picloram0.01～0.02mg/L, 6-BA1～2mg/L, the pH value is 5.7;
(4) adventitious bud rooting medium: MS additional saccharose 20～30g/L, agar A6.5～8.5g/L, IAA are heteroauxin 0.3～0.8mg/L, pH value 5.7.
With above-mentioned medium carry out cathay hickory radicle adventitious bud inducing and the tissue culture method of taking root through the following step:
(1) explant collection and sterilization: choose the young fruit of being tied in 100～120 days after healthy and strong cathay hickory blooms, carry out running water flushing 2h successively, on the superclean bench with 75% ethanol sterilization 3min, aseptic water washing 3～4 times, concentration is 0.525% NaClO sterilization 20min and vacuumizes, and behind the aseptic water washing 5 times, blots surface moisture with sterilization filter paper, and peel off the keratin pericarp, it is stand-by to take out rataria;
(2) initial culture: the rataria that step (1) aseptic condition is taken out down inserts in the aforesaid initial culture base, transfers every day 1 time; Change switching in per 3 days after 3 days into once, all be forwarded in the identical fresh culture at every turn, corotation connects three times, places dark cabinet to cultivate in wherein preceding 7 days, and placing light dark period in back 5 days is to cultivate under the condition of the dark 8h of light 16h/.Cultivation temperature is 25 ± 2 ℃;
(3) radicle adventitious bud inducing: the rataria that radicle growth behind the initial culture is good inserts in the aforesaid radicle adventitious bud induction culture base, through inducing of 30d, obtains indefinite bud.Micro computer time-controlled switch control culturing room light application time every day, every day, light dark period was the dark 8h of light 16h/, intensity of illumination is 40～50 μ mol m
-2s
-1, cultivation temperature is 25 ± 2 ℃;
(4) adventitious bud proliferation is cultivated: the indefinite bud that obtains is carried out enrichment culture on aforesaid adventitious bud proliferation medium, subculture was 1 time in 30 days, and subculture is 3 times altogether, and growth coefficient is 5～8.Illumination cultivation, light dark period are the dark 8h of light 16h/, and intensity of illumination is 40～50 μ mol m
-2s
-1, cultivation temperature is 25 ± 2 ℃;
(5) root induction: the root induction in root media of aforesaid indefinite bud, to cultivate 30 days, rooting rate reaches 50%.Illumination cultivation, light dark period are the dark 8h of light 16h/, and intensity of illumination is 40～50 μ mol m
-2s
-1, cultivation temperature is 25 ± 1 ℃.
The invention has the beneficial effects as follows:
Use cathay hickory radicle adventitious bud inducing and carry out culture of rootage, can make regeneration plant keep maternal merit preferably, improve the reproduction coefficient of cathay hickory effectively, be fast numerous the laying a good foundation of cathay hickory breeding.Compare with seed propagation, tissue culture method can effectively utilize seed, its germination rate height, and growing-seedling period is short, and seed only needs can sprout in 2 weeks under the aseptic condition.Comparing with cottage propagation, is 50～60 times of cottage propagation at the reproduction coefficient of a growth cycle inner tissue cultivation of 6 months by a definite date.
Embodiment
The present invention is described in further detail below in conjunction with embodiment: in this cathay hickory radicle adventitious bud inducing and the medium of taking root, selecting minimal medium for use is the MS medium, this is through all additional saccharose 20～30g/L, agar A 6.5～8.5g/L, Picloram are 4-amino-3 separately with MS, 1/2MS, four kinds of minimal mediums of WPM, DKW, 5,6-trichloropyridine carboxylic acid 0.01mg/L, 6-BA are 6-aminoadenine 1.0～3.0mg/L, pH value 5.7, test, every group is repeated 3 times, and every kind of minimal medium has the comparative trial of 20 explants to draw.After handling 30 days in the MS medium, radicle growth is best, and the indefinite bud number of inducing is maximum, best in quality.
In a word, medium disclosed in this invention and proportioning value thereof, temperature, coagulating agent etc. are also available to be substituted such as glucose, quartzy agar etc., and each proportion of raw materials value of medium also can adopt replacing near value of preferred plan definite value of the present invention respectively.
Given below is most preferred embodiment of the present invention.
1. this cathay hickory radicle adventitious bud inducing and root media comprise not isogeneous induction four kinds of medium in period, and the raw material and the additional amount thereof of various medium are respectively:
(1) initial culture base: with MS is minimal medium, and additional saccharose 20～30g/L, Agar type A are agar A 6.5～8.5g/L, and the pH value of medium is 5.7;
(2) radicle adventitious bud induction culture base: MS additional saccharose 20～30g/L, agar A 6.5～8.5g/L, Picloram are 4-amino-3,5, and 6-trichloropyridine carboxylic acid 0.01mg/L, 6-BA are 6-aminoadenine 1.0～3.0mg/L, pH value 5.7;
(3) adventitious bud proliferation medium: MS additional saccharose 20～30g/L, agar A6.5～8.5g/L, Picloram0.01～0.02mg/L, 6-BA1～2mg/L, the pH value is 5.7;
(4) adventitious bud rooting medium: MS additional saccharose 20～30g/L, agar A6.5～8.5g/L, IAA are heteroauxin 0.3～0.8mg/L, pH value 5.7.
Carry out cathay hickory radicle adventitious bud inducing and rooting method process the following step with above-mentioned medium:
(1) explant collection and sterilization: choose the young fruit of being tied in 100～120 days after healthy and strong cathay hickory blooms, carry out running water flushing 2h successively, on the superclean bench with 75% ethanol sterilization 3min, aseptic water washing 3～4 times, concentration is 0.525% NaClO sterilization 20min and vacuumizes, and behind the aseptic water washing 5 times, blots surface moisture with sterilization filter paper, and peel off the keratin pericarp, it is stand-by to take out rataria;
(2) initial culture: the rataria that step (1) aseptic condition is taken out down inserts in the aforesaid initial culture base, transfers every day 1 time; Change switching in per 3 days after 3 days into once, all be forwarded in the identical fresh culture at every turn, corotation connects three times, places dark cabinet to cultivate in wherein preceding 7 days, and placing light dark period in back 5 days is to cultivate intensity of illumination under the condition of the dark 8h of light 16h/: 40～50 μ mol m
-2s
-1Cultivation temperature is 25 ± 2 ℃, be inoculated in the initial culture base, this medium comprises sodium dihydrogen phosphate 85g/L, White ' s vitamin and MS salt, also have 30g/L sucrose, 500mg/L caseinhydrolysate, 6.5～8.5g/L agar A, adding 1.0～3.0mg/L BA is that benzyl aminoadenine, 0.01mg/LPicloram are 4-amino-3,5,6-trichloropyridine carboxylic acid is formulated.Cultivated 12 days, radicle growth is good;
(3) radicle adventitious bud inducing: the rataria that radicle growth behind the initial culture is good inserts in the MS medium of additional 0.01～0.02mg/L Picloram, 1.0～3.0mg/L 6-BA, sucrose 20～30g/L, agar A 6.5～8.5g/L, pH value 5.7, intensity of illumination: 40～50 μ mol m
-2s
-125 ± 2 ℃ of cultivation temperature, light application time 16h/ days, cultivated 30 days, get well-developed indefinite bud;
(4) adventitious bud proliferation is cultivated: the indefinite bud that obtains is grown to 0.5cm left and right sides Gao Shi, downcut and be inoculated into the aforesaid adventitious bud proliferation medium from radicle and cultivate, subculture was 1 time in 30 days, and subculture is 3 times altogether, and each growth coefficient is 5～8.Illumination cultivation, light dark period are the dark 8h of light 16h/, and intensity of illumination is 40～50 μ mol m
-2s
-1, 25 ± 2 ℃ of cultivation temperature;
(5) root induction: when aforesaid indefinite bud grows to 0.5～1.0cm when high, be cut into simple bud and be inoculated in the aforesaid root media, cultivated 30 days, rooting rate reaches 50%.Illumination cultivation, light dark period are the dark 8h of light 16h/, and intensity of illumination is 40～50 μ molm
-2s
-1, cultivation temperature is 25 ± 2 ℃.
The MS salt is commercially available with White ' s vitamin and other relevant chemicals in the above-mentioned medium, presses the catalogue adapted, and is known for going together.
Claims (2)
1. cathay hickory radicle adventitious bud inducing and root media is characterized in that comprising not isogeneous induction four kinds of medium in period, and the raw material and the additional amount thereof of various medium are respectively:
(1) initial culture base: with MS is minimal medium, and additional saccharose 20～30g/L, Agar type A are agar A 6.5～8.5g/L, and the pH value of medium is 5.7;
(2) radicle adventitious bud induction culture base: MS additional saccharose 20～30g/L, agar A 6.5～8.5g/L, Picloram are 4-amino-3,5, and 6-trichloropyridine carboxylic acid 0.01mg/L, 6-BA are 6-aminoadenine 1.0～3.0mg/L, pH value 5.7;
(3) adventitious bud proliferation medium: MS additional saccharose 20～30g/L, agar A6.5～8.5g/L, Picloram0.01～0.02mg/L, 6-BA1～2mg/L, the pH value is 5.7;
(4) adventitious bud rooting medium: MS additional saccharose 20～30g/L, agar A6.5～8.5g/L, IAA are heteroauxin 0.3～0.8mg/L, pH value 5.7.
2. tissue culture method that carries out cathay hickory radicle adventitious bud inducing and take root with the described medium of claim 1 is characterized in that through the following step:
(1) explant collection and sterilization: choose the young fruit of being tied in 100～120 days after healthy and strong cathay hickory blooms, carry out running water flushing 2h successively, on the superclean bench with 75% ethanol sterilization 3min, aseptic water washing 3～4 times, concentration is 0.525% NaClO sterilization 20min and vacuumizes, and behind the aseptic water washing 5 times, blots surface moisture with sterilization filter paper, and peel off the keratin pericarp, it is stand-by to take out rataria;
(2) initial culture: the rataria that step (1) aseptic condition is taken out down inserts in the aforesaid initial culture base, transfers every day 1 time; Change switching in per 3 days after 3 days into once, all be forwarded in the identical fresh culture at every turn, corotation connects three times, places dark cabinet to cultivate in wherein preceding 7 days, and placing light dark period in back 5 days is to cultivate under the condition of the dark 8h of light 16h/.Cultivation temperature is 25 ± 2 ℃;
(3) radicle adventitious bud inducing: the rataria that radicle growth behind the initial culture is good inserts in the aforesaid radicle adventitious bud induction culture base, through inducing of 30d, obtains indefinite bud.Micro computer time-controlled switch control culturing room light application time every day, every day, light dark period was the dark 8h of light 16h/, intensity of illumination is 40～50 μ mol m
-2s
-1, cultivation temperature is 25 ± 2 ℃;
(4) adventitious bud proliferation is cultivated: the indefinite bud that obtains is carried out enrichment culture on aforesaid adventitious bud proliferation medium, subculture was 1 time in 30 days, and subculture is 3 times altogether, and growth coefficient is 5～8.Illumination cultivation, light dark period are the dark 8h of light 16h/, and intensity of illumination is 40～50 μ mol m
-2s
-1, cultivation temperature is 25 ± 2 ℃;
(5) root induction: the root induction in root media of aforesaid indefinite bud, to cultivate 30 days, rooting rate reaches 50%.Illumination cultivation, light dark period are the dark 8h of light 16h/, and intensity of illumination is 40～50 μ mol m
-2s
-1, cultivation temperature is 25 ± 1 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010191460410A CN101785432B (en) | 2010-02-05 | 2010-02-05 | Adventitious bud inducing and rooting medium for radicle of Carya cathayensis Sarg and tissue culture method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010191460410A CN101785432B (en) | 2010-02-05 | 2010-02-05 | Adventitious bud inducing and rooting medium for radicle of Carya cathayensis Sarg and tissue culture method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101785432A true CN101785432A (en) | 2010-07-28 |
| CN101785432B CN101785432B (en) | 2011-12-21 |
Family
ID=42528936
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2010191460410A Expired - Fee Related CN101785432B (en) | 2010-02-05 | 2010-02-05 | Adventitious bud inducing and rooting medium for radicle of Carya cathayensis Sarg and tissue culture method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101785432B (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102388806A (en) * | 2011-09-15 | 2012-03-28 | 江苏省林业科学研究院 | Carya illinoensis tissue culture propagating method |
| CN102668774A (en) * | 2012-06-05 | 2012-09-19 | 江苏省林业科学研究院 | Seedling method for promoting growing of nursery stock lateral roots |
| CN103004593A (en) * | 2012-12-11 | 2013-04-03 | 浙江农林大学 | Thin-shell pecan tissue culture bud micro grafting method |
| CN104957040A (en) * | 2015-07-16 | 2015-10-07 | 句容市容北茶文化有限公司 | Carya illinoensis cell embryo tissue culture method |
| CN107155894A (en) * | 2017-07-11 | 2017-09-15 | 李国松 | A kind of green ridge walnut somatic embryo method for culturing seedlings |
| CN113179950A (en) * | 2021-05-18 | 2021-07-30 | 广西壮族自治区农业科学院 | Method for sterilizing explants by using reduced pressure |
-
2010
- 2010-02-05 CN CN2010191460410A patent/CN101785432B/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102388806A (en) * | 2011-09-15 | 2012-03-28 | 江苏省林业科学研究院 | Carya illinoensis tissue culture propagating method |
| CN102668774A (en) * | 2012-06-05 | 2012-09-19 | 江苏省林业科学研究院 | Seedling method for promoting growing of nursery stock lateral roots |
| CN103004593A (en) * | 2012-12-11 | 2013-04-03 | 浙江农林大学 | Thin-shell pecan tissue culture bud micro grafting method |
| CN104957040A (en) * | 2015-07-16 | 2015-10-07 | 句容市容北茶文化有限公司 | Carya illinoensis cell embryo tissue culture method |
| CN107155894A (en) * | 2017-07-11 | 2017-09-15 | 李国松 | A kind of green ridge walnut somatic embryo method for culturing seedlings |
| CN113179950A (en) * | 2021-05-18 | 2021-07-30 | 广西壮族自治区农业科学院 | Method for sterilizing explants by using reduced pressure |
| CN113179950B (en) * | 2021-05-18 | 2022-06-07 | 广西壮族自治区农业科学院 | Method for sterilizing explants by using reduced pressure |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101785432B (en) | 2011-12-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103651121B (en) | A kind of bletilla differentiation, strong seedling culture base | |
| CN102369881B (en) | Rapid propagation technique of Dendrobium candidum axillary buds | |
| CN101785432B (en) | Adventitious bud inducing and rooting medium for radicle of Carya cathayensis Sarg and tissue culture method | |
| CN101491215B (en) | Chinese toon tissue-culture quick propagation technique | |
| CN105340747A (en) | Asexual rapid propagation method for radix glycyrrhizae | |
| CN105475130A (en) | Castanopsis hystrix high efficiency isolated culture plant regeneration method | |
| CN103004593A (en) | Thin-shell pecan tissue culture bud micro grafting method | |
| CN103651122B (en) | A kind of bletilla protocorm induction medium | |
| CN103222425A (en) | Efficient and rapid propagation technology suitable for southern highbush blueberry | |
| CN100553446C (en) | A kind of method of European wild black cherry mahogany tissue-culturing rapid propagation | |
| CN102461463A (en) | Paper mulberry seedling tissue culture method suitable for extensive plantation | |
| CN102907326B (en) | Tissue culture propagation method for Medicagao Sativa L. | |
| CN101161055B (en) | Method for exsomatizing screening grassiness salt resistsomatic mutation body | |
| CN104255457A (en) | Rapid propagation method for tissue culture of ilex nitidissima | |
| CN109819892B (en) | Tissue culture method of good single plant of tsaoko | |
| CN102342246B (en) | Rhododendron decorum tissue-culture quick propagation method | |
| CN104082145B (en) | A kind of method of round-pinna maidenhair herb Fast-propagation | |
| CN104488722B (en) | A kind of quick breeding method for tissue culture of South America crutch flower | |
| CN103229721B (en) | Tissue culture propagation method of Gynura formosana | |
| CN102792889B (en) | Chuanminshen violaceum tissue culture rapid propagation technology | |
| CN106165648B (en) | A kind of cercis tissue culture culture medium and cultural method | |
| CN102613089B (en) | High-efficient in-vitro propagation method of 20-year-old schima superba big tree | |
| CN101703003B (en) | Method for quickly propagating grassiness | |
| CN109819939A (en) | A kind of hybridization Cultivating techniques of the small black pig of Yulong | |
| CN101012448A (en) | Method of screening leymus chinensis salt-tolerant somatic mutation |
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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20111221 Termination date: 20150205 |
|
| EXPY | Termination of patent right or utility model |