CN101637127A - Vitro rapid propagation method of kefir lily by using seedling ovaries as explant - Google Patents
Vitro rapid propagation method of kefir lily by using seedling ovaries as explant Download PDFInfo
- Publication number
- CN101637127A CN101637127A CN200910067435A CN200910067435A CN101637127A CN 101637127 A CN101637127 A CN 101637127A CN 200910067435 A CN200910067435 A CN 200910067435A CN 200910067435 A CN200910067435 A CN 200910067435A CN 101637127 A CN101637127 A CN 101637127A
- Authority
- CN
- China
- Prior art keywords
- plant
- callus
- explant
- regeneration
- once
- 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
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention belongs to a plant propagation method, particularly relating to an asexual rapid propagation method of flowers. In the method, technologies of explant inoculation, callus induction, adventitious bud induction, root induction, regeneration plant transplantation and used, ovary longitudinal cutting blocks of seedling inflorescence of clivia miniata are used as explants, so that kefir lilies can generate large quantities of regeneration plants having the entirely same gene background and unified plant type within short time. The dedifferentiation ratio can reach more than 80%, and the differentiation ratio can reach 70%. Calluses induced by one explant can be differentiated into more than 100 cluster buds through repeated regeneration; the rooting rate of the cluster buds can reach 100%, and the survival rate of transplantation in favorable environment can reach 100%; after transplantation, plants can grow favorably, and the cost of each regeneration plant is low. The methodis suitable for the rapid propagation of good and scarce species of kaffir lilies.
Description
Technical field
The invention belongs to method for propagation, being specifically related to is the asexual rapid propagation method of flowers.
Background technology
Plant Tissue Breeding is a plant aseptic culture technology, it is totipotency according to plant cell, the organ (as root, stem, leaf, stem apex, flower, fruit etc.), tissue (as cambium, epidermis, cortex, endosperm etc.) or cell (as megaspore, microspore, somatic cell etc.) and the protoplast that utilize plant corpus to exsomatize, under artificial conditions such as aseptic and suitable synthetic medium and illumination, temperature, can induce callus, indefinite bud, adventive root, form the technology of the other products of complete plant or generation economically valuable.The merit that except extremely low variation, can both keep parent (explant source plant) by the regeneration plant that Plant Tissue Breeding produced substantially.Obtained very big progress since the plant tissue culture technique invention, Plant Tissue Breeding becomes a kind of extensive industrial production method in batch from the laboratory research stage since the sixties in last century.Kaffir lily is that the perennial herb of Amaryllidaceae gentleman Cymbidium is viewed and admired plant.Originate in the South Africa high mountain, property happiness Wen Liang introduces China the beginning of this century via Germany and Japan, and settles down.Be China's rare flower, economic worth is higher.Its plant type dignity, leaf, flower, fruit and beautiful are subjected to domestic and international flowers fan's favor deeply.At present, the modes of reproduction of kaffir lily is mainly seminal propagation and division propagation, and the reproduction speed of this dual mode is all very slow: kaffir lily only opened once and spends in 1 year, and 9 months ability of seed demand maturation; Division propagation was told the 1-3 strain in 1 year at most.Because kaffir lily height heterozygosis, the yield that seminal propagation obtains the treasure kaffir lily is very low, and originally to obtain the probability of treasure still very low even if be father and mother with the treasure kaffir lily, and supply falls short of demand always in the international market at home for the treasure kaffir lily.The breeding kaffir lily can produce the unified kaffir lily of plant type on a large scale to utilize plant tissue culture technique to exsomatize fast, breeds the good rare kind of kaffir lily fast, further satisfies the demand of domestic and international market.Though the kaffir lily tissue culture that forefathers did experiment is tentatively succeedd, break up can only differentiate on the average every explant of best kind and be less than 2.5 young plants, reproduction coefficient is extremely low.This has just limited the application of kaffir lily tissue culture technique in actual production.And but the present invention fruit of Rangoon creeper orchid bred with interior realization fast in 1 year.Explant is renewable to go out the regeneration plant of strain more than 100, is fit to very much batch production production.
Summary of the invention
Produce the individual shortcoming that plant type varies, reproduction speed is slow of kaffir lily for overcoming kaffir lily tradition propagation method; Also in order to overcome the little shortcoming of gain factor in the experiment of forefathers kaffir lily tissue culture.Fruit of Rangoon creeper orchid of the present invention is a large amount of generation regeneration plants that genetic background is identical, plant type is unified in the short relatively time.Ovary rip cutting piece with kafir lily children inflorescence (blue or green bud) is an explant, and dedifferentiation frequency can reach more than 80%, and differentiation rate can reach 70%.Can differentiate the bud of growing thickly of strain more than 100 by the callus of an explant induction through subculture repeatedly.The bud rooting rate of growing thickly can reach 100%, and transplanting survival rate reaches 100% under control environment.
The technical solution adopted in the present invention is:
1. explant is inoculated and callus induction: get kafir lily children inflorescence, put into aseptic super-clean bench, with the alcohol-pickled 30-60 of 75% (volume/volume) second, soaked aseptic water washing 3-6 time 3-10 minute with the mercuric chloride solution of 0.1-0.2% (mass/volume).Young inflorescence ovary is downcut,, be seeded in and contain 6-BA1.0mg/L, NAA1.0mg/L, 2, on the MS medium of 4-D1.0mg/L, sucrose 3.0-3.5% (mass/volume), place cultivation 20 ℃ of-28 ℃ of scattered lights under from middle part rip cutting two cuttves.30-50 days subcultures once just can be induced differentiation when waiting to grow the finer and close callus of yellow.
2. inducing of indefinite bud: the callus of yellow is transferred to inductive differentiation medium: MS+6-BA (0.5-1.0mg/L)+NAA (0.5-4mg/L)+3.0-3.5% sucrose, 30-50 days subcultures once, 20 ℃-28 ℃ 40W fluorescent lamp illumination every day 8-10 hour.This callus differentiation capability is sustainable more than 1 year, can differentiate the regrowth of strain more than 100 from a callus through subculture repeatedly.
3. inducing of root: regeneration plant forwards on the root media of 1/2MS+NAA (0.2-0.5mg/L)+sucrose 2%, 20 ℃ of-28 ℃ of whole day illumination, and 60 days left and right sides subcultures are once.
4. regeneration plant is transplanted: can transplant after plant takes root.121 ℃ of sterilizations of humus soil 30-60 minute.Before transplanting the foreign material such as agar on the plant are rinsed well.Watered once in 7-15 days and permeablely just can survive.
The invention has the beneficial effects as follows: fruit of Rangoon creeper orchid large-scale breeding in the short relatively time goes out the plant that genetic background is identical, outward appearance is neat, and growth conditions is good behind the plantlet of transplant, and the cost of average individual plant regrowth is low; Be suitable for the quick breeding of the good rare kind of kaffir lily.
Embodiment
The present invention is further illustrated below in conjunction with practical operation, but content of the present invention is not limited only to following 3 embodiment.
Embodiment 1:
1. explant inoculation and callus induction: get kafir lily children inflorescence, put into aseptic super-clean bench, with 75% (volume/volume) alcohol-pickled 30 seconds, with the mercuric chloride solution immersion of 0.1% (mass/volume) 3 minutes, aseptic water washing 3 times.Young inflorescence ovary is downcut,, be seeded in and contain 6-BA1.0mg/L, NAA1.0mg/L, 2 from middle part rip cutting two cuttves, 4-D1.0mg/L, on the MS medium of sucrose 3.5% (mass/volume), 20 ℃ of scattered lights are cultivated down, and 30 days subcultures once just can be induced differentiation when waiting to grow the finer and close callus of yellow.
2. inducing of indefinite bud: the callus of yellow is transferred to inductive differentiation medium: MS+6-BA0.5mg/L+NAA0.5mg/L+3.5% sucrose, 30 days subcultures once, 20 ℃ of illumination every day 8 hours.This callus differentiation capability is sustainable more than 1 year, can differentiate the regrowth of strain more than 100 from a callus through subculture repeatedly.
3. inducing of root: regeneration plant forwards on the root media of 1/2MS+NAA0.2mg/L+ sucrose 2%, 20 ℃ of whole day illumination, and 60 days left and right sides subcultures are once.
4. regeneration plant is transplanted: can transplant after plant takes root.121 ℃ of sterilizations of humus soil 30 minutes.Before transplanting the foreign material such as agar on the plant are rinsed well.Watered once in 7 days and permeablely just can survive.
Embodiment 2:
1. explant inoculation and callus induction: get kafir lily children inflorescence, put into aseptic super-clean bench, with 75% (volume/volume) alcohol-pickled 60 seconds, with the mercuric chloride solution immersion of 0.2% (mass/volume) 10 minutes, aseptic water washing 6 times.Young inflorescence ovary is downcut,, be seeded in and contain 6-BA1.0mg/L, NAA1.0mg/L, 2 from middle part rip cutting two cuttves, 4-D1.0mg/L, on the MS medium of sucrose 3% (mass/volume), 28 ℃ of scattered lights are cultivated down, and 50 days subcultures once just can be induced differentiation when waiting to grow the finer and close callus of yellow.
2. inducing of indefinite bud: the callus of yellow is transferred to inductive differentiation medium: MS+6-BA 1mg/L+NAA 4mg/L+3% sucrose, 50 days subcultures once, 28 ℃ of illumination every day 10 hours.This callus differentiation capability is sustainable more than 1 year, can differentiate the regrowth of strain more than 100 from a callus through subculture repeatedly.
3. inducing of root: regeneration plant forwards on the root media of 1/2MS+NAA0.5mg/L+ sucrose 2%, 28 ℃ of whole day illumination, and 60 days left and right sides subcultures are once.
4. regeneration plant is transplanted: can transplant after plant takes root.121 ℃ of sterilizations of humus soil 60 minutes.Before transplanting the foreign material such as agar on the plant are rinsed well.Watered once in 15 days and permeablely just can survive.
Embodiment 3:
1. explant inoculation and callus induction: get kafir lily children inflorescence, put into aseptic super-clean bench, with 75% (volume/volume) alcohol-pickled 40 seconds, with the mercuric chloride solution immersion of 0.15% (mass/volume) 7 minutes, aseptic water washing 4 times.Young inflorescence ovary is downcut,, be seeded in and contain 6-BA1.0mg/L, NAA1.0mg/L, 2 from middle part rip cutting two cuttves, 4-D1.0mg/L, on the MS medium of sucrose 3.2% (mass/volume), 25 ℃ of scattered lights are cultivated down, and 40 days subcultures once just can be induced differentiation when waiting to grow the finer and close callus of yellow.
2. inducing of indefinite bud: the callus of yellow is transferred to inductive differentiation medium: MS+6-BA 0.75mg/L+NAA2mg/L+3.2% sucrose, 40 days subcultures once, 25 ℃ of illumination every day 9 hours.This callus differentiation capability is sustainable more than 1 year, can differentiate the regrowth of strain more than 100 from a callus through subculture repeatedly.
3. inducing of root: regeneration plant forwards on the root media of 1/2MS+NAA0.3mg/L+ sucrose 2%, 25 ℃ of whole day illumination, and 60 days left and right sides subcultures are once.
4. regeneration plant is transplanted: can transplant after plant takes root.121 ℃ of sterilizations of humus soil 40 minutes.Before transplanting the foreign material such as agar on the plant are rinsed well.Watered once in 10 days and permeablely just can survive.
Claims (1)
1, be the method for in-vitro rapid propagation of clivia miniata of explant with the seedling ovaries, it is characterized in that concrete steps are as follows:
1). explant inoculation and callus induction: get kafir lily children inflorescence, put into aseptic super-clean bench, with 75% alcohol-pickled 30-60 second of volume, volume ratio, soaked aseptic water washing 3-6 time 3-10 minute with the mercuric chloride solution of quality, volume ratio 0.1-0.2%; Young inflorescence ovary is downcut,, be seeded in and contain 6-BA1.0mg/L, NAA1.0mg/L, 2, on the MS medium of 4-D1.0mg/L, quality, volume ratio 3.0-3.5% sucrose, place cultivation 20 ℃ of-28 ℃ of scattered lights under from middle part rip cutting two cuttves; 30-50 days subcultures are once induced differentiation when waiting to grow the finer and close callus of yellow;
2). inducing of indefinite bud: the callus of yellow is transferred to inductive differentiation medium: MS+6-BA (0.5-1.0mg/L)+NAA (0.5-4mg/L)+3.0-3.5% sucrose, 30-50 days subcultures once, 20 ℃-28 ℃ 40W fluorescent lamp illumination every day 8-10 hour, this callus differentiation capability continued more than 1 year, differentiated the regrowth of strain more than 100 from a callus through subculture repeatedly;
3). inducing of root: regeneration plant forwards on the root media of 1/2MS+NAA (0.2-0.5mg/L)+sucrose 2%, 20 ℃ of-28 ℃ of whole day illumination, 60 days left and right sides subcultures are once;
4). regeneration plant is transplanted: promptly transplant after plant takes root, 121 ℃ of sterilizations of humus soil 30-60 minute before transplanting are rinsed agar on the plant and foreign material well, water once in 7-15 days permeablely promptly to survive.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910067435A CN101637127A (en) | 2009-08-21 | 2009-08-21 | Vitro rapid propagation method of kefir lily by using seedling ovaries as explant |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910067435A CN101637127A (en) | 2009-08-21 | 2009-08-21 | Vitro rapid propagation method of kefir lily by using seedling ovaries as explant |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101637127A true CN101637127A (en) | 2010-02-03 |
Family
ID=41612451
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200910067435A Pending CN101637127A (en) | 2009-08-21 | 2009-08-21 | Vitro rapid propagation method of kefir lily by using seedling ovaries as explant |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101637127A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101810141A (en) * | 2010-03-18 | 2010-08-25 | 东北师范大学 | Method for in-vitro rapid propagation of clivia miniata by using mature embryos as explants |
| CN101822218A (en) * | 2010-03-17 | 2010-09-08 | 东北师范大学 | Kaffir lily in-vitro propagation method using young plants and young blade cut blocks as explants |
-
2009
- 2009-08-21 CN CN200910067435A patent/CN101637127A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101822218A (en) * | 2010-03-17 | 2010-09-08 | 东北师范大学 | Kaffir lily in-vitro propagation method using young plants and young blade cut blocks as explants |
| CN101810141A (en) * | 2010-03-18 | 2010-08-25 | 东北师范大学 | Method for in-vitro rapid propagation of clivia miniata by using mature embryos as explants |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101647393B (en) | Fast tissue culture reproducing method of actinidia eriantha | |
| CN103583358B (en) | A kind of method of dendrobium cultured in vitro regeneration plant | |
| CN101536674B (en) | Tissue culture reproduction method of Chinese evergreen | |
| CN102577969B (en) | Breeding method of tissue culture seedling of lonicera macranthoides Yulei No.1 | |
| CN101720670B (en) | Rapid breeding method for pinellia tuber tissue culture | |
| CN103931498B (en) | The Ornithogalum caudatum's rapid propagation in vitro method into explant is spent with children | |
| CN101347097A (en) | Method of somatic embryogenesis of cassava and rapid propagation of regenerated plant | |
| CN102870680A (en) | Efficient rapid propagation technique appropriate for detoxified rabbiteye blueberries | |
| CN104885948A (en) | Method for directly regenerating plants by tea-oil tree cotyledonary nodes | |
| CN104719157A (en) | Vaccinium australe tissue culture and rapid propagation method | |
| CN108142283B (en) | Tissue culture rapid propagation method of Acer catalpa Maxim | |
| CN102823505A (en) | Method for high-efficiency cyclic regeneration of blackberry tissue culture seedling leaves | |
| CN104642141A (en) | Gerbera adventitious bud induction and plant regeneration method by using root as explant | |
| CN101637126B (en) | Clivia miniata in-vitro rapid propagation method taking young petals as explants | |
| CN104429941A (en) | In-vitro rapid propagation technique of melaleuca alternifolia | |
| CN102283130A (en) | Method for culturing and fast propagating tissue of chimaera hosta plantaginea plant | |
| CN104094848B (en) | The method of the induction of tung oil tree hypocotyledonery axis callus and highly efficient regeneration plant | |
| KR101531308B1 (en) | Method of propagating Paulownia coreana using tissue culture | |
| CN105519442A (en) | Culture method of Prunus humilis callus regeneration system | |
| CN103858773A (en) | Method for rapid propagation in vitro of ornithogalum saundersiae by taking flower stalk as explant | |
| CN101836589B (en) | Method of rapid propagation of populus | |
| CN101810141B (en) | Method for in-vitro rapid propagation of clivia miniata by using mature embryos as explants | |
| CN102754599B (en) | Method for quickly breeding cymbidium hybridium by use of root inducing protocorm | |
| CN101637127A (en) | Vitro rapid propagation method of kefir lily by using seedling ovaries as explant | |
| CN101707981A (en) | Rubber tree cotyledon embryo high-efficiency embryonic callus induction and regeneration method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Open date: 20100203 |