CN107494277A - A kind of gingko tissue culture and rapid propagation method - Google Patents
A kind of gingko tissue culture and rapid propagation method Download PDFInfo
- Publication number
- CN107494277A CN107494277A CN201710956234.7A CN201710956234A CN107494277A CN 107494277 A CN107494277 A CN 107494277A CN 201710956234 A CN201710956234 A CN 201710956234A CN 107494277 A CN107494277 A CN 107494277A
- Authority
- CN
- China
- Prior art keywords
- culture
- gingko
- root
- illumination
- 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.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
Abstract
The invention discloses a kind of gingko tissue culture and rapid propagation method, maidenhair tree is grown on national most area, and height can be 30 meters, and deciduous tree, trunk is upright, bark grey.Leaf fasciation on brachyplast, in long shoot alternate.Flower unisexuality, dioecism.Seed drupe state, obovate or ellipse, it is faint yellow, by white powder shape wax；Exosper meat, there is foul smell；There is seamed edge endotesta canescence, sclerotin, both sides；Endosperm enriches, cotyledon 2.4 ~ May of florescence, 7 ~ October of fruiting period.Ripening fruits is harvested when autumn and winter, is stacked on the ground, or is immersed in the water, meat exosper is rotted, cleans, dries.The present invention is using stem with bud as explant, by Fiber differentiation, squamous subculture, take root and the process such as acclimatization and transplantses successfully obtains gingko Regeneration in Vitro plant, establish the Vitro Quick Reproduction technical system of gingko, the fast numerous and large-scale production development of gingko breeding is promoted, realizes high yield high yield.
Description
Technical field
The present invention relates to the method for Plant Tissue Breeding in agricultural biotechnologies, specifically, is related to a kind of gingko tissue culture
Quick-breeding method.
Background technology
Gingko also known as：Mental eye, Buddhist refer to mandarin orange, manystem stonecrop leaf.Belong to deciduous tree, it is high up to 30 meters.Trunk is upright, bark grey.
Leaf fasciation on brachyplast, in long shoot alternate.Blade is fan-shaped, and tip centre 2 is shallow to be split, and base portion wedge shape, vein is parallel, fork-shaped branch；
Petiole grows 2 ~ 7 centimetres.Flower unisexuality, dioecism；Male flower is in sagging short catkin, has most stamens, the Room of flower pesticide 2, is born in
The top of short handle；Every 2 ~ 3 consor of female flower are on brachyplast branch, and often flower has a long handle, and pommel two is pitched.Each carpel of life 1, ovule are grown nonparasitically upon another plant
Yu Shang, generally only 1 Ovule Development maturation.Seed drupe state, obovate or ellipse, it is faint yellow, by white powder shape wax；
Exosper meat, there is foul smell；There is seamed edge endotesta canescence, sclerotin, both sides；Endosperm enriches, cotyledon 2.4 ~ May of florescence, fruiting period 7
~ October.Ripening fruits is harvested when autumn and winter, is stacked on the ground, or is immersed in the water, meat exosper is rotted, cleans, dries.In vain
Fruit is grown on national most area, cold in nature, sweet.With sword lung qi, Dingchuan cough, reduce just.At present, gingko on domestic market
Sales volume is big.Therefore, gingko resource is reasonably developed, good ecological benefits, economic benefit and social benefit will be produced.
At present, have no that foreign scholar carries out research report to gingko, it is domestic sprout in seed, biomass etc. has some scattered to grind
Study carefully, be even more to have no report to the research of its vegetative propagation technique.Therefore, to promote gingko to multiply and live, establish gingko vegetative propagation
System is necessary, and the fast numerous and large-scale production development of gingko breeding also has important practical significance imitates with significantly economical
Benefit, social benefit and ecological benefits.
The content of the invention
It is an object of the invention to provide one kind is gone out using stem with bud as explant, by Fiber differentiation, squamous subculture, life
The process such as root and acclimatization and transplantses successfully obtains gingko Regeneration in Vitro plant, establishes the Vitro Quick Reproduction technology of gingko
System, it is pure to reach seed seedling, and breeding is fast, and income is high.
A kind of gingko tissue culture and rapid propagation method of the present invention, it is characterised in that comprise the following steps：
（1）Fiber differentiation：The excellent strain root of the gingko raw tender rudiment bar of children then is gathered, stem section, will be outer as explant at the top of clip
Implant first with liquid detergent soak 5h, while use the pubescence of banister brush brushing surface, then with running water flushing 2h, stem be cut into 0.8cm,
Stem section with 4 axillary buds, on superclean bench, 22min is soaked with sterilizing 32s in 80% ethanol solution, then with 0.1% mercuric chloride,
Aseptic water washing 10 times, it is seeded to progress adventitious bud in inducing culture and lures, daily illumination 16 hours is placed in after inoculation, illumination is strong
Spend for 1900lx, it is culture under conditions of 29 DEG C until forming adventitious bud to be placed in cultivation temperature；
（2）Squamous subculture：By step（1）The tender shoots grown after culture, which is cut from base portion and is transferred on proliferated culture medium, to be bred
Culture, first full light culture 5 days under the conditions of 29 DEG C after inoculation, is subsequently placed in daily illumination 10 hours, intensity of illumination 3500lx,
It is placed under conditions of cultivation temperature is 29 DEG C and cultivates, every 25 days subcultures is once；
（3）Culture of rootage：By step（1）Or（2）Sprout length to 3cm it is high when, cut into simple bud and be inoculated into root media
Upper carry out root induction, is placed in daily illumination 18 hours, intensity of illumination 2900lx after inoculation, cultivation temperature is 29 DEG C of condition
Lower culture is to taking root；
（4）Acclimatization and transplantses：After the semi-open hardening in culturing room of culture bottle cap 5 days of the long rooting tube plantlet to 5cm, training
Support bottle cap and open entirely and take out test tube seedling from blake bottle after natural lighting lower refining seedling 6 days in outdoor, wash root culture off
Base, root is not injured as far as possible, plant into by peat soil:Yellow soil=2：In 1 matrix being mixed into and transplant planting is in plot, transplanting
Air humidity more than 80% is kept in 31 days.
Step（1）Described inducing culture is：MS+8.3mg/L 6-BA+4.6mg/L NAA+32g/L sucrose+7.0g/
L agar, pH 5.9.
Step（2）Described proliferated culture medium is：MS+1.4mg/L NAA+7.0mg/L 6-BA+32g/L sucrose+7.0g/
L agar, pH 5.9.
Step（4）Described root media is：1/2MS+3.1mg/L IBA+2.5mg/L ABT1+ 33g/L sucrose+
7.0g/L agar, pH 5.9.
It is an advantage of the invention that：Gingko is cold in nature, sweet.With sword lung qi, Dingchuan cough, the effect of reducing just.It is white to promote
Fruit multiplies and lives, the necessary Vitro Quick Reproduction technical system for establishing gingko.The present invention is using stem with bud as explant, warp
Cross Fiber differentiation, squamous subculture, take root and the process such as acclimatization and transplantses successfully obtains gingko Regeneration in Vitro plant, establish
The Vitro Quick Reproduction technical system of gingko, had important practical significance for the fast numerous and large-scale production development of gingko breeding,
With with obvious economic benefit, social benefit.
Embodiment
Following examples are the further explanations to the present invention, are not limitations of the present invention.
Embodiment 1
（1）Fiber differentiation：The excellent strain root of the gingko raw tender rudiment bar of children then is gathered, the stem section required for clip is as explant.
Explant is first soaked into 4h with liquid detergent, while uses the pubescence of banister brush brushing surface, then with running water flushing 6h, stem is cut into
4 1.8cm, band axillary buds stem section.On superclean bench, soaked with sterilizing 13s in 70% ethanol solution, then with 0.1% mercuric chloride
15min, aseptic water washing 10 times, is seeded in inducing culture and carries out adventitious bud inducing.It is small that daily illumination 16 is placed in after inoculation
When, intensity of illumination 2500lx, culture under conditions of cultivation temperature is 26 DEG C is placed in until forming adventitious bud, inductivity 69%,
Described inducing culture is：MS+6.5mg/L 6-BA+1.67mg/L NAA+25g/L sucrose+4.4g/L agar, pH 5.7.
（2）Squamous subculture：By step（1）The tender shoots grown after culture is cut from base portion and is transferred on proliferated culture medium and carries out
Multiplying culture, first full light culture 2 days under the conditions of 26 DEG C after inoculation, is subsequently placed in daily illumination 16 hours, intensity of illumination is
1900lx, be placed in cultivation temperature to cultivate under conditions of 26 DEG C, every 23 days subcultures once, proliferation rate 48%.Described propagation training
Foster base is：MS+0.3mg/L NAA+3.9mg/L 6-BA+29g/L sucrose+4.8g/L agar, pH 5.5.
（3）Culture of rootage：By step（1）Or（2）Sprout length to 4cm it is high when, cut into simple bud and be inoculated into training of taking root
Support and carry out root induction on base, daily illumination is placed in after inoculation 16 hours, intensity of illumination 2900lx, cultivation temperature is 26 DEG C
Under the conditions of cultivate to taking root, rooting rate 82%.Described root media is：1/2MS+3.0mg/L IBA+1.5mg/L
ABT1+ 33g/L sucrose+7.0g/L agar, pH 5.5.
（4）Acclimatization and transplantses：By the semi-open hardening in culturing room of culture bottle cap 2 days of the long rooting tube plantlet to 6cm
Afterwards, cultivate bottle cap and open entirely and take out test tube seedling from blake bottle after natural lighting lower refining seedling 3 days in outdoor, wash root off
Culture medium, root is not injured as far as possible, plant into by peat soil:Yellow soil=1：In 2 matrix being mixed into and transplant planting is in ground furrow
In, transplanting keeps air humidity more than 83%, transplanting survival rate 83% in 32 days.
Embodiment 2
（1）Fiber differentiation：The excellent strain root of the gingko raw tender rudiment bar of children then is gathered, the stem section required for clip is as explant.
Explant is first soaked into 5h with liquid detergent, while uses the pubescence of banister brush brushing surface, then with running water flushing 7h, stem is cut into
4 2.5cm, band axillary buds stem section.On superclean bench, soaked with sterilizing 24s in 70% ethanol solution, then with 0.1% mercuric chloride
18min, aseptic water washing 9 times, is seeded in inducing culture and carries out adventitious bud inducing.It is small that daily illumination 17 is placed in after inoculation
When, intensity of illumination 2900lx, culture under conditions of cultivation temperature is 27 DEG C is placed in until forming adventitious bud, inductivity 73%,
Described inducing culture is：MS+7.0mg/L 6-BA+1.4mg/L NAA+29g/L sucrose+4.9g/L agar, pH 5.9.
（2）Squamous subculture：By step（1）The tender shoots grown after culture is cut from base portion and is transferred on proliferated culture medium and carries out
Multiplying culture, first full light culture 1 day under the conditions of 28 DEG C after inoculation, is subsequently placed in daily illumination 15 hours, intensity of illumination is
2900lx, be placed in cultivation temperature to cultivate under conditions of 26 DEG C, every 23 days subcultures once, proliferation rate 39%.Described propagation training
Foster base is：MS+0.9mg/L NAA+4.8mg/L 6-BA+33g/L sucrose+4.8g/L agar, pH 5.5.
（3）Culture of rootage：By step（1）Or（2）Sprout length to 4cm it is high when, cut into simple bud and be inoculated into training of taking root
Support and carry out root induction on base, daily illumination is placed in after inoculation 17 hours, intensity of illumination 3900lx, cultivation temperature is 28 DEG C
Under the conditions of cultivate to taking root, rooting rate 74%.Described root media is：1/2MS+3.4mg/L IBA+1.3mg/L
ABT1+ 32g/L sucrose+7.0g/L agar, pH 5.5.
（4）Acclimatization and transplantses：By culture bottle cap semi-open hardening half a day in culturing room of the long rooting tube plantlet to 5cm
Afterwards, cultivate bottle cap and open entirely and take out test tube seedling from blake bottle after natural lighting lower refining seedling 1 day in outdoor, wash root off
Culture medium, root is not injured as far as possible, plant into by peat soil:Yellow soil=1：In 2 matrix being mixed into and transplant planting is in plot,
Transplanting keeps air humidity more than 83%, transplanting survival rate 78% in 32 days.
Claims (4)
1. a kind of gingko tissue culture and rapid propagation method, it is characterised in that comprise the following steps：
（1）Fiber differentiation：The excellent strain root of the gingko raw tender rudiment bar of children then is gathered, stem section, will be outer as explant at the top of clip
Implant first with liquid detergent soak 5h, while use the pubescence of banister brush brushing surface, then with running water flushing 2h, stem be cut into 0.8cm,
Stem section with 4 axillary buds, on superclean bench, 22min is soaked with sterilizing 32s in 80% ethanol solution, then with 0.1% mercuric chloride,
Aseptic water washing 10 times, it is seeded to progress adventitious bud in inducing culture and lures, daily illumination 16 hours is placed in after inoculation, illumination is strong
Spend for 1900lx, it is culture under conditions of 29 DEG C until forming adventitious bud to be placed in cultivation temperature；
（2）Squamous subculture：By step（1）The tender shoots grown after culture, which is cut from base portion and is transferred on proliferated culture medium, to be bred
Culture, first full light culture 5 days under the conditions of 29 DEG C after inoculation, is subsequently placed in daily illumination 10 hours, intensity of illumination 3500lx,
It is placed under conditions of cultivation temperature is 29 DEG C and cultivates, every 25 days subcultures is once；
（3）Culture of rootage：By step（1）Or（2）Sprout length to 3cm it is high when, cut into simple bud and be inoculated into root media
Upper carry out root induction, is placed in daily illumination 18 hours, intensity of illumination 2900lx after inoculation, cultivation temperature is 29 DEG C of condition
Lower culture is to taking root；
（4）Acclimatization and transplantses：After the semi-open hardening in culturing room of culture bottle cap 5 days of the long rooting tube plantlet to 5cm, training
Support bottle cap and open entirely and take out test tube seedling from blake bottle after natural lighting lower refining seedling 6 days in outdoor, wash root culture off
Base, root is not injured as far as possible, plant into by peat soil:Yellow soil=2：In 1 matrix being mixed into and transplant planting is in plot, transplanting
Air humidity more than 80% is kept in 31 days.
A kind of 2. gingko tissue culture and rapid propagation method according to claim 1, it is characterised in that step（1）Described Fiber differentiation
Base is：MS+8.3mg/L 6-BA+4.6mg/L NAA+32g/L sucrose+7.0g/L agar, pH 5.9.
A kind of 3. gingko tissue culture and rapid propagation method according to claim 1, it is characterised in that step（2）Described Multiplying culture
Base is：MS+1.4mg/L NAA+7.0mg/L 6-BA+32g/L sucrose+7.0g/L agar, pH 5.9.
A kind of 4. gingko tissue culture and rapid propagation method according to claim 1, it is characterised in that step（4）Described culture of rootage
Base is：1/2MS+3.1mg/L IBA+2.5mg/L ABT1+ 33g/L sucrose+7.0g/L agar, pH 5.9.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710956234.7A CN107494277A (en) | 2017-10-15 | 2017-10-15 | A kind of gingko tissue culture and rapid propagation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710956234.7A CN107494277A (en) | 2017-10-15 | 2017-10-15 | A kind of gingko tissue culture and rapid propagation method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107494277A true CN107494277A (en) | 2017-12-22 |
Family
ID=60701678
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710956234.7A Pending CN107494277A (en) | 2017-10-15 | 2017-10-15 | A kind of gingko tissue culture and rapid propagation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107494277A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108782239A (en) * | 2018-05-14 | 2018-11-13 | 山东组培农业发展有限公司 | A kind of ginkgo detoxification method for tissue culture |
| CN109362564A (en) * | 2018-11-25 | 2019-02-22 | 韦宇 | A kind of coriander tissue culture and rapid propagation method |
| CN109392718A (en) * | 2018-11-23 | 2019-03-01 | 张世燊 | A kind of cortex moutan tissue culture and rapid propagation method |
| CN110291989A (en) * | 2019-07-29 | 2019-10-01 | 南京林业大学 | A kind of method for building up in the sterile leaf source for gingkgo callus induction |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104719155A (en) * | 2015-03-05 | 2015-06-24 | 罗焕荣 | Phoebe bournei tissue culture and rapid propagation method |
-
2017
- 2017-10-15 CN CN201710956234.7A patent/CN107494277A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104719155A (en) * | 2015-03-05 | 2015-06-24 | 罗焕荣 | Phoebe bournei tissue culture and rapid propagation method |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108782239A (en) * | 2018-05-14 | 2018-11-13 | 山东组培农业发展有限公司 | A kind of ginkgo detoxification method for tissue culture |
| CN109392718A (en) * | 2018-11-23 | 2019-03-01 | 张世燊 | A kind of cortex moutan tissue culture and rapid propagation method |
| CN109362564A (en) * | 2018-11-25 | 2019-02-22 | 韦宇 | A kind of coriander tissue culture and rapid propagation method |
| CN110291989A (en) * | 2019-07-29 | 2019-10-01 | 南京林业大学 | A kind of method for building up in the sterile leaf source for gingkgo callus induction |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101647393B (en) | Fast tissue culture reproducing method of actinidia eriantha | |
| CN103931492B (en) | The tissue culture fast seedling-cultivating method of apple rootstock M9 | |
| CN104719136B (en) | Rooting culture method of catalpa bungei tissue culture seedling | |
| CN107494277A (en) | A kind of gingko tissue culture and rapid propagation method | |
| CN105815213A (en) | Establishing method for in-vitro regeneration system of Kiwi berry | |
| CN101595824B (en) | Rapid in-vitro seedling raising method by utilizing sandalwood seed embryo | |
| CN104855292B (en) | A kind of method of Cinnamomum kanahirai hay stem segment tissue culture fast breeding | |
| CN105052738A (en) | Tamarix chinensis tissue culture and rapid propagation method | |
| CN104012417A (en) | High-efficiency and rapid micropropagation method for toxicodendron vernicifluum | |
| CN107494275A (en) | A kind of smilax tissue culture and rapid propagation method | |
| CN103141387A (en) | Method for cultivating haworthia maughanii tissue | |
| CN106942053B (en) | A kind of tissue culture and rapid propagation method for Xingan lucid asparagus | |
| CN101717748B (en) | Method for generating macrogametocyte by somatic cell of inducing undaria pinnatifida parthenogenesis for juvenile sporophyte | |
| CN103283504B (en) | Method for grafting pear polyploidy test-tube plantlet outside test tube | |
| CN102613087A (en) | Method for culturing and breeding Correa carmen by using biological tissue | |
| CN102283113B (en) | Method for constructing sweet sorghum high-frequency regeneration system by using young ear as explant | |
| CN104094848A (en) | Induction of tung tree hypocotyls callus and method for efficiently regenerating plants | |
| CN101796921A (en) | Tissue culture rapid breeding technology of betula luminfera leaf bud | |
| CN104488709B (en) | A kind of method of floral leaf tulbaghia violacea bulb tissue cultures | |
| CN101081005A (en) | Exosomatic breeding method of switchgrass | |
| CN110402818A (en) | A kind of method of excellent kind of Chinese chestnut Mature Embryo Tissue culture rapid seedling cultivation | |
| CN106538385B (en) | A kind of extracorporeal culturing method of katsura tree | |
| CN104686336A (en) | Tissue culture rapid propagation method of ailanthus altissima | |
| CN108935100A (en) | A kind of method of apple rootstock T337 tissue-cultured seedling rooting induction | |
| CN108112479A (en) | A kind of stem section of papaya sprout Bud Differentiation vacantly plants leaf promoting root growth method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20171222 |