CN107494283A - A kind of davidia involucrata var. vilmoriniana tissue culture and rapid propagation method - Google Patents
A kind of davidia involucrata var. vilmoriniana tissue culture and rapid propagation method Download PDFInfo
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- CN107494283A CN107494283A CN201710969750.3A CN201710969750A CN107494283A CN 107494283 A CN107494283 A CN 107494283A CN 201710969750 A CN201710969750 A CN 201710969750A CN 107494283 A CN107494283 A CN 107494283A
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- seedling
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- 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
Abstract
A kind of davidia involucrata var. vilmoriniana tissue culture and rapid propagation method, it is made up of following step：A:Explant obtains, B:Sterile stem section obtains, C:The sprouting induction of lateral bud, D:Inducing clumping bud and propagation.The present invention has following notable feature relative to the method bred using prior seed sowing：Reproduction speed is fast, and 4 first quarter moons are only needed from explant to seedling；Breeding coefficient is high, and seedling exponentially increases；It is not affected by the external environment, section can carry out seedling breeding within 1 year four；Seedling uniformity is good, and separation situation occurs less；The all detoxic seedlings of seedling, be advantageous to improve resistance, few pest and disease damage；Small investment, high financial profit, can biological control, solve source of seedling this bottleneck, the wild environment cultivation of tissue-cultured seedling is advantageous to the recovery of wild population, produces good economy, society and ecological benefits.
Description
Technical field
The present invention relates to a kind of davidia involucrata var. vilmoriniana tissue culture and rapid propagation method, belong to biological technical field.
Background technology
Davidia involucrata var. vilmoriniana（Davidia involucrata Baill. var.vilmoriniana）Deciduous tree, it is China
Distinctive branch platymiscium, share two kinds, one is dove tree, and one is davidia involucrata var. vilmoriniana, and two kinds similar, difference be blade face whether by
Hair.The Relict Plant left for the 1000 ten thousand years Pre-Cenozoic Tertiary Period, is influenceed by quaternary glacier period, most area
Davidia Involucrata Population becomes extinct in succession, now only has fragmentary distribution, quilt on China Sichuan, Guizhou, Hubei and other places《Chinese Plants Red Data Book》With
《Chinese rare endangered plants register》Include, be listed in country-level Top-rated protected wild plants.
Davidia involucrata var. vilmoriniana spray is graceful, and as pigeon spreads the wings, florescence kind, the davidia involucrata var. vilmoriniana flower completely set is roamed leisurely in green shade such as pigeon
In, thus pigeon tree of gaining the name.Dove tree likes being grown in the evergreen broad-leaved Deciduous Broad Leaved Mixed Forest of the wetting of height above sea level 1500-2200 rice
In.Soil be born in the dark and damp place of air, that happiness is neutral or subacidity humus is deep more, drying is windy, in place of periods of direct sunlight it is raw
Length is bad, not barren-resistant, not drought-resistant.Growth of seedling is slow, and happiness is dark and damp, and bearing tree tends to enjoy light.
Davidia involucrata var. vilmoriniana tree setting percentage is extremely low under natural conditions, and often completely tree fresh flower dances in the air, and autumn, but difficulty was shown in a fruit, was known as thousand
Spend saying for a fruit.Conventional to be bred using fruit, extremely difficult and seed germination rate of collecting seed often is no more than 10%, causes light
The protection of leaf dove tree does not obtain good implementation always.
The content of the invention
The invention aims to establish a set of davidia involucrata var. vilmoriniana to carry out the asexual technology quickly bred using stem with bud,
So as to propose to build a set of new seedling raising mannerses do not bred by seed merely, using group culturation rapid propagating technology expand it is numerous go out it is de-
Malicious seedling maintains the characteristic of female parent to greatest extent, while can obtain that a large amount of characters are consistent in a short time, the davidia involucrata var. vilmoriniana of detoxification
Tissue-cultured seedling, a kind of new technology branch is provided for the protection of wild davidia involucrata var. vilmoriniana resource, the recovery of group and rational exploitation and utilization
Support.
The present invention can be realized by following technology：
A kind of davidia involucrata var. vilmoriniana tissue culture and rapid propagation method, is made up of following step：
A:Explant obtains：Between month in autumn 9-10, when davidia involucrata var. vilmoriniana blade is turned yellow by green grass or young crops and not fallen leaves, clip annual shoot
Bar, need in gatherer process it is whole moisturizing transport is wrapped with wet newspaper, as explant material source；
B:Sterile stem section obtains：Davidia involucrata var. vilmoriniana stem with bud is intercepted, terminal bud stays 1.5-2cm stem sections, and 0.5cm is stayed in lateral bud upper end, under
1.5-2cm stem sections are stayed at end, soak 20min with 800 times of carbendazim, are then instilled 2 drop tweens, 2-3 drop liquid detergents, are fully stirred evenly,
After cleaning, vessel port is covered with gauze, running water down-flow water is placed in and rinses 2h；Material after flushing is on superclean bench, first
With 75% alcohol-pickled 30s, then directly explant is put into 0.1% mercuric chloride and sterilizes 8min, aseptic water washing 5 times, every time
2min, the moisture remained on material are blotted with the blotting paper after sterilized；Breakage is cut, the part of browning, top is cut away before inoculation
End and each 2mm stem sections in bottom, each stem section to be seeded retain a hibernaculum, are inoculated in transitional culture medium, excessive culture medium is
The WPM basal mediums of any plant growth regulator are not added；Cultivate 18-23 DEG C of room temperature, humidity 50-70%, intensity of illumination
Inducing culture is transferred to after 2200lx, daily light application time 12-14h, 10d；
C:The sprouting induction of lateral bud：By 30d culture, it is good to select mounted blade, sterile, the tender shoots of no variation, removes old
Stem section, individually leave the tender stem newly sprouted, be transferred to callus inducing medium culture, gently press during inoculation explant material with
Material is set fully to be contacted with culture medium；Cultivate 20-25 DEG C of room temperature, humidity 50-70%, intensity of illumination 2200lx, during daily illumination
Between 12-14h, visible white or rice white or rice white Lax callus are grown after 25d, treat that callus grows up in incision
During ring-type, inducing clumping bud medium culture is transferred to；
D:Inducing clumping bud and propagation：Callus is grown fine, it is loose, start seedling that small lateral bud occur, remove complete
The blade of expansion, is inoculated in proliferated culture medium, and inoculation depth can just be stood with seedling and is advisable, and cultivate 20-25 DEG C of room temperature,
Humidity 50-70%, intensity of illumination 2200lx, daily light application time 12h, after cultivating 40d, it is seen that the tender sprouting of lateral bud in part, will
Lateral bud, which is cut, to be continued to breed, and the preferable seedling of growing way is chosen；
E:Strengthening seedling and rooting culture：High 3-5cm is intercepted, the tufted seedling of leaf 3-5 pieces, is inoculated on strengthening seedling and rooting culture medium, culturing room
18-23 DEG C of temperature, humidity 50-70%, intensity of illumination 2200lx, daily light application time 12h, healthy and strong light leaf can be obtained after cultivating 20d
Dove tree rooted seedling.
It is using the beneficial effect of above-mentioned technical proposal：
The present invention has following notable feature relative to the method bred using prior seed sowing：Reproduction speed is fast, from
Explant only needs 4 first quarter moons to seedling；Breeding coefficient is high, and seedling exponentially increases；It is not affected by the external environment, section is equal within 1 year four
Seedling breeding can be carried out；Seedling uniformity is good, and separation situation occurs less；The all detoxic seedlings of seedling, be advantageous to improve resistance,
Few pest and disease damage；Small investment, high financial profit, can biological control, solve source of seedling this bottleneck, the wild environment of tissue-cultured seedling
Cultivation is advantageous to the recovery of wild population, produces good economy, society and ecological benefits.
Embodiment
A kind of davidia involucrata var. vilmoriniana tissue culture and rapid propagation method, is made up of following step：
A:Explant obtains：Between month in autumn 9-10, when davidia involucrata var. vilmoriniana blade is turned yellow by green grass or young crops and not fallen leaves, clip annual shoot
Bar, need in gatherer process it is whole moisturizing transport is wrapped with wet newspaper, as explant material source；
B:Sterile stem section obtains：Davidia involucrata var. vilmoriniana stem with bud is intercepted, terminal bud stays 1.5-2cm stem sections, and 0.5cm is stayed in lateral bud upper end, under
1.5-2cm stem sections are stayed at end, soak 20min with 800 times of carbendazim, are then instilled 2 drop tweens, 2-3 drop liquid detergents, are fully stirred evenly,
After cleaning, vessel port is covered with gauze, running water down-flow water is placed in and rinses 2h；Material after flushing is on superclean bench, first
With 75% alcohol-pickled 30s, then directly explant is put into 0.1% mercuric chloride and sterilizes 8min, aseptic water washing 5 times, every time
2min, the moisture remained on material are blotted with the blotting paper after sterilized；Breakage is cut, the part of browning, top is cut away before inoculation
End and each 2mm stem sections in bottom, each stem section to be seeded retain a hibernaculum, are inoculated in transitional culture medium, excessive culture medium is
The WPM basal mediums of any plant growth regulator are not added；Cultivate 18-23 DEG C of room temperature, humidity 50-70%, intensity of illumination
Inducing culture is transferred to after 2200lx, daily light application time 12-14h, 10d；
C:The sprouting induction of lateral bud：By 30d culture, it is good to select mounted blade, sterile, the tender shoots of no variation, removes old
Stem section, individually leave the tender stem newly sprouted, be transferred to callus inducing medium culture, gently press during inoculation explant material with
Material is set fully to be contacted with culture medium；Cultivate 20-25 DEG C of room temperature, humidity 50-70%, intensity of illumination 2200lx, during daily illumination
Between 12-14h, visible white or rice white or rice white Lax callus are grown after 25d, treat that callus grows up in incision
During ring-type, inducing clumping bud medium culture is transferred to；
D:Inducing clumping bud and propagation：Callus is grown fine, it is loose, start seedling that small lateral bud occur, remove complete
The blade of expansion, is inoculated in proliferated culture medium, and inoculation depth can just be stood with seedling and is advisable, and cultivate 20-25 DEG C of room temperature,
Humidity 50-70%, intensity of illumination 2200lx, daily light application time 12h, after cultivating 40d, it is seen that the tender sprouting of lateral bud in part, will
Lateral bud, which is cut, to be continued to breed, and the preferable seedling of growing way is chosen；
E:Strengthening seedling and rooting culture：High 3-5cm is intercepted, the tufted seedling of leaf 3-5 pieces, is inoculated on strengthening seedling and rooting culture medium, culturing room
18-23 DEG C of temperature, humidity 50-70%, intensity of illumination 2200lx, daily light application time 12h, healthy and strong light leaf can be obtained after cultivating 20d
Dove tree rooted seedling.
In step B：The davidia involucrata var. vilmoriniana explant to be seeded is the stem section of one terminal bud of band or lateral bud.
In step C：Gently pressed with tweezers when gently being pressed during the inoculation.
In step D：When culture is to 25d, callus is grown fine, it is loose, start seedling that small lateral bud occur, remove
The blade being fully deployed, is inoculated in proliferated culture medium, and inoculation depth can just be stood with seedling.
After cultivating 40d, davidia involucrata var. vilmoriniana generally can be grown high to 4-5cm, intercept high 3-5cm, the tufted seedling of leaf 3-5 pieces, be inoculated with
In on strengthening seedling and rooting culture medium.
Scheme institute using culture medium is：
1. transition culture (in terms of 1L culture mediums)：
WPM+1.0g activated carbon+5.2g agar
2. Fiber differentiation (in terms of 1L culture mediums)：
WPM+1.0g activated carbon+5.2g agar+20g sucrose+0.5mg/L NAA+2.0mg/L6-BA
3. adventitious buds proliferation culture：
WPM+0.5g activated carbon+5.2g agar+25g sucrose+0.5mg/L NAA+1.0mg/L6-BA
4. strengthening seedling and rooting culture
WPM+0.5g activated carbon+5.2g agar+25g sucrose+0.5mg/L NAA+0.5mg/LIBA
Efficiently quick breeding is carried out in the present invention to davidia involucrata var. vilmoriniana resource using tissue culture and rapid propagation method, to obtain within a short period of time
It is good to obtain uniformity, the big davidia involucrata var. vilmoriniana tissue-cultured seedling of stock number.Through consulting literatures, existing related scientific research personnel have carried out skewer to dove tree
The researchs of reproduction technique such as slotting, tissue cultures, air layering simultaneously obtain corresponding achievement, but to the group culturation rapid propagating technology of davidia involucrata var. vilmoriniana
There is not been reported for research, in order to improve the structure of davidia involucrata var. vilmoriniana tissue culture rapid propagation system, is proposed first in manual intervention in this research
Under the conditions of, tissue-culturing rapid propagation is carried out using davidia involucrata var. vilmoriniana stem with bud, it is slower using seminal propagation speed so as to solve davidia involucrata var. vilmoriniana,
And by Environmental Factors it is larger the problem of.The present invention with similar or identical technical elements there is not yet report, therefore have one
Fixed novelty and novelty.
Claims (5)
- A kind of 1. davidia involucrata var. vilmoriniana tissue culture and rapid propagation method, it is characterised in that：It is made up of following step：A:Explant obtains：Between month in autumn 9-10, when davidia involucrata var. vilmoriniana blade is turned yellow by green grass or young crops and not fallen leaves, clip annual shoot Bar, need in gatherer process it is whole moisturizing transport is wrapped with wet newspaper, as explant material source；B:Sterile stem section obtains：Davidia involucrata var. vilmoriniana stem with bud is intercepted, terminal bud stays 1.5-2cm stem sections, and 0.5cm is stayed in lateral bud upper end, under 1.5-2cm stem sections are stayed at end, soak 20min with 800 times of carbendazim, are then instilled 2 drop tweens, 2-3 drop liquid detergents, are fully stirred evenly, After cleaning, vessel port is covered with gauze, running water down-flow water is placed in and rinses 2h；Material after flushing is on superclean bench, first With 75% alcohol-pickled 30s, then directly explant is put into 0.1% mercuric chloride and sterilizes 8min, aseptic water washing 5 times, every time 2min, the moisture remained on material are blotted with the blotting paper after sterilized；Breakage is cut, the part of browning, top is cut away before inoculation End and each 2mm stem sections in bottom, each stem section to be seeded retain a hibernaculum, are inoculated in transitional culture medium, excessive culture medium is The WPM basal mediums of any plant growth regulator are not added；Cultivate 18-23 DEG C of room temperature, humidity 50-70%, intensity of illumination Inducing culture is transferred to after 2200lx, daily light application time 12-14h, 10d；C:The sprouting induction of lateral bud：By 30d culture, it is good to select mounted blade, sterile, the tender shoots of no variation, removes old Stem section, individually leave the tender stem newly sprouted, be transferred to callus inducing medium culture, gently press during inoculation explant material with Material is set fully to be contacted with culture medium；Cultivate 20-25 DEG C of room temperature, humidity 50-70%, intensity of illumination 2200lx, during daily illumination Between 12-14h, visible white or rice white or rice white Lax callus are grown after 25d, treat that callus grows up in incision During ring-type, inducing clumping bud medium culture is transferred to；D:Inducing clumping bud and propagation：Callus is grown fine, it is loose, start seedling that small lateral bud occur, remove complete The blade of expansion, is inoculated in proliferated culture medium, and inoculation depth can just be stood with seedling and is advisable, and cultivate 20-25 DEG C of room temperature, Humidity 50-70%, intensity of illumination 2200lx, daily light application time 12h, after cultivating 40d, it is seen that the tender sprouting of lateral bud in part, will Lateral bud, which is cut, to be continued to breed, and the preferable seedling of growing way is chosen；E:Strengthening seedling and rooting culture：High 3-5cm is intercepted, the tufted seedling of leaf 3-5 pieces, is inoculated on strengthening seedling and rooting culture medium, culturing room 18-23 DEG C of temperature, humidity 50-70%, intensity of illumination 2200lx, daily light application time 12h, healthy and strong light leaf can be obtained after cultivating 20d Dove tree rooted seedling.
- A kind of 2. davidia involucrata var. vilmoriniana tissue culture and rapid propagation method according to claims 1, it is characterised in that：In step B：The light Leaf dove tree explant to be seeded is the stem section of one terminal bud of band or lateral bud.
- A kind of 3. davidia involucrata var. vilmoriniana tissue culture and rapid propagation method according to claims 1, it is characterised in that：In step C：It is described to connect Gently pressed with tweezers when gently being pressed during kind.
- A kind of 4. davidia involucrata var. vilmoriniana tissue culture and rapid propagation method according to claims 1, it is characterised in that：In step D：Culture is extremely After 25d, callus is grown fine, it is loose, start seedling that small lateral bud occur, remove the blade being fully deployed, be inoculated in In proliferated culture medium, inoculation depth can just be stood with seedling.
- A kind of 5. davidia involucrata var. vilmoriniana tissue culture and rapid propagation method according to claims 1, it is characterised in that：In step D：Culture After 40d, davidia involucrata var. vilmoriniana generally can be grown high to 4-5cm, intercept high 3-5cm, the tufted seedling of leaf 3-5 pieces, be inoculated in strengthening seedling and rooting and train Support on base.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111406655A (en) * | 2020-05-26 | 2020-07-14 | 陕西理工大学 | Davidia involucrata seedling and Davidia involucrata tissue culture method |
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| CN102919129A (en) * | 2012-11-14 | 2013-02-13 | 四川大学 | Method for acquiring regeneration seedlings of dove trees through tissue culture by taking dove tree leaves as explants |
| CN104255489A (en) * | 2014-09-16 | 2015-01-07 | 毕节市中药研究所 | Rapid vegetative propagation technical method for psammosilene tunicoides with axillary buds and immature stems |
| CN104871970A (en) * | 2015-05-14 | 2015-09-02 | 广西大学 | Tissue culture rapid propagation method of davidia involucrata |
| CN106489731A (en) * | 2016-10-19 | 2017-03-15 | 中国长江三峡集团公司 | A kind of dove tree efficiently induces the tissue culture and rapid propagation method of Multiple Buds |
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2017
- 2017-10-18 CN CN201710969750.3A patent/CN107494283A/en active Pending
Patent Citations (4)
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|---|---|---|---|---|
| CN102919129A (en) * | 2012-11-14 | 2013-02-13 | 四川大学 | Method for acquiring regeneration seedlings of dove trees through tissue culture by taking dove tree leaves as explants |
| CN104255489A (en) * | 2014-09-16 | 2015-01-07 | 毕节市中药研究所 | Rapid vegetative propagation technical method for psammosilene tunicoides with axillary buds and immature stems |
| CN104871970A (en) * | 2015-05-14 | 2015-09-02 | 广西大学 | Tissue culture rapid propagation method of davidia involucrata |
| CN106489731A (en) * | 2016-10-19 | 2017-03-15 | 中国长江三峡集团公司 | A kind of dove tree efficiently induces the tissue culture and rapid propagation method of Multiple Buds |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111406655A (en) * | 2020-05-26 | 2020-07-14 | 陕西理工大学 | Davidia involucrata seedling and Davidia involucrata tissue culture method |
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Application publication date: 20171222 |