CN106472314A - A kind of dead ears Ke's tissue cultured seedling root induction method - Google Patents
A kind of dead ears Ke's tissue cultured seedling root induction method Download PDFInfo
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- CN106472314A CN106472314A CN201610900833.2A CN201610900833A CN106472314A CN 106472314 A CN106472314 A CN 106472314A CN 201610900833 A CN201610900833 A CN 201610900833A CN 106472314 A CN106472314 A CN 106472314A
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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
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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
- A01H4/001—Culture apparatus for tissue culture
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- Engineering & Computer Science (AREA)
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a kind of dead ears Ke's tissue cultured seedling root induction method, choose the dead ears Ke's subculture bud cultivating 35～40d through bud strong in conventional tissue culture, choose further and be inoculated in pre- root media after pruning, proceed in root media again after the pre- root culture of 30～35d and cultivated.Present invention reduces dead ears Ke's subculture bud is taken root the cycle, rooting rate is high, and root system is many, quality is good, and tissue cultured seedling transplanting survival rate of taking root is high, achievable dead ears Ke's tissue culture industrialization nursery, construction for artificial clone woods provides high quality seedling, has preferable economic benefit, social benefit and ecological benefits.
Description
Technical field
The present invention relates to dead ears Ke's vegetative propagation technique, especially a kind of dead ears Ke's tissue cultured seedling root induction method.
Background technology
Dead ears Ke（Lithocarpus craibianus Barn.）, it is Fagaceae（Fagaceae）Lithocarpus
（Lithocarpus）Arbor, arbor, up to 20 meters.Annual shoot, blade back and female inflorescence axle all have brown color or canescence wax squama
Layer.Leaf keratin, avette or ovate-elliptic, long 12-19 centimetre, sometimes shorter or longer, wide 4-7 centimetre, dilute wider, top is long
Point, the short point of base portion to wide wedge shape, full edge, the hypomere of middle arteries is thick on blade face, and vein there are about 8-12 bar, often recessed in splitting channel-shaped on blade face
Fall into, offshoot does not show or mays be seen indistinctly, and after tender leaf is done, often there is glossy gloss on blade face, biennial leaf is then grey dark；Petiole is long 1-2.5 li
Rice.Tassel locusta axil is given birth to, dilute for panicle, up to 15 centimetres；The top of female inflorescence often raw minority male flower, up to 30 lis
Rice, 3-5-7 collection of female flower generates cluster, style long 1-1.2 millimeter.Acorn-cup spheroidal or slightly flat, top is in often mamillary short projection,
Footpath 15-20 millimeter, full bag nut, occasionally there are top cracking warp, therefore nut part exposes, squamella triangle, apex point
Shape, fits perfectly in shell wall, and imbricate arranges, clearly, positioned at the slightly long and narrow of acorn-cup top and to curved under acorn-cup oral area, by yellowish-brown,
Fine platy, the wax squama being slightly close to；The nearly spheroidal of nut, footpath 13-18 millimeter, by very sparse thin FUMAO, the hair at top is closeer, really
Umbilicuss are raised, account for the 1/3 of nut area.The month at florescence 8-9, fruit same period next year is ripe.Produce South-west Sichuan, south of Yunnan and the west and south
Various places.It is born in 700 meters of mountain region shaws of 1 500-2, is more common in and hillside fields is relatively dried, Chang Yin is cut down, therefore generate shrub shape,
It is usually about 10 meters of dungarunga.Laos, Northern Thailand also have.
At present, dead ears Ke resource is still in wild distribution, but with keeping the demand of Plant Diversity, scale people
Work cultivation dead ears Ke's resource is imperative.Tissue culture technique is a kind of fast asexual propagation technology, selects dead ears Ke You respectable family
System or clone are material, breed nursery stock by method for tissue culture, both can meet the quantitative requirement in production, and can protect again
Hold maternal character.But during tissue culture, usually occur that rooting rate is low, disunity of taking root, root quantity are few and root system not
Sturdy problem.
Content of the invention
It is an object of the invention to provide a kind of rooting rate that can improve dead ears Ke's tissue cultured seedling, root quantity and root system are sprouted
Send out dead ears Ke's tissue cultured seedling root induction method of regularity.
In order to realize foregoing invention purpose, technical scheme is as follows：
A kind of dead ears Ke's tissue cultured seedling root induction method, chooses proliferation and subculture dead ears Ke's seedling, first carries out pre- root culture, then
Carry out root culture again, be placed in culture in control environment, obtain band root tissue cultured seedling, main operational steps are as follows：
（1）Draw materials：Choose the dead ears Ke's subculture bud cultivating 35～40d through bud strong in conventional tissue culture, after sterilization bottle appearance, super
In sterilized space on net workbench, choose robust growth, the highly simple bud of 1～2cm in subculture bud clump, at the lower 2～3mm of section
Shearing, removes radical leaves and petiole, standby；
（2）Pre- root culture：By step（1）Simple bud after pruning is inoculated in pre- root media, in specific photo-thermal reaction
In carry out pre- root culture；
（3）Root culture：Treat step（2）In simple bud after the pre- root culture of 30～35d, simple bud is transferred into root culture
Base, carries out root culture in specific photo-thermal reaction.
Above-described its material content of pre- root media is：1/2 modified MS medium+NAA 1.0～2.0mg/L+
Vitamin C 6g/L++VC 15mg/L+ sucrose 25g/L+ agar 5.0g/L.
Its material content of above-described root media is：1/2 modified MS medium+IAA 1.0～2.0mg/L+
ABT2#0.5～2.0 mg/L+ sucrose 25g/L+ agar 5.0g/L.
Basic composition is of above-described modified MS medium：KNO3950 mg/L；NH4NO3790 mg/L；
CaCl2·2H2O 260 mg/L；MgSO4·7H2O 400 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4330 mg/
L；MnSO4·H2O 22.3 mg/L；ZnSO4·7H2O 8.6 mg/L；CuSO4·5H2O 0.025 mg/L；H3BO36.2 mg/
L；Na2MoO4·2H2O 0.025 mg/L；KI 0.83 mg/L；CoCl2·6H2O 0.025 mg/L；Vitamin B11.0 mg/
L；Vitamin B60.5 mg/L；Nicotinic acid 0.5 mg/L；Glycine 2.0 mg/L；Inositol 90 mg/L.
Above step（2）And step（3）Described photo-thermal reaction is：20 ± 1 DEG C of temperature, humidity 40～45%, intensity of illumination
2000～2500lux, illumination 15～16h/d.
The present invention has the advantage that and has the beneficial effect that：
1st, the present invention adopts 1/2 modified MS medium as minimal medium, and emphasis have adjusted the phase taken root with dead ears Ke simultaneously
The trace element closing and organic principle, so that culture medium is more scientific, more targetedly, are more easy to promote dead ears Ke's subculture bud to take root,
Effect is significant.
2nd, the present invention adopts low concentration NAA and ascorbic acid before root induction（VC）Subculture simple bud is carried out with pre- training of taking root
Support, then carry out root culture again, tissue cultured seedling root of hair can be made to unify, root of hair speed is fast, and root system is sturdy and quantity is more.
3rd, the present invention proceeds to root culture in proliferation and subculture simple bud after 30～35d time pre- root culture, you can reach
To pre- root culture effect, turn avoid seedling and grow root system and make later stage root culture root of hair not whole in the pre- root culture stage
Together.
4th, the present invention carry out under specific light and temperature condition pre- take root and root culture, adapt to dead ears Ke's tissue cultured seedling life
Long characteristic, promotes the growth of nursery stock.
5 present invention reduces dead ears Ke's subculture bud is taken root the cycle, and rooting rate is high, and root system is many, and quality is good, tissue cultured seedling of taking root
Transplanting survival rate is high, achievable dead ears Ke's tissue culture industrialization nursery, and the construction for artificial clone woods provides high quality seedling, has
Preferably economic benefit, social benefit and ecological benefits.
Specific embodiment
With reference to specific embodiment, the invention will be further described.
Embodiment 1：
Choose the dead ears Ke's subculture bud cultivating 35～36d through bud strong in conventional tissue culture, after sterilization bottle appearance, in superclean bench
On sterilized space in, choose robust growth, the highly simple bud of 1～2cm in subculture bud clump, in section lower 2～3mm place's shearing, clearly
Except radical leaves and petiole.Simple bud after pruning is inoculated in pre- root media, is placed in 20 ± 1 DEG C of temperature, humidity 40%,
Intensity of illumination 2000lux, carries out pre- root culture under conditions of illumination 16h/d.Wherein said its raw material of pre- root media
Content is：1/2 modified MS medium+NAA 1.0mg/L+ vitamin C 6g/L++VC 15mg/L+ sucrose 25g/L+ agar 5.0g/
L.
Simple bud, after the pre- root culture of 30～35d, simple bud is transferred into root media, is placed in 20 ± 1 DEG C of temperature, wet
Degree 40%, intensity of illumination 2000lux, carry out root culture under conditions of illumination 16h/d.Its raw material of described root media contains
Measure and be：1/2 modified MS medium+IAA 1.0mg/L+ABT2#0.5 mg/L+ sucrose 25g/L+ agar 5.0g/L.
Basic composition is of above-described modified MS medium：KNO3950 mg/L；NH4NO3790 mg/L；
CaCl2·2H2O 260 mg/L；MgSO4·7H2O 400 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4330 mg/
L；MnSO4·H2O 22.3 mg/L；ZnSO4·7H2O 8.6 mg/L；CuSO4·5H2O 0.025 mg/L；H3BO36.2 mg/
L；Na2MoO4·2H2O 0.025 mg/L；KI 0.83 mg/L；CoCl2·6H2O 0.025 mg/L；Vitamin B11.0 mg/
L；Vitamin B60.5 mg/L；Nicotinic acid 0.5 mg/L；Glycine 2.0 mg/L；Inositol 90 mg/L.
Embodiment 2：
Choose the dead ears Ke's subculture bud cultivating 36～37d through bud strong in conventional tissue culture, after sterilization bottle appearance, in superclean bench
On sterilized space in, choose robust growth, the highly simple bud of 1～2cm in subculture bud clump, in section lower 2～3mm place's shearing, clearly
Except radical leaves and petiole.Simple bud after pruning is inoculated in pre- root media, is placed in 20 ± 1 DEG C of temperature, humidity 40%,
Intensity of illumination 2500lux, carries out pre- root culture under conditions of illumination 15h/d.Wherein said its raw material of pre- root media
Content is：1/2 modified MS medium+NAA 1.5mg/L+ vitamin C 6g/L++VC 15mg/L+ sucrose 25g/L+ agar 5.0g/
L.
Simple bud, after the pre- root culture of 30～35d, simple bud is transferred into root media, is placed in 20 ± 1 DEG C of temperature, wet
Degree 40%, intensity of illumination 2500lux, carry out root culture under conditions of illumination 15h/d.Its raw material of described root media contains
Measure and be：1/2 modified MS medium+IAA 1.0mg/L+ABT2#1.0 mg/L+ sucrose 25g/L+ agar 5.0g/L.
Basic composition is of above-described modified MS medium：KNO3950 mg/L；NH4NO3790 mg/L；
CaCl2·2H2O 260 mg/L；MgSO4·7H2O 400 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4330 mg/
L；MnSO4·H2O 22.3 mg/L；ZnSO4·7H2O 8.6 mg/L；CuSO4·5H2O 0.025 mg/L；H3BO36.2 mg/
L；Na2MoO4·2H2O 0.025 mg/L；KI 0.83 mg/L；CoCl2·6H2O 0.025 mg/L；Vitamin B11.0 mg/
L；Vitamin B60.5 mg/L；Nicotinic acid 0.5 mg/L；Glycine 2.0 mg/L；Inositol 90 mg/L.
Embodiment 3：
Choose the dead ears Ke's subculture bud cultivating 37～38d through bud strong in conventional tissue culture, after sterilization bottle appearance, in superclean bench
On sterilized space in, choose robust growth, the highly simple bud of 1～2cm in subculture bud clump, in section lower 2～3mm place's shearing, clearly
Except radical leaves and petiole.Simple bud after pruning is inoculated in pre- root media, is placed in 20 ± 1 DEG C of temperature, humidity 45%,
Intensity of illumination 2000lux, carries out pre- root culture under conditions of illumination 16h/d.Wherein said its raw material of pre- root media
Content is：1/2 modified MS medium+NAA 2.0mg/L+ vitamin C 6g/L++VC 15mg/L+ sucrose 25g/L+ agar 5.0g/
L.
Simple bud, after the pre- root culture of 30～35d, simple bud is transferred into root media, is placed in 20 ± 1 DEG C of temperature, wet
Degree 45%, intensity of illumination 2000lux, carry out root culture under conditions of illumination 16h/d.Its raw material of described root media contains
Measure and be：1/2 modified MS medium+IAA 2.0mg/L+ABT2#1.5 mg/L+ sucrose 25g/L+ agar 5.0g/L.
Basic composition is of above-described modified MS medium：KNO3950 mg/L；NH4NO3790 mg/L；
CaCl2·2H2O 260 mg/L；MgSO4·7H2O 400 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4330 mg/
L；MnSO4·H2O 22.3 mg/L；ZnSO4·7H2O 8.6 mg/L；CuSO4·5H2O 0.025 mg/L；H3BO36.2 mg/
L；Na2MoO4·2H2O 0.025 mg/L；KI 0.83 mg/L；CoCl2·6H2O 0.025 mg/L；Vitamin B11.0 mg/
L；Vitamin B60.5 mg/L；Nicotinic acid 0.5 mg/L；Glycine 2.0 mg/L；Inositol 90 mg/L.
Embodiment 4：
Choose the dead ears Ke's subculture bud cultivating 39～40d through bud strong in conventional tissue culture, after sterilization bottle appearance, in superclean bench
On sterilized space in, choose robust growth, the highly simple bud of 1～2cm in subculture bud clump, in section lower 2～3mm place's shearing, clearly
Except radical leaves and petiole.Simple bud after pruning is inoculated in pre- root media, is placed in 20 ± 1 DEG C of temperature, humidity 45%,
Intensity of illumination 2500lux, carries out pre- root culture under conditions of illumination 15h/d.Wherein said its raw material of pre- root media
Content is：1/2 modified MS medium+NAA 2.0mg/L+ vitamin C 6g/L++VC 15mg/L+ sucrose 25g/L+ agar 5.0g/
L.
Simple bud, after the pre- root culture of 30～35d, simple bud is transferred into root media, is placed in 20 ± 1 DEG C of temperature, wet
Degree 45%, intensity of illumination 2500lux, carry out root culture under conditions of illumination 15h/d.Its raw material of described root media contains
Measure and be：1/2 modified MS medium+IAA 2.0mg/L+ABT2#2.0 mg/L+ sucrose 25g/L+ agar 5.0g/L.
Basic composition is of above-described modified MS medium：KNO3950 mg/L；NH4NO3790 mg/L；
CaCl2·2H2O 260 mg/L；MgSO4·7H2O 400 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4330 mg/
L；MnSO4·H2O 22.3 mg/L；ZnSO4·7H2O 8.6 mg/L；CuSO4·5H2O 0.025 mg/L；H3BO36.2 mg/
L；Na2MoO4·2H2O 0.025 mg/L；KI 0.83 mg/L；CoCl2·6H2O 0.025 mg/L；Vitamin B11.0 mg/
L；Vitamin B60.5 mg/L；Nicotinic acid 0.5 mg/L；Glycine 2.0 mg/L；Inositol 90 mg/L.
Claims (5)
1. a kind of dead ears Ke's tissue cultured seedling root induction method it is characterised in that：Choose proliferation and subculture dead ears Ke's seedling, first carry out pre-
Root culture, then carries out root culture again, is placed in culture in control environment, obtains band root tissue cultured seedling, main operational steps are such as
Under：
（1）Draw materials：Choose the dead ears Ke's subculture bud cultivating 35～40d through bud strong in conventional tissue culture, after sterilization bottle appearance, super
In sterilized space on net workbench, choose robust growth, the highly simple bud of 1～2cm in subculture bud clump, at the lower 2～3mm of section
Shearing, removes radical leaves and petiole, standby；
（2）Pre- root culture：By step（1）Simple bud after pruning is inoculated in pre- root media, in specific photo-thermal reaction
In carry out pre- root culture；
（3）Root culture：Treat step（2）In simple bud after the pre- root culture of 30～35d, simple bud is transferred into root culture
Base, carries out root culture in specific photo-thermal reaction.
2. a kind of dead ears Ke's tissue cultured seedling root induction method according to claim 1 it is characterised in that：Described pre- take root
Its material content of culture medium is：1/2 modified MS medium+NAA 1.0～2.0mg/L+ vitamin C 6g/L++VC 15mg/L+ sugarcane
Sugared 25g/L+ agar 5.0g/L.
3. a kind of dead ears Ke's tissue cultured seedling root induction method according to claim 1 it is characterised in that：Described training of taking root
Its material content of foster base is：1/2 modified MS medium+IAA 1.0～2.0mg/L+ABT2#0.5～2.0 mg/L+ sucrose 25g/
L+ agar 5.0g/L.
4. according to the arbitrary a kind of described dead ears Ke's tissue cultured seedling root induction method of Claims 2 or 3 it is characterised in that：Described
Basic composition is of modified MS medium：KNO3950 mg/L；NH4NO3790 mg/L；CaCl2·2H2O 260 mg/L；
MgSO4·7H2O 400 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4330 mg/L；MnSO4·H2O 22.3 mg/
L；ZnSO4·7H2O 8.6 mg/L；CuSO4·5H2O 0.025 mg/L；H3BO36.2 mg/L；Na2MoO4·2H2O 0.025
mg/L；KI 0.83 mg/L；CoCl2·6H2O 0.025 mg/L；Vitamin B11.0 mg/L；Vitamin B60.5 mg/L；
Nicotinic acid 0.5 mg/L；Glycine 2.0 mg/L；Inositol 90 mg/L.
5. a kind of dead ears Ke's tissue cultured seedling root induction method according to claim 1 it is characterised in that：Step（2）And step
Suddenly（3）Described photo-thermal reaction is：20 ± 1 DEG C of temperature, humidity 40～45%, intensity of illumination 2000～2500lux, illumination 15～
16h/d.
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Citations (4)
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|---|---|---|---|---|
| CN105409686A (en) * | 2015-12-02 | 2016-03-23 | 罗杰 | Reproduction method for grafting Quercus acutissima Carruth container seedlings on Lithocarpus craibianus Barn. |
| CN105409550A (en) * | 2015-12-02 | 2016-03-23 | 黄钱英 | Method for culturing large-scale Lithocarpus craibianus Barn. seedlings |
| CN105475130A (en) * | 2015-11-25 | 2016-04-13 | 华南农业大学 | Castanopsis hystrix high efficiency isolated culture plant regeneration method |
| CN105474999A (en) * | 2015-12-02 | 2016-04-13 | 罗杰 | Asexual rapid reproduction method of lithocarpus craibianus Barn. |
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2016
- 2016-10-17 CN CN201610900833.2A patent/CN106472314A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105475130A (en) * | 2015-11-25 | 2016-04-13 | 华南农业大学 | Castanopsis hystrix high efficiency isolated culture plant regeneration method |
| CN105409686A (en) * | 2015-12-02 | 2016-03-23 | 罗杰 | Reproduction method for grafting Quercus acutissima Carruth container seedlings on Lithocarpus craibianus Barn. |
| CN105409550A (en) * | 2015-12-02 | 2016-03-23 | 黄钱英 | Method for culturing large-scale Lithocarpus craibianus Barn. seedlings |
| CN105474999A (en) * | 2015-12-02 | 2016-04-13 | 罗杰 | Asexual rapid reproduction method of lithocarpus craibianus Barn. |
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Application publication date: 20170308 |