CN106417031A - Rootage induction method of lithocarpus amygdalifolius tissue culture seedlings - Google Patents
Rootage induction method of lithocarpus amygdalifolius tissue culture seedlings Download PDFInfo
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- CN106417031A CN106417031A CN201610903970.1A CN201610903970A CN106417031A CN 106417031 A CN106417031 A CN 106417031A CN 201610903970 A CN201610903970 A CN 201610903970A CN 106417031 A CN106417031 A CN 106417031A
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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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
-
- 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
-
- 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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- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a rootage induction method of lithocarpus amygdalifolius tissue culture seedlings. The method comprises the steps of selecting lithocarpus amygdalifolius subculture buds from strong buds cultured for 35-40d in conventional tissue culture, inoculating to a pre-rootage culture medium after further selecting and trimming, and transferring to a rootage culture medium for culture after pre-rootage culture for 30-35d. The method shortens a rootage cycle of the lithocarpus amygdalifolius subculture buds; the subculture buds have a high rootage rate, many root systems and good quality; the rootage tissue culture seedlings have a high transplanting survival rate; and the method can achieve industrial seedling raising of lithocarpus amygdalifolius tissue culture, provides the high quality seedlings for an artificial clonal forest and has better economic benefits, social benefits and ecologic benefits.
Description
Technical field
The present invention relates to apricot leaf Ke's vegetative propagation technique, especially a kind of apricot leaf Ke's plantlet in vitro root induction method.
Background technology
Apricot leaf Ke（Lithocarpus amygdalifolius (Skan) Hayata）Have another name called apricot leaf lithocarpus glaber, red, ridge
Plum, is Fagaceae（Fagaceae）Lithocarpus（Lithocarpus）Arbor, the shoot that spring and summer extracts out and the tender leaf back side are close yellowish-brown
Look rolls up pubescence, and hair all comes off after autumn.The life of tassel locusta list fringe armpit or many fringes line up panicle, and rachis is close by pubescence；
Every 3 clusters of female flower, sometimes have concurrently single scattered.The nearly spheroidal of acorn-cup, full bag nut, in discontinuous ring-type；The fruit of nut
Wall is slightly thicker than acorn-cup wall, and nut top is by micro-pubescence.It is distributed in Central Taiwan and south, south Fujian, the southeast, Guangdong about northern
Tropic areas to the south, Hainan.Be born in 300 meters of evergreen broadleaf forests of height above sea level 1 100-2 or needle, in Broad-leaved Mixed Forest, Chang Weishang
Layer seeds.
At present, apricot leaf Ke resource is still in wild distribution, but with keeping the demand of Plant Diversity, scale people
Work cultivation apricot leaf Ke's resource is imperative.Tissue culture technique is a kind of fast asexual propagation technology, selects apricot leaf 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 group training, 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 apricot leaf Ke's plantlet in vitro, root quantity and root system are sprouted
Send out apricot leaf Ke's plantlet in vitro root induction method of regularity.
In order to realize foregoing invention purpose, technical scheme is as follows：
A kind of apricot leaf Ke's plantlet in vitro root induction method, chooses proliferation and subculture apricot leaf Ke's seedling, first carries out pre-culture of rootage, then
Carry out culture of rootage again, be placed in control environment cultivation, it is thus achieved that band root plantlet in vitro, main operational steps is as follows：
（1）Draw materials：Choose the apricot leaf Ke's subculture bud cultivating 35～40d through strong bud in the training of routine group, after sterilization bottle appearance, surpassing
In sterilized space on net workbench, choose the simple bud of robust growth, highly 1～2cm in subculture bud clump, at the lower 2～3mm of joint
Shearing, removes radical leaves and petiole, standby；
（2）Pre-culture of rootage：By step（1）Simple bud after pruning is inoculated in pre-root media, at specific photo-thermal reaction
In carry out pre-culture of rootage；
（3）Culture of rootage：Treat step（2）In simple bud after the pre-culture of rootage of 30～35d, transfer simple bud into culture of rootage
Base, carries out culture of rootage 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+
Activated carbon 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+IBA 1.0～2.0mg/L+
ABT1#0.5～2.0 mg/L+ sucrose 25g/L+ agar 5.0g/L.
Basic composition is of above-described modified MS medium：KNO3920 mg/L；NH4NO3880 mg/L；
CaCl2·2H2O 260 mg/L；MgSO4·7H2O 410 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4320 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；Cobastab11.0 mg/
L；Cobastab60.5 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：Temperature 20 ± 1 DEG C, 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 uses 1/2 modified MS medium as minimal medium, and emphasis have adjusted the phase taken root with apricot leaf Ke simultaneously
The trace element closing and organic principle so that culture medium is more scientific, more targetedly, is more easy to promote apricot leaf Ke's subculture blastogenesis root,
Effect is notable.
2nd, the present invention uses low concentration NAA and ascorbic acid before root induction（VC）Carry out pre-training of taking root to subculture simple bud
Supporting, then carrying out culture of rootage again, plantlet in vitro root of hair can be made unified, root of hair speed is fast, and root system is sturdy and quantity is more.
3rd, the present invention proceeds to culture of rootage proliferation and subculture simple bud after 30～35d time pre-culture of rootage, i.e. up to
To pre-culture of rootage effect, turn avoid seedling and grow root system in the pre-culture of rootage stage and make later stage culture of rootage root of hair not whole
Together.
4th, the present invention under specific light and temperature condition, carry out pre-taking root and culture of rootage, adapt to the life of apricot leaf Ke's plantlet in vitro
Long characteristic, promotes the growth of nursery stock.
5th, present invention reduces apricot leaf Ke's subculture blastogenesis root cycle, rooting rate is high, and root system is many, and quality is good, plantlet in vitro of taking root
Transplanting survival rate is high, can realize that apricot leaf Ke organizes training industrialization nursery, and the construction for artificial clone woods provides high quality seedling, has
Preferable economic benefit, social benefit and ecological benefits.
Detailed description of the invention
Below in conjunction with specific embodiment, the invention will be further described.
Embodiment 1：
Choose the apricot leaf Ke's subculture bud cultivating 35～36d through strong bud in the training of routine group, after sterilization bottle appearance, at superclean bench
On sterilized space in, choose the simple bud of robust growth, highly 1～2cm in subculture bud clump, shear in joint lower 2～3mm place, clearly
Except radical leaves and petiole.Simple bud after pruning is inoculated in pre-root media, is placed in temperature 20 ± 1 DEG C, humidity 40%,
Intensity of illumination 2000lux, carries out pre-culture of rootage 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+ activated carbon 6g/L++VC 15mg/L+ sucrose 25g/L+ agar 5.0g/
L.
Simple bud, after the pre-culture of rootage of 30～35d, is transferred into root media by simple bud, is placed in temperature 20 ± 1 DEG C, wet
Degree 40%, intensity of illumination 2000lux, carry out culture of rootage under conditions of illumination 16h/d.Its raw material of described root media contains
Amount is：1/2 modified MS medium+IBA 1.0mg/L+ABT1#0.5 mg/L+ sucrose 25g/L+ agar 5.0g/L.
Basic composition is of above-described modified MS medium：KNO3920 mg/L；NH4NO3880 mg/L；
CaCl2·2H2O 260 mg/L；MgSO4·7H2O 410 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4320 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；Cobastab11.0 mg/
L；Cobastab60.5 mg/L；Nicotinic acid 0.5 mg/L；Glycine 2.0 mg/L；Inositol 90 mg/L.
Embodiment 2：
Choose the apricot leaf Ke's subculture bud cultivating 36～37d through strong bud in the training of routine group, after sterilization bottle appearance, at superclean bench
On sterilized space in, choose the simple bud of robust growth, highly 1～2cm in subculture bud clump, shear in joint lower 2～3mm place, clearly
Except radical leaves and petiole.Simple bud after pruning is inoculated in pre-root media, is placed in temperature 20 ± 1 DEG C, humidity 40%,
Intensity of illumination 2500lux, carries out pre-culture of rootage 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+ activated carbon 6g/L++VC 15mg/L+ sucrose 25g/L+ agar 5.0g/
L.
Simple bud, after the pre-culture of rootage of 30～35d, is transferred into root media by simple bud, is placed in temperature 20 ± 1 DEG C, wet
Degree 40%, intensity of illumination 2500lux, carry out culture of rootage under conditions of illumination 15h/d.Its raw material of described root media contains
Amount is：1/2 modified MS medium+IBA 1.0mg/L+ABT1#1.0 mg/L+ sucrose 25g/L+ agar 5.0g/L.
Basic composition is of above-described modified MS medium：KNO3920 mg/L；NH4NO3880 mg/L；
CaCl2·2H2O 260 mg/L；MgSO4·7H2O 410 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4320 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；Cobastab11.0 mg/
L；Cobastab60.5 mg/L；Nicotinic acid 0.5 mg/L；Glycine 2.0 mg/L；Inositol 90 mg/L.
Embodiment 3：
Choose the apricot leaf Ke's subculture bud cultivating 37～38d through strong bud in the training of routine group, after sterilization bottle appearance, at superclean bench
On sterilized space in, choose the simple bud of robust growth, highly 1～2cm in subculture bud clump, shear in joint lower 2～3mm place, clearly
Except radical leaves and petiole.Simple bud after pruning is inoculated in pre-root media, is placed in temperature 20 ± 1 DEG C, humidity 45%,
Intensity of illumination 2000lux, carries out pre-culture of rootage 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+ activated carbon 6g/L++VC 15mg/L+ sucrose 25g/L+ agar 5.0g/
L.
Simple bud, after the pre-culture of rootage of 30～35d, is transferred into root media by simple bud, is placed in temperature 20 ± 1 DEG C, wet
Degree 45%, intensity of illumination 2000lux, carry out culture of rootage under conditions of illumination 16h/d.Its raw material of described root media contains
Amount is：1/2 modified MS medium+IBA 2.0mg/L+ABT1#1.5 mg/L+ sucrose 25g/L+ agar 5.0g/L.
Basic composition is of above-described modified MS medium：KNO3920 mg/L；NH4NO3880 mg/L；
CaCl2·2H2O 260 mg/L；MgSO4·7H2O 410 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4320 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；Cobastab11.0 mg/
L；Cobastab60.5 mg/L；Nicotinic acid 0.5 mg/L；Glycine 2.0 mg/L；Inositol 90 mg/L.
Embodiment 4：
Choose the apricot leaf Ke's subculture bud cultivating 39～40d through strong bud in the training of routine group, after sterilization bottle appearance, at superclean bench
On sterilized space in, choose the simple bud of robust growth, highly 1～2cm in subculture bud clump, shear in joint lower 2～3mm place, clearly
Except radical leaves and petiole.Simple bud after pruning is inoculated in pre-root media, is placed in temperature 20 ± 1 DEG C, humidity 45%,
Intensity of illumination 2500lux, carries out pre-culture of rootage 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+ activated carbon 6g/L++VC 15mg/L+ sucrose 25g/L+ agar 5.0g/
L.
Simple bud, after the pre-culture of rootage of 30～35d, is transferred into root media by simple bud, is placed in temperature 20 ± 1 DEG C, wet
Degree 45%, intensity of illumination 2500lux, carry out culture of rootage under conditions of illumination 15h/d.Its raw material of described root media contains
Amount is：1/2 modified MS medium+IBA 2.0mg/L+ABT1#2.0 mg/L+ sucrose 25g/L+ agar 5.0g/L.
Basic composition is of above-described modified MS medium：KNO3920 mg/L；NH4NO3880 mg/L；
CaCl2·2H2O 260 mg/L；MgSO4·7H2O 410 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4320 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；Cobastab11.0 mg/
L；Cobastab60.5 mg/L；Nicotinic acid 0.5 mg/L；Glycine 2.0 mg/L；Inositol 90 mg/L.
Claims (5)
1. apricot leaf Ke's plantlet in vitro root induction method, it is characterised in that：Choose proliferation and subculture apricot leaf Ke's seedling, first carry out pre-
Culture of rootage, then carries out culture of rootage again, is placed in control environment cultivation, it is thus achieved that band root plantlet in vitro, main operational steps is such as
Under：
（1）Draw materials：Choose the apricot leaf Ke's subculture bud cultivating 35～40d through strong bud in the training of routine group, after sterilization bottle appearance, surpassing
In sterilized space on net workbench, choose the simple bud of robust growth, highly 1～2cm in subculture bud clump, at the lower 2～3mm of joint
Shearing, removes radical leaves and petiole, standby；
（2）Pre-culture of rootage：By step（1）Simple bud after pruning is inoculated in pre-root media, at specific photo-thermal reaction
In carry out pre-culture of rootage；
（3）Culture of rootage：Treat step（2）In simple bud after the pre-culture of rootage of 30～35d, transfer simple bud into culture of rootage
Base, carries out culture of rootage in specific photo-thermal reaction.
2. a kind of apricot leaf Ke's plantlet in vitro 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+ activated carbon 6g/L++VC 15mg/L+ sugarcane
Sugar 25g/L+ agar 5.0g/L.
3. a kind of apricot leaf Ke's plantlet in vitro root induction method according to claim 1, it is characterised in that：Described training of taking root
Supporting its material content of base is：1/2 modified MS medium+IBA 1.0～2.0mg/L+ABT1#0.5～2.0 mg/L+ sucrose 25g/
L+ agar 5.0g/L.
4. a kind of apricot leaf Ke's plantlet in vitro root induction method according to Claims 2 or 3 is arbitrary, it is characterised in that：Described
Basic composition is of modified MS medium：KNO3920 mg/L；NH4NO3880 mg/L；CaCl2·2H2O 260 mg/L；
MgSO4·7H2O 410 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4320 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；Cobastab11.0 mg/L；Cobastab60.5 mg/L；
Nicotinic acid 0.5 mg/L；Glycine 2.0 mg/L；Inositol 90 mg/L.
5. a kind of apricot leaf Ke's plantlet in vitro root induction method according to claim 1, it is characterised in that：Step（2）And step
Suddenly（3）Described photo-thermal reaction is：Temperature 20 ± 1 DEG C, humidity 40～45%, intensity of illumination 2000～2500lux, illumination 15～
16h/d.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109287491A (en) * | 2018-11-29 | 2019-02-01 | 钟天路 | A kind of almond tree tissue culture and rapid propagation method |
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