CN106613949A - Rooting induction method for tissue culture seedling of cork oak - Google Patents
Rooting induction method for tissue culture seedling of cork oak Download PDFInfo
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- CN106613949A CN106613949A CN201610903974.XA CN201610903974A CN106613949A CN 106613949 A CN106613949 A CN 106613949A CN 201610903974 A CN201610903974 A CN 201610903974A CN 106613949 A CN106613949 A CN 106613949A
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- cork oak
- rootage
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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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- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a rooting induction method for a tissue culture seedling of cork oak. The rooting induction method comprises the steps of selecting cork oak proliferation buds which are subjected to strong bud culture for 35-40 days in a conventional tissue culture process, trimming the cork oak proliferation buds, inoculating the cork oak proliferation buds into a pre-rooting culture medium, carrying out pre-rooting culture for 30-35 days, and transferring the cork oak proliferation buds into a rooting culture medium for culturing. The rooting induction method has the beneficial effects that the rooting cycle of the cork oak proliferation buds is shortened, the rooting rate is high, root systems are well developed with high quality, the transplanting survival rate of rooted tissue culture seedlings is high, tissue culture industrialized seedling of the cork oak can be realized, high-quality seedlings are provided for the construction of artificial clonal forests; and the method has relatively high economic, social and ecological benefits.
Description
Technical field
The present invention relates to cork oak vegetative propagation technique, especially a kind of cork oak plantlet in vitro root induction method.
Background technology
Cork oak（Quercus variabilis Bl）Also known as cork oak, tertia Qinggang, hemp oak, it is Fagaceae
（Fagaceae）Oak belongs to arbor, and tree crown is extensively avette, and trunk is more, taupe, deep lobe, and phellem layer is special thick.Sprig is filbert, male flower
Sequence is born in annual shoot bottom, the life of female flower list or twin and annual shoot axil.May at florescence；The fruit next year 9-10 months are ripe.In being
The important commerical tree species of state.Cork oak enjoys light, and is often born in mountain region tailo, but treelet is preferably having lateral shelter.To weather, soil
Strong adaptability.- 20 DEG C of low temperature is resistant to, in the acidity of pH4-8, there is growth in neutral and calacareous soil, also drought-resistant,
It is barren and with it is deep, fertile, it is appropriate moistening and well-drained loam and sandy loam optimum, intolerant to ponding.Product Liaoning,
It is Hebei, Shanxi, Shaanxi, Gansu, Shandong, Jiangsu, Anhui, Zhejiang, Jiangxi, Fujian, TaiWan, China, Henan, Hubei, Hunan, wide
The provinces and regions such as east, Guangxi, Sichuan, Guizhou, Yunnan.The tailo that North China is generally born in below 800 meters of height above sea level, southwest is reachable
Height above sea level 2000-3000 rice.
At present, the common planting type of cork oak is afforestation seeding.Tissue culture technique is a kind of fast asexual propagation skill
Art, selects cork oak superior families or clone to be material, and by method for tissue culture nursery stock is bred, and can both meet production
On quantitative requirement, maternal character can be kept again.But during tissue culture, usually occur that rooting rate is low, disunity of taking root, root
The problem that coefficient amount is few and root system is not sturdy.
The content of the invention
It is an object of the invention to provide a kind of rooting rate that can improve cork oak plantlet in vitro, root quantity and root system are sprouted
Send out the cork oak plantlet in vitro root induction method of regularity.
In order to realize foregoing invention purpose, technical scheme is as follows：
A kind of cork oak plantlet in vitro root induction method, chooses proliferation and subculture cork oak seedling, first carries out pre- culture of rootage, then
Culture of rootage being carried out again, being placed in control environment and is cultivated, obtain band root plantlet in vitro, main operational steps are as follows：
（1）Draw materials：The cork oak subculture bud that bud 35～40d of culture is strengthened in Jing routine tissue cultures is chosen, after sterilization bottle appearance, super
In sterilized space on net workbench, robust growth in subculture bud clump, the simple bud of 1～2cm of height are chosen, at the lower 2～3mm of section
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, in 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, simple bud is transferred 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 1.0～2.0mg/L+ of+NAA
Activated carbon 6g/L++VC 15mg/L+ sucrose 25g/L+ agar 5.0g/L.
Above-described root media its material content is：1/2 modified MS medium 1.0～2.0mg/L+ of+IAA
ABT1#0.5～2.0 mg/L+ sucrose 25g/L+ agar 5.0g/L.
Above-described modified MS medium basic composition is：KNO3950 mg/L；NH4NO3790 mg/L；
CaCl2·2H2O 260 mg/L；MgSO4·7H2O 386 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4 315 mg/
L；MnSO4·H2O 22.3 mg/L；ZnSO4·7H2O 8.6 mg/L；CuSO4·5H2O 0.025 mg/L；H3BO3 6.2 mg/
L；Na2MoO4·2H2O 0.025 mg/L；KI 0.83 mg/L；CoCl2·6H2O 0.025 mg/L；Cobastab1 1.0 mg/
L；Cobastab60.5 mg/L；The mg/L of nicotinic acid 0.5；The mg/L of glycine 2.0；The mg/L of inositol 90.
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, 15～16h/d of illumination.
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, while emphasis have adjusted the phase taken root with cork oak
The trace element of pass and organic principle so that culture medium is more scientific, more targetedly, is more easy to promote cork oak subculture bud to take root,
Effect is significant.
2nd, the present invention adopts low concentration NAA and ascorbic acid before root induction（VC）Pre- training of taking root is carried out to subculture simple bud
Support, culture of rootage is then carried out again, unify can plantlet in vitro root of hair, root of hair speed is fast, and root system is sturdy and quantity is more.
3rd, the present invention proceeds to culture of rootage in proliferation and subculture simple bud after 30～35d times pre- culture of rootage, you can reach
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, it is of the invention that the pre- life taken root and culture of rootage, adapt to cork oak plantlet in vitro is carried out under specific light and temperature condition
Long characteristic, promotes the growth of nursery stock.
5th, present invention reduces cork oak subculture bud is taken root the cycle, rooting rate is high, and root system is more, and quality is good, plantlet in vitro of taking root
Transplanting survival rate is high, is capable of achieving cork oak tissue culture industrialization nursery, and the construction for artificial clone woods provides high quality seedling, has
Preferable economic benefit, social benefit and ecological benefits.
Specific embodiment
With reference to specific embodiment, the invention will be further described.
Embodiment 1：
The cork oak subculture bud that bud 35～36d of culture is strengthened in Jing routine tissue cultures is chosen, after sterilization bottle appearance, in superclean bench
On sterilized space in, choose robust growth in subculture bud clump, the simple bud of 1～2cm of height, in the lower 2～3mm places shearing of section, clearly
Except radical leaves and petiole.Simple bud after pruning is inoculated in pre- root media, 20 ± 1 DEG C of temperature is placed in, humidity 40%,
Intensity of illumination 2000lux, carries out pre- culture of rootage under conditions of illumination 16h/d.Wherein described 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 transfers simple bud into root media after the pre- culture of rootage of 30～35d, is placed in 20 ± 1 DEG C of temperature, wet
Degree 40%, intensity of illumination 2000lux carries out culture of rootage under conditions of illumination 16h/d.Described root media its raw material contains
Measure and be：1/2 modified MS medium+IAA 1.0mg/L+ABT1#0.5 mg/L+ sucrose 25g/L+ agar 5.0g/L.
Above-described modified MS medium basic composition is：KNO3950 mg/L；NH4NO3790 mg/L；
CaCl2·2H2O 260 mg/L；MgSO4·7H2O 386 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4 315 mg/
L；MnSO4·H2O 22.3 mg/L；ZnSO4·7H2O 8.6 mg/L；CuSO4·5H2O 0.025 mg/L；H3BO3 6.2 mg/
L；Na2MoO4·2H2O 0.025 mg/L；KI 0.83 mg/L；CoCl2·6H2O 0.025 mg/L；Cobastab1 1.0 mg/
L；Cobastab60.5 mg/L；The mg/L of nicotinic acid 0.5；The mg/L of glycine 2.0；The mg/L of inositol 90.
Embodiment 2：
The cork oak subculture bud that bud 36～37d of culture is strengthened in Jing routine tissue cultures is chosen, after sterilization bottle appearance, in superclean bench
On sterilized space in, choose robust growth in subculture bud clump, the simple bud of 1～2cm of height, in the lower 2～3mm places shearing of section, clearly
Except radical leaves and petiole.Simple bud after pruning is inoculated in pre- root media, 20 ± 1 DEG C of temperature is placed in, humidity 40%,
Intensity of illumination 2500lux, carries out pre- culture of rootage under conditions of illumination 15h/d.Wherein described 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 transfers simple bud into root media after the pre- culture of rootage of 30～35d, is placed in 20 ± 1 DEG C of temperature, wet
Degree 40%, intensity of illumination 2500lux carries out culture of rootage under conditions of illumination 15h/d.Described root media its raw material contains
Measure and be：1/2 modified MS medium+IAA 1.0mg/L+ABT1#1.0 mg/L+ sucrose 25g/L+ agar 5.0g/L.
Above-described modified MS medium basic composition is：KNO3950 mg/L；NH4NO3790 mg/L；
CaCl2·2H2O 260 mg/L；MgSO4·7H2O 386 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4 315 mg/
L；MnSO4·H2O 22.3 mg/L；ZnSO4·7H2O 8.6 mg/L；CuSO4·5H2O 0.025 mg/L；H3BO3 6.2 mg/
L；Na2MoO4·2H2O 0.025 mg/L；KI 0.83 mg/L；CoCl2·6H2O 0.025 mg/L；Cobastab1 1.0 mg/
L；Cobastab60.5 mg/L；The mg/L of nicotinic acid 0.5；The mg/L of glycine 2.0；The mg/L of inositol 90.
Embodiment 3：
The cork oak subculture bud that bud 37～38d of culture is strengthened in Jing routine tissue cultures is chosen, after sterilization bottle appearance, in superclean bench
On sterilized space in, choose robust growth in subculture bud clump, the simple bud of 1～2cm of height, in the lower 2～3mm places shearing of section, clearly
Except radical leaves and petiole.Simple bud after pruning is inoculated in pre- root media, 20 ± 1 DEG C of temperature is placed in, humidity 45%,
Intensity of illumination 2000lux, carries out pre- culture of rootage under conditions of illumination 16h/d.Wherein described 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 transfers simple bud into root media after the pre- culture of rootage of 30～35d, is placed in 20 ± 1 DEG C of temperature, wet
Degree 45%, intensity of illumination 2000lux carries out culture of rootage under conditions of illumination 16h/d.Described root media its raw material contains
Measure and be：1/2 modified MS medium+IAA 2.0mg/L+ABT1#1.5 mg/L+ sucrose 25g/L+ agar 5.0g/L.
Above-described modified MS medium basic composition is：KNO3950 mg/L；NH4NO3790 mg/L；
CaCl2·2H2O 260 mg/L；MgSO4·7H2O 386 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4 315 mg/
L；MnSO4·H2O 22.3 mg/L；ZnSO4·7H2O 8.6 mg/L；CuSO4·5H2O 0.025 mg/L；H3BO3 6.2 mg/
L；Na2MoO4·2H2O 0.025 mg/L；KI 0.83 mg/L；CoCl2·6H2O 0.025 mg/L；Cobastab1 1.0 mg/
L；Cobastab60.5 mg/L；The mg/L of nicotinic acid 0.5；The mg/L of glycine 2.0；The mg/L of inositol 90.
Embodiment 4：
The cork oak subculture bud that bud 39～40d of culture is strengthened in Jing routine tissue cultures is chosen, after sterilization bottle appearance, in superclean bench
On sterilized space in, choose robust growth in subculture bud clump, the simple bud of 1～2cm of height, in the lower 2～3mm places shearing of section, clearly
Except radical leaves and petiole.Simple bud after pruning is inoculated in pre- root media, 20 ± 1 DEG C of temperature is placed in, humidity 45%,
Intensity of illumination 2500lux, carries out pre- culture of rootage under conditions of illumination 15h/d.Wherein described 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 transfers simple bud into root media after the pre- culture of rootage of 30～35d, is placed in 20 ± 1 DEG C of temperature, wet
Degree 45%, intensity of illumination 2500lux carries out culture of rootage under conditions of illumination 15h/d.Described root media its raw material contains
Measure and be：1/2 modified MS medium+IAA 2.0mg/L+ABT1#2.0 mg/L+ sucrose 25g/L+ agar 5.0g/L.
Above-described modified MS medium basic composition is：KNO3950 mg/L；NH4NO3790 mg/L；
CaCl2·2H2O 260 mg/L；MgSO4·7H2O 386 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4 315 mg/
L；MnSO4·H2O 22.3 mg/L；ZnSO4·7H2O 8.6 mg/L；CuSO4·5H2O 0.025 mg/L；H3BO3 6.2 mg/
L；Na2MoO4·2H2O 0.025 mg/L；KI 0.83 mg/L；CoCl2·6H2O 0.025 mg/L；Cobastab1 1.0 mg/
L；Cobastab60.5 mg/L；The mg/L of nicotinic acid 0.5；The mg/L of glycine 2.0；The mg/L of inositol 90.
Claims (5)
1. a kind of cork oak plantlet in vitro root induction method, it is characterised in that：Proliferation and subculture cork oak seedling is chosen, is first carried out pre-
Culture of rootage, then carries out again culture of rootage, is placed in control environment and cultivates, and obtains band root plantlet in vitro, and main operational steps are such as
Under：
（1）Draw materials：The cork oak subculture bud that bud 35～40d of culture is strengthened in Jing routine tissue cultures is chosen, after sterilization bottle appearance, super
In sterilized space on net workbench, robust growth in subculture bud clump, the simple bud of 1～2cm of height are chosen, at the lower 2～3mm of section
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, in 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, simple bud is transferred into culture of rootage
Base, carries out culture of rootage in specific photo-thermal reaction.
2. a kind of cork oak plantlet in vitro root induction method according to claim 1, it is characterised in that：It is described pre- to take root
Culture medium its material content is：1/2 modified MS medium+NAA 1.0～2.0mg/L+ activated carbon 6g/L++VC 15mg/L+ sugarcanes
Sugared 25g/L+ agar 5.0g/L.
3. a kind of cork oak plantlet in vitro root induction method according to claim 1, it is characterised in that：Described training of taking root
Foster base its material content is：1/2 modified MS medium+IAA 1.0～2.0mg/L+ABT1#0.5～2.0 mg/L+ sucrose 25g/
L+ agar 5.0g/L.
4. according to the arbitrary a kind of described cork oak plantlet in vitro root induction method of Claims 2 or 3, it is characterised in that：It is described
Modified MS medium basic composition is：KNO3950 mg/L；NH4NO3790 mg/L；CaCl2·2H2O 260 mg/L；
MgSO4·7H2O 386 mg/L；Ca(NO3)2·4H2O 600 mg/L；KH2PO4315 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；
The mg/L of nicotinic acid 0.5；The mg/L of glycine 2.0；The mg/L of inositol 90.
5. a kind of cork oak 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：20 ± 1 DEG C of temperature, humidity 40～45%, 2000～2500lux of intensity of illumination, illumination 15～
16h/d。
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| CN201610903974.XA CN106613949A (en) | 2016-10-17 | 2016-10-17 | Rooting induction method for tissue culture seedling of cork oak |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115735774A (en) * | 2022-12-10 | 2023-03-07 | 河北省洪崖山国有林场(河北雄安新区白洋淀上游规模化林场) | Tissue culture method of quercus mongolica |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1491535A (en) * | 2002-10-25 | 2004-04-28 | 中国科学院沈阳应用生态研究所 | Root inductive method for microbody reproduction of Japan dahurian larch |
| CN105830923A (en) * | 2016-04-12 | 2016-08-10 | 广西壮族自治区林业科学研究院 | A rooting method for tissue-cultured shoots of cinnamomum camphora (L.) presl |
-
2016
- 2016-10-17 CN CN201610903974.XA patent/CN106613949A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1491535A (en) * | 2002-10-25 | 2004-04-28 | 中国科学院沈阳应用生态研究所 | Root inductive method for microbody reproduction of Japan dahurian larch |
| CN105830923A (en) * | 2016-04-12 | 2016-08-10 | 广西壮族自治区林业科学研究院 | A rooting method for tissue-cultured shoots of cinnamomum camphora (L.) presl |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115735774A (en) * | 2022-12-10 | 2023-03-07 | 河北省洪崖山国有林场(河北雄安新区白洋淀上游规模化林场) | Tissue culture method of quercus mongolica |
| CN115735774B (en) * | 2022-12-10 | 2024-03-26 | 河北省洪崖山国有林场(河北雄安新区白洋淀上游规模化林场) | Tissue culture method of Quercus mongolica |
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Application publication date: 20170510 |