CN106359103A - Lithocarpus elmerrillii tissue culture seedling rooting induction method - Google Patents
Lithocarpus elmerrillii tissue culture seedling rooting induction method Download PDFInfo
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- CN106359103A CN106359103A CN201610900826.2A CN201610900826A CN106359103A CN 106359103 A CN106359103 A CN 106359103A CN 201610900826 A CN201610900826 A CN 201610900826A CN 106359103 A CN106359103 A CN 106359103A
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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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Abstract
The invention discloses a lithocarpus elmerrillii tissue culture seedling rooting induction method. The method comprises the steps that lithocarpus elmerrillii subculture buds subjected to bud strengthening culture in conventional tissue culture for 35 d to 40 d are selected and then further selected and pruned, a pre-rooting culture medium is inoculated with the pruned lithocarpus elmerrillii subculture buds, and after pre-rooting culture is conducted for 30 d to 35 d, the buds are transferred into a rooting culture medium to be cultured. According to the method, the rooting cycle of the lithocarpus elmerrillii subculture buds is shortened, the rooting rate is high, more root systems are generated, the quality is good, the transplanting survival rate of rooted tissue culture seedlings is high, lithocarpus elmerrillii tissue culture industrialization seedling raising can be achieved, high-quality seedlings are provided for construction of man-made clonal forests, and the good economic benefit, social benefit and ecological benefit are achieved.
Description
Technical field
The present invention relates to Wanning Ke's vegetative propagation technique, a kind of especially Wanning Ke's tissue cultured seedling root induction method.
Background technology
Wanning Ke (lithocarpus elmerrillii), it is that Fagaceae (fagaceae) Lithocarpus (lithocarpus) is tall
Wood, up to 25 meters, current growth has longitudinal furrow rib, no hair, crineous after doing, 2 or 3 years raw branch scattered canescence hole skins, perula semicircle
Shape or wealthy triangle are glossy glossy after doing.The thin keratin of leaf, oblong or rare ovate-elliptic, long 10-17 centimetre,
Wide 3-6 centimetre, top is tapering, base portion gradually long and narrow point, and along the downward near base portion of petiole, full edge, lateral vein every side 9-12 bar, in nearly leaf
At edge, sharp turn is disappeared upwards, and offshoot is very thin, and blade back has the wax squama layer of consolidation, carries canescence after doing；Petiole is long 2-2.5 centimetre.
Single of female flower scattered on rachises；Infructescence axle thick 6-7 millimeter, acorn-cup consor is in the top of infructescence axle, shallow bowl type, high 6-10 milli
Rice, wide 17-25 millimeter, top is very thin, gradually substantially thickens to bottom, and wooden, base portion is narrow, in very short handle, encloses
Nut base portion, squamella triangle, it is close to shell wall, the imbricate arrangement on top, very little, the usual adhesion of bottom is circlewise；Hard
The nearly spheroidal of fruit and top is sharp, or for oblate, high 20-25 millimeter, high 25-30 millimeter, no hair, Chestnut, have vertical when knowing well
Crackle, areola is located at bottom, deep about 2 millimeters, footpath 14-16 millimeter, and Guo Bi top thickness reaches 2.5 millimeters.The fruiting period 9-10 month.Produce Hainan
Luoshan (is hung) in the middle and south.It is born in height above sea level 500-800 rice evergreen broadleaf forest.
At present, Wanning Ke resource is still in wild distribution, but the demand with protection Plant Diversity, scale people
Work cultivation Wanning Ke's resource is imperative.Tissue culture technique is a kind of fast asexual propagation technology, selects Wanning 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 Wanning Ke's tissue cultured seedling, root quantity and root system are sprouted
Send out Wanning 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 Wanning Ke's tissue cultured seedling root induction method, chooses proliferation and subculture Wanning 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 Wanning 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: the simple bud after step (1) is pruned is inoculated in pre- root media, in specific photo-thermal reaction
In carry out pre- root culture；
(3) root culture: after the simple bud in step (2) 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 improvement ms culture medium+naa 1.0～2.0mg/l+
Vitamin c 6g/l++vc 15mg/l+ Lactose 25g/l+ agar 5.0g/l.
Its material content of above-described root media is: 1/2 improvement ms culture medium+iaa 1.0～2.0mg/l+
Abt2#0.5～2.0 mg/l+ Lactose 25g/l+ agar 5.0g/l.
Basic composition is of above-described improvement ms culture medium: kno31000 mg/l；nh4no3960 mg/l；
cacl2·2h2o 260 mg/l；mgso4·7h2o 350 mg/l；ca(no3)2·4h2o 600 mg/l；kh2po4360 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 80 mg/l.
Photo-thermal reaction described in above step (2) and step (3) 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 improvement ms culture medium as minimal medium, and emphasis have adjusted the phase taken root with Wanning Ke simultaneously
The trace element closing and organic principle, so that culture medium is more scientific, more targetedly, are more easy to promote Wanning Ke's subculture bud to take root,
Effect is significant.
2nd, the present invention adopts low concentration naa and ascorbic acid (vc) that subculture simple bud is carried out with pre- training of taking root before root induction
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 the pre- life taken root and root culture, adapt to Wanning Ke's tissue cultured seedling under specific light and temperature condition
Long characteristic, promotes the growth of nursery stock.
5 present invention reduces Wanning 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, Ke's tissue culture industrialization nursery of achievable Wanning, 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 Wanning 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 improvement ms culture medium+naa 1.0mg/l+ vitamin c 6g/l++vc 15mg/l+ Lactose 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 improvement ms culture medium+iaa 1.0mg/l+abt2#0.5 mg/l+ Lactose 25g/l+ agar 5.0g/l.
Basic composition is of above-described improvement ms culture medium: kno31000 mg/l；nh4no3960 mg/l；
cacl2·2h2o 260 mg/l；mgso4·7h2o 350 mg/l；ca(no3)2·4h2o 600 mg/l；kh2po4360 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 80 mg/l.
Embodiment 2:
Choose the Wanning 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 improvement ms culture medium+naa 1.5mg/l+ vitamin c 6g/l++vc 15mg/l+ Lactose 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 improvement ms culture medium+iaa 1.0mg/l+abt2#1.0 mg/l+ Lactose 25g/l+ agar 5.0g/l.
Basic composition is of above-described improvement ms culture medium: kno31000 mg/l；nh4no3960 mg/l；
cacl2·2h2o 260 mg/l；mgso4·7h2o 350 mg/l；ca(no3)2·4h2o 600 mg/l；kh2po4360 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 80 mg/l.
Embodiment 3:
Choose the Wanning 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 improvement ms culture medium+naa 2.0mg/l+ vitamin c 6g/l++vc 15mg/l+ Lactose 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 improvement ms culture medium+iaa 2.0mg/l+abt2#1.5 mg/l+ Lactose 25g/l+ agar 5.0g/l.
Basic composition is of above-described improvement ms culture medium: kno31000 mg/l；nh4no3960 mg/l；
cacl2·2h2o 260 mg/l；mgso4·7h2o 350 mg/l；ca(no3)2·4h2o 600 mg/l；kh2po4360 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 80 mg/l.
Embodiment 4:
Choose the Wanning 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 improvement ms culture medium+naa 2.0mg/l+ vitamin c 6g/l++vc 15mg/l+ Lactose 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 improvement ms culture medium+iaa 2.0mg/l+abt2#2.0 mg/l+ Lactose 25g/l+ agar 5.0g/l.
Basic composition is of above-described improvement ms culture medium: kno31000 mg/l；nh4no3960 mg/l；
cacl2·2h2o 260 mg/l；mgso4·7h2o 350 mg/l；ca(no3)2·4h2o 600 mg/l；kh2po4360 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 80 mg/l.
Claims (5)
1. a kind of Wanning Ke's tissue cultured seedling root induction method it is characterised in that: choose proliferation and subculture Wanning 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 Wanning 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: the simple bud after step (1) is pruned is inoculated in pre- root media, in specific photo-thermal reaction
In carry out pre- root culture；
(3) root culture: after the simple bud in step (2) 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 Wanning 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 improvement ms culture medium+naa 1.0～2.0mg/l+ vitamin c 6g/l++vc 15mg/l+ breast
Sugared 25g/l+ agar 5.0g/l.
3. a kind of Wanning 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 improvement ms culture medium+iaa 1.0～2.0mg/l+abt2#0.5～2.0 mg/l+ Lactose 25g/
L+ agar 5.0g/l.
4. according to a kind of arbitrary described Wanning Ke's tissue cultured seedling root induction method of Claims 2 or 3 it is characterised in that: described
Basic composition is of improvement ms culture medium: kno31000 mg/l；nh4no3960 mg/l；cacl2·2h2o 260 mg/l；
mgso4·7h2o 350 mg/l；ca(no3)2·4h2o 600 mg/l；kh2po4360 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 80 mg/l.
5. a kind of Wanning Ke's tissue cultured seedling root induction method according to claim 1 it is characterised in that: step (2) and step
Suddenly the photo-thermal reaction described in (3) is: 20 ± 1 DEG C of temperature, humidity 40～45%, intensity of illumination 2000～2500lux, illumination 15～
16h/d.
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| CN201610900826.2A CN106359103A (en) | 2016-10-17 | 2016-10-17 | Lithocarpus elmerrillii tissue culture seedling rooting induction method |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106718935A (en) * | 2017-02-14 | 2017-05-31 | 唐春艳 | The method taken root in tung oil tree tissue-cultured seedling bottle |
| CN106718936A (en) * | 2017-02-14 | 2017-05-31 | 唐春艳 | A kind of yellow wingceltis tissue-cultured seedling rooting induction method |
| CN106797887A (en) * | 2017-02-14 | 2017-06-06 | 唐春艳 | The method taken root in chinquapin tissue-cultured seedling bottle |
| CN106818484A (en) * | 2017-02-14 | 2017-06-13 | 唐春艳 | A kind of method for improving castanopsis concinna rooting rate |
| CN107047297A (en) * | 2017-02-14 | 2017-08-18 | 唐春艳 | A kind of rice sweet oak tissue-cultured seedling rooting induction method |
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| 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 |
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2016
- 2016-10-17 CN CN201610900826.2A patent/CN106359103A/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 (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106718935A (en) * | 2017-02-14 | 2017-05-31 | 唐春艳 | The method taken root in tung oil tree tissue-cultured seedling bottle |
| CN106718936A (en) * | 2017-02-14 | 2017-05-31 | 唐春艳 | A kind of yellow wingceltis tissue-cultured seedling rooting induction method |
| CN106797887A (en) * | 2017-02-14 | 2017-06-06 | 唐春艳 | The method taken root in chinquapin tissue-cultured seedling bottle |
| CN106818484A (en) * | 2017-02-14 | 2017-06-13 | 唐春艳 | A kind of method for improving castanopsis concinna rooting rate |
| CN107047297A (en) * | 2017-02-14 | 2017-08-18 | 唐春艳 | A kind of rice sweet oak tissue-cultured seedling rooting induction method |
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