CN106359106A - Lithocarpus oleaefolius tissue culture seedling rooting inducing method - Google Patents

Lithocarpus oleaefolius tissue culture seedling rooting inducing method Download PDF

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Publication number
CN106359106A
CN106359106A CN201610903991.3A CN201610903991A CN106359106A CN 106359106 A CN106359106 A CN 106359106A CN 201610903991 A CN201610903991 A CN 201610903991A CN 106359106 A CN106359106 A CN 106359106A
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root
culture
bud
lithocarpus
oleaefolius
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黄文卫
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Liuzhou Tinkling Of Pieces Of Jade Leads To Science And Technology Ltd Co
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Liuzhou Tinkling Of Pieces Of Jade Leads To Science And Technology Ltd Co
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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
    • 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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  • Life Sciences & Earth Sciences (AREA)
  • Developmental Biology & Embryology (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Botany (AREA)
  • Environmental Sciences (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

The invention discloses a lithocarpus oleaefolius tissue culture seedling rooting inducing method. The lithocarpus oleaefolius tissue culture seedling rooting inducing method comprises the following steps: selecting lithocarpus oleaefolius subculture buds which are subjected to bud strengthening culture of conventional tissue culture for 35 to 40 days, further selecting a trimmed single bud, inoculating the trimmed single bud into a pre-rooting culture medium, performing pre-rooting culture for 30 to 35 days, and transferring into a rooting culture medium for culturing. According to the lithocarpus oleaefolius tissue culture seedling rooting inducing method, the rooting period of the lithocarpus oleaefolius subculture buds is shortened; the rooting rate is high; the number of root systems is large; the quality is high; the transplanting survival rate of the rooted tissue culture seedlings is high; lithocarpus oleaefolius tissue culture industrialized seedling can be realized; high-quality seedlings can be provided for the construction of an artificial clonal forest; therefore, the lithocarpus oleaefolius tissue culture seedling root inducing method has good economic benefit, social benefit and ecological benefit.

Description

A kind of olive leaf Ke's tissue cultured seedling root induction method
Technical field
The present invention relates to olive leaf Ke's vegetative propagation technique, especially a kind of olive leaf Ke's tissue cultured seedling root induction method.
Background technology
Lan Yeke (lithocarpus oleaefoliusA. camus) also known as olive leaf lithocarpus glaber, olive leaf, it is Fagaceae (fagaceae) Lithocarpus (lithocarpus) arbor, high 8～15 meters, several no hair of perula, annual shoot, petiole, blade back and inflorescence Axle by caducous brown rust or the long pubescence of brown color, the dun black of biennial branch, hole skin is very little, is difficult to examine and sees.Bark dull gray To grey black, do not ftracture, about 8 millimeters of the bark thickness that about 13 meters of the height of tree, endothelium reddish tan, ridge rib is significantly prominent.Fruit contains shallow lake Powder is 50%～60%.It is distributed in Jiangxi, Fujian, Hunan, Guizhou Si Sheng south, Guangdong, Guangxi.It is born in 500～1200 meters of height above sea level In the shaw of mountain region.
At present, olive leaf Ke resource is still in wild distribution, but with keeping the demand of Plant Diversity, scale people Work cultivation olive leaf Ke's resource is imperative.Tissue culture technique is a kind of fast asexual propagation technology, selects olive 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 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 olive leaf Ke's tissue cultured seedling, root quantity and root system are sprouted Send out olive leaf 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 olive leaf Ke's tissue cultured seedling root induction method, chooses proliferation and subculture olive leaf 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 olive leaf 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+ 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 improvement ms culture 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 improvement ms culture medium: kno3940 mg/l；nh4no3790 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；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.
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 Lan Yeke simultaneously The trace element closing and organic principle, so that culture medium is more scientific, more targetedly, are more easy to promote olive leaf 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 under specific light and temperature condition pre- take root and root culture, adapt to olive leaf Ke's tissue cultured seedling life Long characteristic, promotes the growth of nursery stock.
5 present invention reduces olive leaf 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 olive leaf 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 olive leaf 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+ activated carbon 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 improvement ms culture medium+iba 1.0mg/l+abt1#0.5 mg/l+ sucrose 25g/l+ agar 5.0g/l.
Basic composition is of above-described improvement ms culture medium: kno3940 mg/l；nh4no3790 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；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 olive leaf 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+ activated carbon 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 improvement ms culture medium+iba 1.0mg/l+abt1#1.0 mg/l+ sucrose 25g/l+ agar 5.0g/l.
Basic composition is of above-described improvement ms culture medium: kno3940 mg/l；nh4no3790 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；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 olive leaf 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+ activated carbon 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 improvement ms culture medium+iba 2.0mg/l+abt1#1.5 mg/l+ sucrose 25g/l+ agar 5.0g/l.
Basic composition is of above-described improvement ms culture medium: kno3940 mg/l；nh4no3790 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；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 olive leaf 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+ activated carbon 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 improvement ms culture medium+iba 2.0mg/l+abt1#2.0 mg/l+ sucrose 25g/l+ agar 5.0g/l.
Basic composition is of above-described improvement ms culture medium: kno3940 mg/l；nh4no3790 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；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 olive leaf Ke's tissue cultured seedling root induction method it is characterised in that: choose proliferation and subculture olive leaf 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 olive leaf 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 olive leaf 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+ activated carbon 6g/l++vc 15mg/l+ sugarcane Sugared 25g/l+ agar 5.0g/l.
3. a kind of olive leaf 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+iba 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 olive leaf 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: kno3940 mg/l；nh4no3790 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；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 olive leaf 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.
CN201610903991.3A 2016-10-17 2016-10-17 Lithocarpus oleaefolius tissue culture seedling rooting inducing method Pending CN106359106A (en)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105265059A (en) * 2015-11-04 2016-01-27 张莘蔓 Method for promoting seed germination of lithocarpus oleifolius A. camus
CN105359918A (en) * 2015-12-02 2016-03-02 罗杰 Asexual rapid propagation method for lithocarpus oleaefolius
CN105418227A (en) * 2015-12-02 2016-03-23 罗杰 Special compound fertilizer for lithocarpus oleaefolia and preparation method thereof
CN105409549A (en) * 2015-12-02 2016-03-23 黄钱英 Method for culturing large-scale Lithocarpus oleaefolius A. Camus seedings
CN105475130A (en) * 2015-11-25 2016-04-13 华南农业大学 Castanopsis hystrix high efficiency isolated culture plant regeneration method

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105265059A (en) * 2015-11-04 2016-01-27 张莘蔓 Method for promoting seed germination of lithocarpus oleifolius A. camus
CN105475130A (en) * 2015-11-25 2016-04-13 华南农业大学 Castanopsis hystrix high efficiency isolated culture plant regeneration method
CN105359918A (en) * 2015-12-02 2016-03-02 罗杰 Asexual rapid propagation method for lithocarpus oleaefolius
CN105418227A (en) * 2015-12-02 2016-03-23 罗杰 Special compound fertilizer for lithocarpus oleaefolia and preparation method thereof
CN105409549A (en) * 2015-12-02 2016-03-23 黄钱英 Method for culturing large-scale Lithocarpus oleaefolius A. Camus seedings

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Title
巩合德等: "哀牢山多花山矾幼苗在森林及模拟森林光环境条件下的生长特征", 《浙江农林大学学报》 *

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