CN107223565A - Cubeb litsen tree tissue-cultured seedling root media and cultural method - Google Patents
Cubeb litsen tree tissue-cultured seedling root media and cultural method Download PDFInfo
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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
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- A01H4/001—Culture apparatus for tissue culture
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Abstract
The invention provides cubeb litsen tree tissue-cultured seedling root media and cultural method, it is related to cubeb litsen tree tissue-cultured seedling culture of rootage technical field.The invention provides cubeb litsen tree tissue-cultured seedling root media, the root media is that the rooting induction materials such as indolebutyric acid, heteroauxin, methyl α-naphthyl acetate are introduced on the basis of minimal medium, and the coordinated effect using each material and the specific restriction to each material consumption, so that it has significant root induction effect to cubeb litsen tree tissue-cultured seedling, rooting rate reaches more than 95%, transplanting survival rate reaches 100%, improves and lifts the not good technical problem of effect for the rooting rate and transplanting survival rate of tissue-cultured seedling using traditional cubeb litsen tree tissue-cultured seedling root media.Present invention also offers cubeb litsen tree tissue-cultured seedling culture of rootage method, by using above-mentioned cubeb litsen tree tissue-cultured seedling root media and adjustment condition of culture, so that the rooting rate of cubeb litsen tree tissue-cultured seedling is further improved, reliable technical basis is provided for factorial praluction.
Description
Technical field
The present invention relates to cubeb litsen tree tissue-cultured seedling root culture technique field, trained in particular to cubeb litsen tree tissue culture seedling rooting
Support base and cultural method.
Background technology
Cubeb litsen tree (Litsea cubeba (Lour.) Pers.), also known as the fruit of a cubeb litsea tree, Tetranthera citrata etc., belong to Lauraceae
(Lauracea) Litsea (Litsea Lam) machaka or dungarunga, are that the precious natural perfume material for originating in China is planted
Thing.The tissue such as its fruit, leaf, flower can steam and carry essential oil, and lemon aldehyde is up to 60-90% in essential oil, be manufacture superior cosmetics,
The important source material of medicine, food additives, disease and pest control agent, lubricating oil etc..The cubeb litsen tree essential oil market demand is huge, but
It is in the processing of cubeb litsen tree essential oil to utilize wild cubeb litsen tree resource more, and chopped at a tree more with destructive as main Softening, cause
Cubeb litsen tree resource is seriously exhausted, and nurseries' supplies put upon the full stretch.Traditional modes of reproduction is seminal propagation, but cubeb litsen tree germination
Rate is extremely low, and advantageous property can not stablize heredity, have a strong impact on cubeb litsen tree essential oil quality and yield.Can using method for tissue culture
Greatly shorten growing-seedling period, not only breeding potential is high and can keep stock merit, be to solve cubeb litsen tree nursery stock raw material to supply
Answer nervous splendid means.
At present, using common culture medium to the rooting rate of cubeb litsen tree tissue-cultured seedling culture of rootage up to 80% or so, but group
Training rooting rate can directly affect tissue culture production cost, and be related to the height of transplanting survival rate, it is therefore necessary to cubeb litsen tree
Tissue-cultured seedling root media and cultural method make further optimization.
In view of this, it is special to propose the present invention to solve above-mentioned technical problem.
The content of the invention
First purpose of the present invention is to provide a kind of cubeb litsen tree tissue-cultured seedling root media, using the root media
The rooting rate of cubeb litsen tree tissue-cultured seedling can be effectively improved, to improve using existing cubeb litsen tree tissue-cultured seedling root media to cubeb litsen tree group
Train the not good technical problem of the rooting rate lifting effect of seedling.
Second object of the present invention is to provide a kind of cubeb litsen tree tissue-cultured seedling culture of rootage method.
In order to realize the above-mentioned purpose of the present invention, spy uses following technical scheme：
The invention provides a kind of cubeb litsen tree tissue-cultured seedling root media, mainly it is made up of the component of following concentration：1/2MS
Minimal medium, indolebutyric acid (Indolebutyric acid, IBA) 0.7-0.9mg/L, heteroauxin (Indoleacetic
Acid, IAA) 0.2-0.3mg/L, methyl α-naphthyl acetate (Naphthylacetic acid, NAA) 0.1-0.2mg/L, activated carbon
(Activated carbon, AC) 0.5-1.0mg/L, sucrose 19.0-21.0g/L and agar 7.0-8.0g/L, pH are 5.8-
6.0。
Further, the cubeb litsen tree tissue-cultured seedling root media is mainly made up of the component of following concentration：1/2MS is basic
Culture medium, IBA 0.75-0.85mg/L, IAA 0.2-0.3mg/L, NAA 0.1-0.15mg/L, AC 0.75-1.0mg/L, sugarcane
Sugared 19.5-20.5g/L and agar 7.2-7.8g/L, pH are 5.8-6.0.
Further, the cubeb litsen tree tissue-cultured seedling root media is mainly made up of the component of following concentration：1/2MS is basic
Culture medium, IBA 0.8mg/L, IAA 0.2-0.3mg/L, NAA 0.1mg/L, AC 1.0mg/L, sucrose 20.0g/L and agar
7.5g/L, pH are 5.8.
Further, the 1/2MS minimal mediums include a great number of elements, trace element, molysite and organic matter, mainly by
The component composition of following concentration：
A great number of elements：Potassium nitrate 950mg/L, ammonium nitrate 825mg/L, epsom salt 185mg/L, potassium dihydrogen phosphate
85mg/L, calcium chloride dihydrate 220mg/L；
Trace element：Four water manganese sulfate 22.3mg/L, white vitriol 8.6mg/L, boric acid 6.2mg/L, KI
0.83mg/L, Sodium Molybdate Dihydrate 0.25mg/L, cupric sulfate pentahydrate 0.025mg/L, CoCL2 6H2O 0.025mg/L；
Molysite：Ferrous sulfate heptahydrate 27.8mg/L, disodium ethylene diamine tetraacetate 37.3mg/L；
Organic matter：Glycine 2mg/L, puridoxine hydrochloride 0.5mg/L, thiamine hydrochloride 0.1mg/L, nicotinic acid 0.5mg/L, flesh
Alcohol 100mg/L.
Further, culture of rootage is carried out to cubeb litsen tree tissue-cultured seedling using the cubeb litsen tree tissue-cultured seedling root media, it is raw
Root rate is more than or equal to 95%.
Present invention also offers a kind of culture of rootage method of cubeb litsen tree tissue-cultured seedling, given birth to using above-mentioned cubeb litsen tree tissue-cultured seedling
Root culture medium carries out culture of rootage to cubeb litsen tree tissue-cultured seedling.
Further, the condition of the culture of rootage is：Cultivation temperature is 23-27 DEG C, light application time 15-16h, when dark
Between be 8-9h, intensity of illumination is 1800-2200Lux, and incubation time is at least 25 days.
Further, the condition of the culture of rootage is：Cultivation temperature is 23-27 DEG C, light application time 16h, interlunation
For 8h, intensity of illumination is 2000Lux, and incubation time is 25 days.
Further, choose lignifying, be about 2-3cm cubeb litsen tree tissue-cultured seedling progress culture of rootage.
Further, culture of rootage is carried out to cubeb litsen tree tissue-cultured seedling using the cubeb litsen tree tissue-cultured seedling root media, it is raw
Root rate is more than or equal to 95%.
Compared with the prior art, the present invention has the advantages that：
(1) the invention provides a kind of cubeb litsen tree tissue-cultured seedling root media, the root media is in minimal medium
On the basis of introduce indolebutyric acid, heteroauxin, the rooting induction material such as methyl α-naphthyl acetate, and acted on using the coordinated of each material
And passing through the restriction to each material consumption so that the root media has stronger root induction to imitate for cubeb litsen tree tissue-cultured seedling
Should, so that the rooting rate of cubeb litsen tree tissue-cultured seedling reaches more than 95%, transplanting survival rate reaches 100%, effectively improves and adopts
With traditional cubeb litsen tree tissue-cultured seedling root media the not good skill of effect is lifted for the rooting rate and transplanting survival rate of tissue-cultured seedling
Art problem.
(2) the invention provides a kind of culture of rootage method of cubeb litsen tree tissue-cultured seedling, by using above-mentioned cubeb litsen tree tissue culture
Seedling rooting culture medium and adjustment condition of culture so that the rooting rate of cubeb litsen tree tissue-cultured seedling is further improved, and is cubeb litsen tree
Factorial praluction provide more reliable, the stronger technical basis of operability, accelerate its breeding with promoting.
Brief description of the drawings
, below will be to specific in order to illustrate more clearly of the specific embodiment of the invention or technical scheme of the prior art
The accompanying drawing used required in embodiment or description of the prior art is briefly described, it should be apparent that, in describing below
Accompanying drawing is some embodiments of the present invention, for those of ordinary skill in the art, before creative work is not paid
Put, other accompanying drawings can also be obtained according to these accompanying drawings.
Fig. 1 is the tissue culture gone out using the cubeb litsen tree tissue-cultured seedling root media and culture of rootage CMC model in embodiment 10
The situation of taking root of seedling；
Fig. 2 is the tissue culture gone out using the cubeb litsen tree tissue-cultured seedling root media and culture of rootage CMC model in comparative example 14
The situation of taking root of seedling；
Fig. 3 is the tissue culture gone out using the cubeb litsen tree tissue-cultured seedling root media and culture of rootage CMC model in embodiment 10
Transplant survival situation after seedling rooting, three plants of tissue-cultured seedling in same level are same growth period, wherein：(a) transplant
10 days；(b) transplant 30 days；(c) transplant 60 days.
Embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the present invention.It is unreceipted specific in embodiment
Condition person, the condition advised according to normal condition or manufacturer is carried out.Agents useful for same or the unreceipted production firm person of instrument, be
The conventional products that can be obtained by commercially available purchase.
According to an aspect of the invention, there is provided a kind of cubeb litsen tree tissue-cultured seedling root media, mainly by following concentration
Component composition：1/2MS minimal mediums, IBA 0.7-0.9mg/L, IAA 0.2-0.3mg/L, NAA 0.1-0.2mg/L, AC
0.5-1.0mg/L, sucrose 19.0-21.0g/L and agar 7.0-8.0g/L, pH are 5.8-6.0.
The cubeb litsen tree tissue-cultured seedling root media that the present invention is provided, is the introducing indoles fourth on the basis of minimal medium
The rooting induction material such as acid, heteroauxin, methyl α-naphthyl acetate, and acted on and to each material consumption using the coordinated of each material
It is specific to limit so that the root media has stronger root induction effect for cubeb litsen tree tissue-cultured seedling, so that cubeb litsen tree
The rooting rate of tissue-cultured seedling reaches more than 95%, and transplanting survival rate reaches 100%, improves using traditional cubeb litsen tree tissue culture seedling rooting
Culture medium lifts the not good technical problem of effect for the rooting rate and transplanting survival rate of tissue-cultured seedling.
Specifically, being high nutrient and sugar in a kind of instinct of plant, culture medium using Root Absorption nutrition and moisture
Tissue-cultured seedling can be made to produce dependence and be difficult to take root, reducing nutritional ingredient and sugared concentration can stimulate and take root.In addition, culture medium
In some inorganic salts ingredients be unfavorable for the generation of root, therefore suitably to reduce the differentiation that inorganic salt concentration is just conducive to root, therefore
The present invention is in using MS as the culture of rootage of minimal medium, and a great number of elements will drop to 1/2, therefore select 1/2MS to be basic culture
Base.
In tissue cultures, the effect taken root with exogenous hormone of explant is inseparable, wherein the growth of debita spissitudo
Plain class material is very big to Rooting effect, with certain induction of differentiation.Different plants are to auxin required for rooting induction
The species of class material is also different.
In the present invention, mainly rooting induction thing is used as from indolebutyric acid, heteroauxin, methyl α-naphthyl acetate auxin substance
Matter.
Wherein, indolebutyric acid, also known as IBA, its main function are the growths for inducing Cutting rooting, especially adventitious root.
IBA is typical but infinite concentration is 0.7mg/L, 0.72mg/L, 0.74mg/L, 0.76mg/L, 0.78mg/L,
0.80mg/L, 0.82mg/L, 0.84mg/L, 0.86mg/L, 0.88mg/L or 0.90mg/L.
Heteroauxin, also known as IAA, can be by stem, leaf and Root Absorption, and concentration is different, can both play a driving role, and also might be used
Play inhibitory action.Stability is relatively poor, and oxidation Decomposition, in tissue cultures, callus induction are easier under high light conditions
The formation of tissue and root, to accelerate the formation of root and improve the percentage that plant takes root.
IAA is typical but infinite concentration is 0.2mg/L, 0.21mg/L, 0.22mg/L, 0.23mg/L, 0.24mg/L,
0.25mg/L, 0.26mg/L, 0.27mg/L, 0.28mg/L, 0.29mg/L or 0.3mg/L.
Methyl α-naphthyl acetate, also known as NAA, are artificial synthesized auxin substance, and not oxidizable, stability is more in use
By force.
NAA is typical but infinite concentration is 0.1mg/L, 0.11mg/L, 0.12mg/L, 0.13mg/L, 0.14mg/L,
0.15mg/L, 0.16mg/L, 0.17mg/L, 0.18mg/L, 0.19mg/L or 0.2mg/L.
Activated carbon (AC) is added in the medium, and its purpose mainly uses its adsorption capacity, reduces some harmful substances
Adverse effect, it is favourable to certain plants root induction while also create dark situation.
In the present invention, AC is typical but infinite concentration is 0.5mg/L, 0.55mg/L, 0.6mg/L, 0.65mg/L,
0.7mg/L, 0.75mg/L, 0.8mg/L, 0.85mg/L, 0.9mg/L, 095mg/L or 1.0mg/L.
In tissue rapid propagation, the culture being cultured can not carry out photosynthesis mostly, and what can be carried out can not meet it
To the demand of carbohydrate, it is therefore necessary to sugar is added in the medium as carbon source and the energy, while to maintaining culture medium oozing necessarily
Pressure also plays an important role thoroughly.And sucrose is one of most important carbon source.
In the present invention, sucrose is typical but infinite concentration is 19.0mg/L, 19.2mg/L, 19.4mg/L, 19.6mg/
L, 19.8mg/L, 20.0mg/L, 20.2mg/L, 20.4mg/L, 20.6mg/L, 20.8mg/L or 21.0mg/L.
Agar is typical but infinite concentration is 7.0mg/L, 7.1mg/L, 7.2mg/L, 7.3mg/L, 7.4mg/L,
7.5mg/L, 7.6mg/L, 7.7mg/L, 7.8mg/L, 7.9mg/L or 8.0mg/L.
" mainly by ... constitute " of the present invention, it is intended that it can also include other components in addition to the component, this
A little other components assign cubeb litsen tree tissue-cultured seedling root media different characteristics.In addition, it is of the present invention " main
By ... constitute ", may be replaced by enclosed " being " or " by ... constitute ".
In the preferred embodiment of the present invention, the cubeb litsen tree tissue-cultured seedling root media is main by following concentration
Component composition：1/2MS minimal mediums, IBA 0.75-0.85mg/L, IAA 0.2-0.3mg/L, NAA 0.1-0.15mg/
L, AC 0.75-1.0mg/L, sucrose 19.5-20.5g/L and agar 7.2-7.8g/L, pH are 5.8-6.0.
In the preferred embodiment of the present invention, the cubeb litsen tree tissue-cultured seedling root media is main by following concentration
Component composition：1/2MS minimal mediums, IBA 0.8mg/L, IAA 0.2-0.3mg/L, NAA 0.1mg/L, AC 1.0mg/
L, sucrose 20.0g/L, agar 7.5g/L, pH are 5.8.
Tissue-cultured seedling is very sensitive to the species and concentration of the additive in root media, especially rooting induction material,
Pass through each component in reasonable selection and collocation cubeb litsen tree tissue-cultured seedling root media so that the coordinated between each component is made
With stronger, cubeb litsen tree tissue-cultured seedling root media is stronger for the rooting induction effect of tissue-cultured seedling so that rooting rate is further carried
It is high.
In the preferred embodiment of the present invention, the 1/2MS minimal mediums include a great number of elements, micro member
Element, molysite and organic matter, are mainly made up of the component of following concentration：
A great number of elements：Potassium nitrate 950mg/L, ammonium nitrate 825mg/L, epsom salt 185mg/L, potassium dihydrogen phosphate
85mg/L, calcium chloride dihydrate 220mg/L；
Trace element：Four water manganese sulfate 22.3mg/L, white vitriol 8.6mg/L, boric acid 6.2mg/L, KI
0.83mg/L, Sodium Molybdate Dihydrate 0.25mg/L, cupric sulfate pentahydrate 0.025mg/L, CoCL2 6H2O 0.025mg/L；
Molysite：Ferrous sulfate heptahydrate 27.8mg/L, disodium ethylene diamine tetraacetate 37.3mg/L；
Organic matter：Glycine 2mg/L, puridoxine hydrochloride 0.5mg/L, thiamine hydrochloride 0.1mg/L, nicotinic acid 0.5mg/L, flesh
Alcohol 100mg/L.
Compared with MS minimal mediums, a great number of elements in 1/2MS minimal mediums will drop to 1/2, other trace elements,
Molysite, organic concentration are constant.
It should be noted that each component and IBA, IAA, NAA, AC, sucrose, fine jade in above-mentioned 1/2MS minimal mediums
The concentration of each material such as fat is respectively provided with identical calculating benchmark.
In the preferred embodiment of the present invention, using the cubeb litsen tree tissue-cultured seedling root media to cubeb litsen tree group
Train seedling and carry out culture of rootage, rooting rate is more than or equal to 95%.
The cubeb litsen tree tissue-cultured seedling root media of use has stronger rooting induction effect, rooting rate to cubeb litsen tree tissue-cultured seedling
More than or equal to 95%, typical but non-limiting rooting rate be 95%, 95.5%, 96%, 96.5%, 97%, 97.5%,
98%th, 98.5%, 99%, 99.5% or 100%.
According to another aspect of the present invention, a kind of culture of rootage method of cubeb litsen tree tissue-cultured seedling is additionally provided, using upper
State cubeb litsen tree tissue-cultured seedling root media and culture of rootage is carried out to cubeb litsen tree tissue-cultured seedling.
Successful tissue cultures, are largely dependent upon the selection to culture medium.In the training of cubeb litsen tree tissue culture seedling rooting
Above-mentioned cubeb litsen tree tissue-cultured seedling root media is used in the method for supporting, the root media is to utilize IAA, IBA, NAA and 1/2MS base
Basal culture medium coordinated is acted on, and under suitable condition of culture so that the rooting rate of cubeb litsen tree tissue-cultured seedling can reach
More than 95%.
The growth relationship of root is to the height of transplanting survival rate, and high rooting rate contributes to the lifting of transplanting survival rate.Using
The cubeb litsen tree tissue-cultured seedling root media that the present invention is provided, and under suitable condition of culture, rooting rate up to more than 95%,
Transplanting survival rate reaches 100%.
In the preferred embodiment of the present invention, the condition of the culture of rootage is：Cultivation temperature is 23-27 DEG C,
Light application time 15-16h, interlunation is 8-9h, and intensity of illumination is 1800-2200Lux, and incubation time is at least 25 days.
Culture of rootage condition has a significant impact for rooting development and the transplant survival in the later stage tool of tissue-cultured seedling.In this hair
In bright, typical but non-limiting cultivation temperature be 23 DEG C, 23.5 DEG C, 24 DEG C, 24.5 DEG C, 25 DEG C, 25.5 DEG C, 26 DEG C, 26.5
DEG C or 27 DEG C.
Typical but non-limiting light application time is 15h, 15.5h or 16h, and interlunation is 8h, 8.5h or 9h.
Typical but non-limiting intensity of illumination be 1800Lux, 1850Lux, 1900Lux, 1950Lux, 2000Lux,
2050Lux, 2100Lux, 2150Lux or 2200Lux.
In the preferred embodiment of the present invention, the condition of the culture of rootage is：Cultivation temperature is 23-27 DEG C,
Light application time 16h, interlunation is 8h, and intensity of illumination is 2000Lux, and incubation time is 25 days.
By the further restriction to culture of rootage condition, rooting rate and the later stage of cubeb litsen tree tissue-cultured seedling are further improved
Transplanting survival rate.
In the preferred embodiment of the present invention, the cubeb litsen tree tissue-cultured seedling for choose lignifying, being about 2-3cm is given birth to
Root culture.
Mainly by the way that the explant of selection is used after Initial culture, squamous subculture is obtained again in the source of cubeb litsen tree tissue-cultured seedling
Arrive.
Material selection：The cubeb litsen tree semi-lignified band bud branch of health, no disease and pests harm is taken as explant；
Pretreatment：Explant is soaked 16 hours in running water, clip stem with bud 2-3cm is rinsed with running water
Totally, washing powder solution is soaked 10-15 minutes, and flowing water is rinsed 1 hour；
Disinfect：70% alcohol-pickled 20s, 0.1% mercuric chloride sterilization 7min, aseptic water washing 5 times；
Initial culture：It is inoculated in Initial culture base and carries out induction differentiation culture, obtains primary tissue-cultured seedling；
Squamous subculture：Primary tissue-cultured seedling is placed in subculture medium again, the Regenerated plant of stalwartness is obtained.
Lignifying is chosen in obtained Regenerated plant, 2-3cm cubeb litsen tree tissue-cultured seedling progress culture of rootage is about, can be promoted
Cubeb litsen tree is gone out root neat and consistent without offspring, improve rooting rate.
In the preferred embodiment of the present invention, using above-mentioned cubeb litsen tree tissue-cultured seedling root media to cubeb litsen tree group
Train seedling and carry out culture of rootage, rooting rate is more than or equal to 95%.
In the preferred embodiment of the present invention, the cubeb litsen tree tissue-cultured seedling root media is main by following concentration
Component composition：1/2MS minimal mediums, indolebutyric acid 0.7-0.9mg/L, heteroauxin 0.2-0.3mg/L, methyl α-naphthyl acetate 0.1-
0.2mg/L, activated carbon 0.5-1.0mg/L, sucrose 19.0-21.0g/L, agar 7.0-8.0g/L, pH are 5.8-6.0；
It is preferred that, the cubeb litsen tree tissue-cultured seedling root media is mainly made up of the component of following concentration：1/2MS is trained substantially
Support base, indolebutyric acid 0.8mg/L, heteroauxin 0.2-0.3mg/L, methyl α-naphthyl acetate 0.1mg/L, activated carbon 1.0mg/L, sucrose
20.0g/L and agar 7.5g/L, pH are 5.8.
With reference to specific embodiment and comparative example, the invention will be further described.
Material selection：The cubeb litsen tree semi-lignified band bud branch of health, no disease and pests harm is taken as explant；
Pretreatment：Explant is soaked into 16h, clip stem with bud 2-3cm in running water, rinses dry with running water
Only, washing powder solution immersion 10-15min, flowing water rinses 1h；
Disinfect：70% alcohol-pickled 20s, 0.1% mercuric chloride sterilization 7min, aseptic water washing 5 times；
Initial culture：It is inoculated in Initial culture base and carries out induction differentiation culture, obtains primary tissue-cultured seedling, Initial culture base
For MS+BA 2.0mg/L+IBA 0.1mg/L；
Squamous subculture：Primary tissue-cultured seedling is placed in subculture medium again, the Regenerated plant of stalwartness is obtained, subculture medium is
MS+BA 2.0mg/L+IBA 0.1mg/L；
Wherein BA (Benzylaminopurine) is benzyl aminoadenine.
Lignifying is chosen in obtained Regenerated plant, 2-3cm cubeb litsen tree tissue-cultured seedling progress culture of rootage is about, from reality
The cubeb litsen tree tissue-cultured seedling root media and cultural method for applying a 1-14 and comparative example 1-16 carry out culture of rootage.
Embodiment 1
A kind of cubeb litsen tree tissue-cultured seedling root media, is made up of the component of following concentration：1/2MS minimal mediums, IBA
0.7mg/L, IAA 0.2mg/L, NAA 0.1mg/L, AC 1.0mg/L, sucrose 19.0g/L and agar 7.0g/L, pH are 6.0.
Embodiment 2
A kind of cubeb litsen tree tissue-cultured seedling root media, is made up of the component of following concentration：1/2MS minimal mediums, IBA
0.7mg/L, IAA 0.2mg/L, NAA 0.1mg/L, AC 1.0mg/L, sucrose 21.0g/L and agar 8.0g/L, pH are 6.0.
Embodiment 3
A kind of cubeb litsen tree tissue-cultured seedling root media, is made up of the component of following concentration：1/2MS minimal mediums, IBA
0.8mg/L, IAA 0.2mg/L, NAA 0.1mg/L, AC 1.0mg/L, sucrose 20.0g/L and agar 7.5g/L, pH are 5.8.
Embodiment 4
A kind of cubeb litsen tree tissue-cultured seedling root media, is made up of the component of following concentration：1/2MS minimal mediums, IBA
0.8mg/L, IAA 0.3mg/L, NAA 0.1mg/L, AC 1.0mg/L, sucrose 20.0g/L and agar 7.5g/L, pH are 5.8.
Embodiment 5
A kind of cubeb litsen tree tissue-cultured seedling root media, is made up of the component of following concentration：1/2MS minimal mediums, IBA
0.8mg/L, IAA 0.2mg/L, NAA 0.1mg/L, AC 0.5mg/L, sucrose 20.0g/L and agar 7.5g/L, pH are 5.8.
Embodiment 6
A kind of cubeb litsen tree tissue-cultured seedling root media, is made up of the component of following concentration：1/2MS minimal mediums, IBA
0.8mg/L, IAA 0.3mg/L, NAA 0.2mg/L, AC 1.0mg/L, sucrose 20.0g/L and agar 7.5g/L, pH are 5.8.
Embodiment 7
A kind of cubeb litsen tree tissue-cultured seedling root media, is made up of the component of following concentration：1/2MS minimal mediums, IBA
0.9mg/L, IAA 0.25mg/L, NAA 0.15mg/L, AC 0.8mg/L, sucrose 21.0g/L and agar 7.0g/L, pH are 5.9.
Embodiment 8
A kind of culture of rootage method of cubeb litsen tree tissue-cultured seedling, it is raw using the cubeb litsen tree tissue-cultured seedling root media of embodiment 1
The condition of root culture is：Cultivation temperature is 23 DEG C or so, and light application time 15h, interlunation is 9h, and intensity of illumination is 1900Lux,
Incubation time is 25 days.
Embodiment 9
A kind of culture of rootage method of cubeb litsen tree tissue-cultured seedling, it is raw using the cubeb litsen tree tissue-cultured seedling root media of embodiment 2
The condition of root culture is identical with the condition of the culture of rootage in embodiment 8.
Embodiment 10
A kind of culture of rootage method of cubeb litsen tree tissue-cultured seedling, it is raw using the cubeb litsen tree tissue-cultured seedling root media of embodiment 3
The condition of root culture is：Cultivation temperature is 25 DEG C or so, and light application time 16h, interlunation is 8h, and intensity of illumination is 2000Lux,
Incubation time is 25 days.
Embodiment 11
A kind of culture of rootage method of cubeb litsen tree tissue-cultured seedling, it is raw using the cubeb litsen tree tissue-cultured seedling root media of embodiment 4
The condition of root culture is identical with the condition of the culture of rootage of embodiment 10.
Embodiment 12
A kind of culture of rootage method of cubeb litsen tree tissue-cultured seedling, it is raw using the cubeb litsen tree tissue-cultured seedling root media of embodiment 5
The condition of root culture is identical with the condition of the culture of rootage of embodiment 10.
Embodiment 13
A kind of culture of rootage method of cubeb litsen tree tissue-cultured seedling, it is raw using the cubeb litsen tree tissue-cultured seedling root media of embodiment 6
The condition of root culture is identical with the condition of the culture of rootage of embodiment 10.
Embodiment 14
A kind of culture of rootage method of cubeb litsen tree tissue-cultured seedling, it is raw using the cubeb litsen tree tissue-cultured seedling root media of embodiment 7
The condition of root culture is：Cultivation temperature is 27 DEG C or so, and light application time 16h, interlunation is 8h, and intensity of illumination is 2200Lux,
Incubation time is 25 days.
Comparative example 1
This comparative example is the comparative example of embodiment 10, except IBA concentration is 0.50mg/L, the training of cubeb litsen tree tissue culture seedling rooting
The other components and concentration and condition of culture for supporting base are same as in Example 10.
Comparative example 2
This comparative example is the comparative example of embodiment 11, except IBA concentration is 0.50mg/L, the training of cubeb litsen tree tissue culture seedling rooting
The other components for supporting base are identical with embodiment 11 with concentration and condition of culture.
Comparative example 3
A kind of cubeb litsen tree tissue-cultured seedling root media, is made up of the component of following concentration：1/2MS minimal mediums, IBA
0.5mg/L, IAA 0.1mg/L, NAA 0.1mg/L, AC 0.5mg/L, sucrose 20.0g/L and agar 7.5g/L, pH are 5.8；
Use the root media to cubeb litsen tree tissue-cultured seedling carry out culture of rootage condition of culture for：Cultivation temperature is 25 DEG C
Left and right, light application time 16h, interlunation is 8h, and intensity of illumination is 2000Lux, and incubation time is 25 days.
Comparative example 4
A kind of cubeb litsen tree tissue-cultured seedling root media, is made up of the component of following concentration：1/2MS minimal mediums, IBA
0.5mg/L, IAA 0.2mg/L, NAA 0.4mg/L, AC 1.0mg/L, sucrose 20.0g/L and agar 7.5g/L, pH are 5.8；
Use the root media to cubeb litsen tree tissue-cultured seedling carry out culture of rootage condition of culture for：Cultivation temperature is 25 DEG C
Left and right, light application time 16h, interlunation is 8h, and intensity of illumination is 2000Lux, and incubation time is 25 days.
Comparative example 5
A kind of cubeb litsen tree tissue-cultured seedling root media, is made up of the component of following concentration：1/2MS minimal mediums, IBA
0.5mg/L, IAA 0.3mg/L, NAA 0.4mg/L, AC 1.0mg/L, sucrose 20.0g/L and agar 7.5g/L, pH are 5.8.
The condition of culture for carrying out culture of rootage to cubeb litsen tree tissue-cultured seedling using the root media is identical with comparative example 4.
Comparative example 6
This comparative example is the contrast experiment of embodiment 10, except NAA concentration is 0.40mg/L, cubeb litsen tree tissue culture seedling rooting
The other components of culture medium and concentration and condition of culture are same as in Example 10.
Comparative example 7
This comparative example is the contrast experiment of embodiment 11, except NAA concentration is 0.40mg/L, cubeb litsen tree tissue culture seedling rooting
The other components of culture medium are identical with embodiment 11 with concentration and condition of culture.
Comparative example 8
This comparative example is the contrast experiment of embodiment 12, except IAA concentration is 0.10mg/L, cubeb litsen tree tissue culture seedling rooting
The other components of culture medium are identical with embodiment 12 with concentration and condition of culture.
Comparative example 9
A kind of cubeb litsen tree tissue-cultured seedling root media, is made up of the component of following concentration：1/2MS minimal mediums, IBA
0.20mg/L, IAA 0.20mg/L, NAA 0.10mg/L, AC 1.0mg/L, sucrose 19.0g/L and agar 7.0g/L, pH are 5.9；
Use the root media to cubeb litsen tree tissue-cultured seedling carry out culture of rootage condition of culture for：Cultivation temperature is 26 DEG C
Left and right, light application time 16h, interlunation is 8h, and intensity of illumination is 2100Lux, and incubation time is 25 days.
Comparative example 10
A kind of cubeb litsen tree tissue-cultured seedling root media, is made up of the component of following concentration：1/2MS minimal mediums, IBA
0.20mg/L, IAA 0.30mg/L, NAA 0.10mg/L, AC 1.0mg/L, sucrose 19.0g/L and agar 7.0g/L, pH are 5.9；
The condition of culture for carrying out culture of rootage to cubeb litsen tree tissue-cultured seedling using the root media is identical with comparative example 9.
Comparative example 11
A kind of cubeb litsen tree tissue-cultured seedling root media, is made up of the component of following concentration：1/2MS minimal mediums, IBA
0.20mg/L, IAA 0.20mg/L, NAA 0.40mg/L, AC 1.0mg/L, sucrose 19.0g/L and agar 7.0g/L, pH are 5.9；
The condition of culture for carrying out culture of rootage to cubeb litsen tree tissue-cultured seedling using the root media is identical with comparative example 9.
Comparative example 12
A kind of cubeb litsen tree tissue-cultured seedling root media, is made up of the component of following concentration：1/2MS minimal mediums, IBA
0.20mg/L, IAA 0.30mg/L, NAA 0.40mg/L, AC 1.0mg/L, sucrose 19.0g/L and agar 7.0g/L, pH are 5.9；
The condition of culture for carrying out culture of rootage to cubeb litsen tree tissue-cultured seedling using the root media is identical with comparative example 9.
Comparative example 13
A kind of cubeb litsen tree tissue-cultured seedling root media, is made up of the component of following concentration：1/2MS minimal mediums, IBA
0.20mg/L, IAA 0.10mg/L, NAA 0.10mg/L, AC 0.5mg/L, sucrose 21.0g/L and agar 8.0g/L, pH are 6.0；
The condition of culture for carrying out culture of rootage to cubeb litsen tree tissue-cultured seedling using the root media is identical with comparative example 9.
Comparative example 14
A kind of cubeb litsen tree tissue-cultured seedling root media, is made up of the component of following concentration：1/2MS minimal mediums, IBA
0.80mg/L, IAA 0.20mg/L, AC 1.0mg/L, sucrose 20.0g/L and agar 7.5g/L, pH are 5.8.
This comparative example is the contrast experiment of embodiment 10, except being not added with the cubeb litsen tree tissue-cultured seedling root media
Outside NAA, the other components and concentration and condition of culture of cubeb litsen tree tissue-cultured seedling root media are same as in Example 10.
Comparative example 15
A kind of cubeb litsen tree tissue-cultured seedling root media, is made up of the component of following concentration：1/2MS minimal mediums, IBA
0.80mg/L, NAA 0.20mg/L, AC 1.0mg/L, sucrose 20.0g/L and agar 7.5g/L, pH are 5.8；
This comparative example is the contrast experiment of embodiment 10, except being not added with the cubeb litsen tree tissue-cultured seedling root media
Outside IAA, the other components and concentration and condition of culture of cubeb litsen tree tissue-cultured seedling root media are same as in Example 10.
Comparative example 16
A kind of cubeb litsen tree tissue-cultured seedling root media, is made up of the component of following concentration：1/2MS minimal mediums, IAA
0.20mg/L, NAA 0.10mg/L, AC 1.0mg/L, sucrose 20.0g/L and agar 7.5g/L, pH are 5.8；
This comparative example is the contrast experiment of embodiment 10, except being not added with the cubeb litsen tree tissue-cultured seedling root media
Outside IBA, the other components and concentration and condition of culture of cubeb litsen tree tissue-cultured seedling root media are same as in Example 10.
Comparative example 17
A kind of cubeb litsen tree tissue-cultured seedling root media, is made up of the component of following concentration：1/2MS minimal mediums, IBA
0.8mg/L, IAA 0.3mg/L, AC 1.0mg/L, sucrose 20.0g/L and agar 7.5g/L, pH are 5.8；
This comparative example is the contrast experiment of embodiment 11, except being not added with the cubeb litsen tree tissue-cultured seedling root media
Outside NAA, the other components of cubeb litsen tree tissue-cultured seedling root media are identical with embodiment 11 with concentration and condition of culture.
Comparative example 18
A kind of cubeb litsen tree tissue-cultured seedling root media, is made up of the component of following concentration：1/2MS minimal mediums, IAA
0.30mg/L, NAA 0.10mg/L, AC 1.0mg/L, sucrose 20.0g/L and agar 7.5g/L, pH are 5.8；
This comparative example is the contrast experiment of embodiment 11, except being not added with the cubeb litsen tree tissue-cultured seedling root media
Outside IBA, the other components of cubeb litsen tree tissue-cultured seedling root media are identical with embodiment 11 with concentration and condition of culture.
Comparative example 19
This comparative example is the contrast experiment of embodiment 11, in addition to the concentration of sucrose is 17.00mg/L, cubeb litsen tree tissue-cultured seedling
The other components of root media are identical with embodiment 11 with concentration and condition of culture.
It should be noted that the 1/2MS minimal mediums composition all same used in above-mentioned all embodiments and comparative example,
Including a great number of elements, trace element, molysite and organic matter, mainly it is made up of the component of following concentration：
A great number of elements：Potassium nitrate 950mg/L, ammonium nitrate 825mg/L, epsom salt 185mg/L, potassium dihydrogen phosphate
85mg/L, calcium chloride dihydrate 220mg/L；
Trace element：Four water manganese sulfate 22.3mg/L, white vitriol 8.6mg/L, boric acid 6.2mg/L, KI
0.83mg/L, Sodium Molybdate Dihydrate 0.25mg/L, cupric sulfate pentahydrate 0.025mg/L, CoCL2 6H2O 0.025mg/L；
Molysite：Ferrous sulfate heptahydrate 27.8mg/L, disodium ethylene diamine tetraacetate 37.3mg/L；
Organic matter：Glycine 2mg/L, puridoxine hydrochloride 0.5mg/L, thiamine hydrochloride 0.1mg/L, nicotinic acid 0.5mg/L, flesh
Alcohol 100mg/L.
Experimental example 1
Using above-described embodiment 8-14 and comparative example 1-19 cubeb litsen tree tissue-cultured seedling root media in respective culture bar
Root induction experiment is carried out to cubeb litsen tree tissue-cultured seedling under part, 50 plants are taken, to rooting rate, mean elements, average root length and transplanting
Survival rate is measured.Wherein, rooting rate (%)=strain number of taking root/inoculation total strain number × 100%, mean elements=always take root
Number/total strain number, average root length=all overall lengths/number of always taking root, transplanting survival rate=transplant survival strain number/transplanting total strain number ×
100%, it is necessary to which explanation, the transplanting medium all same that each embodiment is used with comparative example, and transplanting survival rate are to move
Counted within 30 days after cultivation, specific measurement result is shown in Table 1：
The situation measurement result of taking root of cubeb litsen tree tissue-cultured seedling in each embodiment of table 1 and comparative example
From table 1 it follows that the cubeb litsen tree tissue-cultured seedling root media that embodiment 8-14 is provided is for cubeb litsen tree tissue culture
The rooting induction effect of seedling is far above the cubeb litsen tree tissue-cultured seedling root media in comparative example 1-19.
Specifically, embodiment 11 is the control experiment of embodiment 10, both differences are cubeb litsen tree tissue culture seedling rooting
IAA concentration is different in culture medium.IAA concentration is different, different for tissue-cultured seedling rooting induction effect.
Embodiment 12 is the control experiment of embodiment 10, and both differences are in cubeb litsen tree tissue-cultured seedling root media
The concentration of activated carbon is different.Within the specific limits, the content of activated carbon is more, more can adsorb the harmful substance in culture medium, from
And it is favourable for cubeb litsen tree tissue-cultured seedling root induction.
Comparative example 1 and comparative example 2 are respectively the contrast experiment of embodiment 10 and embodiment 11, in embodiment 10 and embodiment
IBA concentration is that the IBA concentration in 0.8mg/L, comparative example 1 and comparative example 2 is in cubeb litsen tree tissue-cultured seedling root media in 11
0.5mg/L.It can be seen from the data in Table 1 that cubeb litsen tree tissue-cultured seedling is more sensitive for the concentration of IBA concentration, IBA can promote
Enter the increase of root system bar number, have remarkable effect for the growth of taking root of cubeb litsen tree tissue-cultured seedling.
Comparative example 4 is the contrast experiment of comparative example 1, and both differences are that NAA concentration is different.It is possible thereby to see
Go out, the change of NAA concentration also directly affects the rooting rate of cubeb litsen tree tissue-cultured seedling.
Comparative example 6 and comparative example 7 are respectively the contrast experiment of embodiment 10 and embodiment 11, in embodiment 10 and embodiment
NAA concentration is 0.1mg/L in 11 cubeb litsen tree tissue-cultured seedling root media, and NAA concentration is in comparative example 1 and comparative example 2
0.4mg/L.It can be seen from the data in Table 1 that NAA concentration is bigger, it is unfavorable for the lifting of cubeb litsen tree tissue-cultured seedling rooting rate on the contrary, therefore
NAA concentration should be in suitable scope.
Comparative example 8 is the contrast experiment of embodiment 12, and both differences are in cubeb litsen tree tissue-cultured seedling root media
IAA concentration is different.When IAA concentration exceed this scope of 0.20-0.30mg/L when, cubeb litsen tree tissue-cultured seedling rooting rate under
Drop.
Comparative example 14-16 be in the contrast experiment of embodiment 10, embodiment 10 cubeb litsen tree tissue-cultured seedling root media be
1/2MS+IBA 0.8mg/L+IAA 0.2mg/L+NAA 0.1mg/L+AC 1mg/L+ sucrose 20g/L+ agar 7.5g/L, contrast
Cubeb litsen tree tissue-cultured seedling root media is 1/2MS+IBA 0.8mg/L+IAA 0.2mg/L+AC 1mg/L+ sucrose 20g/ in example 14
Cubeb litsen tree tissue-cultured seedling root media is 1/2MS+IBA 0.8mg/L+NAA 0.1mg/L+ in L+ agar 7.5g/L, comparative example 15
Cubeb litsen tree tissue-cultured seedling root media is 1/2MS+IAA in AC 1mg/L+ sucrose 20g/L+ agar 7.5g/L, comparative example 16
0.2mg/L+NAA 0.1mg/L+AC 1mg/L+ sucrose 20g/L+ agar 7.5g/L, it can be seen that what comparative example 14-16 was used
Rooting induction material is the combination of IBA, IAA and NAA two of which material.From table 1 it follows that using in embodiment 10
Cubeb litsen tree tissue-cultured seedling root media is whether for rooting rate or mean elements, average root length, or from transplant survival
For rate, far superior to comparative example 14-16.
Fig. 1 is the tissue culture gone out using the cubeb litsen tree tissue-cultured seedling root media and culture of rootage CMC model in embodiment 10
The situation of taking root of seedling, Fig. 2 is to be gone out using the cubeb litsen tree tissue-cultured seedling root media and culture of rootage CMC model in comparative example 14
Tissue-cultured seedling situation of taking root, be only that embodiment 10 and comparative example 14 have generation in respective 50 plants of Duplicate Samples shown in Fig. 1 and Fig. 2
One plant of table.It can also intuitively find out very much from Fig. 1 and 2, the cubeb litsen tree tissue culture of culture of rootage is carried out using embodiment 10
Miao Genchang is longer, and twig growing way is preferably, and its situation of taking root is substantially better than the cubeb litsen tree tissue-cultured seedling that use comparative example 14 is cultivated
Take root situation.And from figure 3, it can be seen that shifting of the cubeb litsen tree tissue-cultured seedling in the later stage of culture of rootage is carried out using embodiment 10
Survived during cultivation all right.
Comparative example 17 and comparative example 18 are the contrast experiment of embodiment 11.It is different from embodiment 11, the pheasant of comparative example 17
It is not added with being not added with IBA in NAA, the cubeb litsen tree tissue-cultured seedling root media of comparative example 18 in green pepper tissue-cultured seedling root media.By table 1
Data can be seen that may be up to 100% using the rooting rate of the cubeb litsen tree tissue-cultured seedling root media of embodiment 11, that is, realize
100% takes root, and mean elements and average root length are all in higher level.And use comparative example 17 and the cubeb litsen tree of comparative example 18
The rooting rate of tissue-cultured seedling root media and the mean elements of tissue culture seedling rooting are significantly lower than embodiment 11, rooting rate and average root
It is several, the transplanting survival rate in later stage is directly influenced, therefore use the pheasant of the root media culture of comparative example 17 and comparative example 18
The transplanting survival rate of green pepper tissue-cultured seedling is also far below embodiment 11.
It can thus be seen that coordinated relation is there is between these three materials of IBA, IAA and NAA, and this association
Just it is achieved in specific concentration range with matching relationship.In other words, using these three things of IBA, IAA and NAA
Matter is needed in particular range, could produce the good root induction effect to cubeb litsen tree tissue-cultured seedling.Deviate this particular range
When, rooting rate, mean elements, average root length and transplanting survival rate all decrease.
Comparative example 19 is also the contrast experiment of embodiment 11.Both differences are the concentration of sucrose in root media
It is different.The situation of taking root using the cubeb litsen tree tissue-cultured seedling of the root media of comparative example 19 is inferior to embodiment 11.Sucrose is used as pheasant
Carbon source in green pepper tissue-cultured seedling culture of rootage, concentration is too low, can directly influence the situation of taking root of cubeb litsen tree tissue-cultured seedling, therefore sucrose is dense
Degree should be in the concentration range that the present invention is limited.
In summary, the cubeb litsen tree tissue-cultured seedling root media that the present invention is provided is made by the coordinated between each material
With and for certain concentration restriction so that the rooting rate of cubeb litsen tree tissue-cultured seedling can reach more than 95%, and transplanting survival rate reaches
To 100%, compared to rooting rate, the transplanting survival rate using traditional cubeb litsen tree tissue-cultured seedling root media for cubeb litsen tree tissue-cultured seedling
Have and be obviously improved, provide technical basis more reliable, that operability is stronger for the factorial praluction of cubeb litsen tree, accelerate
It is bred with promoting.
Finally it should be noted that：Various embodiments above is merely illustrative of the technical solution of the present invention, rather than its limitations；To the greatest extent
The present invention is described in detail with reference to foregoing embodiments for pipe, it will be understood by those within the art that：Its according to
The technical scheme described in foregoing embodiments can so be modified, or which part or all technical characteristic are entered
Row equivalent substitution；And these modifications or replacement, the essence of appropriate technical solution is departed from various embodiments of the present invention technology
The scope of scheme.
Claims (10)
1. a kind of cubeb litsen tree tissue-cultured seedling root media, it is characterised in that be mainly made up of the component of following concentration：1/2MS bases
Basal culture medium, indolebutyric acid 0.7-0.9mg/L, heteroauxin 0.2-0.3mg/L, methyl α-naphthyl acetate 0.1-0.2mg/L, activated carbon 0.5-
1.0mg/L, sucrose 19.0-21.0g/L and agar 7.0-8.0g/L, pH are 5.8-6.0.
2. cubeb litsen tree tissue-cultured seedling root media according to claim 1, it is characterised in that the main group by following concentration
It is grouped into：1/2MS minimal mediums, indolebutyric acid 0.75-0.85mg/L, heteroauxin 0.2-0.3mg/L, methyl α-naphthyl acetate 0.1-
0.15mg/L, activated carbon 0.75-1.0mg/L, sucrose 19.5-20.5g/L and agar 7.2-7.8g/L, pH are 5.8-6.0.
3. cubeb litsen tree tissue-cultured seedling root media according to claim 1, it is characterised in that the main group by following concentration
It is grouped into：1/2MS minimal mediums, indolebutyric acid 0.8mg/L, heteroauxin 0.2-0.3mg/L, methyl α-naphthyl acetate 0.1mg/L, activity
Charcoal 1.0mg/L, sucrose 20.0g/L and agar 7.5g/L, pH are 5.8.
4. the cubeb litsen tree tissue-cultured seedling root media according to claim 1-3 any one, it is characterised in that described 1/
2MS minimal mediums include a great number of elements, trace element, molysite and organic matter, are mainly made up of the component of following concentration：
A great number of elements：Potassium nitrate 950mg/L, ammonium nitrate 825mg/L, epsom salt 185mg/L, potassium dihydrogen phosphate 85mg/L,
Calcium chloride dihydrate 220mg/L；
Trace element：Four water manganese sulfate 22.3mg/L, white vitriol 8.6mg/L, boric acid 6.2mg/L, KI 0.83mg/L,
Sodium Molybdate Dihydrate 0.25mg/L, cupric sulfate pentahydrate 0.025mg/L, CoCL2 6H2O 0.025mg/L；
Molysite：Ferrous sulfate heptahydrate 27.8mg/L, disodium ethylene diamine tetraacetate 37.3mg/L；
Organic matter：Glycine 2mg/L, puridoxine hydrochloride 0.5mg/L, thiamine hydrochloride 0.1mg/L, nicotinic acid 0.5mg/L, inositol
100mg/L。
5. the cubeb litsen tree tissue-cultured seedling root media according to claim 1-3 any one, it is characterised in that using described
Cubeb litsen tree tissue-cultured seedling root media carries out culture of rootage to cubeb litsen tree tissue-cultured seedling, and rooting rate is more than or equal to 95%.
6. a kind of culture of rootage method of cubeb litsen tree tissue-cultured seedling, it is characterised in that using as described in claim 1-5 any one
Cubeb litsen tree tissue-cultured seedling root media to cubeb litsen tree tissue-cultured seedling carry out culture of rootage.
7. the culture of rootage method of cubeb litsen tree tissue-cultured seedling according to claim 6, it is characterised in that the culture of rootage
Condition is：Cultivation temperature is 23-27 DEG C, and light application time 15-16h, interlunation is 8-9h, and intensity of illumination is 1800-
2200Lux, incubation time is at least 25 days.
8. the culture of rootage method of cubeb litsen tree tissue-cultured seedling according to claim 6, it is characterised in that the culture of rootage
Condition is：Cultivation temperature is 23-27 DEG C, and light application time 16h, interlunation is 8h, and intensity of illumination is 2000Lux, incubation time
For 25 days.
9. the culture of rootage method of cubeb litsen tree tissue-cultured seedling according to claim 6, it is characterised in that choose lignifying, length
About 2-3cm cubeb litsen tree tissue-cultured seedling carries out culture of rootage.
10. the culture of rootage method of the cubeb litsen tree tissue-cultured seedling according to claim 6-9 any one, it is characterised in that adopt
Culture of rootage is carried out to cubeb litsen tree tissue-cultured seedling with the cubeb litsen tree tissue-cultured seedling root media, rooting rate is more than or equal to 95%.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107980466A (en) * | 2017-12-29 | 2018-05-04 | 福建省林业科学研究院 | A kind of fruit of a cubeb litsea tree tissue-cultured seedling minitype cuttage method |
| CN114410679A (en) * | 2022-03-10 | 2022-04-29 | 中国林业科学研究院亚热带林业研究所 | Genetic transformation method and application of litsea cubeba |
| CN116584381A (en) * | 2023-05-10 | 2023-08-15 | 江西农业大学 | Litsea cubeba embryogenic callus induction medium and induction method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102227976A (en) * | 2011-04-11 | 2011-11-02 | 广西壮族自治区林业科学研究院 | Litsea cubeba pers asexual quick propagation and seedling breeding method |
| CN102668980A (en) * | 2012-05-09 | 2012-09-19 | 中国计量学院 | Tissue culture method for lindera glauca |
| CN103535172A (en) * | 2013-09-23 | 2014-01-29 | 福建万芳生态农林科技有限公司 | Method for cultivation of test-tube seedlings of Litsea cubeba |
| CN104041409A (en) * | 2014-06-04 | 2014-09-17 | 四川省林业科学研究院 | Tissue culturing method of Litsea mollis |
-
2017
- 2017-06-13 CN CN201710441462.0A patent/CN107223565A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102227976A (en) * | 2011-04-11 | 2011-11-02 | 广西壮族自治区林业科学研究院 | Litsea cubeba pers asexual quick propagation and seedling breeding method |
| CN102668980A (en) * | 2012-05-09 | 2012-09-19 | 中国计量学院 | Tissue culture method for lindera glauca |
| CN103535172A (en) * | 2013-09-23 | 2014-01-29 | 福建万芳生态农林科技有限公司 | Method for cultivation of test-tube seedlings of Litsea cubeba |
| CN104041409A (en) * | 2014-06-04 | 2014-09-17 | 四川省林业科学研究院 | Tissue culturing method of Litsea mollis |
Non-Patent Citations (3)
| Title |
|---|
| 刘弘: "《植物组织培养技术》", 31 March 2012, 机械工业出版社 * |
| 裘文达、曹孜义: "《植物组织培养实用技术》", 31 October 1989, 高等教育出版社 * |
| 赵佐敏: "影响山苍子离体快繁效果的主要因素研究", 《安徽农业科学》 * |
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| CN114410679A (en) * | 2022-03-10 | 2022-04-29 | 中国林业科学研究院亚热带林业研究所 | Genetic transformation method and application of litsea cubeba |
| CN114410679B (en) * | 2022-03-10 | 2024-01-09 | 中国林业科学研究院亚热带林业研究所 | Genetic transformation method and application of capsicum annuum |
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