CN106718884B - RADIX PRZEWALSKIAE TANGUTICAE seedling and its method for culturing seedlings - Google Patents
RADIX PRZEWALSKIAE TANGUTICAE seedling and its method for culturing seedlings Download PDFInfo
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- CN106718884B CN106718884B CN201611071300.4A CN201611071300A CN106718884B CN 106718884 B CN106718884 B CN 106718884B CN 201611071300 A CN201611071300 A CN 201611071300A CN 106718884 B CN106718884 B CN 106718884B
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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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Abstract
The present invention provides a kind of RADIX PRZEWALSKIAE TANGUTICAE seedling and its method for culturing seedlings, is related to bioengineering field.The method for culturing seedlings chooses RADIX PRZEWALSKIAE TANGUTICAE, takes its tip of a root, sterilizes, obtains the tip of a root to be seeded；Tip of a root 0.3-0.5mm to be seeded is cut, is accessed in inductive differentiation medium, differentiation culture two months；The tip of a root after differentiation is placed in proliferated culture medium, Multiplying culture one month；The tip of a root after Multiplying culture is placed in root media, in 17-27 DEG C of day temperature, 5-7 DEG C of night temperature, intensity of illumination 1800-2700lx, under conditions of light application time is 10-11h, cultivate 30-35d, rooted seedling, then rooted seedling is transplanted after hardening, this method makes seedling offspring obtained not only inherit the merit of former plant, and resistance is good, and it is adaptable.Another object of the present invention is to provide a kind of RADIX PRZEWALSKIAE TANGUTICAE seedlings, and without virus, stable yield, resistance is good, adaptable.
Description
Technical field
The present invention relates to bioengineering fields, and in particular to a kind of RADIX PRZEWALSKIAE TANGUTICAE seedling and its method for culturing seedlings.
Background technique
RADIX PRZEWALSKIAE TANGUTICAE (Przewalskia tangutica Maxim.) also known as short henbane, are Solanaceae RADIX PRZEWALSKIAE TANGUTICAE platymisciums, are
China Qinghai-Tibet Platean endemic plant；With being grown on 3200-5000 meters of height above sea level of high mountain gravel and aridity grass land.According to " Jingzhubencao "
And " Tibetan medicine will " is recorded, it is that the common medicine in Tibetan medicine, root and seed and herb can be used for medicinal purpose, and has analgesia, spasmolysis, kills
The drug effects such as worm, anti-inflammatory are used for gastrointestinal convulsion pain, diphtheria, anthrax；External application sore, pruitus.Due to tropane class in RADIX PRZEWALSKIAE TANGUTICAE
Alkaloid is high, can be directly as the raw material of such drug of industrial production, in the market not to the demand of RADIX PRZEWALSKIAE TANGUTICAE
Disconnected increase, and only meet the needs of market currently, and the RADIX PRZEWALSKIAE TANGUTICAE breeding master of modern artificial growth by picking wild RADIX PRZEWALSKIAE TANGUTICAE
To pass through RADIX PRZEWALSKIAE TANGUTICAE seed sexual propagation, since under natural breeding condition, seminal propagation progeny variation is more, individual plant difference
Larger, Character instability, therefore, the expansion of excellent variety is numerous to become urgent need.
Summary of the invention
The purpose of the present invention is to provide a kind of RADIX PRZEWALSKIAE TANGUTICAE method for culturing seedlings, use and manually choose excellent Wild plant, lead to
Crossing detoxification organization of root tips culture fast breeding technique combines imitative wild environment to cultivate, and seedling offspring is made not only to inherit the excellent of former plant
Character, without virus, stable yield, and resistance is good, adaptable.
Another object of the present invention is to provide a kind of RADIX PRZEWALSKIAE TANGUTICAE seedlings, inherit the merit of former plant, not in spite of illness
Poison, stable yield, resistance is good, adaptable.
The present invention solves its technical problem and adopts the following technical solutions to realize.
The present invention proposes a kind of RADIX PRZEWALSKIAE TANGUTICAE method for culturing seedlings comprising:
The vigorous wild RADIX PRZEWALSKIAE TANGUTICAE of growing way is chosen, its tip of a root is taken, sterilizes, obtains the tip of a root to be seeded.
It cuts tip of a root 0.3-0.5mm to be seeded, accesses in inductive differentiation medium, be 20 DEG C in temperature, when daily illumination
Between 12h, intensity of illumination be 2000lx under conditions of, differentiation culture two months.
The tip of a root after differentiation is placed in proliferated culture medium, at 20 DEG C of temperature, intensity of illumination 2000lx, light application time 12h's
Under the conditions of, Multiplying culture one month.
The tip of a root after Multiplying culture is placed in root media, in 17-27 DEG C of day temperature, 5-7 DEG C of night temperature, intensity of illumination is
1800-2700lx cultivates 30-35d, obtains rooted seedling, then rooted seedling is moved back through hardening under conditions of light application time is 10-11h
It plants.
The present invention proposes a kind of RADIX PRZEWALSKIAE TANGUTICAE seedling, is made by above-mentioned RADIX PRZEWALSKIAE TANGUTICAE method for culturing seedlings.
The beneficial effect of RADIX PRZEWALSKIAE TANGUTICAE seedling and its method for culturing seedlings provided in an embodiment of the present invention is: this method is chosen using artificial
Wild plant with merit combines imitative wild environment to cultivate, makes to obtain by virus-elimination seedlings tissue culture rapid propagating technology
RADIX PRZEWALSKIAE TANGUTICAE seedling not only inherit the merit of former plant, without virus, stable yield, and resistance is good, adaptable.
Specific embodiment
It in order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below will be in the embodiment of the present invention
Technical solution be clearly and completely described.The person that is not specified actual conditions in embodiment, according to normal conditions or manufacturer builds
The condition of view carries out.Reagents or instruments used without specified manufacturer is the conventional production that can be obtained by commercially available purchase
Product.
The RADIX PRZEWALSKIAE TANGUTICAE seedling and its method for culturing seedlings of the embodiment of the present invention are specifically described below.
A kind of RADIX PRZEWALSKIAE TANGUTICAE method for culturing seedlings comprising:
The vigorous wild RADIX PRZEWALSKIAE TANGUTICAE of growing way is chosen, its tip of a root is taken, sterilizes, obtains the tip of a root to be seeded.
Wherein, the vigorous wild RADIX PRZEWALSKIAE TANGUTICAE of growing way, with good environmental resistance and resistance.Preferably, wild
The careless age of RADIX PRZEWALSKIAE TANGUTICAE is 0.5-1, and careless age is excessively high, is not easy to carry out tissue cultures, careless age is too small, and resistance is bad.
Preferably, tip of a root disinfection carries out in the following manner: taking the tip of a root of 1-1.5cm first to rinse 5-7min with flowing water, then
40-55s is sterilized with 75% ethanol solution, residual ethanol solution is removed with aseptic water washing, is then then at mass fraction
It is impregnated 5 minutes in the mercuric chloride solution of 0.1%-0.15%, removes remaining mercuric chloride solution with aseptic water washing.Within the scope of this, can have
The bacterium on effect removal tip of a root surface, and will not influence the differentiation of the tip of a root to be seeded, the control of concentration and time are reasonable, prevent dense
Spend low, time it is too short caused by the tip of a root to be seeded caused by bactericidal effect is bad or excessive concentration, overlong time activity not
It is good.
Tip of a root 0.3-0.5mm to be seeded is cut, accesses in inductive differentiation medium and is broken up.Preferably, it is in temperature
20 DEG C, daily light application time 12h, intensity of illumination carries out under conditions of being 2000lx.Under this condition, cell differentiation speed is fast,
Differentiation culture two months, the tip of a root after obtaining enough differentiation.
Preferably, the raw material of inductive differentiation medium includes: 100 parts of MS culture medium according to parts by weight, 0.4mg/L's
NAA5-7 parts, mass fraction is 3-10 parts of coconut palm cream of 10%, 0.5-5 parts of the agar of 6g/L, 15-18 parts of the sucrose of 30g/L.It is lured
It is good to lead differentiation effect.
Specifically, using MS culture medium as minimal medium, methyl α-naphthyl acetate (NAA) is plant growth regulator, can promote cell
Division and expand, induced synthesis adventitious root increases, the methyl α-naphthyl acetates (NAA) of 5-7 parts by weight promote root-tip cells continue differentiation with
And promote the speed of differentiation and the direction of differentiation.On the one hand the mass fraction of 3-10 parts by weight provides nutrition for 10% coconut palm cream
Substance, on the other hand for promoting the differentiation of the tip of a root.Sucrose maintains osmotic pressure as carbon source.Agar makes to train as curing agent
Support gelatinization, the differentiation that 0.5-5 parts of agar of the coagulation result of 6g/L makes inductive differentiation medium be more advantageous to the tip of a root.
It is highly preferred that the raw material of inductive differentiation medium further includes the diatom powder of 5-10 parts by weight.Diatom powder has sterilization
Disinfection prevents the effect of mildew.
The tip of a root after differentiation is placed in proliferated culture medium, it is preferable that at 20 DEG C of temperature, intensity of illumination 2000lx, illumination
Between under conditions of 12h, Multiplying culture one month.
Preferably, the raw material of proliferated culture medium includes: 100 parts of MS culture medium according to parts by weight, the NAA 3- of 1.0mg/L
5 parts, 0.5-0.8 parts of the IBA of 0.5mg/L, 1-2 parts of 0.2 mg/L6-BA, mass fraction is 5-7 parts of potato juice of 10%,
2-4 parts of the agar of 5.7g/L, 10-20 parts of the sucrose of 30g/L, 4-8 part of thiocarbamide.
Wherein, the methyl α-naphthyl acetate (NAA) of 3-5 parts of parts by weight promotes the rapid proliferation of differentiated root-tip cells.Indolebutyric acid
(IBA) promote division, promote the proliferation of cell.6- benzyl aminoadenine (6-BA) also known as the basic element of cell division, 1-2 parts by weight
The 6-BA of 0.2mg/L promotes the division growth of cell.
The tip of a root after Multiplying culture is placed in root media.Preferably, in the case where imitative wild environment, specifically
Ground, in 17-27 DEG C of day temperature, 5-7 DEG C of night temperature, intensity of illumination 1800-2700lx is trained under conditions of light application time is 10-11h
30-35d is supported, rooted seedling is obtained.In the case where imitative wild environment, the growth of RADIX PRZEWALSKIAE TANGUTICAE is stimulated, its resistance is improved.
Preferably, the raw material of root media includes: 100 parts of MS culture medium according to parts by weight, the NAA 1- of 0.2mg/L
5 parts, 1-2 parts of the 6-BA of 0.2mg/L, mass fraction is 5-8 parts of coconut palm cream of 10%, 1-3 parts of the agar of 5.7g/L, diatom powder
0.5-1.5 parts, 0.1-0.3 parts of magnesium chloride.
Wherein, the 6-BA of the 0.2mg/L of 1-2 parts by weight promotes the formation of bud, and further induction root is former for indolebutyric acid (IBA)
The formation of body promotes cell differentiation and division, promotes new root to generate the differentiation with fibrovascular system, promotes the formation of adventitious root.
Vulnerable to bacterium infection during RADIX PRZEWALSKIAE TANGUTICAE in vitro culture, and sugar can stimulate bacterium, and bacterial multiplication speed is greater than horse
The urine bubble speed of growth causes RADIX PRZEWALSKIAE TANGUTICAE growing way weak, resistance is low so that the bacterium in growing environment be made to increase rapidly.Invention human hair
It is existing, sugar is substituted using coconut palm cream, while increasing diatom powder and magnesium chloride, proportions are reasonable, can not only guarantee RADIX PRZEWALSKIAE TANGUTICAE
Nutrient needed for own growth can obtain ample supply, moreover it is possible to reduce bacteria breed, improve its survival rate, and finally obtain imitative
The rooted seedling well-grown of wild environment culture, compared with the root media of addition sucrose, through acclimatization and transplants to identical
More adaptable when experimental field, tolerance and resistance to adverse circumstances are more preferably.
Preferably, the raw material of root media further includes the tea root powder of 7-15 parts by weight, for promoting new root breath.
Preferably, the raw material of root media further includes the active carbon of 0.1-1 parts by weight, prevents new root brown stain, is unfavorable for
Observation.
Rooted seedling is transplanted after hardening again.Preferably, the hardening process of rooted seedling includes in-bottle seeding and bottle epimatrix
2 links of hardening, specifically, in-bottle seeding are that the rooted seedling in root media is placed in 17-27 DEG C of temperature day temperature, night temperature 4-
7 DEG C, light intensity 1700-3000lx, illumination 10-11h/d, in the closed environment of relative air humidity 50-60% or so, open bottle
Lid, is allowed to adapt to environment 3-5d, can be completed；And external substrate seeding: after the rooted seedling for completing in-bottle seeding is taken out, plant
In the matrix seedbed that seeding room gets ready, the careless carbon of matrix formulations 1: 1: 1: vermiculite: clay, seeding room's diurnal temperature is in day temperature
17-28 DEG C, bottle can be completed through 15-20d in 3-7 DEG C of night temperature, noon light intensity 2500-3000lx, relative air humidity 40-50%
Epimatrix hardening, i.e. transplanting crop field or field.
Embodiment 1
A kind of method for culturing seedlings of RADIX PRZEWALSKIAE TANGUTICAE takes the tip of a root of 1cm first to be rushed with flowing water firstly, choosing healthy and strong wild RADIX PRZEWALSKIAE TANGUTICAE
5min is washed, then sterilizes 55s with 75% ethanol solution, residual ethanol solution is removed with aseptic water washing, then then at quality
It is impregnated 5 minutes in the mercuric chloride solution that score is 0.15%, removes remaining mercuric chloride solution with aseptic water washing, obtain to be seeded
Point.Then, the tip of a root to be seeded is cut into 0.3mm, accessed in inductive differentiation medium, be 20 DEG C in temperature, intensity of illumination
Under conditions of 2000lx, daily light application time 12h, differentiation culture two months；Wherein, the raw material of inductive differentiation medium is by weight
Number meter includes: 100 parts of MS culture medium, NAA7 part of 0.4mg/L, 8 parts of the coconut palm cream that mass fraction is 10%, the agar 3 of 6g/L
Part, 16 parts of the sucrose of 30g/L, 6 parts of diatom powder.Secondly, the tip of a root after differentiation is placed in proliferated culture medium, at 20 DEG C of temperature, light
According to intensity 2000lx, under conditions of light application time 12h, Multiplying culture one month；Wherein, the raw material of proliferated culture medium is by weight
Number meter includes: 100 parts of MS culture medium, 1 part of the 6-BA of 0.6 part of the IBA of 5 parts of the NAA of 1.0mg/L, 0.5mg/L, 0.2mg/L,
Mass fraction is 6 parts of potato juice of 10%, 3 parts of the agar of 5.7g/L, 16 parts of the sucrose of 30g/L, 5 parts of thiocarbamide.Then, will increase
The tip of a root after growing culture is placed in root media, in 17-27 DEG C of day temperature, 5-7 DEG C of night temperature, and intensity of illumination 1800-2700lx,
Under conditions of light application time is 10h, 30-35d is cultivated, obtains rooted seedling, wherein the raw material of root media wraps according to parts by weight
It includes: 100 parts of MS culture medium, 2 parts of 3 parts of the NAA of 0.2mg/L, 0.5mg/L IBA, 7 parts of the coconut palm cream that mass fraction is 10%,
2 parts of the agar of 5.7g/L, 1.5 parts of diatom powder, 0.3 part of magnesium chloride, 9 parts of-tea root powder, 0.4 part of active carbon.Finally by rooted seedling
It is transplanted after hardening, obtains RADIX PRZEWALSKIAE TANGUTICAE seedling.
Embodiment 2
A kind of method for culturing seedlings of RADIX PRZEWALSKIAE TANGUTICAE takes the tip of a root of 1.5cm first to use flowing water firstly, choosing healthy and strong wild RADIX PRZEWALSKIAE TANGUTICAE
7min is rinsed, then sterilizes 40s with 75% ethanol solution, residual ethanol solution is removed with aseptic water washing, then then at matter
It is impregnated 5 minutes in the mercuric chloride solution that amount score is 0.1%, removes remaining mercuric chloride solution with aseptic water washing, obtain to be seeded
Point.Then, the tip of a root to be seeded is cut into 0.3mm, accessed in inductive differentiation medium, be 20 DEG C in temperature, intensity of illumination
Under conditions of 2000lx, daily light application time 12h, differentiation culture two months；Wherein, the raw material of inductive differentiation medium is by weight
Number meter includes: 100 parts of MS culture medium, NAA5 part of 0.4mg/L, 10 parts of the coconut palm cream that mass fraction is 10%, the agar 5 of 6g/L
Part, 18 parts of the sucrose of 30g/L, 10 parts of diatom powder.Secondly, the tip of a root after differentiation is placed in proliferated culture medium, at 20 DEG C of temperature, light
According to intensity 2000lx, under conditions of light application time 12h, Multiplying culture one month；Wherein, the raw material of proliferated culture medium is by weight
Number meter includes: 100 parts of MS culture medium, 6-BA2 part of 0.5 part of the IBA of 3 parts of the NAA of 1.0mg/L, 0.5mg/L, 0.2mg/L,
Mass fraction is 7 parts of potato juice of 10%, 4 parts of the agar of 5.7g/L, 10 parts of the sucrose of 30g/L, 4 parts of thiocarbamide.Then, will increase
The tip of a root after growing culture is placed in root media, in 17-27 DEG C of day temperature, 5-7 DEG C of night temperature, and intensity of illumination 1800-2700lx,
Under conditions of light application time is 10-11h, 30-35d is cultivated, obtains rooted seedling, wherein the raw material of root media is in parts by weight
Meter includes: 100 parts of MS culture medium, 1.5 parts of the IBA of 5 parts of the NAA of 0.2mg/L, 0.5mg/L, the coconut palm cream that mass fraction is 10%
5 parts, 3 parts of the agar of 5.7g/L, 1.2 parts of diatom powder, 0.15 part of magnesium chloride, 15 parts of tea root powder, 1 part of active carbon.It will finally give birth to
Offspring is transplanted after hardening, obtains RADIX PRZEWALSKIAE TANGUTICAE seedling.
Embodiment 3
A kind of method for culturing seedlings of RADIX PRZEWALSKIAE TANGUTICAE takes the tip of a root of 1.2cm first to use flowing water firstly, choosing healthy and strong wild RADIX PRZEWALSKIAE TANGUTICAE
6min is rinsed, then sterilizes 45s with 75% ethanol solution, residual ethanol solution is removed with aseptic water washing, then then at matter
It is impregnated 5 minutes in the mercuric chloride solution that amount score is 0.12%, removes remaining mercuric chloride solution with aseptic water washing, obtain to be seeded
Point.Then, the tip of a root to be seeded is cut into 0.3mm, accessed in inductive differentiation medium, be 20 DEG C in temperature, intensity of illumination
Under conditions of 2000lx, daily light application time 12h, differentiation culture two months；Wherein, the raw material of inductive differentiation medium is by weight
Number meter includes: 100 parts of MS culture medium, NAA6 part of 0.4mg/L, 5 parts of the coconut palm cream that mass fraction is 10%, the agar of 6g/L
0.5 part, 17 parts of the sucrose of 30g/L, 5 parts of diatom powder.Secondly, the tip of a root after differentiation is placed in proliferated culture medium, at 20 DEG C of temperature,
Under conditions of intensity of illumination 2000lx, light application time 12h, Multiplying culture one month；Wherein, the raw material of proliferated culture medium is by weight
Number meter includes: 100 parts of MS culture medium, 0.8 part of the IBA of 4 parts of the NAA of 1.0mg/L, 0.5mg/L, the 6-BA of 0.2mg/L
1.5 parts, mass fraction is 5 parts of potato juice of 10%, 2.5 parts of the agar of 5.7g/L, 20 parts of the sucrose of 30g/L, 6 parts of thiocarbamide.
Then, the tip of a root after Multiplying culture is placed in root media, in 17-27 DEG C of day temperature, 5-7 DEG C of night temperature, intensity of illumination is
1800-2700lx cultivates 30-35d, obtains rooted seedling, wherein the original of root media under conditions of light application time is 10-11h
Material includes: 100 parts of MS culture medium according to parts by weight, 1.7 parts of the IBA of 4 parts of the NAA of 0.2mg/L, 0.5mg/L, mass fraction
8 parts newborn, 2.5 parts of the agar of 5.7g/L for 10% coconut palm, 0.8 part of diatom powder, 0.2 part of magnesium chloride, 7 parts of tea root powder, active carbon 0.1
Part.Rooted seedling is transplanted after hardening finally, obtains RADIX PRZEWALSKIAE TANGUTICAE seedling.
Embodiment 4
A kind of method for culturing seedlings of RADIX PRZEWALSKIAE TANGUTICAE takes the tip of a root of 1.3cm first to use flowing water firstly, choosing healthy and strong wild RADIX PRZEWALSKIAE TANGUTICAE
Rinse 5.5min, then with 75% ethanol solution sterilize 50s, with aseptic water washing removal residual ethanol solution, then then at
It is impregnated 5 minutes in the mercuric chloride solution that mass fraction is 0.14%, removes remaining mercuric chloride solution with aseptic water washing, obtain to be seeded
The tip of a root.Then, the tip of a root to be seeded is cut into 0.3mm, accessed in inductive differentiation medium, be 20 DEG C in temperature, intensity of illumination
Under conditions of 2000lx, daily light application time 12h, differentiation culture two months；Wherein, the raw material of inductive differentiation medium is by weight
Number meter includes: 100 parts of MS culture medium, NAA5.5 part of 0.4mg/L, 7 parts of the coconut palm cream that mass fraction is 10%, the agar of 6g/L
2 parts, 15 parts of the sucrose of 30g/L, 8 parts of diatom powder.Secondly, the tip of a root after differentiation is placed in proliferated culture medium, at 20 DEG C of temperature, light
According to intensity 2000lx, under conditions of light application time 12h, Multiplying culture one month；Wherein, the raw material of proliferated culture medium is by weight
Number meter includes: 100 parts of MS culture medium, 0.65 part of the IBA of 3.5 parts of the NAA of 1.0mg/L, 0.5mg/L, 0.2mg/L 6-BA's
1.6 parts, mass fraction be 5.5 parts of 10% potato juice, 2 parts of 5.7g/L agar, 18 parts of the sucrose of 30g/L, 8 parts of thiocarbamide.Then,
The tip of a root after Multiplying culture is placed in root media, in 17-27 DEG C of day temperature, 5-7 DEG C of night temperature, intensity of illumination 1800-
2700lx cultivates 30-35d, obtains rooted seedling under conditions of light application time is 10-11h, wherein the raw material of root media is by weight
Measuring number meter includes: 100 parts of MS culture medium, NAA2 part of 0.2mg/L, 1.8 parts of the IBA of 0.5mg/L, mass fraction 10%
Coconut palm cream 6 parts, 1 part of the agar of 5.7g/L, 1 part of diatom powder, 0.2 part of magnesium chloride, 10 parts of tea root powder.Finally by rooted seedling through refining
It is transplanted after seedling, obtains RADIX PRZEWALSKIAE TANGUTICAE seedling.
Embodiment 5
A kind of method for culturing seedlings of RADIX PRZEWALSKIAE TANGUTICAE takes the tip of a root of 1.4cm first to use flowing water firstly, choosing healthy and strong wild RADIX PRZEWALSKIAE TANGUTICAE
6min is rinsed, then sterilizes 54s with 75% ethanol solution, residual ethanol solution is removed with aseptic water washing, then then at matter
It is impregnated 5 minutes in the mercuric chloride solution that amount score is 0.11%, removes remaining mercuric chloride solution with aseptic water washing, obtain to be seeded
Point.Then, the tip of a root to be seeded is cut into 0.3mm, accessed in inductive differentiation medium, be 20 DEG C in temperature, intensity of illumination
Under conditions of 2000lx, daily light application time 12h, differentiation culture two months；Wherein, the raw material of inductive differentiation medium is by weight
Number meter includes: 100 parts of MS culture medium, NAA6.5 part of 0.4mg/L, 4 parts of the coconut palm cream that mass fraction is 10%, the agar of 6g/L
4 parts, 16.5 parts of the sucrose of 30g/L.Secondly, the tip of a root after differentiation is placed in proliferated culture medium, at 20 DEG C of temperature, intensity of illumination
Under conditions of 2000lx, light application time 12h, Multiplying culture one month；Wherein, the raw material of proliferated culture medium wraps according to parts by weight
It includes: 100 parts of MS culture medium, NAA4.5 part of 1.0mg/L, 6-BA1.4 part of 0.55 part of the IBA of 0.5mg/L, 0.2mg/L, matter
Measuring score is 6.5 parts of potato juice of 10%, 3.5 parts of the agar of 5.7g/L, 14 parts of the sucrose of 30g/L, 7 parts of thiocarbamide.Then, will
The tip of a root after Multiplying culture is placed in root media, in 17-27 DEG C of day temperature, 5-7 DEG C of night temperature, and intensity of illumination 1800-
2700lx cultivates 30-35d, obtains rooted seedling under conditions of light application time is 10-11h, wherein the raw material of root media is by weight
Measuring number meter includes: 100 parts of MS culture medium, 2 parts of the IBA of 1 part of the NAA of 0.2mg/L, 0.5mg/L, and mass fraction is 10%
6.5 parts of coconut palm cream, 1.5 parts of the agar of 5.7g/L, 0.5 part of diatom powder, 0.1 part of magnesium chloride.Finally rooted seedling is transplanted after hardening,
Obtain RADIX PRZEWALSKIAE TANGUTICAE seedling.
Test example
The tip of a root after 1 Multiplying culture of embodiment is equally divided into 3 groups, i.e. embodiment 1, comparative example 1 and comparative example 2.By three
Group is respectively placed in root media.Wherein, the only original of the root media of culture of rootage based raw material and embodiment 1 in comparative example 1
Material is compared, and more sucrose, culture of rootage based raw materials include: 100 parts of MS culture medium according to parts by weight, the NAA 3 of 0.2mg/L
Part, 2 parts of the IBA of 0.5mg/L, 7 parts of the coconut palm cream that mass fraction is 10%, 2 parts of the agar of 5.7g/L, 9 parts of tea root powder, activity
0.4 part of charcoal, 12 parts of sucrose.The raw material of wherein comparative example 2, root media is same as Example 1, but it is in steady temperature
20 DEG C, intensity of illumination 2000lx, under conditions of light application time is 12h, cultivate 30-35d.By embodiment 1, comparative example 1 and
RADIX PRZEWALSKIAE TANGUTICAE seedling made from comparative example 2 is transplanted after same Trained Transition Methods of Chinese Dwarf into same experimental field.Qualified rooted seedling rate is
Refer to the percentage for the qualified rooted seedling and total rooted seedling that root media obtains, qualified rooted seedling refer to growing way it is luxuriant with
And leaf is green non-yellowing, not by the healthy plant of bacterium infection.Transplanting to experimental field survival rate refer to 100 comparative example rooted seedlings with
100 embodiment rooted seedlings are in same environment, after same breeding method is cultivated 1 month, the number of the rooted seedling of survival
Measure the mass percent with 100.Compare to obtain 1 data of table.
1 comparing result of table
| Qualified rooted seedling rate | It transplants to experimental field survival rate | |
| Comparative example 1 | 66% | 89% |
| Comparative example 2 | 72% | 67% |
| Embodiment 1 | 68% | 96% |
It can be obtained by table 1, in imitative field environment, the rooted seedling rate for adding the root media qualification of sucrose, which is greater than, to be not added with
The root media culture of sucrose, and when these rooted seedlings move to crop field, the survival rate for the rooted seedling that embodiment 1 provides are greater than pair
The rooted seedling that ratio 1 provides.And its Comprehensive Traits of the rooted seedling of embodiment 1 are better than comparative example 2, therefore the life that embodiment 1 provides
The adaptive faculty of offspring is more excellent.
The RADIX PRZEWALSKIAE TANGUTICAE seedling and its method for culturing seedlings of the offer of the embodiment of the present invention, use and manually choose excellent Wild plant,
It combines imitative wild environment to cultivate by virus-elimination seedlings tissue culture rapid propagating technology, seedling offspring is made not only to inherit the excellent of former plant
Benign shape, without virus, stable yield, and resistance is good, adaptable.
Embodiments described above is a part of the embodiment of the present invention, instead of all the embodiments.Reality of the invention
The detailed description for applying example is not intended to limit the range of claimed invention, but is merely representative of selected implementation of the invention
Example.Based on the embodiments of the present invention, obtained by those of ordinary skill in the art without making creative efforts
Every other embodiment, shall fall within the protection scope of the present invention.
Claims (6)
1. a kind of RADIX PRZEWALSKIAE TANGUTICAE method for culturing seedlings, characterized in that it comprises:
The vigorous wild RADIX PRZEWALSKIAE TANGUTICAE of growing way is chosen, its tip of a root is taken, sterilizes, obtains the tip of a root to be seeded；
It cuts the tip of a root 0.3-0.5mm to be seeded, accesses in inductive differentiation medium, be 20 DEG C in temperature, when daily illumination
Between 12h, intensity of illumination be 2000lx under conditions of, differentiation culture two months；
The tip of a root after differentiation is placed in proliferated culture medium, at 20 DEG C of temperature, intensity of illumination 2000lx, the condition of light application time 12h
Under, Multiplying culture one month；
The tip of a root after Multiplying culture is placed in root media, in 17-27 DEG C of day temperature, 5-7 DEG C of night temperature, intensity of illumination is
1800-2700lx cultivates 30-35d, obtains rooted seedling, then by the rooted seedling through hardening under conditions of light application time is 10-11h
After transplant；
The raw material of the root media includes: 100 parts of MS culture medium according to parts by weight, 1-5 parts of the NAA of 0.2mg/L,
1-2 parts of the IBA of 0.5mg/L, mass fraction are 5-8 parts of coconut palm cream of 10%, 1-3 parts of the agar of 5.7g/L, diatom powder 0.5-1.5
Part, 0.1-0.3 parts of magnesium chloride；
The raw material of the inductive differentiation medium includes: 100 parts of MS culture medium according to parts by weight, NAA5-7 part of 0.4mg/L,
Mass fraction is 3-10 parts of coconut palm cream of 10%, 0.5-5 parts of the agar of 6g/L, 15-18 parts of the sucrose of 30g/L；
The raw material of the proliferated culture medium includes: 100 parts of MS culture medium according to parts by weight, 3-5 parts of the NAA of 1.0mg/L,
1-2 parts of the 6-BA of 0.5-0.8 parts of the IBA of 0.5mg/L, 0.2mg/L, mass fraction are 5-7 parts of potato juice of 10%,
2-4 parts of the agar of 5.7g/L, 10-20 parts of the sucrose of 30g/L, 4-8 parts of thiocarbamide.
2. RADIX PRZEWALSKIAE TANGUTICAE method for culturing seedlings according to claim 1, which is characterized in that the raw material of the root media further includes
The tea root powder of 7-15 parts by weight.
3. RADIX PRZEWALSKIAE TANGUTICAE method for culturing seedlings according to claim 1, which is characterized in that the raw material of the root media further includes
The activated carbon of 0.1-1 parts by weight.
4. RADIX PRZEWALSKIAE TANGUTICAE method for culturing seedlings according to claim 1, which is characterized in that the raw material of the inductive differentiation medium is also
Diatom powder including 5-10 parts by weight.
5. RADIX PRZEWALSKIAE TANGUTICAE method for culturing seedlings according to claim 1, which is characterized in that tip of a root disinfection in the following manner into
Row: taking the tip of a root of 1-1.5cm first to rinse 5-7min with flowing water, then 40-55s is sterilized with 75% ethanol solution, with nothing
Bacterium water washes off the remaining ethanol solution, then impregnates 5 in the mercuric chloride solution that mass fraction is 0.1%-0.15%
Minute, with the remaining mercuric chloride solution of aseptic water washing removal.
6. RADIX PRZEWALSKIAE TANGUTICAE method for culturing seedlings according to claim 1, which is characterized in that it is 0.5- that the wild RADIX PRZEWALSKIAE TANGUTICAE, which is careless age,
1 year wild RADIX PRZEWALSKIAE TANGUTICAE.
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| CN102893866A (en) * | 2012-10-15 | 2013-01-30 | 长阳勤劳农夫农产品有限公司 | Strawberry root tip detoxification and tissue culture method |
| CN104823852A (en) * | 2015-05-08 | 2015-08-12 | 长阳勤劳农夫农产品有限公司 | Dendrobium officinale rapid breeding method through root tip tissue culture |
| CN104855287A (en) * | 2015-05-11 | 2015-08-26 | 中国科学院西北高原生物研究所 | Establishment method of rapid propagation system of Tibetan drug przewalskia tangutica maxim |
| CN104855288A (en) * | 2015-05-11 | 2015-08-26 | 中国科学院西北高原生物研究所 | Establishment method of Tibetan drug przewalskia tangutica maxim autonomous rooting system |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102893866A (en) * | 2012-10-15 | 2013-01-30 | 长阳勤劳农夫农产品有限公司 | Strawberry root tip detoxification and tissue culture method |
| CN104823852A (en) * | 2015-05-08 | 2015-08-12 | 长阳勤劳农夫农产品有限公司 | Dendrobium officinale rapid breeding method through root tip tissue culture |
| CN104855287A (en) * | 2015-05-11 | 2015-08-26 | 中国科学院西北高原生物研究所 | Establishment method of rapid propagation system of Tibetan drug przewalskia tangutica maxim |
| CN104855288A (en) * | 2015-05-11 | 2015-08-26 | 中国科学院西北高原生物研究所 | Establishment method of Tibetan drug przewalskia tangutica maxim autonomous rooting system |
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