CN106717260A - The method for culturing seedlings of flower of Radix Gentianae and the implantation methods of flower of Radix Gentianae - Google Patents
The method for culturing seedlings of flower of Radix Gentianae and the implantation methods of flower of Radix Gentianae Download PDFInfo
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- CN106717260A CN106717260A CN201611072300.6A CN201611072300A CN106717260A CN 106717260 A CN106717260 A CN 106717260A CN 201611072300 A CN201611072300 A CN 201611072300A CN 106717260 A CN106717260 A CN 106717260A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G9/00—Cultivation in receptacles, forcing-frames or greenhouses; Edging for beds, lawn or the like
- A01G9/14—Greenhouses
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/25—Greenhouse technology, e.g. cooling systems therefor
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Abstract
The present invention relates to the method for culturing seedlings and the implantation methods of flower of Radix Gentianae of flower of Radix Gentianae,Belong to plant production field,It is removed the peel and is rinsed with the liquor natrii hypochloritis that mass concentration is 10% 20%,It is 0.1 0.5mg/L heteroauxins that peeling seed is inoculated in containing perlite and mass concentration,The 6 benzyl aminoadenines of 0.5 1mg/L,In the MS culture mediums of the EM bacterium of 0.01 0.05mg/L,15 25 DEG C of cultivation temperature,Irradiating 10 12h in 2000 3000lx intensities of illumination daily carries out Initial culture,The aseptic seedling that will be obtained is inoculated in subculture medium,To obtain being transferred in root media without offspring again,Obtain flower of Radix Gentianae seedling,After flower of Radix Gentianae seedling is trained,Training seedling is cultivated in greenhouse,Daily 10 12h are irradiated under 2000 3000lx illumination,Coordinate field management,Obtain ripe flower of Radix Gentianae,Its emergence rate and survival rate of seedling are high.
Description
Technical field
The present invention relates to a kind of plant production field, and more particularly to a kind of flower of Radix Gentianae method for culturing seedlings and the kind of flower of Radix Gentianae
Plant method.
Background technology
Flower of Radix Gentianae, another name：Ground courage head, felwort, Premna microphylla Turez etc..In flower of Radix Gentianae big bitter, Great Cold, it is only second among Chinese medicine bitter taste
Vomiting nut, is better than the coptis, occupies second, multiplex to upset one's stomach, and is difficult entrance, helps with Radix Glycyrrhizae, to adjust taste, is usually used in treatment obstinate
Property antimigraine, head eczema, hypertension, acute conjunctivitis, rhinitis etc..
Because flower of Radix Gentianae has splendid medical value, in the market constantly increases the demand of flower of Radix Gentianae, and
The demand in market is only met by plucking wild flower of Radix Gentianae, not only harvesting amount does not reach demand, and excessive harvesting also results in rough gentian
Colored existence is dangerous.Furthermore, flower of Radix Gentianae is grown in height above sea level domain higher, and the condition requirement of its growth is harsh, artificial breeding, cultivation
Difficulty is larger, and further result in the yield of flower of Radix Gentianae cannot meet the demand in market.
The content of the invention
It is an object of the invention to provide a kind of method for culturing seedlings of flower of Radix Gentianae, after its seed peeling by flower of Radix Gentianae, artificially
Help seed the step of complete broken skin, shorten the time of seed germination；Perlite is added in Initial culture base to ensure
Seed can carry out good breathing in Initial culture base, and the pH of Initial culture base is kept suitable acidity, more added with
Beneficial to emerging for seed；EM bacterium make aseptic seedling more sturdy, increase survival rate；By after squamous subculture and culture of rootage, obtaining
The flower of Radix Gentianae seedling for arriving, its incubation time from seed to seedling is short, and can effectively lift emergence rate and survival rate.
Another object of the present invention is to provide a kind of implantation methods of flower of Radix Gentianae, by the seedling of flower of Radix Gentianae by domestication
Afterwards, it is easier to adapt to planting environment, the survival rate of flower of Radix Gentianae seedling is improved, lifts the plantation yield of flower of Radix Gentianae, meet market
Demand, slows down the demand pressure of wild flower of Radix Gentianae, and the extinction of flower of Radix Gentianae is avoided to a certain extent.
The present invention is solved its technical problem and is realized using following technical scheme.
A kind of method for culturing seedlings of flower of Radix Gentianae, its be by the seed of flower of Radix Gentianae be soaked in mass concentration for 10%-20% time
Sodium chlorate solution is removed the peel, and is rinsed with sterilized water after peeling, obtains removing the peel seed；Peeling seed is inoculated into primary
In culture medium, Initial culture is carried out, obtain aseptic seedling；Using the cotyledon of aseptic seedling and hypocotyl as explant, and be inoculated in after
In for culture medium, squamous subculture is carried out, obtained without offspring；To turn to plant in root media without offspring, carry out culture of rootage, obtain
To flower of Radix Gentianae seedling；The temperature of Initial culture is 15-25 DEG C, and intensity of illumination is 2000-3000lx, and light application time is 10-12h,
Initial culture base is that to be added with perlite and mass concentration be heteroauxin, the 6- benzyl ammonia of 0.5-1mg/L of 0.1-0.5mg/L
The MS culture mediums of the EM bacterium of base adenine and 0.01-0.05mg/L.
A kind of implantation methods of flower of Radix Gentianae, it is to cultivate flower of Radix Gentianae seedling according to the method for culturing seedlings of above-mentioned flower of Radix Gentianae to tame and docile
Change, obtain taming seedling；Domestication seed is planted in greenhouse, controlling the light application time in greenhouse for 10-12h, intensity of illumination is
2000-3000lx, coordinates field management, to the flower of Radix Gentianae for obtaining maturation.
The beneficial effect of the method for culturing seedlings of flower of Radix Gentianae provided in an embodiment of the present invention and the implantation methods of flower of Radix Gentianae is：Pass through
Peeling treatment to rough gentian flower seed, accelerates the speed of flower of Radix Gentianae seed germination, and by adding pearl in Initial culture base
Rock, can be conducive to the breathing of seed, while in turn ensure that the suitable acidity of Initial culture base pH, further speed up going out for seed
Bud；EM bacterium can make aseptic seedling more sturdy, increase the survival rate of aseptic seedling；In a series of of squamous subculture and culture of rootage
In the presence of tissue cultures, the emergence rate and survival rate of rough gentian flower seed are lifted, and strengthen the adaptability of flower of Radix Gentianae seedling；
By flower of Radix Gentianae seedling by being planted again after domestication, the survival rate of flower of Radix Gentianae seedling can be further lifted, increase flower of Radix Gentianae
Yield, meet the demand in market, reduce the harvesting amount of wild flower of Radix Gentianae, it is to avoid the danger of extinction occurs in flower of Radix Gentianae.
Specific embodiment
To make the purpose, technical scheme and advantage of the embodiment of the present invention clearer, below will be in the embodiment of the present invention
Technical scheme be clearly and completely described.Unreceipted actual conditions person, builds according to normal condition or manufacturer in embodiment
The condition of view is carried out.Agents useful for same or the unreceipted production firm person of instrument, are the conventional product that can be obtained by commercially available purchase
Product.
The method for culturing seedlings of flower of Radix Gentianae and the implantation methods of flower of Radix Gentianae to the embodiment of the present invention are specifically described below.
The embodiment of the present invention provides a kind of method for culturing seedlings of flower of Radix Gentianae, and it is the seed to flower of Radix Gentianae with liquor natrii hypochloritis
Removed the peel, be rinsed with sterilized water after peeling, obtain removing the peel seed；Above-mentioned peeling seed by Initial culture, after
It is commissioned to train after foster and culture of rootage, you can obtain flower of Radix Gentianae seedling.
The embodiment of the present invention also provides a kind of implantation methods of flower of Radix Gentianae, and it is by domestication by above-mentioned flower of Radix Gentianae seedling
Afterwards, obtain taming seedling；Domestication seed is planted in greenhouse, controlling the light application time in greenhouse for 10-12h, intensity of illumination is
2000-3000lx, coordinates field management, to the flower of Radix Gentianae for obtaining maturation.
Specifically, the implantation methods of the method for culturing seedlings of flower of Radix Gentianae and flower of Radix Gentianae, are carried out according to following steps：
S1 peeling steps：
The seed of flower of Radix Gentianae is put into the container that can be sealed, such as：The plastic bottle of the PVC materials with bottle cap, then
To the liquor natrii hypochloritis that mass concentration is 10%-20% is poured into the container for having filled seed, said vesse is sealed, use the matter
Amount concentration is liquor natrii hypochloritis's immersion seed 5-10min of 10%-20%.Liquor natrii hypochloritis can be soft de- kind of skin
Fall, in addition to it can be removed the peel to seed, can also be sterilized to seed disinfection, it is ensured that tissue culture procedures after can be into
The carrying out of work(, it is ensured that seed emergence rate higher.
Preferably, during being soaked, seed can have been filled and liquor natrii hypochloritis's container shakes to above-mentioned
Swing, under the double action of earthquake and liquor natrii hypochloritis, the aeration level of the crust of seed can increased.
Further, before pouring into liquor natrii hypochloritis in said vesse, several stones can also be in a reservoir put into
Son, after liquor natrii hypochloritis being filled with container and starts concussion, the stone of the inside also can together vibrate with vibration force, stone
In the collision of son and seed, part kind skin can be made to loosen faster, the work that the kind skin after being easy to is removed completely.
Specifically, soak after completing, the liquor natrii hypochloritis in removing container, and the seed for completing will be soaked and be placed in net
In sieve, while using aseptic water washing seed, while seed is pressed against on mesh screen being rubbed, the crust of seed is set quickly to take off
Fall, and after the crust of seed comes off, aseptic water washing 20-30min is used in continuation, obtains removing the peel seed.With sterilized water to secondary
The seed that sodium chlorate solution soaked carries out the flushing of long period, can be effectively prevented from sodium hypochlorite to seed belt to damage
Wound, it is ensured that bud ratio higher.
It is highly preferred that the seed for completing can also be removed the peel being laid on blotting paper, and send into ultraviolet-sterilization case and carry out purple
Outer induction, the ultraviolet wavelength of above-mentioned ultraviolet induction is, for example, 250-254nm, and the time control of ultraviolet induction is 2-3h.Pass through
Above-mentioned ultraviolet induction, not only can further give the peeling degerming sterilization of seed, can also further improve the emergence rate of seed.
S2 Initial cultures:
The seed of peeling is inoculated in Initial culture base, Initial culture is carried out, aseptic seedling is obtained.
Above-mentioned Initial culture base is that addition perlite and mass concentration are the indoles of 0.1-0.5mg/L in MS culture mediums
The 6- benzyl aminoadenines (GA of acetic acid (IAA), 0.5-1mg/L3), the EM bacterium of 0.01-0.05mg/L prepare, above-mentioned treasure
The addition of Zhu Yan be every 1L above-mentioned MS culture mediums in add 10-20g, and the particle diameter of the perlite is 6-8mm；Above-mentioned MS
Culture medium is the commercially available MS culture mediums containing sucrose and agar.Heteroauxin can stimulate the division of cambial cell, stimulate branch
Cell elongation, suppress the cell growth of root, adjust the morphogenesis of callus, make an addition to and can promote in MS culture mediums
Skin seed quickly sprouts, and lifts the bud ratio of peeling seed, but the assurance to the amount of the heteroauxin for adding is very heavy
Will, if the amount of the heteroauxin of addition can excessively suppress sprouting for seed on the contrary；6- benzyls aminoadenine can improve differentiation
And resistance, promote sprouting for seed, the amount of the amount more than heteroauxin of the 6- benzyl aminoadenines added in MS culture mediums
Can further promote to remove the peel sprouting for seed；The beneficial mushroom such as Bacillus, saccharomycete and lactic acid bacteria, energy are included in EM bacterium
Enough strengthen the metabolism of plant, promote aseptic seedling more sturdy, improve survival rate；Make an addition to the perlite in culture medium, energy
It is enough to provide enough oxygen to the seed for being inoculated in culture medium, it is ensured that seed can carry out enough exhaling in being inoculated in culture medium
Inhale, if seed cannot carry out normal respiration, also result in the reduction of germination rate, additionally it is possible to keep Initial culture base
PH acidity, further promotes the rudiment of seed.Heteroauxin, 6- benzyls aminoadenine and when being used in mixed way of EM bacterium, every kind of thing
The influence power of confrontation seed germination rate is improved, and increases the survival rate of aseptic seedling, obtains the germination rate of peeling seed
To further being lifted.
Cultivation temperature during Initial culture is 15-25 DEG C, and intensity of illumination is 2000-3000lx, and light application time is 10-12h,
And ambient humidity during culture is 55%-75%.Because the growth of flower of Radix Gentianae is higher to humidity and the requirement of the condition of illumination,
When Initial culture is carried out in order to improve the bud ratio of seed, by the conditional regulatory of culture into the weather bar for meeting its growth
Warmer during part, i.e. Initial culture, illumination is weaker, and light application time is more long, and ambient humidity is higher, lifts the rudiment of seed
Rate.
S3 squamous subcultures:
After by Initial culture, the cotyledon of the aseptic seedling for obtaining and hypocotyl are inoculated in subculture medium as explant,
Squamous subculture is carried out, is obtained without offspring.
It is that the cotyledon of the aseptic seedling of Initial culture is cut into blade tip, leaves petiole, and by petiole be cut into 0.3cm ×
The fritter of 0.3cm；The hypocotyl of aseptic seedling is cut into the segment of 0.3cm；By above-mentioned cotyledon piece and hypocotyl section be inoculated in after
For on culture medium, squamous subculture is carried out.To again be trained after the cotyledon of the aseptic seedling from seed development out and hypocotyl cutting
Support, can effectively increase the yield of flower of Radix Gentianae seedling, a seed is just cultivated many seedlings of high-quality, greatly
Lift the yield of seedling.
The culture medium of squamous subculture for example can be that the 6- benzyls that mass concentration is 0.1-0.5mg/L are added in MS culture mediums
Gibberellin (the GA of aminoadenine (6-BA), 1-1.5mg/L3) and the heteroauxin (IAA) of 0.5-1mg/L after obtain, its
In, above-mentioned MS culture mediums are the commercially available MS culture mediums containing sucrose and agar.6- benzyls aminoadenine can promote cell division,
Promote cell increase getting fat, promote the elongation growth of cauline leaf, suppress antibody Monoclonal ability that is aging, improving plant etc., in explant
Culture in, addition 6-BA can effectively lift differentiation and the resistance of explant, can both improve the emergence rate of explant；
IAA can stimulate the division of cambial cell, stimulate the cell elongation of branch, suppress the cell growth of root, regulation callus
Morphogenesis, make an addition to can promote in MS culture mediums it is quick formed without offspring, lift emergence rate；GA3Accelerate cell division, promote
Enter the elongation growth of stem, leaf, accelerate the growth without offspring.In the presence of above-mentioned three Plant Hormone, can act synergistically
In emerging for explant, the emergence rate of explant is further lifted.
The cultivation temperature of squamous subculture is preferably 15-25 DEG C, and intensity of illumination is 2000-3000lx, and light application time is 10-
12h, and the ambient humidity of culture is 55%-75%, its temperature, illumination condition and ambient humidity are close to the original producton location of flower of Radix Gentianae
Warm, dim light, the environmental condition of relatively moistening, ensure that the emergence rate of explant, and lift the nothing that explant is cultivated
The resistance of offspring, makes that the growing way without offspring is stronger, and survival rate is higher.
S4 culture of rootage：
Planted what squamous subculture was obtained in root media without offspring turn, culture of rootage is carried out, until obtaining flower of Radix Gentianae
Seedling.
Squamous subculture is that the methyl α-naphthyl acetate (NAA) and 0.5-1mg/L that mass concentration is 1-1.5mg/L are added in MS culture mediums
Kinetin (KT) after obtain, above-mentioned MS culture mediums are similarly the commercially available MS culture mediums containing sucrose and agar.NAA is one
The plant growth regulator for promoting plant root growth is planted, with the effect for promoting cell division with expand, can be made without offspring
Quickly grow adventitious root.KT can not only promote cell division, with the differentiation of induced bud and development and can increase stomata
Aperture, so as to lift Phytoextraction carbon dioxide, increases the photosynthesis of plant, to have more Nutrients uptakes without offspring
Growth and root of hair.
Temperature during culture of rootage is 15-25 DEG C, and before being sent out roots without offspring, carries out shading treatment, is conducive to nothing
Offspring is taken root, and ambient humidity is 55%-75% during culture of rootage；Meanwhile, relatively low temperature and humidity condition is close to flower of Radix Gentianae
The height above sea level in original producton location environment higher, not only contributes to taking root without offspring, can also lift the resistance of seedling, increases seedling
Survival rate.
The plantation of S5 flower of Radix Gentianae：
After the flower of Radix Gentianae seedling cultivated according to the method described above is by domestication, obtain taming seedling, then domestication seed is planted
In in greenhouse, by a series of field management, the flower of Radix Gentianae of maturation is obtained.
Above-mentioned domestication refers to by flower of Radix Gentianae seedling cultivation in illumination box, intensity of illumination to be set into 2000-
3000lx, and light application time is 10-12h, in whole nurturing period of the seedling in illumination box, the humidity of illumination box with
1h is interval, is alternately set by 70% and 50%.By carrying out to seedling the alternately cultivation of high humidity environment and low-humidity environment
Resistance trains, and enables flower of Radix Gentianae while the of a relatively high growing environment of the humidity for adapting to low altitude area and high altitude localities
The relatively low growing environment of humidity, the survival ability that further lifting flower of Radix Gentianae seedling enters after greenhouse gardening.And it is above-mentioned
Seedling is transplanted in illumination condition of the illumination condition close to flower of Radix Gentianae original producton location, it is ensured that the vigorous growth gesture of flower of Radix Gentianae seedling, lifting
Survival rate, also promote flower of Radix Gentianae seedling can preferably adapt to different humidity resistance train.Have passed through under different humidity
Flower of Radix Gentianae after resistance training to transplant its ratio of stock after greenhouse higher, so as to lift the yield of flower of Radix Gentianae.
When flower of Radix Gentianae is planted in greenhouse, intensity of illumination is preferably arranged to 2000-3000lx, and light application time is 10-12h,
And the growth that flower of Radix Gentianae is more vigorous can be enable with the use of farm manure, the work such as cut weeds, its yield can be carried
Rise.
Method for culturing seedlings and the implantation methods of flower of Radix Gentianae with reference to embodiments to flower of Radix Gentianae of the invention makees further
Describe in detail.
Embodiment 1
Rough gentian flower seed is fitted into PVC plastic bottle, and a small amount of stone is put into above-mentioned PVC plastic bottle, be then injected into
Mass concentration is 10% liquor natrii hypochloritis, covers tightly bottle cap, after soaking and shaking 10min, liquor natrii hypochloritis is removed, by bottle
In seed, pour into mesh screen, with aseptic water washing, rinse and rubbed while seed is pressed against on mesh screen, until will plant
Skin is all removed, and continues to use aseptic water washing 20min.
The seed that the peeling of water will be blotted is put into ultraviolet-sterilization case, and 3h is processed under the wavelength of 250nm.
Peeling seed is inoculated in and contains 0.1mg/L heteroauxins, the 6- benzyls aminoadenine of 0.5mg/L, 0.05mg/L
EM bacterium and 10g, size for 6mm perlite Initial culture base in, temperature be 25 DEG C, intensity of illumination be 2000lx, light
It is that 10h and humidity are to be cultivated under conditions of 55% according to the time, until obtaining aseptic seedling.
The cotyledon of aseptic seedling is cut into the fritter of 0.3cm × 0.3cm, hypocotyl it is cut into the segment of 0.3cm and is inoculated with respectively
In the subculture medium of 6- benzyls aminoadenine, 1mg/L gibberellin and 0.5mg/L heteroauxins containing 0.1mg/L, in temperature
It is 15 DEG C, intensity of illumination are 2000lx, light application time is 12h and culture ambient humidity to cultivate under the conditions of 55% to spend, directly
To obtaining without offspring.
By in the root media of the kinetin transferred in the methyl α-naphthyl acetate containing 1mg/L and 0.5mg/L without offspring, in temperature
It it is 15 DEG C, ambient humidity is 55%, is cultivated under conditions of shading, until obtaining flower of Radix Gentianae seedling.
Flower of Radix Gentianae seedling is cultivated in illumination box, intensity of illumination is 2000lx, light application time is 12h, and light
Humidity according to incubator is interval with 1h, is alternately set according to 70% and 50%, until obtaining taming seedling.
Domestication seed is planted in greenhouse, it is ensured that the intensity of illumination in greenhouse reaches 2000lx, and light application time reaches 10h,
And the flower of Radix Gentianae of maturation is planted out in the work such as compounding application farm manure and weeding.
Embodiment 2
Rough gentian flower seed is fitted into PVC plastic bottle, and a small amount of stone is put into above-mentioned PVC plastic bottle, be then injected into
Mass concentration is 20% liquor natrii hypochloritis, covers tightly bottle cap, after soaking and shaking 5min, liquor natrii hypochloritis is removed, by bottle
In seed, pour into mesh screen, with aseptic water washing, rinse and rubbed while seed is pressed against on mesh screen, until will plant
Skin is all removed, and continues to use aseptic water washing 30min.
The seed that the peeling of water will be blotted is put into ultraviolet-sterilization case, and 2h is processed under the wavelength of 254nm.
Seed will be removed the peel and be inoculated in and contain 0.5mg/L heteroauxins, the 6- benzyls aminoadenine of 1mg/L, 0.01mg/L
During EM bacterium and 20g, size are for the Initial culture base of the perlite of 8mm, temperature be 15 DEG C, intensity of illumination be 3000lx, illumination
Time is that 12h and humidity are to be cultivated under conditions of 75%, until obtaining aseptic seedling.
The cotyledon of aseptic seedling is cut into the fritter of 0.3cm × 0.3cm, hypocotyl it is cut into the segment of 0.3cm and is inoculated with respectively
In the subculture medium of 6- benzyls aminoadenine, 1.5mg/L gibberellin and 1mg/L heteroauxins containing 0.5mg/L, in temperature
It is 25 DEG C, intensity of illumination are 3000lx, light application time is 10h and culture ambient humidity to cultivate under the conditions of 75% to spend, directly
To obtaining without offspring.
By in the root media of the kinetin transferred in the methyl α-naphthyl acetate containing 1.5mg/L and 1mg/L without offspring, in temperature
It it is 25 DEG C, ambient humidity is 75%, is cultivated under conditions of shading, until obtaining flower of Radix Gentianae seedling.
Flower of Radix Gentianae seedling is cultivated in illumination box, intensity of illumination is 3000lx, light application time is 10h, and light
Humidity according to incubator is interval with 1h, is alternately set according to 70% and 50%, until obtaining taming seedling.
Domestication seed is planted in greenhouse, it is ensured that the intensity of illumination in greenhouse reaches 3000lx, and light application time reaches 12h,
And the flower of Radix Gentianae of maturation is planted out in the work such as compounding application farm manure and weeding.
Embodiment 3
Rough gentian flower seed is fitted into PVC plastic bottle, and a small amount of stone is put into above-mentioned PVC plastic bottle, be then injected into
Mass concentration is 15% liquor natrii hypochloritis, covers tightly bottle cap, after soaking and shaking 6min, liquor natrii hypochloritis is removed, by bottle
In seed, pour into mesh screen, with aseptic water washing, rinse and rubbed while seed is pressed against on mesh screen, until will plant
Skin is all removed, and continues to use aseptic water washing 25min.
The seed that the peeling of water will be blotted is put into ultraviolet-sterilization case, and 2.5h is processed under the wavelength of 252nm.
Peeling seed is inoculated in and contains 0.3mg/L heteroauxins, the 6- benzyls aminoadenine of 0.8mg/L, 0.03mg/L
EM bacterium and 15g, size for 7mm perlite Initial culture base in, temperature be 20 DEG C, intensity of illumination be 2500lx, light
It is that 11h and humidity are to be cultivated under conditions of 65% according to the time, until obtaining aseptic seedling.
The cotyledon of aseptic seedling is cut into the fritter of 0.3cm × 0.3cm, hypocotyl it is cut into the segment of 0.3cm and is inoculated with respectively
In the subculture medium of 6- benzyls aminoadenine, 1.2mg/L gibberellin and 0.8mg/L heteroauxins containing 0.3mg/L,
Temperature is 20 DEG C, intensity of illumination is that 2500lx, light application time are 11h and the ambient humidity of culture is to cultivate under the conditions of 65%,
Until obtaining without offspring.
By in the root media of the kinetin transferred in the methyl α-naphthyl acetate containing 1.2mg/L and 0.7mg/L without offspring, in temperature
It is 20 DEG C to spend, and ambient humidity is 65%, is cultivated under conditions of shading, until obtaining flower of Radix Gentianae seedling.
Flower of Radix Gentianae seedling is cultivated in illumination box, intensity of illumination is 2500lx, light application time is 11h, and light
Humidity according to incubator is interval with 1h, is alternately set according to 70% and 50%, until obtaining taming seedling.
Domestication seed is planted in greenhouse, it is ensured that the intensity of illumination in greenhouse reaches 2500lx, and light application time reaches 11h,
And the flower of Radix Gentianae of maturation is planted out in the work such as compounding application farm manure and weeding.
Embodiment 4
Rough gentian flower seed is fitted into PVC plastic bottle, and a small amount of stone is put into above-mentioned PVC plastic bottle, be then injected into
Mass concentration is 16% liquor natrii hypochloritis, covers tightly bottle cap, after soaking and shaking 9min, liquor natrii hypochloritis is removed, by bottle
In seed, pour into mesh screen, with aseptic water washing, rinse and rubbed while seed is pressed against on mesh screen, until will plant
Skin is all removed, and continues to use aseptic water washing 24min.
The seed that the peeling of water will be blotted is put into ultraviolet-sterilization case, and 2.2h is processed under the wavelength of 251nm.
Peeling seed is inoculated in and contains 0.2mg/L heteroauxins, the 6- benzyls aminoadenine of 0.8mg/L, 0.02mg/L
EM bacterium and 12g, size for 6mm perlite Initial culture base in, temperature be 18 DEG C, intensity of illumination be 2700lx, light
It is that 12h and humidity are to be cultivated under conditions of 68% according to the time, until obtaining aseptic seedling.
The cotyledon of aseptic seedling is cut into the fritter of 0.3cm × 0.3cm, hypocotyl it is cut into the segment of 0.3cm and is inoculated with respectively
In the subculture medium of 6- benzyls aminoadenine, 1.3mg/L gibberellin and 0.8mg/L heteroauxins containing 0.2mg/L,
Temperature is 22 DEG C, intensity of illumination is that 2700lx, light application time are 11.5h and the ambient humidity of culture is to train under the conditions of 68%
Support, until obtaining without offspring.
By in the root media of the kinetin transferred in the methyl α-naphthyl acetate containing 1.4mg/L and 0.7mg/L without offspring, in temperature
It is 22 DEG C to spend, and ambient humidity is 68%, is cultivated under conditions of shading, until obtaining flower of Radix Gentianae seedling.
Flower of Radix Gentianae seedling is cultivated in illumination box, intensity of illumination is 2700lx, light application time is 12h, and light
Humidity according to incubator is interval with 1h, is alternately set according to 70% and 50%, until obtaining taming seedling.
Domestication seed is planted in greenhouse, it is ensured that the intensity of illumination in greenhouse reaches 2700lx, and light application time reaches 12h,
And the flower of Radix Gentianae of maturation is planted out in the work such as compounding application farm manure and weeding.
The Initial culture bud ratio of comparing embodiment 1, embodiment 2, embodiment 3, embodiment 4 and control group 1, wherein compareing
The processing mode of group 1 is similar with embodiment, only difference is that, without addition EM bacterium in the Initial culture base of control group 1.
The survival rate of aseptic seedling=(quantity of the quantity/seed of aseptic seedling) × 100%, above-mentioned 5 groups for the treatment of are according to 100 seed meters
Calculate.Comparing result is shown in Table 1.
The emergence rate of the seed of table 1
| Group number | Embodiment 1 | Embodiment 2 | Embodiment 3 | Embodiment 4 | Control group 1 |
| Emergence rate | 90% | 93% | 91% | 89% | 68% |
It can be seen from comparing result, by 4 groups of emergence rates of embodiment of peeling treatment apparently higher than not carrying out at peeling
The control group 1 of reason.Illustrate, when nursery is carried out, EM bacterium to be with the addition of in Initial culture base, can significantly improve aseptic seedling
Survival rate.
The Initial culture bud ratio of comparing embodiment 1, embodiment 2, embodiment 3, embodiment 4 and control group 2, wherein compareing
The processing mode of group 2 is similar with embodiment, only difference is that, perlite is not added with Initial culture base.Emergence rate=
(quantity of the quantity/seed of aseptic seedling) × 100%, above-mentioned 5 groups for the treatment of are calculated according to 100 seeds.Comparing result is shown in Table
2。
The emergence rate of the seed of table 2
| Group number | Embodiment 1 | Embodiment 2 | Embodiment 3 | Embodiment 4 | Control group 2 |
| Emergence rate | 90% | 93% | 91% | 89% | 72% |
It can be seen from comparing result, the emergence rate that perlite seed is with the addition of in Initial culture base is higher than to be not added with pearl
The control group 2 of rock.Illustrate that perlite is with the addition of in Initial culture base can be conducive to the respiration of seed, raising is emerged
Rate.
The seedling cultivation survival rate of comparing embodiment 1, embodiment 2, embodiment 3, embodiment 4 and control group 3, wherein compareing
The processing mode of group 3 is similar with embodiment, only difference is that, seedling cultivation was not being carried out into difference before greenhouse
The training of ambient humidity.Survival rate=(transplanting the transplanting quantity of the seedling quantity/total of survival) × 100%, above-mentioned 5 groups for the treatment of are pressed
Calculated according to 100 seedling.Comparing result is shown in Table 3.
The survival rate of the seedling of table 3
| Group number | Embodiment 1 | Embodiment 2 | Embodiment 3 | Embodiment 4 | Control group 3 |
| Survival rate | 88% | 89% | 91% | 93% | 64% |
It can be seen from comparing result, the survival rate of 4 groups of seedling of embodiment in greenhouse is higher than the survival rate of control group 3,
Illustrate to be carried out before by seedling replanting to greenhouse the training of different humidity, the survival rate of seedling replanting can be lifted.
In sum, the implantation methods of the method for culturing seedlings of the flower of Radix Gentianae of the embodiment of the present invention and flower of Radix Gentianae, by rough gentian
The peeling treatment of flower seed, accelerates the speed of flower of Radix Gentianae seed germination, and by adding perlite in Initial culture base, can
Be conducive to the breathing of seed, and keep the acidity of Initial culture base pH, further speed up sprouting for seed；EM bacterium can make nothing
Vaccine is more sturdy, increases the survival rate of aseptic seedling；And in squamous subculture and a series of work of tissue cultures of culture of rootage
Under, the emergence rate and survival rate of rough gentian flower seed are lifted, and strengthen the adaptability of flower of Radix Gentianae seedling；By flower of Radix Gentianae seedling
By being planted again after domestication, the survival rate of flower of Radix Gentianae seedling can be further lifted, increase the yield of flower of Radix Gentianae, meet city
The demand of field, reduces the harvesting amount of wild flower of Radix Gentianae, it is to avoid the danger of extinction occurs in flower of Radix Gentianae.
Embodiments described above is a part of embodiment of the invention, rather than whole embodiments.Reality of the invention
The detailed description for applying example is not intended to limit the scope of claimed invention, but is merely representative of selected implementation of the invention
Example.Based on the embodiment in the present invention, what those of ordinary skill in the art were obtained under the premise of creative work is not made
Every other embodiment, belongs to the scope of protection of the invention.
Claims (10)
1. a kind of method for culturing seedlings of flower of Radix Gentianae, it is characterised in that the seed of flower of Radix Gentianae is soaked in mass concentration for 10%-20%
Liquor natrii hypochloritis removed the peel, be rinsed with sterilized water after peeling, obtain remove the peel seed；The peeling seed is connect
Plant in Initial culture base, carry out Initial culture, obtain aseptic seedling；Using the cotyledon of the aseptic seedling and hypocotyl as explant
Body, and be inoculated in subculture medium, squamous subculture is carried out, obtain without offspring；Turn to plant in root media without offspring by described
In, culture of rootage is carried out, obtain flower of Radix Gentianae seedling；The temperature of the Initial culture is 15-25 DEG C, and intensity of illumination is 2000-
3000lx, light application time is 10-12h, and the Initial culture base is 0.1-0.5mg/L to be added with perlite and mass concentration
Heteroauxin, the 6- benzyls aminoadenine of 0.5-1mg/L and 0.01-0.05mg/L EM bacterium MS culture mediums.
2. the method for culturing seedlings of flower of Radix Gentianae according to claim 1, it is characterised in that with the sterilized water be rinsed when
Between be 20-30min.
3. the method for culturing seedlings of flower of Radix Gentianae according to claim 1, it is characterised in that also including being connect to the peeling seed
The step of carrying out ultraviolet induction to before the Initial culture base is planted, the ultraviolet induction is that the peeling seed is put into wavelength
To process 2-3h under the ultraviolet of 250-254nm.
4. the method for culturing seedlings of flower of Radix Gentianae according to claim 1, it is characterised in that the temperature of the squamous subculture is 15-
25 DEG C, intensity of illumination is 2000-3000lx, and light application time is 10-12h.
5. the method for culturing seedlings of flower of Radix Gentianae according to claim 1, it is characterised in that the temperature of the culture of rootage is 15-
25 DEG C, and to turning to plant to carry out shading treatment without offspring in the described of the root media.
6. the method for culturing seedlings of flower of Radix Gentianae according to claim 1, it is characterised in that the Initial culture, subculture training
Support and the ambient humidity of the culture of rootage is 55%-75%.
7. the method for culturing seedlings of flower of Radix Gentianae according to claim 1, it is characterised in that gone with the liquor natrii hypochloritis
Skin is carried out in the following manner：The seed 5-10min is soaked with the liquor natrii hypochloritis.
8. the method for culturing seedlings of flower of Radix Gentianae according to claim 1, it is characterised in that the subculture medium is to add to have matter
Amount concentration is the heteroauxin of the 6- benzyls aminoadenine, the gibberellin of 1mg-1.5mg/L and 0.5-1mg/L of 0.1-0.5mg/L
MS culture mediums；The root media be added with mass concentration be 1mg-1.5mg/L methyl α-naphthyl acetate and 0.5-1mg/L swash
The MS culture mediums of therbligs.
9. a kind of implantation methods of flower of Radix Gentianae, it is characterised in that by the nursery side of the flower of Radix Gentianae as described in claim any one of 1-8
The flower of Radix Gentianae seedling domestication that method is cultivated, obtains taming seedling；The domestication seed is planted in greenhouse, is controlled in the greenhouse
Light application time is 10-12h, and intensity of illumination is 2000-3000lx, coordinates field management, to the flower of Radix Gentianae for obtaining maturation.
10. implantation methods of flower of Radix Gentianae according to claim 9, it is characterised in that the domestication is by the flower of Radix Gentianae
In illumination box, the intensity of illumination of the illumination box is 2000-3000lx to seedling cultivation, and light application time is 10-
12h, the humidity in the illumination box was interval with one hour, was alternately set by 70% and 50%.
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