CN107155882A - A kind of medicinal bletilla striata aseptic seeding fast seedling-cultivating method - Google Patents
A kind of medicinal bletilla striata aseptic seeding fast seedling-cultivating method Download PDFInfo
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- CN107155882A CN107155882A CN201710316735.9A CN201710316735A CN107155882A CN 107155882 A CN107155882 A CN 107155882A CN 201710316735 A CN201710316735 A CN 201710316735A CN 107155882 A CN107155882 A CN 107155882A
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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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- 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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Abstract
The invention discloses a kind of medicinal bletilla striata aseptic seeding fast seedling-cultivating method, it is desirable to provide a kind of that the method for culturing seedlings of the medicinal bletilla striata can be cultivated with rapid scale and commercially producing, its drip irrigation device is, including following incubation step：S1：Explant is chosen：Explant is used as from the fissureless Fruit pod in surface；S2：Explant is sterilized；S3：Primary culture, induces naked embryonic development and rudiment；S4：Complete Primary culture；S5：Strong seedling culture, the naked embryo of sprouting is inoculated in solid strong seedling culture base, naked embryo is further developed and is grown true leaf；S6：Culture of rootage, the unrooted tissue-cultured seedling Jing Guo strong seedling culture is scooped out with spoon, is laid in root media and is cultivated, completes to take root.
Description
Technical field
The present invention relates to technical field of plant propagation, more particularly to a kind of medicinal bletilla striata aseptic seeding fast seedling-cultivating method.
Background technology
The bletilla striata is one kind of orchid family bletilla striata category, also referred to as bletilla.Perennial herb, high 20~50 centimetres, 4~5 pieces of leaf, base portion
Set builds up bulbous mutually, and scape is extracted in center out.Raceme has several flowers；Flower purple or pale red, about 5 centimetres of diameter, by 3 pieces
The lip composition of sepal, 2 pieces of petals and 1 piece of specialization；Lip 3 splits, and has typhloscle piece above；Stamen forms with style symphysis and closes stamen
There is a flower pesticide on post, gynostemium top, and there is a column cap depression front upper place.There is the root-like stock of thick thickness underground, and such as chicken head shape, richness is viscous
Property, colloid containing bletilla, i.e. bletilla mannan, can hyoscine, have hemostasis tonifying lung, the effect of myogenic pain relieving, also be available for making thickener.
At present, notification number discloses a kind of method of fast culture bletilla striata seedling for CN106416972A Chinese patent,
It is included the seed of the bletilla striata after sprouting culture, protocorm differentiation culture, bulb Fiber differentiation and domestication, obtains bletilla striata seedling
Method.
Although the method regularity of this fast culture bletilla striata seedling is preferably, variation probability is small, and it is mainly by cuing open
The sterile modeling fruit of the bletilla striata is opened, so that take out seed directly carries out culture so as to obtain bletilla striata protocorm on fluid nutrient medium is sprouted,
But this planting patterns is by seed being placed in culture medium so as to carry out Propogation and culture, but be difficult to carry out on a large scale
Plantation is more difficult for cultivation is bred in batch production, commercialization.
The content of the invention
It is an object of the invention to provide a kind of medicinal bletilla striata aseptic seeding fast seedling-cultivating method, it, which has, can meet the bletilla striata
Large-scale plantation to business seedling breeding need, by using above-mentioned seedling raising mannerses, can cause bletilla striata nursery breed batch production,
The advantage of scale.
The present invention above-mentioned technical purpose technical scheme is that：
A kind of medicinal bletilla striata aseptic seeding fast seedling-cultivating method, including following incubation step：
S1：Explant is chosen：From solid bletilla striata Fruit pod, harvested before Fruit pod maturation cracking, from the fissureless fruit in surface
Pod is used as explant；
S2：Explant is sterilized, and the dust on explant surface is wiped using alcohol swab, explant is soaked using disinfectant,
Immersion 30~60 minutes, is rinsed after the completion of soaking disinfection using sterilized water to explant, is rinsed 3~4 times；
S3：Primary culture, the explant after sterilization is cut along middle part, is cut into 1cm~2cm section, is clamped with tweezers
Explant gently shaken above culture medium bottleneck it is several under, the powdered naked embryo of yellowly in Fruit pod is inoculated into primary culture medium
In, it is transferred to culturing room and is cultivated；Illumination is carried out to culturing room, after culture 4~8 weeks, dormancy axillary bud sprouting, naked embryo is by yellow
When switching to green and expanding individual, proceed culture 4~10 weeks so that the naked embryo of rudiment continues to increase, and completes Primary culture；
S4：Strong seedling culture, the naked embryo of sprouting is inoculated in solid strong seedling culture base, and then light culture 2~7 days gives illumination,
After culture 8~16 weeks, naked embryo is further developed and grows true leaf；
S5：Culture of rootage, the unrooted tissue-cultured seedling Jing Guo strong seedling culture is scooped out with spoon, is laid in root media and is cultivated,
Light culture 2~7 days, gives illumination, cultivates 8~16 weeks, completes to take root.
By using above-mentioned technical proposal, conventional seedbed system mode is to lean on low root-like stock division propagation, and breeding coefficient is low, numerous
Grow speed slow, it is impossible to meet large-scale plantation demand；Its seed is not endospermous naked embryo, without survival under natural conditions
Power, therefore seed is directly placed on culture medium by prior art, so as to provide seed nutritional so that seed growth is former into the bletilla striata
Bulb, is cultivated by seed, and the present invention is by the way that seed is splitted, so that naked embryo is directly contacted with primary culture medium,
So as to pass through the cultivation to naked embryo so that naked embryo rudiment, primary culture medium can quickly break naked embryo dormancy, promote naked embryo to sprout
Hair, while can effectively promote the generation of callus, strong seedling culture base is put into after culture a period of time by the naked embryo of rudiment
In so that the naked embryo of rudiment grows true leaf, and strong seedling culture base can ensure that the naked growth of the embryo of rudiment is artificially controllable, be grown tall in seedling,
Suppress root growth while growing true leaf, be easy to follow-up rolling bottle plant division to operate, easily dehydration after being removed due to the seed of the bletilla striata
It is shrivelled, so that seed shattering vitality, is contacted by naked embryo with the direct of primary culture medium, so that emergence rate reaches
100%, while rudiment fast-growth can be caused to emerge, emerge faster, seedling more grows, therefore can be quickly big using this method
The culture bletilla striata of scale, can be with batch production, scale, and in cultivating process so that cultivating the bletilla striata, explant first
Selection should select the end of spring and the beginning of summer artificial pollination then and the solid Fruit pod of success, be adopted before mid or late September Fruit pod maturation cracking
Receive, choose Fruit pod greatly, sagging, the Fruit pod not ftractureed is explant, it is to avoid the explant of cracking intrudes into inside by bacterium etc.,
Secondly explant is carried out disinfection, it is ensured that culture medium etc. will not be corroded by the germ entrained by explant in incubation, from
And further started again, strong sprout and culture of rootage.
Further set：The thimerosal includes following component：1 part and 4 parts sterilized waters of bleaching water.
By using above-mentioned technical proposal, it is necessary to carry out effective sterilization, the disease that explant surface is carried to explant
The profound removal of bacterium so that reduced when the later stage cultivates and occur the possibility failed occur because of being corroded by germ.
Further set：The culture indoor temperature in the Primary culture stage is 25 DEG C, intensity of illumination 2000LUX, illumination
16 hours, light culture 8 hours was cultivated 6 weeks, when naked embryo switchs to green by yellow and expands individual, continued to cultivate 8 weeks, completion startup
Culture.
By using above-mentioned technical proposal, in the Primary culture stage, it is necessary to which naked embryo dormancy will be broken by suitable environment,
Promote naked embryo germination, acted on by illumination and cause naked embryo to carry out photosynthesis, and light culture is then plumule progress respiration, from
And by controlling time and the intensity of illumination of photoperiod, the photoperiod of naked embryonic development rudiment the most suitable is selected, so that
Naked embryo rudiment, until being changed into green from yellow expands individual.
Further set：Culture environment is 25 DEG C of temperature in the S5, and light culture 5 days gives illumination, intensity of illumination
1500LUX, periodicity of illumination is daily illumination 16 hours, and light culture 8 hours is cultivated 12 weeks.
By using above-mentioned technical proposal, when the naked embryo of rudiment needs to carry out strong sprout, it is put into strong seedling culture base, gives
Illumination, makes its fast-growth, respiration and photosynthetic continuous alternating, so that the seedling grown is more tall and big, simultaneously
Overall well-grown.
Further set：The unrooted tissue-cultured seedling is 1~1.5cm of height strong sprout, is scooped out with spoon, is laid in training of taking root
Support in base and cultivate, then light culture 5 days gives illumination, intensity of illumination 2000LUX, and periodicity of illumination is daily illumination in 16 hours, 8
Hour light culture, environment temperature is controlled at 25 DEG C, is cultivated 12 weeks, and completion is taken root by using above-mentioned technical proposal, to unrooted group
Training seedling is moved into root media after being cultivated is taken root.
Further set：The primary culture medium includes following component：MS culture mediums are added in every liter of water：4.7g/L；6-
Benzyl aminoadenine：1.0~3.5mg/L；Sucrose：15~45g/L；Agar：5.5~7.0g/L.
By using above-mentioned technical proposal, MS culture mediums are characterized in that inorganic salts and ion concentration are higher, are more stable
Ionic equilibrium solution, its nitrate content is high, and the quantity and ratio of its nutrient are suitable, can meet the nutrition and life of plant cell
Reason needs, with its minimal medium mainly as culture medium, and 6- benzyls aminoadenine has efficiently, stably, inexpensively and be easy to
The features such as using, the main function of 6- benzyl aminoadenines is the formation for promoting bud, can also evoked callus occur, and add
Enter the solid medium of the agar advantage compared with fluid nutrient medium and be easy to operate, ventilation problem is easily solved, and is easy to often
Observational study, and sucrose supplies the nutrient required for cell growth then as the standard carbon source of culture medium.
Further set：The strong seedling culture base includes following component：MS culture mediums are added in every liter of water：4.7g/L；6-
Benzyl aminoadenine：0.5~1.5mg/L；Sucrose：20~30g/L；Agar：5.0~7.0g/L.
By using above-mentioned technical proposal, the formula can ensure that the naked growth of the embryo of rudiment is artificially controllable, be grown tall in seedling, and
Suppress root growth while growing true leaf, be easy to follow-up rolling bottle plant division to operate, allow its extensive by the operation of plant division
Production.
Further set：The root media mainly includes following component：MS culture mediums are added in every liter of water：4.7g/
L；Methyl α-naphthyl acetate：1~3.0mg/L；Activated carbon：0.5~1.0g/L；Sucrose：15~30g/L；Agar：5.0~7.0g/L.
By using above-mentioned technical proposal, formula and mode of operation can ensure that seedling is fully taken root, and root system all positions
In media surface, it is easy to the seedling taking when transplanting, reduction damage, promotion is quickly survived, activated carbon is due to itself being adsorptivity
Stronger adsorbent, can preferably mitigate the browning phenomenon of explant.
In summary, the invention has the advantages that：The bletilla striata can be met by above-mentioned planting patterns to plant on a large scale
The need for planting to business seedling breeding, and cultivated by naked embryo and cause rooting rate to reach 100%, far above prior art, effectively
Meet the cultivation of bletilla striata batch production, scale.
Embodiment
The present invention is described in further detail below.
Embodiment 1：A kind of medicinal bletilla striata aseptic seeding fast seedling-cultivating method, including following incubation step：
S1：Explant is chosen：From solid bletilla striata Fruit pod, harvested before Fruit pod maturation cracking, from the fissureless fruit in surface
Pod is used as explant；
S2：Explant is sterilized, and the dust on explant surface is wiped using alcohol swab, explant is soaked using disinfectant,
Immersion 30~60 minutes, is rinsed after the completion of soaking disinfection using sterilized water to explant, is rinsed 3~4 times, and nothing is put in taking-up
Bright water is blotted on bacterium filter paper standby；
S3：Primary culture, the explant after sterilization is cut along middle part, is cut into 1cm~2cm section, is clamped with tweezers
Explant gently shaken above culture medium bottleneck it is several under, the powdered naked embryo of yellowly in Fruit pod is inoculated into bud Primary culture
In base, it is transferred to culturing room and is cultivated, primary culture medium includes following component：MS culture mediums are added in every liter of water：4.7g/L；6-
Benzyl aminoadenine：3.0mg/L；Sucrose：30g/L；Agar：6.0g/L；Illumination is carried out to culturing room, culture indoor temperature is 25
DEG C, intensity of illumination 2000LUX, illumination 16 hours, light culture 8 hours is cultivated 6 weeks, naked embryo switchs to green by yellow and expands individual
When, continue to cultivate 8 weeks so that the naked embryo of rudiment continues to increase, and completes Primary culture；
S4：Strong seedling culture, the naked embryo of sprouting is inoculated in solid strong seedling culture base, and strong seedling culture base includes following component：Often
Rise and MS culture mediums are added in water：4.7g/L；6- benzyl aminoadenines：1.0mg/L；Sucrose：20g/L；Agar：6.0g/L, temperature
25 DEG C, light culture 5 days gives illumination, intensity of illumination 1500LUX, periodicity of illumination is daily illumination 16 hours, and light culture 8 is small
When, cultivate 12 weeks, strong sprout is grown true leaf；
S5：Culture of rootage, chooses the strong sprout that unrooted tissue-cultured seedling is height 1cm, is scooped out with spoon, be laid in root media and train
Support, root media mainly includes following component：MS culture mediums are added in every liter of water：4.7g/L；Methyl α-naphthyl acetate：2.0mg/L；Activity
Charcoal： 0.8g/L；Sucrose：20g/L；Agar：Then 6.0g/L, light culture 5 days gives illumination, intensity of illumination 2000LUX, illumination
Cycle is daily illumination in 16 hours, and 8 hours light cultures, environment temperature is controlled at 25 DEG C, cultivates and completes within 12 weeks to take root.
Embodiment 2：A kind of medicinal bletilla striata aseptic seeding fast seedling-cultivating method, including following incubation step：
S1：Explant is chosen：From solid bletilla striata Fruit pod, harvested before Fruit pod maturation cracking, from the fissureless fruit in surface
Pod is used as explant；
S2：Explant is sterilized, and the dust on explant surface is wiped using alcohol swab, explant is soaked using disinfectant,
Immersion 30~60 minutes, is rinsed after the completion of soaking disinfection using sterilized water to explant, is rinsed 3~4 times, and nothing is put in taking-up
Bright water is blotted on bacterium filter paper standby；
S3：Primary culture, the explant after sterilization is cut along middle part, is cut into 1cm~2cm section, is clamped with tweezers
Explant gently shaken above culture medium bottleneck it is several under, the powdered naked embryo of yellowly in Fruit pod is inoculated into bud Primary culture
In base, it is transferred to culturing room and is cultivated, primary culture medium includes following component：MS culture mediums are added in every liter of water：4.7g/L；6-
Benzyl aminoadenine：3.5mg/L；Sucrose：45g/L；Agar：7.0g/L；Illumination is carried out to culturing room, culture indoor temperature is 25
DEG C, intensity of illumination 2000LUX, illumination 16 hours, light culture 8 hours is cultivated 6 weeks, naked embryo switchs to green by yellow and expands individual
When, continue to cultivate 8 weeks so that the naked embryo of rudiment continues to increase, and completes Primary culture；
S4：Strong seedling culture, will sprout naked embryo and is inoculated in solid strong seedling culture base, strong seedling culture base includes following component：Every liter
MS culture mediums are added in water：4.7g/L；6- benzyl aminoadenines：1.5mg/L；Sucrose：30g/L；Agar：7.0g/L, temperature 25
DEG C, light culture 5 days gives illumination, intensity of illumination 1500LUX, and periodicity of illumination is daily illumination 16 hours, light culture 8 hours, training
Support 12 weeks, strong sprout is grown true leaf；
S5：Culture of rootage, chooses the strong sprout that unrooted tissue-cultured seedling is height 1cm, is scooped out with spoon, be laid in root media and train
Support, root media mainly includes following component：MS culture mediums are added in every liter of water：4.7g/L；Methyl α-naphthyl acetate：1~3.0mg/L；
Activated carbon：1.0g/L；Sucrose：30g/L；Agar：Then 7.0g/L, light culture 5 days gives illumination, intensity of illumination 2000LUX,
Periodicity of illumination is daily illumination in 16 hours, and 8 hours light cultures, environment temperature is controlled at 25 DEG C, cultivates and completes within 12 weeks to take root.
Embodiment 3：A kind of medicinal bletilla striata aseptic seeding fast seedling-cultivating method, including following incubation step：
S1：Explant is chosen：From solid bletilla striata Fruit pod, harvested before Fruit pod maturation cracking, from the fissureless fruit in surface
Pod is used as explant；
S2：Explant is sterilized, and the dust on explant surface is wiped using alcohol swab, explant is soaked using disinfectant,
Immersion 30~60 minutes, is rinsed after the completion of soaking disinfection using sterilized water to explant, is rinsed 3~4 times, and nothing is put in taking-up
Bright water is blotted on bacterium filter paper standby；
S3：Primary culture, the explant after sterilization is cut along middle part, is cut into 1cm~2cm section, is clamped with tweezers
Explant gently shaken above culture medium bottleneck it is several under, the powdered naked embryo of yellowly in Fruit pod is inoculated into bud Primary culture
In base, it is transferred to culturing room and is cultivated, primary culture medium includes following component：MS culture mediums are added in every liter of water：4.7g/L；6-
Benzyl aminoadenine：1.0mg/L；Sucrose：15g/L；Agar：5.5g/L；Illumination is carried out to culturing room, culture indoor temperature is 25
DEG C, intensity of illumination 2000LUX, illumination 16 hours, light culture 8 hours is cultivated 6 weeks, naked embryo switchs to green by yellow and expands individual
When, continue to cultivate 8 weeks so that the naked embryo of rudiment continues to increase, and completes Primary culture；
S4：Strong seedling culture, the naked embryo of sprouting is inoculated in solid strong seedling culture base, and strong seedling culture base includes following component：Often
Rise and MS culture mediums are added in water：4.7g/L；6- benzyl aminoadenines：0.50mg/L；Sucrose：20g/L；Agar：5.0g/L, temperature
25 DEG C, light culture 5 days gives illumination, intensity of illumination 1500LUX, periodicity of illumination is daily illumination 16 hours, and light culture 8 is small
When, cultivate 12 weeks, strong sprout is grown true leaf；
S5：Culture of rootage, chooses the strong sprout that unrooted tissue-cultured seedling is height 1cm, is scooped out with spoon, be laid in root media and train
Support, root media mainly includes following component：MS culture mediums are added in every liter of water：4.7g/L；Methyl α-naphthyl acetate：1.0mg/L；Activity
Charcoal：0.5g/L；Sucrose：15g/L；Agar：Then 5.0g/L, light culture 5 days gives illumination, intensity of illumination 2000LUX, illumination
Cycle is daily illumination in 16 hours, and 8 hours light cultures, environment temperature is controlled at 25 DEG C, cultivates and completes within 12 weeks to take root.
Embodiment 4：A kind of medicinal bletilla striata aseptic seeding fast seedling-cultivating method, including following incubation step：
S1：Explant is chosen：From solid bletilla striata Fruit pod, harvested before Fruit pod maturation cracking, from the fissureless fruit in surface
Pod is used as explant；
S2：Explant is sterilized, and the dust on explant surface is wiped using alcohol swab, explant is soaked using disinfectant,
Immersion 30~60 minutes, is rinsed after the completion of soaking disinfection using sterilized water to explant, is rinsed 3~4 times, and taking-up puts sterile
Bright water is blotted on filter paper standby；
S3：Primary culture, the explant after sterilization is cut along middle part, is cut into 1cm~2cm section, is clamped with tweezers
Explant gently shaken above culture medium bottleneck it is several under, the powdered naked embryo of yellowly in Fruit pod is inoculated into bud Primary culture
In base, it is transferred to culturing room and is cultivated, primary culture medium includes following component：MS culture mediums are added in every liter of water：4.7g/L；6-
Benzyl aminoadenine：3.0mg/L；Sucrose：30g/L；Agar：6.0g/L；Illumination is carried out to culturing room, culture indoor temperature is 25
DEG C, intensity of illumination 2000LUX, illumination 16 hours, light culture 8 hours is cultivated 4 weeks, naked embryo switchs to green by yellow and expands individual
When, continue to cultivate 4 weeks so that the naked embryo of rudiment continues to increase, and completes Primary culture；
S4：Strong seedling culture, the naked embryo of sprouting is inoculated in solid strong seedling culture base, and strong seedling culture base includes following component：Often
Rise and MS culture mediums are added in water：4.7g/L；6- benzyl aminoadenines：1.0mg/L；Sucrose：20g/L；Agar：6.0g/L, temperature
25 DEG C, light culture 2 days gives illumination, intensity of illumination 1500LUX, periodicity of illumination is daily illumination 12 hours, and light culture 12 is small
When, cultivate 8 weeks, strong sprout is grown true leaf；
S5：Culture of rootage, chooses the strong sprout that tissue-cultured seedling is height 1cm, is scooped out with spoon, be laid in root media and cultivate,
Root media mainly includes following component：MS culture mediums are added in every liter of water：4.7g/L；Methyl α-naphthyl acetate：2.0mg/L；Activated carbon：
0.8g/L；Sucrose：20g/L；Agar：Then 6.0g/L, light culture 5 days gives illumination, intensity of illumination 2000LUX, periodicity of illumination
For daily illumination in 12 hours, 12 hours light cultures, environment temperature is controlled at 25 DEG C, is cultivated and is completed within 8 weeks to take root.
Embodiment 5：A kind of medicinal bletilla striata aseptic seeding fast seedling-cultivating method, including following incubation step：
S1：Explant is chosen：From solid bletilla striata Fruit pod, harvested before Fruit pod maturation cracking, from the fissureless fruit in surface
Pod is used as explant；
S2：Explant is sterilized, and the dust on explant surface is wiped using alcohol swab, explant is soaked using disinfectant,
Immersion 30~60 minutes, is rinsed after the completion of soaking disinfection using sterilized water to explant, is rinsed 3~4 times, and nothing is put in taking-up
Bright water is blotted on bacterium filter paper standby；
S3：Primary culture, the explant after sterilization is cut along middle part, is cut into 1cm~2cm section, is clamped with tweezers
Explant gently shaken above culture medium bottleneck it is several under, the powdered naked embryo of yellowly in Fruit pod is inoculated into bud Primary culture
In base, it is transferred to culturing room and is cultivated, primary culture medium includes following component：MS culture mediums are added in every liter of water：4.7g/L；6-
Benzyl aminoadenine：3.0mg/L；Sucrose：30g/L；Agar：6.0g/L；Illumination is carried out to culturing room, culture indoor temperature is 25
DEG C, intensity of illumination 2000LUX, illumination 20 hours, light culture 4 hours is cultivated 8 weeks, naked embryo switchs to green by yellow and expands individual
When, continue to cultivate 10 weeks so that the naked embryo of rudiment continues to increase, and completes Primary culture；
S4：Strong seedling culture, the naked embryo of sprouting is inoculated in solid strong seedling culture base, and strong seedling culture base includes following component：Often
Rise and MS culture mediums are added in water：4.7g/L；6- benzyl aminoadenines：1.0mg/L；Sucrose：20g/L；Agar：6.0g/L, temperature
25 DEG C, light culture 7 days gives illumination, intensity of illumination 1500LUX, periodicity of illumination is daily illumination 20 hours, and light culture 4 is small
When, cultivate 16 weeks, strong sprout is grown true leaf；
S5：Culture of rootage, chooses the strong sprout that tissue-cultured seedling is height 1.5cm, is scooped out with spoon, be laid in root media and train
Support,
Root media mainly includes following component：MS culture mediums are added in every liter of water：4.7g/L；Methyl α-naphthyl acetate：2.0mg/L；It is living
Property charcoal： 0.8g/L；Sucrose：20g/L；Agar：Then 6.0g/L, light culture 5 days gives illumination, intensity of illumination 2000LUX, light
It is daily illumination in 20 hours according to the cycle, 4 hours light cultures, environment temperature is controlled at 25 DEG C, cultivates and complete within 16 weeks to take root.
Above-described embodiment 1 to embodiment 5 is subjected to Viral diagnosis after being sterilized to explant, opened while observing naked embryo and being put into
The naked growth of the embryo situation of rudiment is observed after dynamic culture medium, the true leaf growing state in strong seedling culture base is put into, is put into culture of rootage
Height of seedling situation and coefficient of taking root after base carry out analysis contrast, as a result as shown in the table：
It can be drawn from upper table, by above-mentioned modes of reproduction and with suitable primary culture medium, strong seedling culture base and training of taking root
Support base, can quick reproducing bletilla striata, rooting rate reaches 100%, while effectively healthy and strong root 1~3, root 20~40mm of length, root thick 1
~2mm, 3~5cm of height of seedling, the factorial praluction of the entirely appropriate bletilla striata are conducive to mass producing high-quality lime tree seedling.
The above embodiments are only explanation of the invention, and it is not limitation of the present invention, people in the art
Member can make the modification without creative contribution to the present embodiment as needed after this specification is read, but as long as at this
All protected in the right of invention by Patent Law.
Claims (8)
1. a kind of medicinal bletilla striata aseptic seeding fast seedling-cultivating method, it is characterised in that：Including following incubation step：
S1：Explant is chosen：From solid bletilla striata Fruit pod, harvested before Fruit pod maturation cracking, from the fissureless fruit in surface
Pod is used as explant；
S2：Explant is sterilized, and the dust on explant surface is wiped using alcohol swab, explant is soaked using disinfectant,
Immersion 30~60 minutes, is rinsed after the completion of soaking disinfection using sterilized water to explant, is rinsed 3~4 times；
S3：Primary culture, the explant after sterilization is cut along middle part, is cut into 1cm~2cm section, is clamped with tweezers
Explant gently shaken above culture medium bottleneck it is several under, the powdered naked embryo of yellowly in Fruit pod is inoculated into primary culture medium
In, it is transferred to culturing room and is cultivated；Illumination is carried out to culturing room, after culture 4~8 weeks, dormancy axillary bud sprouting, naked embryo is by yellow
When switching to green and expanding individual, proceed culture 4~10 weeks so that the naked embryo of rudiment continues to increase, and completes Primary culture；
S4：Strong seedling culture, the naked embryo of sprouting is inoculated in solid strong seedling culture base, and then light culture 2~7 days gives illumination,
After culture 8~16 weeks, naked embryo is further developed and grows true leaf；
S5：Culture of rootage, the unrooted tissue-cultured seedling Jing Guo strong seedling culture is scooped out with spoon, is laid in root media and is cultivated,
Light culture 2~7 days, gives illumination, cultivates 8~16 weeks, completes to take root.
2. a kind of medicinal bletilla striata aseptic seeding fast seedling-cultivating method according to claim 1, it is characterised in that：The sterilization
Liquid includes following component：1 part and 4 parts sterilized waters of bleaching water.
3. a kind of medicinal bletilla striata aseptic seeding fast seedling-cultivating method according to claim 1, it is characterised in that：It is described to start
The culture indoor temperature of cultivation stage is 25 DEG C, intensity of illumination 2000LUX, illumination 16 hours, and light culture 8 hours is cultivated 6 weeks,
When naked embryo switchs to green by yellow and expands individual, continue to cultivate 8 weeks, completion Primary culture.
4. a kind of medicinal bletilla striata aseptic seeding fast seedling-cultivating method according to claim 1, it is characterised in that：In the S5
Culture environment is 25 DEG C of temperature, and light culture 5 days gives illumination, intensity of illumination 1500LUX, and periodicity of illumination is that daily illumination 16 is small
When, light culture 8 hours is cultivated 12 weeks.
5. a kind of medicinal bletilla striata aseptic seeding fast seedling-cultivating method according to claim 1, it is characterised in that：The unrooted
Tissue-cultured seedling is 1~1.5cm of height strong sprout, is scooped out with spoon, is laid in root media and cultivates, and then light culture 5 days is given
Illumination, intensity of illumination 2000LUX are given, periodicity of illumination is daily illumination in 16 hours, and 8 hours light cultures, environment temperature is controlled 25
DEG C, cultivate 12 weeks, complete to take root.
6. a kind of medicinal bletilla striata aseptic seeding fast seedling-cultivating method according to claim 1, it is characterised in that：It is described to start
Culture medium includes following component：MS culture mediums are added in every liter of water：4.7g/L；6- benzyl aminoadenines：1.0~3.5 mg/L；
Sucrose：15~45g/L；Agar：5.5~7.0g/L.
7. a kind of medicinal bletilla striata aseptic seeding fast seedling-cultivating method according to claim 1, it is characterised in that：The strong sprout
Culture medium includes following component：MS culture mediums are added in every liter of water：4.7g/L；6- benzyl aminoadenines：0.5~1.5 mg/L；
Sucrose：20~30g/L；Agar：5.0~7.0g/L.
8. a kind of medicinal bletilla striata aseptic seeding fast seedling-cultivating method according to claim 1, it is characterised in that：It is described to take root
Culture medium mainly includes following component：MS culture mediums are added in every liter of water：4.7g/L；Methyl α-naphthyl acetate:1~3.0mg/L；Activated carbon：
0.5~1.0g/L；Sucrose：15~30g/L；Agar：5.0~7.0g/L.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107896851A (en) * | 2017-12-12 | 2018-04-13 | 遵义医学院 | A kind of live fast numerous method of bletilla earth culture |
| CN109076923A (en) * | 2018-08-28 | 2018-12-25 | 河南云帮农业科技有限公司 | A kind of bletilla striata aseptic seeding special culture media |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105325302A (en) * | 2015-12-11 | 2016-02-17 | 电子科技大学 | Method for bletilla striata seedling production based on liquid medium |
| CN105766645A (en) * | 2016-03-31 | 2016-07-20 | 广西医科大学 | Efficient reproduction method of rhizoma bletillae tissue culture seedlings |
-
2017
- 2017-05-05 CN CN201710316735.9A patent/CN107155882A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105325302A (en) * | 2015-12-11 | 2016-02-17 | 电子科技大学 | Method for bletilla striata seedling production based on liquid medium |
| CN105766645A (en) * | 2016-03-31 | 2016-07-20 | 广西医科大学 | Efficient reproduction method of rhizoma bletillae tissue culture seedlings |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107896851A (en) * | 2017-12-12 | 2018-04-13 | 遵义医学院 | A kind of live fast numerous method of bletilla earth culture |
| CN109076923A (en) * | 2018-08-28 | 2018-12-25 | 河南云帮农业科技有限公司 | A kind of bletilla striata aseptic seeding special culture media |
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