CN109362568A - A kind of tissue culture and rapid propagation method of clematis " cherry lip " - Google Patents
A kind of tissue culture and rapid propagation method of clematis " cherry lip " Download PDFInfo
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- CN109362568A CN109362568A CN201811462522.8A CN201811462522A CN109362568A CN 109362568 A CN109362568 A CN 109362568A CN 201811462522 A CN201811462522 A CN 201811462522A CN 109362568 A CN109362568 A CN 109362568A
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- clematis
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- cherry lip
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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
- 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
- A01G22/60—Flowers; Ornamental 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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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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
Abstract
The invention discloses a kind of tissue culture and rapid propagation methods of clematis " cherry lip ", belong to field of plant tissue culture technique.The above method includes: step 1: the selection of material；Step 2: sterilization；Step 3: induction of callus；Step 4: the Multiplying culture of callus；Step 5: the culture of callus induction bud；Step 6: the culture of rootage without offspring.The present invention makes finally to fall below minimum, increase clematis survival rate applied to the viruliferous possibility of callus of induced bud differentiation by callus induction and proliferation.
Description
Technical field
The present invention relates to field of plant tissue culture technique, and in particular to a kind of tissue-culturing rapid propagation side of clematis " cherry lip "
Method.
Background technique
Clematis (Clematis florida Thunb) belongs to a kind of vine flowers and plants of Ranunculaceae Clematis.Because of it
Pattern is changeable, form is colourful, and the good reputation of " liana queen " is enjoyed in gardens.Again very because of performances such as its drought resisting, cold-resistants
By force, deep to be favored by flower amateur all over the world.There are many clematis type, existing, also there is polyphyll
's；Existing petal development type completely, also has as small bell class.The wherein cherry lip clematis of small bell class, petal symphysis, top picture
Just like the cherry microcheilia of maiden, ruddy and high resilience.Thus it gains the name " cherry lip ".This year is chased after by the fanaticism for liking clematis deeply
It holds in both hands.The traditional modes of reproduction of clematis is mainly cuttage and seminal propagation.Clematis internode is longer, and the branch of semi-lignified is
It is easy to cutting survival.So the survival rate of cuttage is generally very low.The quantity of the seed of clematis is relatively fewer, and need to be by certain
Low-temperature treatment could germinate.And the clematis of some kinds can not form seed.Therefore, the proliferative speed of clematis
It is very low, market is far from satisfying for the demand of clematis, limits the gardens of clematis and the application that family's cultivation is ornamental.
Currently, plant tissue culture technique is relatively mature, the successful incubation and quickly numerous on many plants
It grows.The quick breeding that clematis is carried out with tissue culture method is a feasible and effective method.
Patent CN104206281A discloses a kind of method for tissue culture of clematis, comprising the following steps: (1) obtains nothing
Vaccine material；(2) Initial culture；(3) Multiplying culture；(4) Rooting and hardening-off culture；When root long is 2-4cm, domestication shifting can be carried out
It plants.Clematis method for tissue culture provided by the invention, the breeding cycle is short, and breeding coefficient is high, easily takes root, the root system of plant is thick
Strong, newly slightly growing way is good, and stalk is sturdy, and domestication easily survives.
Patent CN104396750 A discloses the tissue culture propagation method of a kind of mao of leaf clematis, mainly by being just commissioned to train
It supports, Multiplying culture, culture of rootage and the step of acclimatization and transplants, has reached that growth rate is fast, stabilization characteristics of genetics, breeding coefficient
High, the eugonic excellent results of test tube seedling, the rate of sprouting of the callus of hair leaf clematis have reached 98%, proliferation times
5.7 times are reached, rooting rate has reached 100%, and final survival rate has reached 96%.
Patent CN 103202229A provides a kind of green colored polyphyll clematis tissue cultivation rapid breeding method, step include: take it is green
The stem apex of flower polyphyll clematis children's stem, uses aseptic water washing, 75% alcohol immersion treatment 20~30 seconds, then use on the super-clean bench
0.1~0.2% mercuric chloride soaking disinfection 6~10 minutes is inoculated into clump bud inducement cultivation base after aseptic water washing is clean and carries out just
For Fiber differentiation；2) Multiple Buds that Fiber differentiation primary obtains are divided into single budlet, be inoculated into clump bud proliferated culture medium into
Row squamous subculture；3) when sprout it is long to 2~3cm when be inoculated into strengthening seedling and rooting culture medium and carry out culture of rootage.The inventive method
Green colored polyphyll clematis can be quickly bred, can not only save and cultivate seedling cost, improve production efficiency, but also not by the external world
The limitation of condition, the four seasons can all carry out, and save nursery occupied area, can form a large amount of excellent test tube seedlings in a short time, be advised
Modelling, the factorial production.
In the prior art by different processing methods and selection different culture medium, the quick breeding of clematis is realized.
Summary of the invention
The object of the present invention is to provide a kind of tissue culture and rapid propagation method of clematis " cherry lip ", by callus induction and
Proliferation falls below the viruliferous possibility of callus finally broken up applied to induced bud minimum, increases clematis and survives
Rate.
In order to solve the above technical problems, present invention offer technical solution is as follows:
The present invention provides a kind of tissue culture and rapid propagation method of clematis " cherry lip ", comprising:
Step 1: the selection of material
Choose healthy and strong, no disease and pests harm shoot, the stem section of clip belt segment；
Step 2: sterilization
On superclean bench, stem section is cut into the about 0.8-1cm long of one section of band；It first uses aseptic water washing 1 time, then with 70%
Alcohol impregnates 30 seconds, then with 10% sodium hypochlorite 10-15 minute, finally use is again with 0.1% mercuric chloride immersion 7-8 minutes, during which otherwise
It is disconnected to shake, so that it is sterilized uniform and complete, then uses aseptic water washing 4-5 times；
Step 3: induction of callus
Above-mentioned stem section is taken, Fiber differentiation in the culture medium of evoked callus is disposed across, is generated largely after 15 days in stem section base portion
Jade-green callus；
Evoked callus culture medium: 1/4 improvement MS+NAA 1.0mg/L+2,4-D 0.5mg/L+6-BA 0.5mg/L+10% is fragrant
Any of several broadleaf plants mud+active carbon 1g/L；
Step 4: the Multiplying culture of callus
The callus induced is taken, the fritter of 0.4-0.6cm is divided into, is inoculated into swallow liquid culture base and carries out callus
Multiplying culture；
Proliferated culture medium are as follows: 3/4 improvement MS+NAA 1.0mg/L+2,4-D 0.5mg/L+6-BA 0.5mg/L+ active carbon 1g/L；
Step 5: the culture of callus induction bud
Callus is taken, the fritter of 1cm or so is cut into, is inoculated into the induction for carrying out bud in differential medium；
Culture medium is swallow liquid culture base, formula are as follows: 3/4 improvement MS+6-BA 1.5mg/L+TDZ 0.5mg/L+NAA
0.01mg/L+ active carbon 1.5g/L；
Step 6: the culture of rootage without offspring
When Multiple Buds grow to 3-5cm, clip branch is inoculated into root induction in root media；
Root media are as follows: 1/2 improvement MS+NAA 0.5mg/L+IBA 0.1mg/L+ active carbon 0.5g/L.
Further, further include step 7: the rooting culture of test tube seedling: first moving to greenhouse for test tube seedling, then takes out seedling
The culture medium for cleaning root attachment, is transplanted to substrate composition are as follows: 1 part of sandy soil, 2 parts of vermiculites: 2 parts of perlites: 5 parts of garden mould part mixing
It is cultivated under natural conditions in matrix.
Further, in the step 1, when clip stem section, every three sections were one section, and the stem section of clip is first in flowing water undershoot
It washes 3-4 hour, it is then primary with aseptic water washing.
Further, in the step 4, appreciation rate is 2-3 times in the Multiplying culture of callus.
Further, VC is added in the mixed-matrix and is mixed thoroughly with the sprinkling of 800 times of carbendazim.
Further, the VC additional amount is 2.5g/L.
Further, the modified MS medium are as follows: by a great number of elements KNO of MS culture medium3Dosage be changed to 2500mg/
L, with (NH4)2SO4Instead of NH4NO3, dosage 200mg/L, other bases are identical as MS.
Further, the sucrose of 25g/L, the agar of 7.5g/L, pH 5.8 be also added in the modified MS medium.
Further, in the step 5 and step 6, temperature is 22-26 DEG C, intensity of illumination 1500-2000Lx, relatively
Humidity is 65-75%.
Compared with prior art, the invention has the following advantages:
The tissue culture and rapid propagation method of clematis " cherry lip " provided by the invention, by tissue cultures can quick Propagated Plantlets, it is full
Sufficient market needs.Meanwhile by callus induction and proliferation, make the callus band virus finally applied to induced bud differentiation
A possibility that fall below it is minimum.By the test tube seedling of the bud culture of callus induction differentiation, largely reduce in spite of illness
A possibility that malicious.So carefulness management, the probability of forth infection virus will be greatly reduced after test tube seedling rooting culture.This is just
High benefit is brought for production, while ecological, environmental protective can be reinforced.
The present invention also adjusts the nutritional ingredient of culture medium simultaneously, it is made to be more suitable for the quick breeding of clematis, reduces callus
A possibility that tissue is to virus increases survival rate.
Specific embodiment
To keep the technical problem to be solved in the present invention, technical solution and advantage clearer, specific embodiment will be used below
It is described in detail, but the present invention is limited to absolutely not these examples.The following is only the preferred embodiment of the present invention, is only used to
Explain the present invention, it cannot be construed as a limitation to the scope of the present invention.It should be pointed out that all of the invention
Any modifications, equivalent replacements, and improvements etc. done within spirit and principle, should all be included in the protection scope of the present invention.
Therefore, the scope of protection of the patent of the invention shall be subject to the appended claims.
Except specified otherwise, component used in the present invention is commercial product.
The present invention provides a kind of tissue culture and rapid propagation method of clematis " cherry lip ", and specific embodiment is as follows.
Embodiment 1
A kind of tissue culture and rapid propagation method of clematis " cherry lip ", comprising:
Step 1: the selection of material
Healthy and strong, no disease and pests harm shoot is chosen, the stem section of clip belt segment, every three sections are one section, and the stem section of clip is first flowing
It rinses under water 4 hours, it is then primary with aseptic water washing；
Step 2: on superclean bench, stem section sterilization: being cut into the about 1cm long of one section of band；First use aseptic water washing 1
It is secondary, it is then impregnated 30 seconds with 70% alcohol, then impregnated 10 minutes with 10% sodium hypochlorite, finally with again with 8 points of 0.1% mercuric chloride immersion
During which clock will constantly shake, it is made to sterilize uniform and complete, then use aseptic water washing 5 times；
Step 3: induction of callus: taking above-mentioned stem section, is disposed across Fiber differentiation in the culture medium of evoked callus, and 15
A large amount of jade-green callus is generated in stem section base portion after it；
Evoked callus culture medium: 1/4 improvement MS+NAA 1.0mg/L+2,4-D 0.5mg/L+6-BA 0.5mg/L+10% is fragrant
Any of several broadleaf plants mud+active carbon 1g/L；
Step 4: the Multiplying culture of callus: taking the callus induced, is divided into the fritter of 0.4-0.6cm, is inoculated into liquid
The Multiplying culture of callus is carried out in body shallow-layer culture medium, appreciation rate is 2 times in the Multiplying culture of callus, is carried out next
Step culture；Proliferated culture medium are as follows: 3/4 improvement MS+NAA 1.0mg/L+2,4-D 0.5mg/L+6-BA 0.5mg/L+ active carbon
1g/L；
Step 5: the culture of callus induction bud: taking callus, is cut into the fritter of 1cm or so, is inoculated into differentiation training
Support the induction that bud is carried out in base；It is trained under conditions of temperature is 22 DEG C, intensity of illumination 2000Lx, relative humidity are 70%
It supports；Culture medium is swallow liquid culture base, formula are as follows: 3/4 improvement MS+6-BA 1.5mg/L+TDZ 0.5mg/L+NAA
0.01mg/L+ active carbon 1.5g/L.
Step 6: the culture of rootage of no offspring: when Multiple Buds grow to 3cm, clip branch is inoculated into root media and lures
It leads and takes root；Root media are as follows: 1/2 improvement MS+NAA 0.5mg/L+IBA 0.1mg/L+ active carbon 0.5g/L.
Step 7: the rooting culture of test tube seedling: first moving to greenhouse for test tube seedling, then takes out seedling and cleans root attachment
Culture medium is transplanted to substrate composition are as follows: 1 part of sandy soil, 2 parts of vermiculites: 2 parts of perlites: in natural item in 5 parts of garden mould part mixed-matrixes
It cultivates under part, VC is added in the mixed-matrix and is mixed thoroughly with the sprinkling of 800 times of carbendazim, the VC additional amount is 2.5g/L.
The modified MS medium are as follows: by a great number of elements KNO of MS culture medium3Dosage be changed to 2500mg/L, with (NH4)2SO4Instead of NH4NO3, dosage 200mg/L, other bases are identical as MS.It also added in the modified MS medium
The sucrose of 25g/L, the agar of 7.5g/L, pH 5.8.
Rooting rate of the present embodiment without offspring is 98.4%, and the survival rate for taming 25 days clematis is 94.6%.
Embodiment 2
A kind of tissue culture and rapid propagation method of clematis " cherry lip ", comprising:
Step 1: the selection of material
Healthy and strong, no disease and pests harm shoot is chosen, the stem section of clip belt segment, every three sections are one section, and the stem section of clip is first flowing
It rinses under water 3 hours, it is then primary with aseptic water washing；
Step 2: on superclean bench, stem section sterilization: being cut into the about 0.8cm long of one section of band；First use aseptic water washing 1
It is secondary, it is then impregnated 30 seconds with 70% alcohol, then impregnated 10 minutes with 10% sodium hypochlorite, finally with again with 8 points of 0.1% mercuric chloride immersion
During which clock will constantly shake, it is made to sterilize uniform and complete, then use aseptic water washing 5 times；
Step 3: induction of callus: taking above-mentioned stem section, is disposed across Fiber differentiation in the culture medium of evoked callus, and 15
A large amount of jade-green callus is generated in stem section base portion after it；Evoked callus culture medium: 1/4 improvement MS+NAA
1.0mg/L+2,4-D 0.5mg/L+6-BA 0.5mg/L+10% banana puree+active carbon 1g/L；
Step 4: the Multiplying culture of callus: taking the callus induced, is divided into the fritter of 0.4cm, it is shallow to be inoculated into liquid
The Multiplying culture of callus is carried out in layer culture medium, appreciation rate is 3 times in the Multiplying culture of callus, carries out next step training
It supports；Proliferated culture medium are as follows: 3/4 improvement MS+NAA 1.0mg/L+2,4-D 0.5mg/L+6-BA 0.5mg/L+ active carbon 1g/L；
Step 5: the culture of callus induction bud: taking callus, is cut into the fritter of 1cm or so, is inoculated into differentiation training
Support the induction that bud is carried out in base；It is trained under conditions of temperature is 22 DEG C, intensity of illumination 2000Lx, relative humidity are 70%
It supports；Culture medium is swallow liquid culture base, formula are as follows: 3/4 improvement MS+6-BA 1.5mg/L+TDZ 0.5mg/L+NAA
0.01mg/L+ active carbon 1.5g/L；
Step 6: the culture of rootage of no offspring: when Multiple Buds grow to 5cm, clip branch is inoculated into root media and induces life
Root；Root media are as follows: 1/2 improvement MS+NAA 0.5mg/L+IBA 0.1mg/L+ active carbon 0.5g/L；
Step 7: the rooting culture of test tube seedling: first moving to greenhouse for test tube seedling, then takes out the culture that seedling cleans root attachment
Base is transplanted to substrate composition are as follows: 1 part of sandy soil, 2 parts of vermiculites: 2 parts of perlites: in 5 parts of garden mould part mixed-matrixes under natural conditions
It cultivates, VC is added in the mixed-matrix and is mixed thoroughly with the sprinkling of 800 times of carbendazim, the VC additional amount is 2.5g/L.
The modified MS medium are as follows: by a great number of elements KNO of MS culture medium3Dosage be changed to 2500mg/L, with (NH4)2SO4Instead of NH4NO3, dosage 200mg/L, other bases are identical as MS.It also added in the modified MS medium
The sucrose of 25g/L, the agar of 7.5g/L, pH 5.8.
Rooting rate of the present embodiment without offspring is 99.2%, and the survival rate for taming 25 days clematis is 96.8%.
Embodiment 3
A kind of tissue culture and rapid propagation method of clematis " cherry lip ", comprising:
Step 1: the selection of material
Healthy and strong, no disease and pests harm shoot is chosen, the stem section of clip belt segment, every three sections are one section, and the stem section of clip is first flowing
It rinses under water 4 hours, it is then primary with aseptic water washing；
Step 2: on superclean bench, stem section sterilization: being cut into the about 1cm long of one section of band；First use aseptic water washing 1
It is secondary, it is then impregnated 30 seconds with 70% alcohol, then impregnated 12 minutes with 10% sodium hypochlorite, finally with again with 8 points of 0.1% mercuric chloride immersion
During which clock will constantly shake, it is made to sterilize uniform and complete, then use aseptic water washing 5 times；
Step 3: induction of callus: taking above-mentioned stem section, is disposed across Fiber differentiation in the culture medium of evoked callus, and 15
A large amount of jade-green callus is generated in stem section base portion after it；
Evoked callus culture medium: 1/4 improvement MS+NAA 1.0mg/L+2,4-D 0.5mg/L+6-BA 0.5mg/L+10% is fragrant
Any of several broadleaf plants mud+active carbon 1g/L；
Step 4: the Multiplying culture of callus: taking the callus induced, is divided into the fritter of 0.5cm, it is shallow to be inoculated into liquid
The Multiplying culture of callus is carried out in layer culture medium, appreciation rate is 3 times in the Multiplying culture of callus, carries out next step training
It supports；Proliferated culture medium are as follows: 3/4 improvement MS+NAA 1.0mg/L+2,4-D 0.5mg/L+6-BA 0.5mg/L+ active carbon 1g/L；
Step 5: the culture of callus induction bud: taking callus, is cut into the fritter of 1cm or so, is inoculated into differentiation training
Support the induction that bud is carried out in base；It is trained under conditions of temperature is 26 DEG C, intensity of illumination 1500Lx, relative humidity are 65%
It supports；Culture medium is swallow liquid culture base, formula are as follows: 3/4 improvement MS+6-BA 1.5mg/L+TDZ 0.5mg/L+NAA
0.01mg/L+ active carbon 1.5g/L；
Step 6: the culture of rootage of no offspring: when Multiple Buds grow to 4cm, clip branch is inoculated into root media and induces life
Root；Root media are as follows: 1/2 improvement MS+NAA 0.5mg/L+IBA 0.1mg/L+ active carbon 0.5g/L；
Step 7: the rooting culture of test tube seedling: first moving to greenhouse for test tube seedling, then takes out the culture that seedling cleans root attachment
Base is transplanted to substrate composition are as follows: 1 part of sandy soil, 2 parts of vermiculites: 2 parts of perlites: in 5 parts of garden mould part mixed-matrixes under natural conditions
It cultivates, VC is added in the mixed-matrix and is mixed thoroughly with the sprinkling of 800 times of carbendazim, the VC additional amount is 2.5g/L.
The modified MS medium are as follows: by a great number of elements KNO of MS culture medium3Dosage be changed to 2500mg/L, with (NH4)2SO4Instead of NH4NO3, dosage 200mg/L, other bases are identical as MS.It also added in the modified MS medium
The sucrose of 25g/L, the agar of 7.5g/L, pH 5.8.
Rooting rate of the present embodiment without offspring is 98.9%, and the survival rate for taming 25 days clematis is 95.7%.
The advantages of tissue culture and rapid propagation method to further illustrate clematis " cherry lip " of the invention, only it is with embodiment 2
Example building comparative example is as follows.
Comparative example 1
Modified MS medium in embodiment 3 is replaced with into MS culture medium, remaining condition is same as Example 2.
The rooting rate for the clematis that the method for the comparative example obtains is 81.2%, and taming 25 days survival rates is 75.8%.
Comparative example 2
In this comparative example, step 1-2 is same as Example 2；
Step 3: inducing clumping bud culture: taking the stem section disinfected to be inoculated into and carry out just being commissioned to train in the culture medium of induction Multiple Buds
It supports；The induced medium of Multiple Buds: 3/4 improvement MS+NAA 0.5mg/L+ZT 0.5mg/L+6-BA 2.0mg/L+ active carbon
1g/L；
Step 4: the Multiplying culture of Multiple Buds: the Multiple Buds induced in step 3 being cut into single bud, are inoculated into subculture medium
The middle proliferation of propagation culture for carrying out Multiple Buds；The subculture medium of Multiple Buds: improvement MS+NAA 0.5mg/L+6-BA 2.0mg/L
+ GA 3.0mg/L active carbon 1g/L；
Step 5: the culture of rootage of no offspring: when Multiple Buds grow to 3-5cm, clip branch is inoculated into root media and induces
It takes root；Root media are as follows: 1/2 improvement MS+NAA 0.5mg/L+IBA 0.1mg/L+ active carbon 0.5g/L；
Above-mentioned steps 3-5 is cultivated under conditions of temperature is 22 DEG C, intensity of illumination 2000Lx, relative humidity are 70%；
Follow-up cultivation is same as Example 2.
The rooting rate of this comparative example clematis is 90.1%, and taming 25 days survival rates is 81.4%.
By above-described embodiment and comparative example it is found that the tissue culture and rapid propagation method of clematis provided by the invention " cherry lip ", leads to
Callus induction and proliferation are crossed, falls below the viruliferous possibility of callus finally applied to induced bud differentiation most
It is low.By the test tube seedling of the bud culture of callus induction differentiation, viruliferous possibility is largely reduced.So
After test tube seedling rooting culture, carefulness management, the probability of forth infection virus be will be greatly reduced, and increase survival rate.
The above is a preferred embodiment of the present invention, it is noted that for those skilled in the art
For, without departing from the principles of the present invention, it can also make several improvements and retouch, these improvements and modifications
It should be regarded as protection scope of the present invention.
Claims (9)
1. a kind of tissue culture and rapid propagation method of clematis " cherry lip " characterized by comprising
Step 1: the selection of material
Choose healthy and strong, no disease and pests harm shoot, the stem section of clip belt segment；
Step 2: sterilization
On superclean bench, stem section is cut into the about 0.8-1cm long of one section of band；It first uses aseptic water washing 1 time, then with 70%
Alcohol impregnates 30 seconds, then with 10% sodium hypochlorite 10-15 minute, finally use is again with 0.1% mercuric chloride immersion 7-8 minutes, during which otherwise
It is disconnected to shake, so that it is sterilized uniform and complete, then uses aseptic water washing 4-5 times；
Step 3: induction of callus
Above-mentioned stem section is taken, Fiber differentiation in the culture medium of evoked callus is disposed across, is generated largely after 15 days in stem section base portion
Jade-green callus；
Evoked callus culture medium: 1/4 improvement MS+NAA 1.0mg/L+2,4-D 0.5mg/L+6-BA 0.5mg/L+10% is fragrant
Any of several broadleaf plants mud+active carbon 1g/L；
Step 4: the Multiplying culture of callus
The callus induced is taken, the fritter of 0.4-0.6cm is divided into, is inoculated into swallow liquid culture base and carries out callus
Multiplying culture；
Proliferated culture medium are as follows: 3/4 improvement MS+NAA 1.0mg/L+2,4-D 0.5mg/L+6-BA 0.5mg/L+ active carbon 1g/L；
Step 5: the culture of callus induction bud
Callus is taken, the fritter of 1cm or so is cut into, is inoculated into the induction for carrying out bud in differential medium；
Culture medium is swallow liquid culture base, formula are as follows: 3/4 improvement MS+6-BA 1.5mg/L+TDZ 0.5mg/L+NAA
0.01mg/L+ active carbon 1.5g/L；
Step 6: the culture of rootage without offspring
When Multiple Buds grow to 3-5cm, clip branch is inoculated into root induction in root media；
Root media are as follows: 1/2 improvement MS+NAA 0.5mg/L+IBA 0.1mg/L+ active carbon 0.5g/L.
2. the tissue culture and rapid propagation method of clematis " cherry lip " according to claim 1, which is characterized in that further include step 7:
The rooting culture of test tube seedling: first moving to greenhouse for test tube seedling, then takes out the culture medium that seedling cleans root attachment, is transplanted to base
Matter proportion are as follows: 1 part of sandy soil, 2 parts of vermiculites: 2 parts of perlites: cultivated under natural conditions in 5 parts of garden mould part mixed-matrixes.
3. the tissue culture and rapid propagation method of clematis " cherry lip " according to claim 1, which is characterized in that in the step 1,
Every three sections are one section when clip stem section, and the stem section of clip first rinses 3-4 hour under flowing water, then use aseptic water washing one
It is secondary.
4. the tissue culture and rapid propagation method of clematis " cherry lip " according to claim 1, which is characterized in that in the step 4,
Appreciation rate is 2-3 times in the Multiplying culture of callus.
5. the tissue culture and rapid propagation method of clematis " cherry lip " according to claim 2, which is characterized in that the mixed-matrix
Middle addition VC is simultaneously mixed thoroughly with the sprinkling of 800 times of carbendazim.
6. the tissue culture and rapid propagation method of clematis " cherry lip " according to claim 5, which is characterized in that the VC additional amount
For 2.5g/L.
7. the tissue culture and rapid propagation method of clematis " cherry lip " according to claim 1, which is characterized in that the improvement MS training
Support base are as follows: by a great number of elements KNO of MS culture medium3Dosage be changed to 2500mg/L, with (NH4)2SO4Instead of NH4NO3, dosage
For 200mg/L, other bases are identical as MS.
8. the tissue culture and rapid propagation method of clematis " cherry lip " according to claim 7, which is characterized in that the improvement MS training
Support the sucrose that also added 25g/L in base, the agar of 7.5g/L, pH 5.8.
9. the tissue culture and rapid propagation method of clematis described in -8 " cherry lip " according to claim 1, which is characterized in that the step 5
With in step 6, temperature is 22-26 DEG C, intensity of illumination 1500-2000Lx, relative humidity 65-75%.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110637722A (en) * | 2019-10-24 | 2020-01-03 | 江苏农林职业技术学院 | Induction method of clematis warrior dance callus |
| CN110651713A (en) * | 2019-10-29 | 2020-01-07 | 上海植物园 | Tissue culture method of clematis' Fuji blue |
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| CN104396750A (en) * | 2014-11-18 | 2015-03-11 | 罗翼 | Tissue culture propagation method for clematis lanuginosa |
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| CN104396750A (en) * | 2014-11-18 | 2015-03-11 | 罗翼 | Tissue culture propagation method for clematis lanuginosa |
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| 魏淑云等: "‘里昂城’铁线莲组培体系的建立", 《上海交通大学学报（农业科学版）》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110637722A (en) * | 2019-10-24 | 2020-01-03 | 江苏农林职业技术学院 | Induction method of clematis warrior dance callus |
| CN110651713A (en) * | 2019-10-29 | 2020-01-07 | 上海植物园 | Tissue culture method of clematis' Fuji blue |
| CN110651713B (en) * | 2019-10-29 | 2022-09-13 | 上海植物园 | Tissue culture method of clematis' Fuji blue |
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