CN110432152A - A kind of method of ice dish tissue fast breeding - Google Patents
A kind of method of ice dish tissue fast breeding Download PDFInfo
- Publication number
- CN110432152A CN110432152A CN201910861616.0A CN201910861616A CN110432152A CN 110432152 A CN110432152 A CN 110432152A CN 201910861616 A CN201910861616 A CN 201910861616A CN 110432152 A CN110432152 A CN 110432152A
- Authority
- CN
- China
- Prior art keywords
- culture
- tissue
- seedling
- illumination
- temperature
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- 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
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The present invention relates to a kind of methods of ice dish tissue fast breeding, belong to ice dish technical field of cultivation, and the present invention includes culture, callus tissue culture, adventitious buds differentiation, the subculture numerous, strong plantlets and rootage of expansion and the transplanting of aseptic seedling；The present invention utilizes the mesophyll tissue of aseptic seedling, is cut using blade, contacts exposed palisade tissue and spongy tissue with culture medium；The present invention is bred using aseptic seedling is cultivated, and is reduced pollution rate, the melting brown rate of callus, is improved the survival rate of test, can promote the industrial seedling rearing of ice dish by tissue rapid propagation by this method to obtain a large amount of ice vegetable seedling.
Description
Technical field
The invention belongs to ice dish technical field of cultivation, specifically, are related to a kind of method of ice dish tissue fast breeding.
Background technique
Ice plant (scientific name: Mesembr yanthemum crystallinum Linn.) is Aizoaceae, flower in day
The annual or biennial herb belonged to is a kind of novel health-care vegetable, has ice crystal on blade and show unique characteristics, mesophyll is big,
It is suitable for people of all ages edible without fibr tissue.Ice dish resistance, pest and disease damage is few, quality is high, be increasing peasant income, social synergy it is good
Selection.Ice dish economic benefit is higher, but China's introducing time is short, and cultivation technique is immature.
Currently, the propagation method of traditional ice dish mainly has sowing, plant division, grafting and press strip etc., but ice colza is careful small, out
Seedling is slow, emergence rate is low, and transplanting survival rate is only 11%-20%, it is difficult to be mass produced, the industrialization, scale, mark to ice dish
Standardization kind is implanted with certain limitation.With the development of tissue culture technology, tissue culture quickly breeds one with the factorial production as ice dish
New way.Researcher has had carried out some research the tissue culture propagation of ice dish, passes through seed and stem section
Explant culture obtains regeneration plant, and carries out large-scale production and need largely to foster promising men, and provides stem section explant for it,
But conventional at present seed seed, stem section Explant Propagation there are pollution rates it is high, reproductive efficiency is low the problems such as, be not able to satisfy reality
Production needs.And the big feature of ice dish one is mesophyll hypertrophy, well-grown one plant of ice dish can provide 15 bottles of mesophyll tissue, benefit
Inexpensive, high usage ice dish tissue cultures can be carried out with mesophyll tissue, it is numerous to realize that efficiently large-scale ice dish expands.But mesh
It is preceding not have also for mesophyll tissue's culture of ice dish and rapid propagation method.
Summary of the invention
In order to solve the problems, such as background technique, the present invention provides a kind of method of ice dish tissue fast breeding,
Using the feature of ice dish leaf meat hypertrophy, tissue rapid propagation is carried out using a large amount of mesophyll tissue, reduce callus pollution rate,
Melting brown rate effectively improves its survival rate, can obtain a large amount of ice vegetable seedling by this method tissue rapid propagation, promote the factory of ice dish
Change nursery.
To achieve the above object, the present invention is achieved through the following technical solutions:
The method of the ice dish tissue fast breeding, it is characterised in that: the following steps are included: 1) callus tissue culture；
2) adventitious buds differentiation；3) subculture expands numerous；4) strong plantlets and rootage；5) it transplants；Wherein, callus tissue culture base includes: MS+6-BA
1.0-2.0mg/L+NAA 0.15-0.5mg/L；Adventitious bud differentiation and culture base includes: MS+6-BA 0.2mg/L+NAA 0.3mg/
L；It includes: MS+6-BA 2.0mg/L+NAA 0.1mg/L that subculture, which expands numerous induced medium,；Rooting and hardening-off culture base includes: MS+
NAA 0.01mg/L+IBA 0.02mg/L。
The method of the ice dish tissue fast breeding, specifically includes the following steps:
1) callus tissue culture: mesophyll tissue 1-2cm of no vein without stem section is cut from aseptic seedling, is seeded to callus group
The induction that callus is carried out on culture medium is knitted, when cultivating 45 days, media surface grows 90% callus；
2) adventitious buds differentiation: the callus that step 1) is obtained is transferred in adventitious bud differentiation and culture base, evoking adventive bud
Generation, Fiber differentiation 35 days；
3) subculture expands numerous: the adventitious bud that step 2) is obtained cuts its emerald green blade, is inoculated into induced medium, training
It supports 60 days, obtains proliferation adventitious bud clump；
4) strong plantlets and rootage: Rooting and hardening-off culture is forwarded to after proliferation adventitious bud clump obtained in step 3) is divided into single plant
It is cultivated on base, cultivates 35-40 days, obtain tissue-cultured seedling；
5) it transplants: tissue-cultured seedling obtained in step 4) being transferred to progress hardening culture in greenhouse, later takes tissue-cultured seedling
Out, the culture medium of root system attachment is rinsed with water completely, is transplanted in culture substrate after natural drying to moisture.
Preferably, the culture of aseptic seedling is the following steps are included: a. prepares 1/2 MS culture medium, addition matter in step 1)
The agar that score is 32% is measured, after all dissolutions, culture medium is stand-by through high-temperature sterilization；B. seed is put into vacuum pump with 10%
Liquor natrii hypochloritis impregnate 1min, handle 3 times, each 10-15s；Then it will be rinsed 3-4 times in vacuum pump with sterile water；
C. the seed of sterilizing is put on culture medium, sealing culture.Condition of culture: temperature (23 ± 2) DEG C, intensity of illumination 1500lux, often
Its illumination 14h/d；Wait grow 3 true leaves, and length of blade 5-6cm, aseptic seedling is obtained.
Preferably, in step 1), callus tissue culture condition: dark processing 48h after inoculation；Temperature (25 ± 2) DEG C, light
According to intensity 1500lux, light application time for 24 hours/d.
Preferably, in step 2), adventitious bud differentiation and culture condition: temperature (25 ± 2) DEG C, intensity of illumination 1500-
2000lux, light application time 12h/d；Temperature (18 ± 2) DEG C, intensity of illumination 0lux, interlunation 12h/d.
Preferably, subculture expands numerous inducing culturing condition: dark processing 48h after inoculation in step 3), temperature is 25 DEG C；
Temperature (25 ± 2) DEG C, intensity of illumination 1500-2000lux, light application time 15h/d；Temperature (18 ± 2) DEG C, intensity of illumination 0lux,
Interlunation 9h/d.
Preferably, in step 4), Rooting and hardening-off culture condition: temperature (25 ± 2) DEG C, intensity of illumination 2000lux, illumination
Time 13h/d；Temperature (18 ± 2) DEG C, intensity of illumination 0lux, interlunation 11h/d.
Preferably, in step 5) during hardening culture, net gradually being taken off using sunshade net and carries out day illumination, the refining of lid lid
Seedling 7 days or so, the lid for opening culture bottle 20% carried out hardening 2 days, opens culture bottle cover 50% and carries out hardening 2 days, opens
After cultivating the progress of bottle cover 70% hardening 2 days, tissue-cultured seedling is placed in the arched shed for keeping shady and cool ventilation, opens lid hardening 23 completely
It；It suitably sprays water during hardening, improves the adaptability of tissue-cultured seedling.
Preferably, culture substrate includes perlite, peat and river sand in step 5), three's weight ratio is 1:1.5:2；
Culture substrate dries 30min in 100 DEG C of baking ovens and carries out high-temperature sterilization, is irrigated with water after cooling with being fitted into seedling basin.
Preferably, the seedling basin equipped with culture medium is put into plastic greenhouse after tissue culture transplantation of seedlings, daily using spraying
Watering, keeps certain humidity, temperature is at 25-30 DEG C；After about ten days, transplanted seedling is set to receive sufficient illumination, and normal
Watering, fertilizing.
Beneficial effects of the present invention:
The present invention utilizes the mesophyll tissue of aseptic seedling, is cut using blade, makes exposed palisade tissue and sponge group as far as possible
It knits and is contacted with culture medium.The present invention utilizes the feature of ice dish leaf meat hypertrophy, carries out tissue rapid propagation, drop using a large amount of mesophyll tissue
The low pollution rate of callus, melting brown rate, improve survival rate.It, can be with based on culture medium provided by the invention, condition of culture
Promote the industrial seedling rearing of ice dish by tissue rapid propagation to obtain a large amount of ice vegetable seedling.
The present invention is the tissue rapid propagation carried out to the plant of Aizoaceae, Mesembryanthemum, is ground at present to the tissue culture of Mesembryanthemum
Study carefully even rare.Therefore thinking can be provided for the research of Mesembryanthemum.
Detailed description of the invention
Fig. 1 is ice dish tissue fast breeding procedure chart of the present invention；
Wherein, a is the reset condition of mesophyll tissue；B is callus；C is adventitious bud；D is Multiple Buds；E is Multiple Buds
Single plant bud after segmentation；F is intact plant.
Specific embodiment
It, below will be to preferred reality of the invention in order to keep the purpose of the present invention, technical scheme and beneficial effects clearer
It applies example to be described in detail, to facilitate the technical staff to understand.
Embodiment 1
The present embodiment carries out tissue fast breeding by taking Shandong Shouguang (ice vegetable kind) as an example, specifically includes the following steps:
1) culture of aseptic seedling, comprising the following steps: 7.5g agar (mass fraction is added in the MS culture medium of a. preparation 1/2
For 32%), after all dissolutions, culture medium and pertinent instruments are stand-by through high-temperature sterilization；B. seed is put into vacuum pump with 10%
Liquor natrii hypochloritis impregnate 3 times, each 10-15s, it is then multiple with aseptic water washing；C. in superclean bench by seed
It is put on culture medium, sealing culture.Condition of culture: temperature (23 ± 2) DEG C, intensity of illumination 1500lux, daily illumination 14h/d；To
3 true leaves, and length of blade 5-6cm are grown, aseptic seedling is obtained.
2) callus tissue culture: callus tissue culture base includes: MS+6-BA 1.0mg/L+NAA 0.15mg/L.Ultra-clean
On workbench, mesophyll 1-2cm of no vein without stem section palisade tissue and spongy tissue exposure is rapidly cut from aseptic seedling,
Be seeded on callus tissue culture base carry out callus induction, 3 pieces of callus of every bottle of inoculation, totally 60 bottles；Condition of culture:
Dark processing 48h after inoculation；Temperature (25 ± 2) DEG C, intensity of illumination 1500lux, light application time for 24 hours/d, cultivate 45 days, formation is cured
Injured tissue.Callus browning rate 10%, pollution rate 24%, survival rate 45%.
3) adventitious buds differentiation: adventitious bud differentiation and culture base includes: MS+6-BA 0.2mg/L+NAA 0.3mg/L.By step
2) callus obtained is transferred in adventitious bud differentiation and culture base, the generation of evoking adventive bud, and every bottle is inoculated with 2, is inoculated with 60 bottles.
Condition of culture: temperature (25 ± 2) DEG C, intensity of illumination 1500-2000lux, light application time 12h/d；Temperature (18 ± 2) DEG C, illumination
Intensity 0lux, interlunation 12h/d observe the induction situation of adventitious bud and count proliferation rate 45% after Fiber differentiation 35 days.
4) subculture expands numerous: it includes: MS+6-BA2.0 mg/L+NAA 0.1mg/L that subculture, which expands numerous induced medium,.By step
3) adventitious bud obtained cuts its emerald green blade, is inoculated into induced medium.2 buds of every bottle of inoculation are inoculated with 25 bottles.Culture
Condition: dark processing 48h after inoculation, temperature are 25 DEG C；Temperature (25 ± 2) DEG C, intensity of illumination 1500-2000lux, light application time
15h/d；Temperature (18 ± 2) DEG C, intensity of illumination 0lux, interlunation 9h/d, the quantity that Multiple Buds are counted after culture 60 days are
65%.
5) strong plantlets and rootage: Rooting and hardening-off culture base includes: MS+NAA 0.01mg/L+IBA0.02mg/L.It will be in step 4)
Obtained proliferation adventitious bud clump is divided into after single plant to be forwarded on Rooting and hardening-off culture base and cultivate, condition of culture: temperature (25 ± 2)
DEG C, intensity of illumination 2000lux, light application time 13h/d；Temperature (18 ± 2) DEG C, intensity of illumination 0lux, interlunation 11h/d, training
It supports 35-40 days, obtains tissue-cultured seedling.Melting brown rate 14%, pollution rate 13%, survival rate 25%.
6) it transplants: tissue-cultured seedling obtained in step 5) being transferred to progress hardening culture in greenhouse, gradually using sunshade net
It takes off net and carries out day illumination, lid lid hardening 7 days or so, then carry out hardening of partially uncapping, after 12 days, tissue-cultured seedling is placed in holding
In the arched shed of shady and cool ventilation, lid hardening is opened completely 23 days；It suitably sprays water during hardening, improves the adaptability of tissue-cultured seedling.
The preparation of culture substrate: culture substrate includes perlite, peat and river sand, and three's weight ratio is 1:1.5:2；Culture
Matrix dries 30min in 100 DEG C of baking ovens, is irrigated with tap water after cooling with being fitted into seedling basin.
Tissue-cultured seedling is carefully taken out with tweezers, the culture medium of root system attachment is rinsed well with tap water, is dried in the air naturally to moisture
It is transplanted in culture substrate after dry.After tissue culture transplantation of seedlings, the seedling basin equipped with culture medium is put into plastic greenhouse, daily
Using spray sprinkler, humidity 55% is kept, temperature is at 25-30 DEG C；After about ten days, transplanted seedling is made to receive sufficient illumination, and
And normal watering, fertilizing.
Embodiment 2
The present embodiment carries out tissue fast breeding by taking Shandong Shouguang (ice vegetable kind) as an example, specifically includes the following steps:
1) culture of aseptic seedling, step is the same as embodiment 1.
2) callus tissue culture: callus tissue culture base includes: MS+6-BA 2.0mg/L+NAA 0.5mg/L.Ultra-clean
On workbench, mesophyll 1-2cm of no vein without stem section palisade tissue and spongy tissue exposure is rapidly cut from aseptic seedling,
Be seeded on callus tissue culture base carry out callus induction, 3 pieces of callus of every bottle of inoculation, totally 60 bottles；Condition of culture:
Dark processing 48h after inoculation；Temperature (25 ± 2) DEG C, intensity of illumination 150lux, light application time for 24 hours/d, cultivate 45 days, formation is cured
Injured tissue.Callus browning rate 3%, pollution rate 58.8%, survival rate 27.5%.
3) adventitious buds differentiation: adventitious bud differentiation and culture base includes: MS+6-BA 0.2mg/L+NAA 0.3mg/L.By step
2) callus obtained is transferred in adventitious bud differentiation and culture base, the generation of evoking adventive bud, and every bottle is inoculated with 2, is inoculated with 60 bottles.
Condition of culture: temperature (25 ± 2) DEG C, intensity of illumination 1500-2000lux, light application time 12h/d；Temperature (18 ± 2) DEG C, illumination
Strong 0lux, interlunation 12h/d observe the induction situation of adventitious bud and count proliferation rate 43% after Fiber differentiation 35 days.
4) subculture expands numerous: it includes: MS+6-BA2.0 mg/L+NAA 0.1mg/L that subculture, which expands numerous induced medium,.By step
3) adventitious bud obtained cuts its emerald green blade, is inoculated into induced medium.2 buds of every bottle of inoculation are inoculated with 25 bottles.Culture
Condition: dark processing 48h after inoculation, temperature are 25 DEG C；Temperature (25 ± 2) DEG C, intensity of illumination 1500-2000lux, light application time
15h/d；Temperature (18 ± 2) DEG C, intensity of illumination 0lux, interlunation 9h/d, the quantity that Multiple Buds are counted after culture 60 days are
65%.
5) strong plantlets and rootage: Rooting and hardening-off culture base includes: MS+NAA 0.01mg/L+IBA0.02mg/L.It will be in step 4)
Obtained proliferation adventitious bud clump is divided into after single plant to be forwarded on Rooting and hardening-off culture base and cultivate, condition of culture: temperature (25 ± 2)
DEG C, intensity of illumination 2000lux, light application time 13h/d；Temperature (18 ± 2) DEG C, intensity of illumination 0lux, interlunation 11h/d, training
It supports 35-40 days, obtains tissue-cultured seedling.Melting brown rate 23.3%, pollution rate 21.7%, survival rate 35%.
6) it transplants: with embodiment 1,.
Experimental analysis
1, influence of the 6-BA and NAA Different adding amount to ice dish callus growth in callus tissue culture base
Using the method for embodiment 1, other steps are all the same, in callus tissue culture, according to the difference of 6-BA and NAA
Setting 5 processing of additive amount, the results are shown in Table 1.
Table 1
It tests and compares between 5 processing, it can be seen that emergence rate processing 1, processing are 2 low；It is higher to handle 5 melting brown rates；And it handles
3, the survival rate of processing 4 is relatively high, wherein processing 3 is obvious to be better than other.And pollution rate largely with it is artificial lack of standardization
It operates related.
2, the influence that 6-BA and NAA Different adding amount grows ice dish adventitious bud in adventitious bud culture base
Using the method for embodiment 1, other steps are all the same, in Adventitious bud culture, added according to the difference of 6-BA and NAA
Setting 3 processing of dosage, the results are shown in Table 2.
Table 2
Test 3 processing between compare, it can be seen that survival rate processing 2 be higher than other；Handle 1 melting brown rate, the death rate compared with
It is high；Pollution rate is largely related with artificial non-standard operation.
3, subculture expands the influence that 6-BA and NAA Different adding amount grows ice dish Multiple Buds in breeding culture medium
Using the method for embodiment 1, other steps are all the same, expand in numerous culture in subculture, according to the difference of 6-BA and NAA
Setting 3 processing of additive amount, the results are shown in Table 3.
Table 3
Test 3 processing between compare, it can be seen that survival rate processing 3 be higher than other；Handle 1,2 melting brown rates, the death rate compared with
It is high；Pollution rate is largely related with artificial non-standard operation.
4, the influence that 6-BA and NAA Different adding amount grows ice dish adventitious bud in Rooting and hardening-off culture base
Using the method for embodiment 1, other steps are all the same, in Rooting and hardening-off culture, according to the difference of 6-BA and NAA
Setting 3 processing of additive amount, the results are shown in Table 4.
Table 4
Test 3 processing between compare, it can be seen that survival rate processing 2 be higher than other；Handle 1,3 melting brown rates, the death rate compared with
It is high；Pollution rate is largely related with artificial non-standard operation.
The present invention utilizes the mesophyll tissue of aseptic seedling, is cut using blade, makes exposed palisade tissue and sponge group as far as possible
It knits and is contacted with culture medium.The present invention utilizes the feature of ice dish leaf meat hypertrophy, carries out tissue rapid propagation, drop using a large amount of mesophyll tissue
The low pollution rate of callus, melting brown rate, improve survival rate.It, can be with based on culture medium provided by the invention, condition of culture
Promote the industrial seedling rearing of ice dish by tissue rapid propagation to obtain a large amount of ice vegetable seedling.The present invention is spent to Aizoaceae, in day
The tissue rapid propagation that the plant of category carries out, it is even rare to the group training research of Mesembryanthemum at present.Therefore it can grinding for Mesembryanthemum
Offer thinking is provided.
Finally, it is stated that preferred embodiment above is only used to illustrate the technical scheme of the present invention rather than limits, although logical
It crosses above preferred embodiment the present invention is described in detail, however, those skilled in the art should understand that, can be
Various changes are made to it in form and in details, without departing from claims of the present invention limited range.
Claims (10)
1. a kind of method of ice dish tissue fast breeding, it is characterised in that: the following steps are included: 1) callus tissue culture；2) not
Normal bud differentiation；3) subculture expands numerous；4) strong plantlets and rootage；5) it transplants；
Wherein, callus tissue culture base includes: MS+6-BA 1.0-2.0mg/L+NAA 0.15-0.5mg/L；Adventitious buds differentiation training
Feeding base includes: MS+6-BA 0.2mg/L+NAA 0.3mg/L；It includes: 2.0 mg/L+ of MS+6-BA that subculture, which expands numerous induced medium,
NAA 0.1mg/L；Rooting and hardening-off culture base includes: MS+NAA 0.01mg/L+IBA 0.02mg/L.
2. a kind of method of ice dish tissue fast breeding according to claim 1, it is characterised in that: specifically include following step
It is rapid:
1) callus tissue culture: cutting mesophyll tissue 1-2cm of no vein without stem section from aseptic seedling, is seeded to callus training
The induction that callus is carried out on base is supported, when cultivating 45 days, media surface grows 90% callus；
2) adventitious buds differentiation: the callus that step 1) is obtained is transferred in adventitious bud differentiation and culture base, the production of evoking adventive bud
It is raw, Fiber differentiation 35 days；
3) subculture expands numerous: the adventitious bud that step 2 is obtained cuts its emerald green blade, is inoculated into induced medium, culture 60
It, obtains proliferation adventitious bud clump；
4) strong plantlets and rootage: proliferation adventitious bud clump obtained in step 3) is divided into after single plant and is forwarded on Rooting and hardening-off culture base
Culture cultivates 35-40 days, obtains tissue-cultured seedling；
5) it transplants: tissue-cultured seedling obtained in step 4) being transferred to progress hardening culture in greenhouse, is later taken out tissue-cultured seedling, root
The culture medium of system's attachment is rinsed with water completely, is transplanted in culture substrate after natural drying to moisture.
3. a kind of method of ice dish tissue fast breeding according to claim 2, it is characterised in that: sterile in step 1)
The culture of seedling the following steps are included:
A. 1/2 MS culture medium is prepared, the agar that mass fraction is 32% is added, after all dissolutions, culture medium is through high-temperature sterilization
For use；
B. seed is put into vacuum pump and impregnates 1min with 10% liquor natrii hypochloritis, handled 3 times, each 10-15s；Then it will use
Sterile water rinses 3-4 times in vacuum pump；
C. the seed of sterilizing is put on culture medium, sealing culture；Condition of culture: temperature (23 ± 2) DEG C, intensity of illumination
1500lux, daily 14 h/d of illumination；Wait grow 3 true leaves, and length of blade 5-6cm, aseptic seedling is obtained.
4. a kind of method of ice dish tissue fast breeding according to claim 2, it is characterised in that: in step 1), callus
Conditions of tissue culture: dark processing 48h after inoculation；Temperature (25 ± 2) DEG C, intensity of illumination 1500lux, light application time for 24 hours/d.
5. a kind of method of ice dish tissue fast breeding according to claim 2, it is characterised in that: indefinite in step 2
Bud breaks up condition of culture: temperature (25 ± 2) DEG C, intensity of illumination 1500-2000lux, 12 h/d of light application time；Temperature (18 ± 2)
DEG C, 0 lux of intensity of illumination, 12 h/d of interlunation.
6. a kind of method of ice dish tissue fast breeding according to claim 2, it is characterised in that: in step 3), subculture
Expand numerous inducing culturing condition: dark processing 48h after inoculation, temperature are 25 DEG C；Temperature (25 ± 2) DEG C, intensity of illumination 1500-
2000lux, 15 h/d of light application time；Temperature (18 ± 2) DEG C, intensity of illumination 0 lux, interlunation 9h/d.
7. a kind of method of ice dish tissue fast breeding according to claim 2, it is characterised in that: in step 4), take root
Strong seedling culture condition: temperature (25 ± 2) DEG C, intensity of illumination 2000lux, 13 h/d of light application time；Temperature (18 ± 2) DEG C, illumination
Intensity 0 lux, interlunation 11h/d.
8. a kind of method of ice dish tissue fast breeding according to claim 2, it is characterised in that: in step 5), hardening
During culture, using sunshade net gradually take off net carry out day illumination, lid lid hardening 7 days or so, open culture bottle 20% lid into
It row hardening 2 days, opens culture bottle cover 50% and carries out hardening 2 days, after opening culture bottle cover 70% carries out hardening 2 days, by tissue culture
Seedling is placed in the arched shed for keeping shady and cool ventilation, opens lid hardening completely 23 days；It suitably sprays water during hardening, improves the suitable of tissue-cultured seedling
It should be able to power.
9. according to a kind of described in any item methods of ice dish tissue fast breeding of claim 2 or 8, it is characterised in that: step
5) in, culture substrate includes perlite, peat and river sand, and three's weight ratio is 1:1.5:2；Culture substrate is in 100 DEG C of baking ovens
It dries 30min and carries out high-temperature sterilization, irrigated with water after cooling with being fitted into seedling basin.
10. -9 a kind of described in any item methods of ice dish tissue fast breeding according to claim 1, it is characterised in that: tissue culture
After transplantation of seedlings, the seedling basin equipped with culture medium is put into plastic greenhouse, spray sprinkler is used daily, keeps certain humidity,
Temperature is at 25-30 DEG C；After 10-12 days, transplanted seedling is made to receive sufficient illumination, and normal watering, fertilizing.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910861616.0A CN110432152A (en) | 2019-09-12 | 2019-09-12 | A kind of method of ice dish tissue fast breeding |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910861616.0A CN110432152A (en) | 2019-09-12 | 2019-09-12 | A kind of method of ice dish tissue fast breeding |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110432152A true CN110432152A (en) | 2019-11-12 |
Family
ID=68440108
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910861616.0A Pending CN110432152A (en) | 2019-09-12 | 2019-09-12 | A kind of method of ice dish tissue fast breeding |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110432152A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112335544A (en) * | 2020-10-30 | 2021-02-09 | 天津农学院 | Method for improving seed germination rate and seedling survival rate of ice vegetable seeds and application |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2005330877A1 (en) * | 2005-04-15 | 2006-10-26 | Saga University | Method of constructing transgenic ice plant |
| CN1887064A (en) * | 2006-07-14 | 2007-01-03 | 云南农业大学 | Tissue culture method for Arenaria orbiculata |
| CN103392599A (en) * | 2013-07-26 | 2013-11-20 | 云南农业大学 | Bergenia purpurascens(hook.f.et Thoms.)Engl. tender leaf tissue culture rapid propagation method |
| CN106489735A (en) * | 2016-11-02 | 2017-03-15 | 天津农学院 | Ice colza and the method for stem section tissue culture |
| CN107198704A (en) * | 2016-03-18 | 2017-09-26 | 百岳特生物科技（上海）有限公司 | Frost flower callus extract is in the purposes for postponing Skin Cell aging, conditioning skin, prevention and treatment cutaneum carcinoma |
| WO2018116164A1 (en) * | 2016-12-20 | 2018-06-28 | Evogene Ltd. | Methods of increasing specific plants traits by over-expressing polypeptides in a plant |
-
2019
- 2019-09-12 CN CN201910861616.0A patent/CN110432152A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2005330877A1 (en) * | 2005-04-15 | 2006-10-26 | Saga University | Method of constructing transgenic ice plant |
| US20090217410A1 (en) * | 2005-04-15 | 2009-08-27 | Saga University | Method of Constructing Transgenic Ice Plant |
| CN1887064A (en) * | 2006-07-14 | 2007-01-03 | 云南农业大学 | Tissue culture method for Arenaria orbiculata |
| CN103392599A (en) * | 2013-07-26 | 2013-11-20 | 云南农业大学 | Bergenia purpurascens(hook.f.et Thoms.)Engl. tender leaf tissue culture rapid propagation method |
| CN107198704A (en) * | 2016-03-18 | 2017-09-26 | 百岳特生物科技（上海）有限公司 | Frost flower callus extract is in the purposes for postponing Skin Cell aging, conditioning skin, prevention and treatment cutaneum carcinoma |
| CN106489735A (en) * | 2016-11-02 | 2017-03-15 | 天津农学院 | Ice colza and the method for stem section tissue culture |
| WO2018116164A1 (en) * | 2016-12-20 | 2018-06-28 | Evogene Ltd. | Methods of increasing specific plants traits by over-expressing polypeptides in a plant |
Non-Patent Citations (2)
| Title |
|---|
| MICHAEL S. MEINERS等: "Regeneration of multiple shoots and plants from Mesembryanthemum crystallinum", 《PLANT CELL REPORTS》 * |
| 陈思等: "非洲冰草快速繁殖及试管苗玻璃化的影响因子", 《植物生理学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112335544A (en) * | 2020-10-30 | 2021-02-09 | 天津农学院 | Method for improving seed germination rate and seedling survival rate of ice vegetable seeds and application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102696487B (en) | Method for building leaf in vitro regeneration system of begonia rex | |
| CN101322474B (en) | Method for breeding polyploid royal paulownia by combination of in vitro culture and colchicine treatment | |
| CN104604687A (en) | Method for inducing clustered shoots to rapidly propagate butterfly orchid by utilizing pedicle stem segments after bud cutting | |
| CN101124889B (en) | Method for inducing new strain of tetraploid dendrobium with seeds protocorm | |
| CN101238795A (en) | Artificial breeding method for spathiphyllum floribundum | |
| CN103444552A (en) | Method for inducing eggplant anther to regenerate haplobiont | |
| CN103688855B (en) | A kind of leaflet red bean isolated seed embryo and plant regeneration method | |
| CN111616052A (en) | Rapid propagation and sugar-free rooting culture method and application of apple rootstock catalpa bungei | |
| CN100456922C (en) | Method for producing detoxified sprout by cultivating garlic stem tip combined with cold treatment | |
| CN104322372A (en) | Tissue culture rapid propagation method of plumbago auriculata | |
| CN102499086B (en) | Method for breeding locust | |
| CN103651137A (en) | Rhynchostylis protocorm rapid breeding method | |
| CN107278891B (en) | A kind of apricot plum quick breeding method for tissue culture | |
| CN102144543A (en) | Tissue culture and rapid propagation technology for Clematis 'Arabella' | |
| CN101855995B (en) | Tissue culture propagation method of Primula mallophylla Balf.f. | |
| CN103004609B (en) | Tissue culture method for Primula beesiana var. leucantha (Balf. f. et Forr.) Fletcher | |
| CN108142281A (en) | A kind of Cortex Eucommiae method for tissue culture | |
| CN110432152A (en) | A kind of method of ice dish tissue fast breeding | |
| CN104542285B (en) | A kind of method of hemerocailis middendorffi leaf tissue culture | |
| CN108260531B (en) | Tissue culture rapid propagation method for regeneration of taxus cuspidata stem induced plants | |
| CN107980612A (en) | A kind of blue quick breeding of short distance groove and tongue and bloom controlling method | |
| CN108812327B (en) | Culture solution and culture method for water culture propagation of chrysosporium | |
| CN112293252A (en) | Artificial efficient clonal propagation method of dendrobium santalinum | |
| CN104686333B (en) | A kind of ripple leaf Stranvaesia Davidiana Dcne tissue cultivation rapid breeding method | |
| CN109526748A (en) | A kind of Spathiphyllum kochii inflorescence method for tissue culture |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20191112 |
|
| WD01 | Invention patent application deemed withdrawn after publication |