CN107258548A - A kind of method of Hibiscus hamabo tissue culture regeneration - Google Patents
A kind of method of Hibiscus hamabo tissue culture regeneration Download PDFInfo
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- CN107258548A CN107258548A CN201710734257.3A CN201710734257A CN107258548A CN 107258548 A CN107258548 A CN 107258548A CN 201710734257 A CN201710734257 A CN 201710734257A CN 107258548 A CN107258548 A CN 107258548A
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- hibiscus hamabo
- culture
- hibiscus
- hamabo
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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
Abstract
The invention discloses a kind of method of Hibiscus hamabo tissue culture regeneration, comprise the following steps：(1) the non-lignifying branch raw then using Hibiscus hamabo is originated as explant, after surface sterilization, is cut into the segment of suitable size；(2) it is inoculated in the culture medium added with variety classes and concentration plant growth regulator, in the sprouting, propagation, the elongation of adventitious bud and the rooting induction culture that carry out axillary bud under suitable illumination and temperature conditionss successively；(3) the healthy and strong Hibiscus hamabo regeneration plant of root system is obtained.The advantage of the invention is that：Not only regeneration efficiency is high, growth coefficient is big and can keep the good characteristic of maternal plant; laid a good foundation for the rapid scale breeding of Hibiscus hamabo elite plant strain, while carrying out genetic improvement to Hibiscus hamabo for later-stage utilization molecular biology method provides important technical support.
Description
Technical field
The present invention relates to biological technical field, more particularly to a kind of method of Hibiscus hamabo tissue culture regeneration.
Background technology
Hibiscus hamabo (Hibiscus hamabo Sieb.et Zucc.) is Malvaceae Hibiscus defoliation small arbor, is originated in
The regions following the line of the sea such as Zhoushan Of Zhejiang Province, Ningbo, its tree crown is dense, spends golden yellow, and flower-shape is big and beautiful in colour, and the florescence is long, autumn has set in posterior lobe
Piece reddens, and substantially, and branch is rich in toughness, resistance to trimming for Seasonal dynamics change, is that a kind of very precious sight flower sees leaf Evergreen garden plant
Kind；In addition, Hibiscus hamabo also has surprising anti-adversity ability, not only can Salt And Alkali Tolerance and seawater invasion and attack, also with very it is strong it is drought-enduring,
Impoverishment tolerant and wind loading rating, are a kind of quite potential Shelter-woods, have in terms of environmental improvement and alkaline land improving
Highly important effect.
In recent decades, because of many reasons such as reclaim fields from the sea, artificial destruction is serious, and Hibiscus hamabo is in extinction border in imminent danger
Ground, is listed in Zhejiang rare and endangered tree species.At present in production, Hibiscus hamabo mainly uses seed propagation, but due to beach wood
Rose of Sharon seed kind shell is hard, and water suction is difficult, causes seed sprout time under natural conditions to postpone and perishable, germination rate is only 10%
Left and right；In addition, using also there are problems that during seed seedling-raising trait segregation and, it is difficult to keep the excellent of maternal strain
Character.Though the rose of Sharon breeds the application of aspect and has been reported that it is cumbersome to there is reproductive process, by season by the sea for cuttage and graft technology
Limitation, and the low problem of reproductive efficiency.Plant tissue culture technique has breeding cycle short because of it, and breeding coefficient is high, required
The advantage such as explant material is few, played an important role in the breeding of endangered plants.
At present, the research bred about Hibiscus hamabo tissue culture still belongs to space state.In order to meet large-scale planting and kind
Improve to the demand of Hibiscus hamabo high quality seedling, be badly in need of exploitation it is a kind of it is simple, stably, the side of efficient Hibiscus hamabo tissue culture regeneration
Method.
The content of the invention
It is an object of the invention to overcome the deficiencies of the prior art and provide it is a kind of it is simple, stably, efficient Hibiscus hamabo
The method of tissue culture regeneration.
The present invention is achieved by the following technical solutions：A kind of method of Hibiscus hamabo tissue culture regeneration, including following step
Suddenly：
(1) the non-lignifying branch raw then using Hibiscus hamabo is originated as explant, after surface sterilization, is cut into suitable big
Small segment；
(2) be inoculated in the culture medium added with variety classes and concentration plant growth regulator, in suitable illumination and
Sprouting, propagation, the elongation of adventitious bud and the rooting induction culture of axillary bud are carried out under temperature conditionss successively；
(3) the healthy and strong Hibiscus hamabo regeneration plant of root system is obtained.
As one of preferred embodiment of the present invention, Hibiscus hamabo is that the having for field that have drawn from is excellent in the step (1)
The perennial Hibiscus hamabo plant of economical character.
One of preferred embodiment as the present invention, the perennial Hibiscus hamabo plant with excellent economical character refers to tool
There are the perennial Hibiscus hamabo plant strains of one or more of characters in high yield, high-quality, good stress resistance.
One of preferred embodiment as the present invention, surface sterilization refers to giving birth to Hibiscus hamabo then in the step (1)
Branch rinses 10-30min through flowing water, after being wiped with 74-76% absolute ethyl alcohols, and the anhydrous second of 74-76% is used in aseptic operating platform
Alcohol sterilizes 15-50s；Then after the hydrogenperoxide steam generator sterilization 2-8min through 10-20%, then 0.05-0.15% mercuric chloride solutions are used
2-5min is sterilized, finally with aseptic water washing 5-6 times.
One of preferred embodiment as the present invention, the segment that suitable size is cut into the step (1) refers to after sterilization
Hibiscus hamabo stem section be cut into the segments of a length of 0.5-1.0cm sizes at least provided with 1 axillary bud, it is standby.
One of preferred embodiment as the present invention, is adjusted in the step (2) added with variety classes and concentration plant growth
The culture medium for saving agent is respectively to be used for the inducing culture that stem segment with axillary buds sprouts induction, the Multiplying culture bred for stem segment with axillary buds
Base, the elongation medium for Elongation of adventitious bud and the root media for adventitious bud rooting.
As one of preferred embodiment of the present invention, the inducing culture be specially added with 0.1-3.0mg/L KT,
The WPM culture mediums of 0.05-0.5mg/L GA3,30g/L sucrose and 7.0g/L agar.
As one of preferred embodiment of the present invention, the proliferated culture medium be specially added with 0.5-2.0mg/L KT,
The WPM culture mediums of 0.1-1.0mg/L TDZ, 30g/L sucrose and 7.0g/L agar；The elongation medium is specially to be added with
0.2-1.0mg/L 6-BA, 0.1-0.5mg/L IBA, the 1/ of 0.05-0.5mg/L GA3,30g/L sucrose and 7.0g/L agar
2WPM culture mediums.
As one of preferred embodiment of the present invention, the root media be specially added with 0.5-2.0mg/L IAA,
The 1/2WPM culture mediums of 0.2-4.0mg/L IBA, 20g/L sucrose and 7.0g/L agar.
One of preferred embodiment as the present invention, suitable illumination and temperature conditionss refer to cultivate in the step (2)
Thing is placed in temperature for 22-26 DEG C, and intensity of illumination is 2000-2500lx, and illumination/dark cycle enters for 16/10h incubated room
Row culture.
One of preferred embodiment as the present invention, the step (1), step (2) and step (3) are aseptically entered
OK.
The advantage of the present invention compared with prior art is：First, convenient material drawing, material are abundant, and it is not subject to seasonal restrictions, it is full
The demand of sufficient Hibiscus hamabo large-scale planting；Second, regeneration efficiency is high, breeding coefficient is big, and axillary bud sprouting rate is up to 100%, armpit
Bud is after Multiplying culture, and proliferation rate is up to 96.8%, and average each explant can produce 7.4 adventitious buds, and the later stage is indefinite
The rooting rate of bud is up to 100%；Third, the stem section of tissue culture regeneration seedling to be obtained can be realized further to sea as explant
The expanding propagation of shore rose of Sharon plant.Therefore, the method for a kind of Hibiscus hamabo tissue culture regeneration provided by the present invention, not only beach
The quick breeding and large-scale production of the rose of Sharon provide technical support, while also entering for later-stage utilization molecular biology method to it
Row genetic improvement provides important technical support.
Brief description of the drawings
Fig. 1 is the Hibiscus hamabo stem section figure being inoculated in inducing culture in embodiment 1；
Fig. 2 is the Hibiscus hamabo stem segment with axillary buds sprouting figure in embodiment 1；
Fig. 3 is the Hibiscus hamabo shoot proliferation figure in embodiment 1；
Fig. 4 is the Hibiscus hamabo Elongation of adventitious bud figure in embodiment 1；
Fig. 5 is the Hibiscus hamabo tissue culture seedling rooting figure in embodiment 1.
Embodiment
Embodiments of the invention are elaborated below, the present embodiment is carried out lower premised on technical solution of the present invention
Implement, give detailed embodiment and specific operating process, but protection scope of the present invention is not limited to following implementations
Example.
Embodiment 1
A kind of method of Hibiscus hamabo tissue culture regeneration of the present embodiment, aseptically carries out, comprises the following steps：
(1) being chosen from field has excellent economical character (one or more of characters in possessing high yield, high-quality, good stress resistance)
Perennial Hibiscus hamabo plant strains, clip non-lignifying branch raw then as explant source；
(2) after branch defoliation, the stem section of 5cm sizes is cut into, surface sterilization is carried out；After surface sterilization, by stem section two ends
Location of necrosis is cut off, then stem section is cut into the segment of a length of 0.7cm sizes at least provided with 1 axillary bud, standby；Wherein, surface
The mode of sterilization is：The current-year branch of Hibiscus hamabo is rinsed into 20min through flowing water, after being wiped with 75% absolute ethyl alcohol, Yu Wu
In bacterium operating desk 30s is sterilized with 75% absolute ethyl alcohol；Then after sterilizing 4min through 15% hydrogenperoxide steam generator, then with 0.1%
Mercuric chloride solution sterilizes 3min, finally with aseptic water washing 6 times；
(3) obtained Hibiscus hamabo stem section segment is inoculated in WPM+1.5mg/L KT, 0.25mg/LGA3+30g/L vertically
Illumination cultivation in the sprouting induction (Fig. 1) of stem segment with axillary buds, incubated room is carried out in the inducing culture of sucrose+7.0g/L agar
After (24 DEG C of temperature, intensity of illumination 2200lx, illumination/dark cycle 16/10h) 2 weeks, the germination rate of axillary bud is up to 100% (figure
2)；
(4) after after axillary bud sprouting, by stem section transfer in WPM+1.0mg/L KT+0.5mg/L TDZ+30g/L sucrose+
The Multiplying culture of axillary bud is carried out in the proliferated culture medium of 7.0g/L agar；Illumination cultivation (24 DEG C of temperature, illumination in incubated room
Intensity 2200lx, illumination/dark cycle 16/10h) after 3 weeks, the proliferation rate of axillary bud is up to 96.8%, and average each explant
Produce 7.4 adventitious buds (Fig. 3)；
(5) treat that adventitious bud length, to 1.0cm, adventitious bud is transferred in 1/2WPM+0.6mg/L 6-BA+, 0.35mg/L IBA+
The elongation culture of adventitious bud is carried out in the elongation medium of 0.3mg/L GA3+30g/L sucrose+7.0g/L agar；Incubated room
Middle illumination cultivation (24 DEG C of temperature, intensity of illumination 2200lx, illumination/dark cycle 16/10h) is after 2 weeks, Elongation of adventitious bud to 2.0-
3.0cm and with 2-6 pieces true leaf (Fig. 4)；
(6) adventitious bud of elongation is transferred in 1/2WPM+1.0mg/L IAA+2.0mg/L IBA+20g/L sucrose+7.0g/
The rooting induction of adventitious bud is carried out in the root media of L agar；(24 DEG C of temperature, illumination is strong for illumination cultivation in incubated room
Spend 2200lx, illumination/dark cycle 16/10h) after 3 weeks, the Hibiscus hamabo intact plant with white healthy and strong root system is obtained, it is raw
Root rate is up to 100% and average each explant produces 6.2 adventitious roots (Fig. 5).
Embodiment 2
A kind of method of Hibiscus hamabo tissue culture regeneration of the present embodiment, aseptically carries out, comprises the following steps：
(1) being chosen from field has excellent economical character (one or more of characters in possessing high yield, high-quality, good stress resistance)
Perennial Hibiscus hamabo plant strains, clip non-lignifying branch raw then as explant source；
(2) after branch defoliation, the stem section of 5cm sizes is cut into, surface sterilization is carried out；After surface sterilization by stem section be cut into
The segment of a length of 0.5cm sizes less with 1 axillary bud, it is standby；Wherein, the mode of surface sterilization is：By working as Hibiscus hamabo
Year, raw branch rinsed 10min through flowing water, after being wiped with 74% absolute ethyl alcohol, was sterilized in aseptic operating platform with 74% absolute ethyl alcohol
15s；Then after sterilizing 2min through 10% hydrogenperoxide steam generator, then with 0.05% mercuric chloride solution sterilization 2min, finally with sterile
Water is rinsed 5 times；
(3) obtained Hibiscus hamabo stem section segment is inoculated in WPM+0.1mg/L KT, 0.05mg/LGA3+30g/L vertically
In the inducing culture of sucrose+7.0g/L agar, and illumination cultivation (22 DEG C of the temperature, intensity of illumination in incubated room
2000lx, illumination/dark cycle 16/10h) 1 week, carry out the sprouting induction of stem segment with axillary buds；
(4) after after axillary bud sprouting, by stem section transfer in WPM+0.5mg/L KT+0.1mg/L TDZ+30g/L sucrose+
In the proliferated culture medium of 7.0g/L agar, and illumination cultivation (22 DEG C of the temperature, intensity of illumination 2000lx, light in incubated room
According to/dark cycle 16/10h) 2 weeks, carry out the Multiplying culture of axillary bud；
(5) treat that to 1.0cm or so, adventitious bud is transferred in 1/2WPM+0.2mg/L 6-BA+, 0.1mg/L for adventitious bud length
In the elongation medium of IBA+0.05mg/L GA3+30g/L sucrose+7.0g/L agar, and the illumination cultivation in incubated room
(22 DEG C of temperature, intensity of illumination 2000lx, illumination/dark cycle 16/10h) 1 week, carries out the elongation culture of adventitious bud；
(6) adventitious bud of elongation is transferred in 1/2WPM+0.5mg/L IAA+0.2mg/L IBA+20g/L sucrose+7.0g/
In the root media of L agar, and illumination cultivation (22 DEG C of temperature, intensity of illumination 2000lx, the illumination/black in incubated room
Dark cycle 16/10h) 2 weeks, the rooting induction of progress adventitious bud；It is final to obtain complete with the Hibiscus hamabo of white healthy and strong root system
Plant.
Embodiment 3
A kind of method of Hibiscus hamabo tissue culture regeneration of the present embodiment, aseptically carries out, comprises the following steps：
(1) being chosen from field has excellent economical character (one or more of characters in possessing high yield, high-quality, good stress resistance)
Perennial Hibiscus hamabo plant strains, clip non-lignifying branch raw then as explant source；
(2) after branch defoliation, the stem section of 5cm sizes is cut into, surface sterilization is carried out；After surface sterilization by stem section be cut into
The segment of a length of 1.0cm sizes less with 1 axillary bud, it is standby；Wherein, the mode of surface sterilization is：By working as Hibiscus hamabo
Year, raw branch rinsed 30min through flowing water, after being wiped with 76% absolute ethyl alcohol, was sterilized in aseptic operating platform with 76% absolute ethyl alcohol
50s；Then after sterilizing 8min through 20% hydrogenperoxide steam generator, then with 0.15% mercuric chloride solution sterilization 5min, finally with sterile
Water is rinsed 6 times；
(3) obtained Hibiscus hamabo stem section segment is inoculated in WPM+3.0mg/L KT, 0.5mg/L GA3+30g/L vertically
In the inducing culture of sucrose+7.0g/L agar, and illumination cultivation (26 DEG C of the temperature, intensity of illumination in incubated room
2500lx, illumination/dark cycle 16/10h) 3 weeks, carry out the sprouting induction of stem segment with axillary buds；
(4) after after axillary bud sprouting, by stem section transfer in WPM+2.0mg/L KT+1.0mg/L TDZ+30g/L sucrose+
In the proliferated culture medium of 7.0g/L agar, and illumination cultivation (26 DEG C of the temperature, intensity of illumination 2500lx, light in incubated room
According to/dark cycle 16/10h) 4 weeks, carry out the Multiplying culture of axillary bud；
(5) treat that to 1.0cm or so, adventitious bud is transferred in 1/2WPM+1.0mg/L 6-BA+, 0.5mg/L for adventitious bud length
In the elongation medium of IBA+0.5mg/L GA3+30g/L sucrose+7.0g/L agar, and the illumination cultivation in incubated room
(26 DEG C of temperature, intensity of illumination 2500lx, illumination/dark cycle 16/10h) 3 weeks, carries out the elongation culture of adventitious bud；
(6) adventitious bud of elongation is transferred in 1/2WPM+2.0mg/L IAA+4.0mg/L IBA+20g/L sucrose+7.0g/
In the root media of L agar, and illumination cultivation (26 DEG C of temperature, intensity of illumination 2500lx, the illumination/black in incubated room
Dark cycle 16/10h) 4 weeks, the rooting induction of progress adventitious bud；It is final to obtain complete with the Hibiscus hamabo of white healthy and strong root system
Plant.
Embodiment 4
The present embodiment tests influence of the stem section growth conditions to Hibiscus hamabo axillary bud sprouting and propagation, and specific steps are such as
Under：
(1) being chosen from field has excellent economical character (one or more of characters in possessing high yield, high-quality, good stress resistance)
Perennial Hibiscus hamabo plant strains, the branch of clip non-lignifying then, semi-lignified and lignifying is as outer respectively
The source of implant；
(2) after branch defoliation, the stem section of 5cm sizes is cut into, surface sterilization is carried out；After surface sterilization, by stem section two ends
Location of necrosis is cut off, then stem section is cut into the segment of a length of 0.7cm sizes at least provided with 1 axillary bud, standby；Wherein, surface
The mode of sterilization is：The current-year branch of Hibiscus hamabo is rinsed into 10min through flowing water, after being wiped with 75% absolute ethyl alcohol, Yu Wu
In bacterium operating desk 15s is sterilized with 75% absolute ethyl alcohol；Then after sterilizing 2min through 10% hydrogenperoxide steam generator, then with 0.1%
Mercuric chloride solution sterilizes 2min, finally with aseptic water washing 5 times；
(3) obtained Hibiscus hamabo stem section segment is inoculated in WPM+1.5mg/L KT, 0.25mg/L GA3+30g/ vertically
In the inducing culture of L sucrose+7.0g/L agar, illumination cultivation in incubated room (24 DEG C of temperature, intensity of illumination 2200lx,
Illumination/dark cycle 16/10h) 3 weeks, carry out the sprouting induction of stem segment with axillary buds；
(4) after after axillary bud sprouting, by stem section transfer in WPM+1.0mg/L KT+0.5mg/L TDZ+30g/L sucrose+
In the proliferated culture medium of 7.0g/L agar, illumination cultivation (24 DEG C of temperature, intensity of illumination 2200lx, illumination/black in incubated room
Dark cycle 16/10h) 3 weeks, the Multiplying culture of progress axillary bud；
(5) treat that adventitious bud length, to 1.0cm, adventitious bud is transferred in 1/2WPM+0.6mg/L 6-BA+, 0.35mg/L IBA+
In the elongation medium of 0.3mg/L GA3+30g/L sucrose+7.0g/L agar, illumination cultivation in incubated room (24 DEG C of temperature,
Intensity of illumination 2200lx, illumination/dark cycle 16/10h) 3 weeks, carry out the elongation culture of adventitious bud；
(6) adventitious bud of elongation is transferred in 1/2WPM+1.0mg/L IAA+2.0mg/L IBA+20g/L sucrose+7.0g/
In the root media of L agar, illumination cultivation (24 DEG C of temperature, intensity of illumination 2200lx, illumination/dark week in incubated room
Phase 16/10h) 3 weeks, carry out the rooting induction of adventitious bud.
Influence of the different growth conditions of stem section to Hibiscus hamabo axillary bud sprouting and propagation is counted, as a result as shown in table 1.
Influence of the stem section growth conditions of table 1 to Hibiscus hamabo axillary bud sprouting and propagation
Note：Data are average value, and each processing includes 120 explants, and each processing is in triplicate.
The research of table 1 shows：Stem section difference growth conditions have on Hibiscus hamabo axillary bud sprouting without influence, but to the propagation of axillary bud
There is certain influence.Using the stem section of lignifying as explant, the cultivation effect of axillary bud is worst, and proliferation rate is only 43.4%, and average
Each explant can only produce 1.5 adventitious buds.And using the Hibiscus hamabo stem section in non-lignifying and semi-lignified as explant
Body, the cultivation effect of axillary bud is preferable, and no significant difference.Wherein using the stem section in non-lignifying state as explant, axillary bud
Proliferation rate be up to 90.5%, and average each explant can produce 4.7 adventitious buds.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
Any modifications, equivalent substitutions and improvements made within refreshing and principle etc., should be included in the scope of the protection.
Claims (10)
1. a kind of method of Hibiscus hamabo tissue culture regeneration, it is characterised in that comprise the following steps：
(1) the non-lignifying branch raw then using Hibiscus hamabo is originated as explant, after surface sterilization, is cut into suitable size
Segment；
(2) it is inoculated in the culture medium added with variety classes and concentration plant growth regulator, in suitable illumination and temperature
Under the conditions of carry out sprouting, propagation, the elongation of adventitious bud and the rooting induction culture of axillary bud successively；
(3) the healthy and strong Hibiscus hamabo regeneration plant of root system is obtained.
2. the method for Hibiscus hamabo tissue culture regeneration according to claim 1, it is characterised in that beach in the step (1)
The rose of Sharon is the perennial Hibiscus hamabo plant with excellent economical character in field of having drawn from.
3. the method for Hibiscus hamabo tissue culture regeneration according to claim 1, it is characterised in that surface in the step (1)
Sterilization refers to the current-year branch of Hibiscus hamabo rinsing 10-30min through flowing water, after being wiped with 74-76% absolute ethyl alcohols, Yu Wu
In bacterium operating desk 15-50s is sterilized with 74-76% absolute ethyl alcohols；Then the hydrogenperoxide steam generator through 10-20% sterilizes 2-8min
Afterwards, then with 0.05-0.15% mercuric chloride solutions 2-5min is sterilized, finally with aseptic water washing 5-6 times.
4. the method for Hibiscus hamabo tissue culture regeneration according to claim 1, it is characterised in that be cut into the step (1)
The segment of suitable size refers to the Hibiscus hamabo stem section after sterilization being cut into a length of 0.5-1.0cm at least provided with 1 axillary bud big
Small segment, it is standby.
5. the method for Hibiscus hamabo tissue culture regeneration according to claim 1, it is characterised in that added in the step (2)
Have variety classes and concentration plant growth regulator culture medium be respectively be used for stem segment with axillary buds sprout induction inducing culture,
Proliferated culture medium, the elongation medium for Elongation of adventitious bud and the taking root for adventitious bud rooting bred for stem segment with axillary buds
Culture medium.
6. the method for Hibiscus hamabo tissue culture regeneration according to claim 5, it is characterised in that the inducing culture is specific
For the WPM culture mediums added with 0.1-3.0mg/L KT, 0.05-0.5mg/L GA3,30g/L sucrose and 7.0g/L agar.
7. the method for Hibiscus hamabo tissue culture regeneration according to claim 5, it is characterised in that the proliferated culture medium is specific
For the WPM culture mediums added with 0.5-2.0mg/L KT, 0.1-1.0mg/L TDZ, 30g/L sucrose and 7.0g/L agar；It is described
Elongation medium is specially to be added with 0.2-1.0mg/L 6-BA, 0.1-0.5mg/L IBA, 0.05-0.5mg/L GA3,30g/L
The 1/2WPM culture mediums of sucrose and 7.0g/L agar.
8. the method for Hibiscus hamabo tissue culture regeneration according to claim 5, it is characterised in that the root media is specific
For the 1/2WPM culture mediums added with 0.5-2.0mg/L IAA, 0.2-4.0mg/L IBA, 20g/L sucrose and 7.0g/L agar.
9. the method for Hibiscus hamabo tissue culture regeneration according to claim 1, it is characterised in that suitable in the step (2)
Illumination and temperature conditionss refer to culture being placed in temperature for 22-26 DEG C, intensity of illumination is 2000-2500lx, illumination/dark
Cycle is cultivated for 16/10h incubated room.
10. the method for the Hibiscus hamabo tissue culture regeneration according to claim any one of 1-9, it is characterised in that the step
(1), step (2) and step (3) are aseptically carried out.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107711514A (en) * | 2017-11-24 | 2018-02-23 | 中国科学院昆明植物研究所 | A kind of nonirrigated farmland rose of Sharon purple flower system fine individual plant tissue culture and rapid propagation method |
| CN107896990A (en) * | 2017-11-24 | 2018-04-13 | 中国科学院昆明植物研究所 | A kind of nonirrigated farmland rose of Sharon seed asepsis sprouting and rapid propagation method |
| CN109258475A (en) * | 2018-11-30 | 2019-01-25 | 四川七彩林业开发有限公司 | A kind of method of rose of Sharon Propogation and culture |
| CN112931217A (en) * | 2021-04-08 | 2021-06-11 | 中国科学院合肥物质科学研究院 | Tissue culture regeneration method of Denmark shrubalthea |
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| CN107896990B (en) * | 2017-11-24 | 2020-04-21 | 中国科学院昆明植物研究所 | Sterile germination and rapid propagation method for hibiscus syriacus seeds in dry land |
| CN109258475A (en) * | 2018-11-30 | 2019-01-25 | 四川七彩林业开发有限公司 | A kind of method of rose of Sharon Propogation and culture |
| CN109258475B (en) * | 2018-11-30 | 2022-04-05 | 四川七彩林科股份有限公司 | Method for breeding and cultivating hibiscus syriacus |
| CN112931217A (en) * | 2021-04-08 | 2021-06-11 | 中国科学院合肥物质科学研究院 | Tissue culture regeneration method of Denmark shrubalthea |
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