CN107258548B - 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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- CN107258548B CN107258548B CN201710734257.3A CN201710734257A CN107258548B CN 107258548 B CN107258548 B CN 107258548B CN 201710734257 A CN201710734257 A CN 201710734257A CN 107258548 B CN107258548 B CN 107258548B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
The invention discloses a kind of method of Hibiscus hamabo tissue culture regeneration, include the following steps: that (1) using the current year of Hibiscus hamabo raw non-lignifying branch as explant source, after surface sterilization, is cut into the dissection of suitable size；(2) it is inoculated in the culture medium added with different kinds and concentrations plant growth regulator, sprouting, proliferation, the elongation of adventitious bud and the rooting induction culture of axillary bud is successively carried out under the conditions of the illumination and temperature of Yu Shiyi；(3) the Hibiscus hamabo regeneration plant of root system stalwartness is obtained.The present invention has the advantages that not only regeneration efficiency is high, growth coefficient is big and is able to maintain the good characteristic of maternal plant; it lays a good foundation for the rapid scale breeding of Hibiscus hamabo elite plant strain, while genetic improvement is carried out to Hibiscus hamabo for later-stage utilization molecular biology method and provides important technical support.
Description
Technical field
The present invention relates to field of biotechnology more particularly to a kind of methods of Hibiscus hamabo tissue culture regeneration.
Background technique
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, tree crown is dense, spends golden yellow, and flower-shape is big and beautiful in colour, and the florescence is long, enters leaf after autumn
Piece reddens, and Seasonal dynamics change is obvious, and branch, rich in toughness, resistance to trimming is a kind of very precious sight Hua Guanye Evergreen garden plant
Kind；In addition, Hibiscus hamabo also has surprising anti-adversity ability, not only can saline-alkali tolerant and seawater invasion, also have it is very 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 equal many reasons of reclaiming fields from the sea, artificial destruction is serious, and Hibiscus hamabo has been 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, leads under natural conditions seed sprout time delay and perishable, germination rate is only 10%
Left and right；In addition, being difficult to keep the excellent of maternal strain using the problems such as there is also trait segregation and hereditary variations when seed seedling-raising
Character.Though the application that the rose of Sharon breeds aspect by the sea of cuttage and graft technology has been reported that it is cumbersome that there are reproductive processes, by season
Limitation, and the problems such as reproductive efficiency is low.For plant tissue culture technique because it is short with breeding cycle, breeding coefficient is high, required
The advantages such as explant material is few, play an important role in the breeding of endangered plants.
Currently, the research in relation to the breeding of Hibiscus hamabo tissue culture still belongs to space state.In order to meet large-scale planting and kind
The demand to Hibiscus hamabo high quality seedling is improved, is badly in need of developing a kind of side of simple, stable, efficient Hibiscus hamabo tissue culture regeneration
Method.
Summary of the invention
It is an object of the invention to overcome the deficiencies of the prior art and provide a kind of simple, stable, efficient Hibiscus hamabos
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 walks as follows
It is rapid:
(1) using the current year of Hibiscus hamabo raw non-lignifying branch as explant source, after surface sterilization, it is suitable for big for being cut into
Small dissection；
(2) be inoculated in the culture medium added with different kinds and concentrations plant growth regulator, the illumination of Yu Shiyi and
Sprouting, proliferation, the elongation of adventitious bud and the rooting induction culture of axillary bud are successively carried out under the conditions of temperature；
(3) the Hibiscus hamabo regeneration plant of root system stalwartness is obtained.
As one of preferred embodiment of the invention, in the step (1) Hibiscus hamabo be have drawn from field have it is excellent
The perennial Hibiscus hamabo plant of economical character.
As one of preferred embodiment of the invention, the perennial Hibiscus hamabo plant with excellent economical character refers to tool
There are high yield, high-quality, one or more of characters in good stress resistance perennial Hibiscus hamabo plant strains.
As one of preferred embodiment of the invention, surface sterilization refers to the current year life of Hibiscus hamabo in the step (1)
Branch rinses 10-30min through flowing water, and after the wiping of 74-76% dehydrated alcohol, the anhydrous second of 74-76% is used in aseptic operating platform
Alcohol sterilizes 15-50s；Then after the hydrogenperoxide steam generator of 10-20% sterilizes 2-8min, then with 0.05-0.15% mercuric chloride solution
2-5min is sterilized, is finally used aseptic water washing 5-6 times.
As one of preferred embodiment of the invention, the dissection of suitable size is cut into the step (1), and refer to will be after disinfection
Hibiscus hamabo stem section be cut into the dissection of a length of 0.5-1.0cm size at least provided with 1 axillary bud, it is spare.
As one of preferred embodiment of the invention, different kinds and concentrations plant growth tune is added in the step (2)
The culture medium of section agent is respectively to be used to stem segment with axillary buds sprout the induced medium of induction, for the Multiplying culture of stem segment with axillary buds proliferation
Base, the elongation medium for Elongation of adventitious bud and the root media for adventitious bud rooting.
As one of preferred embodiment of the invention, the induced medium be specially be added with 0.1-3.0mg/L KT,
The WPM culture medium of 0.05-0.5mg/L GA3,30g/L sucrose and 7.0g/L agar.
As one of preferred embodiment of the invention, the proliferated culture medium be specially be added with 0.5-2.0mg/L KT,
The WPM culture medium 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, 0.05-0.5mg/L GA3,30g/L sucrose and 7.0g/L agar 1/
2WPM culture medium.
As one of preferred embodiment of the invention, the root media be specially be added with 0.5-2.0mg/L IAA,
The 1/2WPM culture medium of 0.2-4.0mg/L IBA, 20g/L sucrose and 7.0g/L agar.
As one of preferred embodiment of the invention, the illumination and temperature condition for being suitable in the step (2) refer to and will cultivate
Object be placed in temperature be 22-26 DEG C, intensity of illumination 2000-2500lx, illumination/dark cycle be 16/10h constant temperature incubation room into
Row culture.
As one of preferred embodiment of the invention, the step (1), step (2) and step (3) aseptically into
Row.
The present invention compared with prior art the advantages of be: first, materials are convenient, material is abundant, and be 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 period is indefinite
The rooting rate of bud is up to 100%；Third, can further realize using the stem section of tissue culture regeneration seedling obtained as explant to sea
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 rose of Sharon it is quick breeding and large-scale production provide technical support, while also for later-stage utilization molecular biology method to its into
Row genetic improvement provides important technical support.
Detailed description of the invention
Fig. 1 is the Hibiscus hamabo stem section figure being inoculated in induced medium 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.
Specific embodiment
It elaborates below to the embodiment of the present invention, the present embodiment carries out under the premise of the technical scheme of the present invention
Implement, the detailed implementation method and specific operation process are given, but protection scope of the present invention is not limited to following implementation
Example.
Embodiment 1
A kind of method of Hibiscus hamabo tissue culture regeneration of the present embodiment, aseptically carries out, includes the following steps:
(1) choosing from field has excellent economical character (having high yield, one or more of characters in high-quality, good stress resistance)
Perennial Hibiscus hamabo plant strains, source of the clip current year raw non-lignifying branch as explant；
(2) after branch defoliation, the stem section of 5cm size is cut into, carries out surface sterilization；After surface sterilization, by stem section both ends
Location of necrosis is cut off, then stem section is cut into the dissection of a length of 0.7cm size at least provided with 1 axillary bud, spare；Wherein, surface
The mode of disinfection are as follows: the current-year branch of Hibiscus hamabo is rinsed into 20min through flowing water, after the wiping of 75% dehydrated alcohol, Yu Wu
30s is sterilized with 75% dehydrated alcohol in bacterium station；Then after 15% hydrogenperoxide steam generator sterilizes 4min, then with 0.1%
Mercuric chloride solution sterilizes 3min, finally uses aseptic water washing 6 times；
(3) obtained Hibiscus hamabo stem section dissection is inoculated in WPM+1.5mg/L KT, 0.25mg/LGA3+30g/L vertically
The sprouting that stem segment with axillary buds is carried out in the induced medium of sucrose+7.0g/L agar induces (Fig. 1), illumination cultivation in constant temperature incubation room
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 axillary bud sprouting, by stem section switching 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 in constant temperature incubation room (24 DEG C of temperature, illumination
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
Generate 7.4 adventitious buds (Fig. 3)；
(5) long to 1.0cm to adventitious bud, 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；Constant temperature incubation 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 piece 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 constant temperature incubation 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 generates 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, includes the following steps:
(1) choosing from field has excellent economical character (having high yield, one or more of characters in high-quality, good stress resistance)
Perennial Hibiscus hamabo plant strains, source of the clip current year raw non-lignifying branch as explant；
(2) after branch defoliation, the stem section of 5cm size is cut into, carries out surface sterilization；After surface sterilization by stem section be cut into
The dissection of a length of 0.5cm size with 1 axillary bud less, it is spare；Wherein, the mode of surface sterilization are as follows: by working as Hibiscus hamabo
Year life branch rinses 10min through flowing water, after the wiping of 74% dehydrated alcohol, is sterilized in aseptic operating platform with 74% dehydrated alcohol
15s；Then through 10% hydrogenperoxide steam generator sterilize 2min after, then with 0.05% mercuric chloride solution disinfection 2min, finally with sterile
Water rinses 5 times；
(3) obtained Hibiscus hamabo stem section dissection is inoculated in WPM+0.1mg/L KT, 0.05mg/LGA3+30g/L vertically
In the induced medium of sucrose+7.0g/L agar, and in constant temperature incubation room illumination cultivation (22 DEG C of temperature, intensity of illumination
2000lx, illumination/dark cycle 16/10h) 1 week, carry out the sprouting induction of stem segment with axillary buds；
(4) after axillary bud sprouting, by stem section switching in WPM+0.5mg/L KT+0.1mg/L TDZ+30g/L sucrose+
In the proliferated culture medium of 7.0g/L agar, and in constant temperature incubation room illumination cultivation (22 DEG C of temperature, intensity of illumination 2000lx, light
According to/dark cycle 16/10h) 2 weeks, carry out the Multiplying culture of axillary bud；
(5) to adventitious bud length to 1.0cm or so, adventitious bud is transferred in 1/2WPM+0.2mg/L 6-BA+, 0.1mg/L
In the elongation medium of IBA+0.05mg/L GA3+30g/L sucrose+7.0g/L agar, and the illumination cultivation in constant temperature incubation room
(22 DEG C of temperature, intensity of illumination 2000lx, illumination/dark cycle 16/10h) 1 week, carry 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 in constant temperature incubation room illumination cultivation (22 DEG C of temperature, intensity of illumination 2000lx, illumination/black
Dark period 16/10h) 2 weeks, carry out the rooting induction of adventitious bud；Final acquisition is 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, includes the following steps:
(1) choosing from field has excellent economical character (having high yield, one or more of characters in high-quality, good stress resistance)
Perennial Hibiscus hamabo plant strains, source of the clip current year raw non-lignifying branch as explant；
(2) after branch defoliation, the stem section of 5cm size is cut into, carries out surface sterilization；After surface sterilization by stem section be cut into
The dissection of a length of 1.0cm size with 1 axillary bud less, it is spare；Wherein, the mode of surface sterilization are as follows: by working as Hibiscus hamabo
Year life branch rinses 30min through flowing water, after the wiping of 76% dehydrated alcohol, is sterilized in aseptic operating platform with 76% dehydrated alcohol
50s；Then through 20% hydrogenperoxide steam generator sterilize 8min after, then with 0.15% mercuric chloride solution disinfection 5min, finally with sterile
Water rinses 6 times；
(3) obtained Hibiscus hamabo stem section dissection is inoculated in WPM+3.0mg/L KT, 0.5mg/L GA3+30g/L vertically
In the induced medium of sucrose+7.0g/L agar, and in constant temperature incubation room illumination cultivation (26 DEG C of temperature, intensity of illumination
2500lx, illumination/dark cycle 16/10h) 3 weeks, carry out the sprouting induction of stem segment with axillary buds；
(4) after axillary bud sprouting, by stem section switching in WPM+2.0mg/L KT+1.0mg/L TDZ+30g/L sucrose+
In the proliferated culture medium of 7.0g/L agar, and in constant temperature incubation room illumination cultivation (26 DEG C of temperature, intensity of illumination 2500lx, light
According to/dark cycle 16/10h) 4 weeks, carry out the Multiplying culture of axillary bud；
(5) to adventitious bud length to 1.0cm or so, adventitious bud is transferred in 1/2WPM+1.0mg/L 6-BA+, 0.5mg/L
In the elongation medium of IBA+0.5mg/L GA3+30g/L sucrose+7.0g/L agar, and the illumination cultivation in constant temperature incubation room
(26 DEG C of temperature, intensity of illumination 2500lx, 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+2.0mg/L IAA+4.0mg/L IBA+20g/L sucrose+7.0g/
In the root media of L agar, and in constant temperature incubation room illumination cultivation (26 DEG C of temperature, intensity of illumination 2500lx, illumination/black
Dark period 16/10h) 4 weeks, carry out the rooting induction of adventitious bud；Final acquisition is 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 proliferation, and specific steps are such as
Under:
(1) choosing from field has excellent economical character (having high yield, one or more of characters in high-quality, good stress resistance)
Perennial Hibiscus hamabo plant strains, the branch of the non-lignifying of clip current year, semi-lignified and lignifying is as outer respectively
The source of implant；
(2) after branch defoliation, the stem section of 5cm size is cut into, carries out surface sterilization；After surface sterilization, by stem section both ends
Location of necrosis is cut off, then stem section is cut into the dissection of a length of 0.7cm size at least provided with 1 axillary bud, spare；Wherein, surface
The mode of disinfection are as follows: the current-year branch of Hibiscus hamabo is rinsed into 10min through flowing water, after the wiping of 75% dehydrated alcohol, Yu Wu
15s is sterilized with 75% dehydrated alcohol in bacterium station；Then after 10% hydrogenperoxide steam generator sterilizes 2min, then with 0.1%
Mercuric chloride solution sterilizes 2min, finally uses aseptic water washing 5 times；
(3) obtained Hibiscus hamabo stem section dissection is inoculated in WPM+1.5mg/L KT, 0.25mg/L GA3+30g/ vertically
In the induced medium of L sucrose+7.0g/L agar, illumination cultivation in constant temperature incubation 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 axillary bud sprouting, by stem section switching in WPM+1.0mg/L KT+0.5mg/L TDZ+30g/L sucrose+
In the proliferated culture medium of 7.0g/L agar, and illumination cultivation in constant temperature incubation room (24 DEG C of temperature, intensity of illumination 2200lx, illumination/black
Dark period 16/10h) 3 weeks, carry out the Multiplying culture of axillary bud；
(5) long to 1.0cm to adventitious bud, 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 constant temperature incubation 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, (24 DEG C of temperature, intensity of illumination 2200lx, illumination/dark is all for illumination cultivation in constant temperature incubation room
Phase 16/10h) 3 weeks, carry out the rooting induction of adventitious bud.
Influence of the stem section difference growth conditions to Hibiscus hamabo axillary bud sprouting and proliferation is counted, the results are shown in Table 1.
Influence of the 1 stem section growth conditions of table to Hibiscus hamabo axillary bud sprouting and proliferation
Note: data are average value, and each processing includes 120 explants, and each processing is in triplicate.
Table 1 research shows that: stem section difference growth conditions on Hibiscus hamabo axillary bud sprouting without influence, but to the proliferation of axillary bud have
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 only can produce 1.5 adventitious buds.And using the Hibiscus hamabo stem section in non-lignifying and semi-lignified as explant
The cultivation effect of body, 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 in essence of the invention
Made any modifications, equivalent replacements, and improvements etc., should all be included in the protection scope of the present invention within mind and principle.
Claims (6)
1. a kind of method of Hibiscus hamabo tissue culture regeneration, which comprises the steps of:
(1) using the current year of Hibiscus hamabo raw non-lignifying branch as explant source, after surface sterilization, it is cut into suitable size
Dissection；
(2) it is inoculated in the culture medium added with different kinds and concentrations plant growth regulator, the illumination and temperature of Yu Shiyi
Under the conditions of successively carry out sprouting, proliferation, the elongation of adventitious bud and the rooting induction culture of axillary bud；
(3) the Hibiscus hamabo regeneration plant of root system stalwartness is obtained；
Culture medium in the step (2) added with different kinds and concentrations plant growth regulator is respectively to be used for stem segment with axillary buds
Sprout induction induced medium, for stem segment with axillary buds proliferation proliferated culture medium, for the elongation medium of Elongation of adventitious bud
And the root media for adventitious bud rooting；
The induced medium is specially 0.1-3.0mg/L KT, 0.05-0.5mg/L GA3,30g/L sucrose and 7.0g/L agar
WPM culture medium；
The proliferated culture medium is specially 0.5-2.0mg/L KT, 0.1-1.0mg/L TDZ, 30g/L sucrose and 7.0g/L agar
WPM culture medium；The elongation medium is specially 0.2-1.0mg/L 6-BA, 0.1-0.5mg/L IBA, 0.05-0.5mg/L
The 1/2WPM culture medium of GA3,30g/L sucrose and 7.0g/L agar；
The root media is specially 0.5-2.0mg/L IAA, 0.2-4.0mg/L IBA, 20g/L sucrose and 7.0g/L agar
1/2WPM culture medium.
2. the method for Hibiscus hamabo tissue culture regeneration according to claim 1, which is characterized 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, which is characterized in that surface in the step (1)
Disinfection, which refers to, rinses 10-30min through flowing water for the current-year branch of Hibiscus hamabo, after the wiping of 74-76% dehydrated alcohol, Yu Wu
15-50s is sterilized with 74-76% dehydrated alcohol in bacterium station；Then the hydrogenperoxide steam generator through 10-20% sterilizes 2-8min
Afterwards, then with 0.05-0.15% mercuric chloride solution 2-5min is sterilized, finally used aseptic water washing 5-6 times.
4. the method for Hibiscus hamabo tissue culture regeneration according to claim 1, which is characterized in that be cut into the step (1)
It is suitable for that the dissection of size refers to that the Hibiscus hamabo stem section after disinfection is cut into a length of 0.5-1.0cm at least provided with 1 axillary bud is big
Small dissection, it is spare.
5. the method for Hibiscus hamabo tissue culture regeneration according to claim 1, which is characterized in that be suitable in the step (2)
Illumination and temperature condition refer to by culture be placed in temperature be 22-26 DEG C, intensity of illumination 2000-2500lx, illumination/dark
Period is that the constant temperature incubation room of 16/10h is cultivated.
6. the method for Hibiscus hamabo tissue culture regeneration according to claim 1-5, which is characterized in that the step
(1), step (2) and step (3) aseptically carry out.
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| CN107711514B (en) * | 2017-11-24 | 2020-02-21 | 中国科学院昆明植物研究所 | Tissue culture and rapid propagation method for excellent individual plant of hibiscus purpureus in dry land |
| 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 |
| 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 |
| CN112931217B (en) * | 2021-04-08 | 2022-04-22 | 中国科学院合肥物质科学研究院 | Tissue culture regeneration method of Denmark shrubalthea |
| CN117243123B (en) * | 2023-10-18 | 2024-07-19 | 连云港市农业科学院 | Tissue culture and rapid propagation method for hibiscus syriacus |
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