CN105706933A - Chuzhou chrysanthemum strain in-vitro regeneration method - Google Patents
Chuzhou chrysanthemum strain in-vitro regeneration method Download PDFInfo
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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
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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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Abstract
The invention discloses a Chuzhou chrysanthemum strain in-vitro regeneration method. Chuzhou chrysanthemum regenerated strains are obtained by taking newly-growing and non-lignified stem segments, with axillary buds, of field Chuzhou chrysanthemum strains as explants, disinfecting the surfaces of the explants, inoculating the stem segments in an adventitious bud induction medium, conducting illumination cultivation for 1-2 weeks, waiting for stem segment axillary bud germination, conducting induction around the axillary buds to obtain adventitious bud clusters, conducting cultivation for 2-3 weeks, transplanting the stem segments with the adventitious bud clusters to a proliferation medium for proliferation and elongation culture of adventitious buds, transplanting the stem segments to a rooting culture medium for rooting when the buds grow to 2-3 cm, conducting illumination culture for 3-4 weeks, and conducting seedling hardening and transplanting when young seedlings grow to 3-4 cm and have 2-4 main leaves. The method is high in reproduction speed and coefficient, each explant at least generates dozens of cluster buds on average after each time of amplified cultivation, excellent characteristics of Chuzhou chrysanthemum maternal plants are kept, and technical support is provided for the rapid and large-scale reproduction and genetic improvement of excellent Chuzhou chrysanthemum strains.
Description
Technical field
The invention belongs to plant biotechnology field, a kind of method being specifically related to Flos Chrysanthemi strain Regeneration in Vitro.
Background technology
Flos Chrysanthemi (DendranthemamorifoliumCV. ' chuju '), is Compositae Chrysanthemum herbaceos perennial, mainly originates in Anhui Chuzhou, be therefore called again Anhui Flos Chrysanthemi.It is used as medicine with flower, belongs to the big famous genuine medicinal materials in Anhui Province four, rank first of the four big medicine chrysanthemums of the whole nation, belong to medicine, tea dual-purpose good merchantable brand, there is high medicinal, health value.
Modern pharmacology detection analysis shows, Flos Chrysanthemi is rich in flavone, volatile oil and 8 kinds of essential amino acids;Additionally, rich in 10 kinds of trace element such as zinc and selenium in Flos Chrysanthemi, wherein the content of " selenium " is significantly higher than other Dendranthema morifolium Varieties.Owing to zinc and selenium are the requisite two kinds of trace element of human body, participating in the multiple metabolic process of body, removing human free radical, protecting cardiovascular, prevention of arterial is atherosis, regulates the aspect such as blood glucose and the liver protecting and has important function.Owing to Flos Chrysanthemi is rich in above-mentioned effective ingredient, long-term drink Flos Chrysanthemi can heat-clearing and toxic substances removing, relaxing muscles and tendons to promote blood circulation, hepatoprotective improving eyesight, enhancing body immunity;Simultaneously to hypertension, coronary heart disease, arteriosclerosis is evident in efficacy.
Research also finds that SRAS virus, cancer (especially hepatocarcinoma) etc. are had good preventive effect by Flos Chrysanthemi, also have obvious therapeutical effect to diabetes.In recent years, along with Flos Chrysanthemi is medicinal and the enforcement of deepen continuously research and the national standard " Products of Local Geographical Indication Flos Chrysanthemi " of health value, market is increasing to the demand of Flos Chrysanthemi so that the industrialization cultivation development of Flos Chrysanthemi is faced with unprecedented opportunities.But lack efficient high quality seedling raising technology, become the bottleneck restricting its industrialization development.
On producing, the breeding of Flos Chrysanthemi mainly adopts traditional asexual propagation means such as plant division, press strip and grafting.It is limited that reproductive process is respectively present reproductive number, management inconvenience and the problem such as reproductive process is loaded down with trivial details;Simultaneously because field cultivation for many years, Flos Chrysanthemi is made to be faced with variety deterioration, yield reduces, quality deteriorates the problems such as serious with pest and disease damage.Adopt conventional breeding methods that Flos Chrysanthemi is carried out breed improvement, far from being suitable for the chrysanthemum agriculture requirement to variety yield and quality.And adopt group culturation rapid propagating technology, good seed substantial amounts of, that genetic background is consistent can be obtained in a short time, to meet the implantation in large scale demand to Flos Chrysanthemi high quality seedling.It addition, it is also extremely urgent that Flos Chrysanthemi improved seeds carry out corresponding genetic improvement research on this basis.
Less about the research of Flos Chrysanthemi tissue culture regeneration at present, mainly to utilize blade for outer implant, Flos Chrysanthemi carried out numerous soon or cultivating detoxification Flos Chrysanthemi by Apex propagation by indirect induction callus, it is common to there is the problem that breeding coefficient is low.Therefore, for problems existing in current Flos Chrysanthemi reproductive process, a kind of method being badly in need of developing Flos Chrysanthemi strain Regeneration in Vitro, to meet the demand of the Fast-propagation to Flos Chrysanthemi elite plant strain and breed improvement.
Summary of the invention
It is an object of the invention to use plant tissue culture technique, it is provided that a kind of method of Flos Chrysanthemi strain Regeneration in Vitro.To achieve these goals, the present invention adopts the following technical scheme that a kind of method of Flos Chrysanthemi strain Regeneration in Vitro, comprises the following steps:
(1) the not lignified stem segment with axillary bud of new life taking field Flos Chrysanthemi strain is outer implant, to its surface disinfection;
(2) stem section is inoculated in adventitious bud induction culture base, carries out 1~2 week illumination cultivation, treat that stem segment with axillary buds is sprouted, and induce adventitious bud clump at axillalry bud periphery;
(3), after cultivating 2~3 weeks, the propagation carrying out adventitious bud in proliferated culture medium of the stem section with adventitious bud clump being transferred and elongation are cultivated;
(4) treat that bud length is to 2~3cm, goes in root media and takes root;
(5) illumination cultivation 3~4 weeks, during until seedling length to 3~4cm and with 2~4 true leaves, carry out seedling exercising and transplant, it is thus achieved that Flos Chrysanthemi regeneration plant.
Preferably, the stem section of described step (1) is the newborn non-lignifying branch of Flos Chrysanthemi elite plant strain that Field Screening goes out that has drawn from.
Preferably, the stem segment with axillary bud described in described step (1) be outer implant after preliminarily pasteurized, in aseptic operating platform, carry out surface sterilization.
Preferably, described preliminarily pasteurized is sub-packed in 500ml beaker after referring to stem section defoliation, rinses 10~20min under flowing water, and period adds 1~2ml cleaning mixture and rinses, and constantly rocks.
Preferably, the sterilization of described stem section surface refers to that stem section cuts into 4~6cm size, in aseptic operating platform, through aseptic water washing 2~3 times;Surface sterilization 10~30s, aseptic water washing 3~5 times is carried out again with 75% ethanol;Afterwards then through the HgCl of 0.1%2Sterilization 1~3min, finally uses aseptic water washing 6~8 times, ceaselessly shakes during sterilization.
Preferably, in described step (2), the stem section for inoculating refers to that stem section cuts two ends sterilization damage location before inoculation, and the cutting being then cut into 0.3~0.6cm size is standby, and each cutting band is at least provided with an axillalry bud.
Preferably, in described step (2), stem section vaccination ways is for being vertically placed in adventitious bud induction culture base.
Preferably, in described step (2), described inducing culture includes: MS+0.2~2.0mg/L6-BA+30g/L sucrose+7.0g/L agar, pH=5.8.
Preferably, in described step (3), described proliferated culture medium includes: MS+0.1~0.75mg/L6-BA+0.0~0.5mg/LNAA+30g/L sucrose+7.0g/L agar, pH=5.8.
Preferably, in described step (3), described root media includes: 1/2MS+0.0~1.0mg/LIBA+20g/L sucrose+7.0g/L agar, pH=5.8.
The method of a kind of Flos Chrysanthemi strain Regeneration in Vitro provided by the present invention, is specifically related to the non-lignifying stem section of Flos Chrysanthemi field elite plant strain current-year branch for outer implant, by the regeneration of sprouting of direct induced bundle, Flos Chrysanthemi elite plant strain is carried out efficient Fast-propagation.
The present invention has advantage highlighted below:
One, stem section is the not lignified branch of new life of field perennial Flos Chrysanthemi elite plant strain of having drawn from, and can keep the Specific character of maternal plant on the one hand;Another aspect is drawn materials conveniently, material enriches;What can realize field Flos Chrysanthemi fine individual plant is numerous soon, it is achieved breed of variety;
Its two, regenerative process China and foreign countries implant disinfectant program is simple, and disinfecting time is short, and the murder by poisoning of external implant is less;
Its three, Multiple Buds regeneration efficiency is high, and inductivity is up to 100%;
Its four, reproduction speed is fast, and breeding coefficient is high, and can realize the aseptic seedling stem sections of acquisition is carried out further expanding propagation, and Fast-propagation and genetic improvement for excellent Flos Chrysanthemi strain provide important technical support.
Therefore, the method for Flos Chrysanthemi strain Regeneration in Vitro provided by the present invention, the Fast-propagation and the large-scale production that are not only Flos Chrysanthemi elite plant strain provide technical support, are also that the breed improvement of later stage Flos Chrysanthemi is laid a good foundation simultaneously.
Accompanying drawing explanation
The sprouting of Fig. 1 Flos Chrysanthemi stem segment with axillary buds
The induction of Fig. 2 stem segment with axillary buds place adventitious bud clump
The propagation of Fig. 3 adventitious bud bud and elongation
Fig. 4 adventitious bud rooting
The seedling exercising of Fig. 5 Flos Chrysanthemi regeneration plant, domestication
Detailed description of the invention
Below in conjunction with embodiment, the present invention is described in further details.
Embodiment 1
A kind of method of Flos Chrysanthemi strain Regeneration in Vitro, concrete operations are as follows:
1, well-grown perennial Flos Chrysanthemi elite plant strain is chosen from land for growing field crops, the not lignified branch of the 20d seedling age that clip is newborn;
2, after defoliation, it is sub-packed in 500ml beaker and adds 1ml detergent, in running water 10min;Then cut into the stem section of 4cm size, in aseptic operating platform, with after aseptic water washing 3 times with 75% alcohol disinfecting 20s, aseptic water washing 5 times, sterilize 1min with the mercuric chloride of 0.1% afterwards, finally with aseptic water washing 6 times, ceaselessly shake during sterilization.After sterilization, stem section two ends location of necrosis is excised, then the cutting that stem section is cut into about 0.3cm is standby;
3, stem section cutting is vertically inoculated in adventitious bud induction culture base and carries out the sprouting of axillalry bud and the induction of adventitious bud, after the illumination cultivation of the 1st week, stem segment with axillary buds is sprouted, again illumination cultivation after 1 week axillalry bud periphery induce adventitious bud clump, axillary bud sprouting rate is 100%, and the inductivity of adventitious bud is 85.7%;Adventitious bud induction culture base is MS+0.2mg/L6-BA+30g/L sucrose+7.0g/L agar, pH=5.8;
4, after continuing cultivation 2 weeks, the propagation carrying out adventitious bud in adventitious bud proliferation culture medium of being transferred by adventitious bud clump and elongation are cultivated, and illumination cultivation obtains the adventitious bud clump of a large amount of elongation, average each outer implant 5.8 adventitious buds of generation after 3 weeks;Adventitious bud proliferation culture medium is: MS+0.1mg/L6-BA+30g/L sucrose+7.0g/L agar, pH=5.8.
5, the adventitious bud being highly 2~3cm is gone in root media and cultivate 2 weeks, it is thus achieved that with the complete regenerated plant of elongated root system, Flos Chrysanthemi aseptic seedling rooting rate is up to 100%, and root media is: 1/2MS+20g/L sucrose+7.5g/L agar, pH=5.8;
6, illumination cultivation 3~4 weeks, during until seedling length to 3~4cm and with 2~4 true leaves, carry out seedling exercising and transplanting, it is thus achieved that healthy and strong Flos Chrysanthemi regeneration plant.
Embodiment 2
A kind of method of Flos Chrysanthemi strain Regeneration in Vitro, concrete operations are as follows:
1, well-grown perennial Flos Chrysanthemi elite plant strain is chosen from land for growing field crops, the not lignified branch of the 30d seedling age that clip is newborn;
2, after defoliation, it is sub-packed in 500ml beaker and adds 2ml detergent, in running water 15min;Then cut into the stem section of 5cm size, in aseptic operating platform, with after aseptic water washing 3 times with 75% alcohol disinfecting 10s, aseptic water washing 3 times, sterilize 2min with the mercuric chloride of 0.1% afterwards, finally with aseptic water washing 7 times, ceaselessly shake during sterilization.After sterilization, stem section two ends location of necrosis is excised, then the cutting that stem section is cut into about 0.4cm is standby;
3, stem section cutting is vertically inoculated in adventitious bud induction culture base and carries out the sprouting of axillalry bud and the induction of adventitious bud, after the illumination cultivation of the 1st week, stem segment with axillary buds sprouts (Fig. 1), again illumination cultivation after 1 week axillalry bud periphery induce adventitious bud clump (Fig. 2), axillary bud sprouting rate is 100%, and the inductivity of adventitious bud is 100%;Adventitious bud induction culture base is MS+1.0mg/L6-BA+30g/L sucrose+7.0g/L agar, pH=5.8;
4, after continuing cultivation 2 weeks, the propagation carrying out adventitious bud in adventitious bud proliferation culture medium of being transferred by adventitious bud clump and elongation are cultivated, and illumination cultivation obtains the adventitious bud clump (Fig. 3) of a large amount of elongation, average each outer implant 11.5 adventitious buds of generation after 3 weeks;Adventitious bud proliferation culture medium is: MS+0.5mg/L6-BA+0.2mg/LNAA+30g/L sucrose+7.0g/L agar, pH=5.8;
5, the adventitious bud being highly 2~3cm is gone in root media and cultivate 2 weeks, obtaining complete regeneration plant (Fig. 4), Flos Chrysanthemi aseptic seedling rooting rate is 100%, root system length and healthy and strong, root media is: 1/2MS+0.2mg/LIBA+20g/L sucrose+7.5g/L agar, pH=5.8;
6, illumination cultivation 3~4 weeks, during until seedling length to 3~4cm and with 2~4 true leaves, carry out seedling exercising and transplanting, it is thus achieved that healthy and strong Flos Chrysanthemi regeneration plant (Fig. 5).
Embodiment 3
A kind of method of Flos Chrysanthemi strain Regeneration in Vitro, concrete operations are as follows:
1, well-grown perennial Flos Chrysanthemi elite plant strain is chosen from land for growing field crops, the not lignified branch of the 45d seedling age that clip is newborn;
2, after defoliation, it is sub-packed in 500ml beaker and adds 2ml detergent, in running water 20min;Then cut into the stem section of 6cm size, in aseptic operating platform, with after aseptic water washing 2 times with 75% alcohol disinfecting 30s, aseptic water washing 4 times, sterilize 3min with the mercuric chloride of 0.1% afterwards, finally with aseptic water washing 8 times, ceaselessly shake during sterilization.After sterilization, stem section two ends location of necrosis is excised, then the cutting that stem section is cut into about 0.6cm is standby;
3, stem section cutting is vertically inoculated in adventitious bud induction culture base and carries out the sprouting of axillalry bud and the induction of adventitious bud, after the illumination cultivation of the 1st week, stem segment with axillary buds is sprouted, again illumination cultivation after 1 week axillalry bud periphery induce adventitious bud clump, axillary bud sprouting rate is 100%, and the inductivity of adventitious bud is 80%;Adventitious bud induction culture base is MS+2.0mg/L6-BA+30g/L sucrose+7.0g/L agar, pH=5.8;
4, after continuing cultivation 2 weeks, the propagation carrying out adventitious bud in adventitious bud proliferation culture medium of being transferred by adventitious bud clump and elongation are cultivated, and illumination cultivation obtains the adventitious bud clump of a large amount of elongation, average each outer implant 7.6 adventitious buds of generation after 3 weeks;Adventitious bud proliferation culture medium is: MS+0.75mg/L6-BA+0.5mg/LNAA+30g/L sucrose+7.0g/L agar, pH=5.8;
5, the adventitious bud being highly 2~3cm is gone in root media and cultivate 2 weeks, obtaining complete regeneration plant, Flos Chrysanthemi aseptic seedling rooting rate is 100%, and root is many and short and thick, root media is: 1/2MS+1.0mg/LIBA+20g/L sucrose+7.5g/L agar, pH=5.8;
6, illumination cultivation 3~4 weeks, during until seedling length to 3~4cm and with 2~4 true leaves, carry out seedling exercising and transplanting, it is thus achieved that healthy and strong Flos Chrysanthemi regeneration plant.
Embodiment 4
A kind of method of Flos Chrysanthemi strain Regeneration in Vitro, concrete operations are as follows:
1, taking the stem section of the seedling age Flos Chrysanthemi elite plant strain group training aseptic seedling obtained, be cut into 0.4cm size, each stem section is at least with an axillalry bud;
2, being vertically inoculated in adventitious bud induction culture base by stem section cutting and carry out the sprouting of axillalry bud and the induction of adventitious bud, through the illumination cultivation of 2 weeks, axillary bud sprouting also induced adventitious bud clump at axillalry bud periphery, and the inductivity of adventitious bud is 100%;Adventitious bud induction culture base is MS+1.0mg/L6-BA+30g/L sucrose+7.0g/L agar, pH=5.8;
3, after continuing cultivation 2 weeks, the propagation carrying out adventitious bud in adventitious bud proliferation culture medium of being transferred by adventitious bud clump and elongation are cultivated, and illumination cultivation obtains the adventitious bud of a large amount of elongation, average each outer implant 8.0 adventitious buds of product after 3 weeks;Adventitious bud proliferation culture medium is: MS+0.5mg/L6-BA+0.2mg/LNAA+30g/L sucrose+7.0g/L agar, pH=5.8;
The adventitious bud being highly 2~3cm is gone in root media and cultivate 2 weeks, obtaining complete regeneration plant, Flos Chrysanthemi aseptic seedling rooting rate is 100%, root system length and healthy and strong, root media is: 1/2MS+0.2mg/LIBA+20g/L sucrose+7.5g/L agar, pH=5.8;
5, the stem section of above-mentioned Flos Chrysanthemi aseptic seedling being cut into 0.5cm size, each stem section, with at least one axillalry bud, is inoculated in the induction and propagation that carry out adventitious bud in above-mentioned Induce aerosor and proliferated culture medium, and average each stem section can produce about 12 adventitious buds;And the adventitious bud after propagation is inoculated in the elongation and the root culture that carry out adventitious bud in above-mentioned identical elongation and root media;
6, illumination cultivation 3~4 weeks, during until seedling length to 3~4cm and with 2~4 true leaves, carry out seedling exercising and transplanting, it is thus achieved that healthy and strong Flos Chrysanthemi regeneration plant.
The foregoing is only presently preferred embodiments of the present invention, not in order to limit the present invention, all any amendment, equivalent replacement and improvement etc. made within the spirit and principles in the present invention, should be included within protection scope of the present invention.
Claims (10)
1. the method for a Flos Chrysanthemi strain Regeneration in Vitro, it is characterised in that: comprise the following steps:
(1) the not lignified stem segment with axillary bud of new life taking field Flos Chrysanthemi strain is outer implant, to its surface disinfection;
(2) stem section is inoculated in adventitious bud induction culture base, carries out 1~2 week illumination cultivation, treat that stem segment with axillary buds is sprouted, and induce adventitious bud clump at axillalry bud periphery;
(3), after cultivating 2~3 weeks, the propagation carrying out adventitious bud in proliferated culture medium of the stem section with adventitious bud clump being transferred and elongation are cultivated;
(4) treat that bud length is to 2~3cm, goes in root media and takes root;
(5) illumination cultivation 3~4 weeks, during until seedling length to 3~4cm and with 2~4 true leaves, carry out seedling exercising and transplant, it is thus achieved that Flos Chrysanthemi regeneration plant.
2. the method for a kind of Flos Chrysanthemi strain Regeneration in Vitro according to claim 1, it is characterised in that: the stem section of described step (1) is the newborn non-lignifying branch of Flos Chrysanthemi elite plant strain that Field Screening goes out that has drawn from.
3. the method for a kind of Flos Chrysanthemi strain Regeneration in Vitro according to claim 1, it is characterised in that the stem segment with axillary bud described in described step (1) be outer implant after preliminarily pasteurized, in aseptic operating platform, carry out surface sterilization.
4. the method for a kind of Flos Chrysanthemi strain Regeneration in Vitro according to claim 3, it is characterized in that, described preliminarily pasteurized is sub-packed in 500ml beaker after referring to stem section defoliation, rinses 10~20min under flowing water, period adds 1~2ml cleaning mixture and rinses, and constantly rocks.
5. the method for a kind of Flos Chrysanthemi strain Regeneration in Vitro according to claim 3, it is characterised in that the sterilization of described stem section surface refers to that stem section cuts into 4~6cm size, in aseptic operating platform, through aseptic water washing 2~3 times;Surface sterilization 10~30s, aseptic water washing 3~5 times is carried out again with 75% ethanol;Afterwards then through the HgCl of 0.1%2Sterilization 1~3min, finally uses aseptic water washing 6~8 times, ceaselessly shakes during sterilization.
6. the method for a kind of Flos Chrysanthemi strain Regeneration in Vitro according to claim 1, it is characterized in that: in described step (2), stem section for inoculating refers to that stem section cuts two ends sterilization damage location before inoculation, the cutting being then cut into 0.3~0.6cm size is standby, and each cutting band is at least provided with an axillalry bud.
7. the method for a kind of Flos Chrysanthemi strain Regeneration in Vitro according to claim 1, it is characterised in that: in described step (2), stem section vaccination ways is for being vertically placed in adventitious bud induction culture base.
8. the method for a kind of Flos Chrysanthemi strain Regeneration in Vitro according to claim 1, it is characterised in that: in described step (2), described inducing culture includes: MS+0.2~2.0mg/L6-BA+30g/L sucrose+7.0g/L agar, pH=5.8.
9. the method for a kind of Flos Chrysanthemi strain Regeneration in Vitro according to claim 1, it is characterized in that: in described step (3), described proliferated culture medium includes: MS+0.1~0.75mg/L6-BA+0.0~0.5mg/LNAA+30g/L sucrose+7.0g/L agar, pH=5.8.
10. the method for a kind of Flos Chrysanthemi strain Regeneration in Vitro according to claim 1, it is characterised in that: in described step (3), described root media includes: 1/2MS+0.0~1.0mg/LIBA+20g/L sucrose+7.0g/L agar, pH=5.8.
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CN108849512A (en) * | 2018-07-16 | 2018-11-23 | 中国科学院合肥物质科学研究院 | A kind of method of spun gold emperor chrysanthemum tissue culture regeneration |
CN114916440A (en) * | 2022-01-14 | 2022-08-19 | 亳州兴禾农业发展有限公司 | Tissue culture medium and tissue culture method for Bo-Chrysanthemum |
CN115868409A (en) * | 2022-11-10 | 2023-03-31 | 上海纳米技术及应用国家工程研究中心有限公司 | Tissue culture medium and tissue culture method for Tibet echinacea |
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Cited By (5)
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CN106386512A (en) * | 2016-12-19 | 2017-02-15 | 庆阳敦博科技发展有限公司 | Tissue culture and rapid culture method for inula flowers |
CN108849512A (en) * | 2018-07-16 | 2018-11-23 | 中国科学院合肥物质科学研究院 | A kind of method of spun gold emperor chrysanthemum tissue culture regeneration |
CN114916440A (en) * | 2022-01-14 | 2022-08-19 | 亳州兴禾农业发展有限公司 | Tissue culture medium and tissue culture method for Bo-Chrysanthemum |
CN115868409A (en) * | 2022-11-10 | 2023-03-31 | 上海纳米技术及应用国家工程研究中心有限公司 | Tissue culture medium and tissue culture method for Tibet echinacea |
CN115868409B (en) * | 2022-11-10 | 2024-03-15 | 上海纳米技术及应用国家工程研究中心有限公司 | Tissue culture medium and tissue culture method of Tibet chamomile |
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