CN105918119A - Method for in-vitro high-efficiency regeneration of leaf of Chuzhou chrysanthemum - Google Patents
Method for in-vitro high-efficiency regeneration of leaf of Chuzhou chrysanthemum Download PDFInfo
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- CN105918119A CN105918119A CN201610262030.9A CN201610262030A CN105918119A CN 105918119 A CN105918119 A CN 105918119A CN 201610262030 A CN201610262030 A CN 201610262030A CN 105918119 A CN105918119 A CN 105918119A
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- callus
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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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- 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 method for in-vitro high-efficiency regeneration of the leaf of Chuzhou chrysanthemum. The method comprises the following steps: 1) material selection: selecting the leaf of Chuzhou chrysanthemum as explants; 2) slicing; 3) induction of callus; 4) proliferation of callus; 5) differentiation of callus; 6) proliferation and elongation of adventitious buds; and 7) rooting: carrying out rooting culture when the adventitious buds extend to 2 to 3 cm and accompanied with 2 to 3 totally unfolded leaves. The method provided by the invention has the advantages of easy and wide availability of materials, high regeneration efficiency, high callus inductivity of 100%, a high differentiation rate of 95%, a high rooting rate of 100%, a fast regeneration speed, a high propagation coefficient, and capacity of realizing further enlarged breeding of obtained aseptic seedling leaves and providing crucial technical support for rapid breeding and breed improvement of excellent Chuzhou chrysanthemum strains.
Description
Technical field
The present invention relates to biological technical field, a kind of method particularly relating in vitro highly efficient regeneration of Flos Chrysanthemi blade.
Background technology
Flos Chrysanthemi (Dendranthema morifolium CV. ' chuju '), for the distinctive traditional product of Chuzhou City, attaches most importance to
The Chinese herbal medicine wanted and industrial crops, have the title of Chuzhou Flos Chrysanthemi.Belong to medicine, tea dual-purpose good merchantable brand, there is high medicine
With and health value, at the cultivation history of China's existing centuries.Long-term drink Flos Chrysanthemi have heat-clearing and toxic substances removing,
The functions such as relaxing muscles and tendons to promote blood circulation, liver protection and eyesight, enhancing body immunity, show coronary heart disease, hypertension curative effect simultaneously
Write.Flos Chrysanthemi ranks first of Anhui Province's " four is big " famous genuine medicinal materials and first of China " four is big " medicine chrysanthemum.Mesh
Before, Flos Chrysanthemi has become as locality, Chuzhou pillar industry, at area, Chuzhou establishing in large scale.
But at present on producing, the asexual propagation means using cuttage and plant division etc. traditional due to Flos Chrysanthemi for a long time are entered
Row breeding so that Flos Chrysanthemi generally exists variety deterioration, pest and disease damage breed improvement serious, relevant and renewal speed
The problem such as slow, greatly limit and cultivate on a large scale using Flos Chrysanthemi as medicine, tea dual-purpose good merchantable brand and industrialization
Application.Therefore, strengthening its breed improvement, cultivating high yield, high-quality and pest-resistant Flos Chrysanthemi improved seeds is current sending out
The task that exhibition Flos Chrysanthemi industry is the most urgent.At present about the research of Flos Chrysanthemi breeding, it is concentrated mainly on utilization hybridization
Means are carried out, far from adapting to chrysanthemum agriculture to variety yield and the requirement of quality.
Owing to utilizing tissue culture technique can obtain substantial amounts of aseptic seedling in a short time, Flos Chrysanthemi can realized
While fine germplasm resources large-scale production so that the breed improvement process accelerating Flos Chrysanthemi is possibly realized.Mesh
Front less about the research of Flos Chrysanthemi Regeneration in Vitro, wherein utilizing Flos Chrysanthemi blade is that outer implant carries out Regeneration in Vitro
It has been reported that but generally there is breeding coefficient difference low, repeatable, the variable black death of outer implant and divide in research
The problems such as change process vitrification phenomenon is serious, seriously limit blade application in Flos Chrysanthemi rapid propagation in vitro.Pin
Problems in the presence of planting above-mentioned Flos Chrysanthemi, simultaneously in order to meet implantation in large scale and breed improvement pair
The demand of Flos Chrysanthemi high quality seedling, a kind of method being badly in need of developing in vitro highly efficient regeneration of Flos Chrysanthemi blade.
Summary of the invention
It is an object of the invention to use plant tissue culture technique, it is provided that a kind of Flos Chrysanthemi blade is in vitro the most again
Raw method.To achieve these goals, the present invention adopts the following technical scheme that
A kind of method of in vitro highly efficient regeneration of Flos Chrysanthemi blade, comprises the following steps:
1) draw materials: choose Flos Chrysanthemi blade as outer implant；
2) section: by step 1) in Flos Chrysanthemi blade cut into slices；
3) induction of callus: by step 2) in section be inoculated in callus inducing medium and carry out more
The inducing culture of injured tissue；
4) propagation of callus: by step 3) in induction section callus be inoculated in proliferated culture medium
In carry out the propagation of callus；
5) differentiation of callus: by step 4) in propagation after callus stripping and slicing and transfer in differentiation training
Support the differentiation carrying out callus in base；
6) propagation of adventitious bud and elongation: by step 5) in differentiate the callus of adventitious bud and transfer in not
The propagation and the elongation that carry out adventitious bud in bud multiplication and elongation medium are cultivated；
7) take root: treat that Elongation of adventitious bud, to 2～3cm, and is given birth to during with the leaf of 2～3 full extension
Root is cultivated.
Preferably, described step 1) Flos Chrysanthemi blade be the leaf that field Flos Chrysanthemi elite plant strain gives birth to edible tender branch then
The blade of the Flos Chrysanthemi aseptic seedling full extension that sheet or Regeneration in Vitro are obtained.
Preferably, the method for the described in vitro highly efficient regeneration of Flos Chrysanthemi blade also includes surface sterilization；Surface sterilization:
By step 2) in have drawn from the dehydrated alcohol wiping one time of the vanes 75% giving birth to edible tender branch then in field
After, with aseptic water washing 2～3 times in aseptic working platform；Then with 75% dehydrated alcohol sterilize 10～25s
After, with sterile water wash 2～3 times；Then with 0.1%HgCl2 solution disinfection 1～3min, aseptic water washing
5～6 times.
Preferably, described step 3) in callus inducing medium include MS+0.1～3.0mg/L TDZ
+ 0.01～0.05mg/L 6-BA+30g/L sucrose+7.0g/L agar, pH=5.8.
Preferably, after illumination cultivation 14～16d, by step 3) in induction Callus of Leaf be inoculated in increasing
Grow the propagation carrying out callus in culture medium；Wherein callus proliferation medium is MS+0.2～2.0
Mg/L TDZ+30g/L sucrose+7.0g/L agar, pH=5.8.
Preferably, after illumination cultivation 10～14d, by step 4) in propagation after callus be cut into (0.4～
0.5) × (0.4～0.5)) cm2The stripping and slicing of size is also transferred and is carried out the differentiation of callus in division culture medium；
Wherein the division culture medium of callus is MS+0.2～1.0mg/L TDZ+0.05～0.2mg/L 6-BA
+ 0.1～1.0mg/L NAA+30g/L sucrose+7.0g/L agar, pH=5.8.
Preferably, after illumination cultivation 21～28d, by step 5) in differentiate adventitious bud callus switching
The propagation and the elongation that carry out adventitious bud in adventitious bud proliferation and elongation medium are cultivated；Wherein adventitious bud proliferation
It is MS+0.5～2.0mg/L 6-BA+0.05～0.2mg/L TDZ+0.2～2.0 with elongation medium
Mg/L NAA+30g/L sucrose+7.0g/L agar, pH=5.8.
Preferably, described step 7) in root media be 1/2MS+0.05～0.3mg/L IBA+
0.05～0.5mg/L IAA+20g/L sucrose+7.0g/L agar, pH=5.8.
It is an advantage of the current invention that: one, convenience of drawing materials, material enrich；Its two, regeneration efficiency is high, more
Injured tissue inductivity can be up to 100%, and differentiation rate can be up to 95%, and rooting rate can be up to 100%；Its three,
Reproduction speed is fast, and breeding coefficient is high, and can realize further being expanded the tests for sterility obtained
Breeding, Fast-propagation and breed improvement for excellent Flos Chrysanthemi strain provide important technical support.Therefore,
The method of a kind of in vitro highly efficient regeneration of Flos Chrysanthemi blade provided by the present invention, the most only Flos Chrysanthemi elite plant strain is fast
Speed breeding and large-scale production provide technical support, and the most also base has been established in the breed improvement for later stage Flos Chrysanthemi
Plinth.
Accompanying drawing explanation
Fig. 1 is the induction of Flos Chrysanthemi Callus of Leaf
Fig. 2 is the propagation of callus
Fig. 3 is the differentiation of callus
Fig. 4 is propagation and the elongation of adventitious bud
Fig. 5 is taking root of adventitious bud
Detailed description of the invention
Below in conjunction with accompanying drawing, the present invention is described in detail.
Embodiment 1
A kind of method of the in vitro highly efficient regeneration of Flos Chrysanthemi blade, concrete operations are as follows:
1, choose well-grown perennial Flos Chrysanthemi elite plant strain from land for growing field crops, choose the raw children of Flos Chrysanthemi tissue regeneration promoting then tender
The blade of branch is outer implant.
2, after 75% dehydrated alcohol wiping 1 time, in aseptic operating platform, with aseptic water washing 3 times, so
Afterwards with 75% alcohol disinfecting 10s, aseptic water washing 3 times, sterilize 1min with the mercuric chloride of 0.1% afterwards,
Use aseptic water washing 6 times afterwards.After sterilization, blade is cut into 0.3 × 0.3cm2The stripping and slicing of size is standby.
3, leafcutting proximal ends is upwards inoculated in callus inducing medium carries out luring of callus
Lead, through the illumination cultivation of 15d, at paddle cutout, induce substantial amounts of light green callus, and wound healing
The inductivity of tissue is 83.7%；Wherein the inducing culture of callus is MS+0.1mg/L TDZ+
0.01mg/L 6-BA+30g/L sucrose+7.0g/L agar, pH=5.8；
4, the blade inducing callus is transferred carry out in proliferated culture medium callus propagation training
Support, after illumination cultivation 12d, obtain the open-textured callus of substantial amounts of green；The propagation training of callus
Foster base is MS+0.2mg/L TDZ+30g/L sucrose+7.0g/L agar, pH=5.8；
5, by the callus after propagation, it is cut into 0.4 × 0.4cm2The stripping and slicing of size is also transferred in differentiation culture
Base carries out the differentiation culture of callus；After illumination cultivation 24d, callus differentiate substantial amounts of green
Color adventitious bud point, and the differentiation rate of callus is 89.4%；Wherein the division culture medium of callus is MS
+ 0.2mg/L TDZ+0.05mg/L 6-BA+0.1mg/L NAA+30g/L sucrose+7.0g/L fine jade
Fat, pH=5.8；
6, the callus of adventitious bud clump will be differentiated, transfer to enter in the culture medium of adventitious bud proliferation and elongation
The propagation of row adventitious bud and elongation；The adventitious bud of substantial amounts of elongation, wherein adventitious bud is obtained after illumination cultivation 16d
Propagation and elongation medium are MS+0.5mg/L 6-BA+0.05mg/L TDZ+0.2mg/L NAA+30
G/L sucrose+7.0g/L agar, pH=5.8；
7, treat that Elongation of adventitious bud to 2cm, and during with the leaf of 2 full extension, proceeds in root media
Carry out root culture；After illumination cultivation 16d, the base portion of adventitious bud grows the most elongated adventitious root, and indefinite
The inductivity of root is up to 100%；Wherein root media is 1/2MS+0.05mg/L IBA+0.05mg/L
IAA+20g/L sucrose+7.0g/L agar, pH=5.8.
Embodiment 2
A kind of method of the in vitro highly efficient regeneration of Flos Chrysanthemi blade, concrete operations are as follows:
1, choose well-grown perennial Flos Chrysanthemi elite plant strain from land for growing field crops, choose the raw children of Flos Chrysanthemi tissue regeneration promoting then tender
The blade of branch is outer implant.
2, after 75% dehydrated alcohol wiping 1 time, in aseptic operating platform, with aseptic water washing 3 times, so
Afterwards with 75% alcohol disinfecting 15s, aseptic water washing 2 times, sterilize 2min with the mercuric chloride of 0.1% afterwards,
Use aseptic water washing 6 times afterwards.After sterilization, blade is cut into 0.3 × 0.5cm2The stripping and slicing of size is standby.
3, leafcutting proximal ends is upwards inoculated in callus inducing medium carries out luring of callus
Lead, through the illumination cultivation of 14d, at paddle cutout, induce substantial amounts of light green callus, and wound healing
The inductivity of tissue is 100% (Fig. 1)；Wherein the inducing culture of callus is MS+1.5mg/L TDZ
+ 0.03mg/L 6-BA+30g/L sucrose+7.0g/L agar, pH=5.8；
4, the blade inducing callus is transferred carry out in proliferated culture medium callus propagation training
Support, after illumination cultivation 10d, obtain the open-textured callus of substantial amounts of green (Fig. 2)；Wherein wound healing group
The proliferated culture medium knitted is MS+1.0mg/L TDZ+30g/L sucrose+7.0g/L agar, pH=5.8；
5, by the callus after propagation, it is cut into 0.4 × 0.5cm2The stripping and slicing of size is also transferred in differentiation culture
Base carries out the differentiation of callus；After illumination cultivation 21d, callus stripping and slicing differentiate substantial amounts of green
Color adventitious bud point (Fig. 3), and the differentiation rate of callus is 97.4%；The division culture medium of callus is
MS+0.5mg/L TDZ+0.1mg/L 6-BA+0.5mg/L NAA+30g/L sucrose+7.0g/L
Agar, pH=5.8；
6, by the callus after differentiation, transfer in the culture medium of adventitious bud proliferation and elongation, carry out adventitious bud
Propagation and elongation, obtain the adventitious bud (Fig. 4) of substantial amounts of elongation after illumination cultivation 14d；Wherein adventitious bud
Propagation and elongation medium are MS+1.0mg/L 6-BA+0.1mg/L TDZ+0.5mg/L NAA+30
G/L sucrose+7.0g/L agar, pH=5.8；
7, treat that Elongation of adventitious bud to 2cm, and during with the leaf of 2 full extension, proceeds in root media
Carry out root culture；After illumination cultivation 14d, the base portion of adventitious bud grows the most sturdy adventitious root (Fig. 5),
And the inductivity of adventitious root is up to 100%；Wherein root media is 1/2MS+0.1mg/L IBA+0.1
Mg/L IAA+20g/L sucrose+7.0g/L agar, pH=5.8.
Embodiment 3
A kind of method of the in vitro highly efficient regeneration of Flos Chrysanthemi blade, concrete operations are as follows:
1, choose well-grown perennial Flos Chrysanthemi elite plant strain from land for growing field crops, choose the raw branch of Flos Chrysanthemi tissue regeneration promoting then
Young leaflet tablet be outer implant.
2, after 75% dehydrated alcohol wiping 1 time, in aseptic operating platform, with aseptic water washing 2 times, then
With 75% alcohol disinfecting 25s, aseptic water washing 3 times, sterilize 3min with the mercuric chloride of 0.1% afterwards, finally use
Aseptic water washing 5 times.After sterilization, blade is cut into 0.5 × 0.5cm2The stripping and slicing of size is standby.
3, leafcutting proximal ends is upwards inoculated in callus inducing medium carries out luring of callus
Lead, through the illumination cultivation of 16d, at paddle cutout, induce substantial amounts of callus, and callus
Inductivity is 100%；The inducing culture of callus is MS+3.0mg/L TDZ+0.05mg/L 6-BA
+ 30g/L sucrose+7.0g/L agar, pH=5.8；
4, the blade inducing callus is transferred carry out in proliferated culture medium callus propagation training
Support, after illumination cultivation 14d, obtain the substantial amounts of green close callus of quality；The propagation training of callus
Foster base is MS+2.0mg/L TDZ+30g/L sucrose+7.0g/L agar, pH=5.8；
5, by the callus after propagation, it is cut into 0.5 × 0.5cm2The stripping and slicing of size is also transferred in differentiation culture
Base carries out the differentiation of callus；After illumination cultivation 28d, callus differentiates substantial amounts of adventitious bud
Point, the differentiation rate of callus is 79.4%；The division culture medium of callus is MS+1.0mg/L TDZ
+ 0.2mg/L 6-BA+1.0mg/L NAA+30g/L sucrose+7.0g/L agar, pH=5.8；
6, by the callus after differentiation, transfer in the culture medium of adventitious bud proliferation and elongation, carry out adventitious bud
Propagation and elongation, after illumination 21d, it is thus achieved that the adventitious bud of substantial amounts of elongation；Adventitious bud proliferation and elongation are cultivated
Base is MS+2.0mg/L 6-BA+0.2mg/L TDZ+2.0mg/L NAA+30g/L sucrose+7.0
G/L agar, pH=5.8；
7, treat that Elongation of adventitious bud to 3cm, and during with the leaf of 3 full extension, proceeds in root media
Carrying out root culture, after illumination cultivation 21d, the base portion of adventitious bud grows the most short and thick adventitious root, and indefinite
The inductivity of root is up to 97.6%；Wherein root media is 1/2MS+0.3mg/L IBA+0.5mg/L
IAA+20g/L sucrose+7.0g/L agar, pH=5.8.
Embodiment 4
A kind of method of the in vitro highly efficient regeneration of Flos Chrysanthemi blade, concrete operations are as follows:
Test the impact on Flos Chrysanthemi blade in-vitro regeneration efficiency of the blade inoculation mode: only change leafcutting
Vaccination ways, by the leafcutting after sterilization respectively by proximal ends and distal shaft end upwards in the way of, be inoculated in more
In injured tissue inducing culture, cultivate under light.Remaining step is with embodiment 2.After illumination cultivation 14-16d,
The induction situation of statistics callus.Result shows the induction to callus of the vaccination ways of (table 1) blade
Play an important role.Blade proximal ends vaccination ways upwards is more conducive to induction and the later stage of callus
Differentiation.
The impact on Flos Chrysanthemi blade Regeneration in Vitro of table 1 vaccination ways
Note: data are average ± standard error, each process comprises 30 outer implant, and each process is repeated 3 times.
Embodiment 5
A kind of method of the in vitro highly efficient regeneration of Flos Chrysanthemi blade, concrete operations are as follows:
Test the source impact on Flos Chrysanthemi in-vitro regeneration efficiency of blade: only change leafcutting source,
To derive from the leafcutting after the sterilization of field elite plant strain and the Flos Chrysanthemi tests for sterility of fast numerous acquisition, all with
Proximal ends mode upwards, is inoculated in callus inducing medium respectively, cultivates under light.Remaining step
Rapid with embodiment 2.After illumination cultivation 14-16d, the induction situation of statistics callus.Result shows (table 2)
The source of the blade induction no significant difference to callus.
The impact on Flos Chrysanthemi blade Regeneration in Vitro of the table 2 blade source
Note: data are average ± standard error, each process comprises 30 outer implant, and each process is repeated 3 times.
Above-described embodiment only for technology design and the feature of the present invention are described, its object is to allow and is familiar with this field
The personage of technology will appreciate that present invention and is carried out, and can not limit the protection model of the present invention with this
Enclose.All equivalence changes made according to spirit of the invention or modification, all should contain the protection in the present invention
In the range of.
Claims (8)
1. the method for the in vitro highly efficient regeneration of Flos Chrysanthemi blade, it is characterised in that: comprise the following steps:
1) draw materials: choose Flos Chrysanthemi blade as outer implant；
2) section: by step 1) in Flos Chrysanthemi blade cut into slices；
3) induction of callus: by step 2) in section be inoculated in callus inducing medium and carry out more
The inducing culture of injured tissue；
4) propagation of callus: by step 3) in induction section callus be inoculated in proliferated culture medium
In carry out the propagation of callus；
5) differentiation of callus: by step 4) in propagation after callus stripping and slicing and transfer in differentiation training
Support the differentiation carrying out callus in base；
6) propagation of adventitious bud and elongation: by step 5) in differentiate the callus of adventitious bud and transfer in not
The propagation and the elongation that carry out adventitious bud in bud multiplication and elongation medium are cultivated；
7) take root: treat that Elongation of adventitious bud, to 2～3cm, and is given birth to during with the leaf of 2～3 full extension
Root is cultivated.
The method of a kind of in vitro highly efficient regeneration of Flos Chrysanthemi blade the most according to claim 1, it is characterised in that:
Described step 1) Flos Chrysanthemi blade be blade or the Regeneration in Vitro that field Flos Chrysanthemi elite plant strain gives birth to edible tender branch then
The blade of the Flos Chrysanthemi aseptic seedling full extension obtained.
The method of a kind of in vitro highly efficient regeneration of Flos Chrysanthemi blade the most according to claim 2, it is characterised in that:
The method of the described in vitro highly efficient regeneration of Flos Chrysanthemi blade also includes surface sterilization；Surface sterilization: by step 2) in take
Material after the dehydrated alcohol wiping one time of the vanes 75% giving birth to edible tender branch then in field, aseptic working platform
Middle aseptic water washing 2～3 times；Then with after the dehydrated alcohol sterilization 10～25s of 75%, clear with sterilized water
Wash 2～3 times；Then with 0.1%HgCl2 solution disinfection 1～3min, aseptic water washing 5～6 times.
The method of a kind of in vitro highly efficient regeneration of Flos Chrysanthemi blade the most according to claim 1, it is characterised in that:
Described step 3) in callus inducing medium include MS+0.1～3.0mg/L TDZ+0.01～0.05
Mg/L 6-BA+30g/L sucrose+7.0g/L agar, pH=5.8.
The method of a kind of in vitro highly efficient regeneration of Flos Chrysanthemi blade the most according to claim 1, it is characterised in that:
After illumination cultivation 14～16d, by step 3) in induction Callus of Leaf be inoculated in proliferated culture medium
The propagation of row callus；Wherein callus proliferation medium is MS+0.2～2.0mg/L TDZ+30
G/L sucrose+7.0g/L agar, pH=5.8.
The method of a kind of in vitro highly efficient regeneration of Flos Chrysanthemi blade the most according to claim 1, it is characterised in that:
After illumination cultivation 10～14d, by step 4) in propagation after callus be cut into (0.4～0.5) × (0.4～
0.5))cm2The stripping and slicing of size is also transferred and is carried out the differentiation of callus in division culture medium；Wherein wound healing group
The division culture medium knitted be MS+0.2～1.0mg/L TDZ+0.05～0.2mg/L 6-BA+0.1～
1.0mg/L NAA+30g/L sucrose+7.0g/L agar, pH=5.8.
The method of a kind of in vitro highly efficient regeneration of Flos Chrysanthemi blade the most according to claim 1, it is characterised in that:
After illumination cultivation 21～28d, by step 5) in differentiate the callus of adventitious bud and transfer in adventitious bud proliferation
Cultivate with the propagation and elongation carrying out adventitious bud in elongation medium；Wherein adventitious bud proliferation and elongation medium
For MS+0.5～2.0mg/L 6-BA+0.05～0.2mg/L TDZ+0.2～2.0mg/L NAA+30
G/L sucrose+7.0g/L agar, pH=5.8.
The method of a kind of in vitro highly efficient regeneration of Flos Chrysanthemi blade the most according to claim 1, it is characterised in that:
Described step 7) in root media be 1/2MS+0.05～0.3mg/L IBA+0.05～0.5mg/L
IAA+20g/L sucrose+7.0g/L agar, pH=5.8.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106804426A (en) * | 2016-12-26 | 2017-06-09 | 中国科学院华南植物园 | Promote the box set and method of Companumoea root vitro proliferation |
| CN108901850A (en) * | 2018-07-26 | 2018-11-30 | 中国科学院合肥物质科学研究院 | A kind of method of spun gold emperor chrysanthemum stem apex Regeneration in Vitro |
| CN111869569A (en) * | 2020-08-18 | 2020-11-03 | 仲恺农业工程学院 | Culture system for in vitro culture of hedychium japonicum flowers and application thereof |
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| CN111869569A (en) * | 2020-08-18 | 2020-11-03 | 仲恺农业工程学院 | Culture system for in vitro culture of hedychium japonicum flowers and application thereof |
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| CN105918119B (en) | 2017-12-05 |
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