CN108849522A - A kind of abductive approach of the callus of the aseptic seedling by the preparation of amomum viosum rhizomes bud - Google Patents
A kind of abductive approach of the callus of the aseptic seedling by the preparation of amomum viosum rhizomes bud Download PDFInfo
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- CN108849522A CN108849522A CN201810896401.8A CN201810896401A CN108849522A CN 108849522 A CN108849522 A CN 108849522A CN 201810896401 A CN201810896401 A CN 201810896401A CN 108849522 A CN108849522 A CN 108849522A
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- rhizomes
- bud
- aseptic seedling
- callus
- amomum viosum
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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
Abstract
The present invention provides a kind of abductive approach of the callus of aseptic seedling by the preparation of amomum viosum rhizomes bud, using the aseptic seedling of the rhizomes bud of amomum viosum as explant, by sterilizing, inoculation and Fiber differentiation, obtain its callus, the induced medium of the Fiber differentiation is MS+1.0~2.5mg/L 6-BA+0.5~1.5mg/L 2,4-D+0.5~1.5mg/L NAA；The light application time of culture environment is 10h, and intensity of illumination 2000Lx, cultivation temperature is 25 DEG C.The stem and the tip of a root that the present invention uses the rhizomes bud aseptic seedling of amomum viosum are as explant, the complete pharmacological property of parent can be retained using vegetative propagation, it is not likely to produce variation, so that pharmacological property is stablized, it can guarantee the effect of drug therapy, and the acquisition of explant material is also easy to, and can be drawn materials at any time and be carried out breeding culture.
Description
Technical field
The present invention relates to a kind of abductive approach of callus, belong to technical field of tissue culture.
Background technique
Amomum viosum (Amomum villosum Lour.) is Zingiber Amomum (Amomum Roxb.) perennial evergreen draft
Plant also known as " fructus amomi ", is famous one of the four great Nan medicines in China, had more than 1300 with the fruit medicine of drying and ripening
The applicating history in year.Fructus amomi is warm-natured, acrid flavour, has dampness elimination appetizing, warming spleen and stopping diarrha, regulating the flow of vital energy and preventing miscarriage and other effects.Amomum viosum is main
The ground such as Guangdong, Guangxi, Yunnan are distributed in, wherein best with Guangdong spring produced fructus amomi drug effect.The demand of fructus amomi is larger, often
The demand in year is estimated to exceed 2.2 × 106The problems such as kg, there are fruit drop rate height, low output due to amomum viosum, lead to fructus amomi
Yield is far from satisfying the demand in market, seriously hinders the development of fructus amomi industry.In addition, traditional breeding method needs
A large amount of human and material resources and time are spent, the low status of current amomum viosum specific yield cannot be also changed.Therefore, by science
Technology cultivates the provenance of high yield and high quality, improves the yield of current amomum viosum, be it is current urgently can not to research work.Tissue cultures
Research is the previous work for cultivating high yield and high quality amomum viosum provenance.
Plant Tissue Breeding is called in vitro culture, refers to isolated organ, tissue, cell by sterile working plant
Even protoplast, the general name of all tissue culture techniques of growth and development under the artificial environmental condition for providing and controlling.
By tissue culture technique, can effective expanding propagation coefficient in a short time, to obtain a large amount of, consistent test tube seedling, from
And overcome the defect of traditional breeding method.Research about amomum viosum tissue cultures at present, there are two main classes, and one kind is with amomum viosum root
It is numerous that shape stem eye or growing point are that explant is expanded in vitro, not by During Callus Induction；It is another kind of be with seed or
The aseptic seedlings that seed is sprouted carry out callus induction and regeneration as explant.Preceding a kind of work is because without passing through callus
Tissue induction, breeding coefficient is lower, and the further research such as genetic transformation can not be carried out using callus.Latter class work
Make because sprouting seedling with seed or seed is explant, and the acquisition of seed is limited by season, uses limited time.
Summary of the invention
For this purpose, the present invention first prepares aseptic seedling with the rhizomes bud of crop field plant amomum viosum, then with the stem section of aseptic seedling and
The tip of a root is explant, carries out the induction of callus, establishes amomum viosum callus induction system, obtains callus, passes through
The amomum viosum germplasm of the methods of cell fusion cultivation good quality and high output.Amomum viosum crop field plant material quantity is more, is easy to get, therefore
The present invention is not limited by the time.
It is turned out the purpose of the present invention is to provide a kind of by tissue cultures means with the preparation of amomum viosum rhizomes bud
The callus of aseptic seedling provides basis to be divided into amomum viosum intact plant.
The present invention is achieved through the following technical solutions：
A kind of abductive approach of the callus of the aseptic seedling by the preparation of amomum viosum rhizomes bud, with amomum viosum rhizomes bud
Aseptic seedling obtain the callus group of amomum viosum rhizomes bud aseptic seedling by sterilizing, inoculation and Fiber differentiation as explant
It knits；The induced medium that the Fiber differentiation uses is MS+1.0~2.5mg/L 6-BA+0.5~2.0mg/L 2,4-D+0.5
~1.5mg/L NAA.I.e.：1.0~2.5mg/L 6-BA, 0.5 are added in the MS culture medium (preparation method is shown in Table 1) of each liter
~1.5mg/L 2,4-D and 0.5~1.5mg/L NAA.
Further, the explant is the stem of the aseptic seedling of amomum viosum rhizomes bud.
Further, the stem of the aseptic seedling for using amomum viosum rhizomes bud as explant induced medium for：
MS+1.5mg/L 6-BA+1.0mg/L 2,4-D+0.5mg/L NAA。
Further, the stem of the aseptic seedling of the amomum viosum rhizomes bud is prepared using following methods：First by amomum viosum root
Shape stem eye carries out disinfection, and aseptic seedling is obtained through in vitro culture, then the stem of the aseptic seedling is cut into the segment of 2~3cm, by stem section
Vertical place is cultivated in the culture medium for being inoculated in and having prepared and passed through sterilization.
Further, the tip of a root of the aseptic seedling for amomum viosum rhizomes bud.
Further, induced medium of the tip of a root of the aseptic seedling using amomum viosum rhizomes bud as explant
For：MS+2.0mg/L 6-BA+1.0mg/L 2,4-D+1.0mg/L NAA.
Further, the tip of a root of the aseptic seedling of the amomum viosum rhizomes bud is prepared using following methods：First by amomum viosum
Rhizomes bud carries out disinfection, and obtains aseptic seedling through in vitro culture, then the tip of a root of the aseptic seedling is cut into the segment of 1~2cm, will
Tip of a root traverse is inoculated in respectively have been prepared and by being cultivated in the culture medium of sterilization.
Further, the environmental Kuznets Curves of the Fiber differentiation are temperature：22 DEG C~25 DEG C；Relative air humidity：50~
80%；The light control of Fiber differentiation is 10~20h, and intensity of illumination is 1500~2000Lx.
Preferably, the intensity of illumination is 2000Lx.
Further, the aseptic seedling by the preparation of amomum viosum rhizomes bud is 30 as the Fiber differentiation period of explant
~50 days.
Preferably, the Fiber differentiation period is 30 days.
Table 1：MS culture medium mother liquor preparation method
Above-mentioned MS be MS culture medium, BA be 6- benzyl aminoadenine, 2,4-D 2,4- dichlorphenoxyacetic acid, NAA is naphthalene
Acetic acid.
Aseptic seedling its preparation process that the present invention is prepared with amomum viosum rhizomes bud is：The amomum viosum rhizomes bud that will have been adopted
It is first rinsed with water 0.5-1.0h, is then washed one time, is placed on the superclean bench after sterilizing through ultraviolet lamp with sterile pond；
By sterile working, impregnate 30s with 75% ethyl alcohol, after use 0.1%HgCl210min is impregnated, then with aseptic water washing 5-6 times, most
After be transferred on sterilized culture dish it is stand-by.Cut it is sterile-processed after amomum viosum rhizomes bud be about 3cm, be inoculated in training
It supports and is cultivated on base (MS), when terminal bud is newly grown to about 0.5cm, then be transferred on the culture medium of MS+NAA (0.1mol/L) and take root
Culture, to cultivate aseptic seedling.Cultivation temperature is 25 DEG C；Fluorescent lamp lighting, light application time 10h/d, illuminance 1000-
1500Ix。
The beneficial effects of the present invention are：The present invention using aseptic seedling prepared by amomum viosum rhizomes bud as explant, into
Row callus induction enriches the source of explant material.The previous general aseptic seedlings sprouted using seed or seed are made
For explant, but since seed is sexual propagation, by maternal and male parent hybridization, inhereditary feature is easy to happen variation, it is difficult to
Guarantee that it is completely hereditary to the character of parent, therefore, the phenomenon that medicinal material failure often occur, and that the present invention uses is asexual numerous
The complete pharmacological property that can retain parent is grown, variation is not likely to produce, so that pharmacological property is stablized, can guarantee the effect of drug therapy, this
It is critically important factor for Chinese medicine.And tissue training is carried out as explant using the aseptic seedlings that seed or seed are sprouted
It supports, because the acquisition of seed is limited by season, so there is certain limitation, and the present invention is in terms of explant materials'use,
It will no longer be limited by season and environmental condition, and the acquisition of explant material is also easy to, can draw materials and be bred at any time
Culture.
Detailed description of the invention
Fig. 1 is the aseptic seedling picture of reason amomum viosum rhizomes bud of the present invention preparation.
Fig. 2 is the effect picture that the callus from stem segment of 1 amomum viosum rhizomes bud aseptic seedling of the embodiment of the present invention induces.
Fig. 3 is the effect picture that the callus from stem segment of 2 amomum viosum rhizomes bud aseptic seedling of the embodiment of the present invention induces.
Fig. 4 is the effect picture that the callus from stem segment of 3 amomum viosum rhizomes bud aseptic seedling of the embodiment of the present invention induces.
Fig. 5 is the effect picture of the tip of a root callus induction of 4 amomum viosum rhizomes bud aseptic seedling of the embodiment of the present invention.
Fig. 6 is the effect picture of the tip of a root callus induction of 5 amomum viosum rhizomes bud aseptic seedling of the embodiment of the present invention.
Fig. 7 is the effect picture of the tip of a root callus induction of 6 amomum viosum rhizomes bud aseptic seedling of the embodiment of the present invention.
Fig. 8 is the effect picture that the callus from stem segment of 1 amomum viosum rhizomes bud aseptic seedling of comparative example of the present invention induces.
Fig. 9 is the effect picture of the tip of a root callus induction of 2 amomum viosum rhizomes bud aseptic seedling of comparative example of the present invention.
Specific embodiment
For more concise displaying technical solution of the present invention, objects and advantages, combined with specific embodiments below
And attached drawing is described in further detail the present invention.
Embodiment 1
A kind of abductive approach of the callus of the aseptic seedling by the preparation of amomum viosum rhizomes bud, includes the following steps：It will
The stem of amomum viosum aseptic seedling is cut into the segment of 2cm, and by its stem section after sterile working, disinfection, vertical placement, which is inoculated in, has matched
It makes and passes through in the good culture medium of sterilization and cultivated, be inoculated with 5 bottles, 1 piece every bottle, observed, recorded within every 3 days.Pass through
Well-grown callus is screened in culture, and carries out squamous subculture to the callus successfully induced, and subculture is primary within 30 days.
The environmental condition wherein cultivated is：Temperature：22℃；Relative air humidity：50%；Light control was 10 small time
According to intensity of illumination 1500Lx.
Culture medium prescription is：MS+1.0mg/L 6-BA+0.5mg/L 2,4-D+0.5mg/L NAA
Embodiment 2
The abductive approach step and implementation of the callus of the aseptic seedling by the preparation of amomum viosum rhizomes bud of the present embodiment
Example 1 is identical.
Wherein the culture environment condition of the present embodiment is：Temperature：25℃；Relative air humidity：80%；Light control is 15
Hour illumination, intensity of illumination 2000Lx.
Culture medium prescription is：MS+2.5mg/L 6-BA+1.5mg/L 2,4-D+1.5mg/L NAA
Embodiment 3
The abductive approach step and implementation of the callus of the aseptic seedling by the preparation of amomum viosum rhizomes bud of the present embodiment
Example 1 is identical.
Wherein the culture environment condition of the present embodiment is：Temperature：25℃；Relative air humidity：80%；Light control is 10
Hour illumination, intensity of illumination 2000Lx.
Culture medium prescription is：MS+1.5mg/L 6-BA+1.0mg/L 2,4-D+0.5mg/L NAA
Embodiment 4
A kind of abductive approach of the callus of the aseptic seedling by the preparation of amomum viosum rhizomes bud, includes the following steps：It will
The tip of a root of amomum viosum aseptic seedling is cut into the segment of 1cm, and by its tip of a root after sterile working, disinfection, traverse, which is inoculated in, have been prepared
And by being cultivated in the good culture medium of sterilization, 5 bottles are inoculated with, 5 pieces every bottle, observed, recorded within every 3 days.Pass through training
The well-grown callus of screening is supported, and squamous subculture is carried out to the callus successfully induced, subculture is primary within 30 days.
The environmental condition wherein cultivated is：Temperature：22℃；Relative air humidity：50%；Light control was 10 small time
According to intensity of illumination 1500Lx.
Culture medium prescription is：MS+1.0mg/L 6-BA+0.5mg/L 2,4-D+0.5mg/L NAA
Embodiment 5
The abductive approach step and implementation of the callus of the aseptic seedling by the preparation of amomum viosum rhizomes bud of the present embodiment
Example 4 is identical.
Wherein the culture environment condition of the present embodiment is：Temperature：25℃；Relative air humidity：80%；Light control is 15
Hour illumination, intensity of illumination 2000Lx.
Culture medium prescription is：MS+2.5mg/L 6-BA+1.5mg/L 2,4-D+1.5mg/L NAA
Embodiment 6
The abductive approach step and implementation of the callus of the aseptic seedling by the preparation of amomum viosum rhizomes bud of the present embodiment
Example 4 is identical.
Wherein the culture environment condition of the present embodiment is：Temperature：25℃；Relative air humidity：80%；Light control is 10
Hour illumination, intensity of illumination 2000Lx.
The culture medium prescription of the present embodiment is：MS+2.0mg/L 6-BA+1.0mg/L 2,4-D+1.0mg/L NAA
Comparative example 1
A kind of abductive approach of the callus of the aseptic seedling by the preparation of amomum viosum rhizomes bud, includes the following steps：It will
The stem of amomum viosum aseptic seedling is cut into the segment of 2cm, and by its stem section after sterile working, disinfection, vertical placement, which is inoculated in, has matched
It makes and passes through in the good culture medium of sterilization and cultivated, be inoculated with 5 bottles, observed, recorded within every 3 days.It is screened by culture
Well-grown callus, and squamous subculture is carried out to the callus successfully induced, subculture is primary within 30 days.
Wherein the culture environment condition of the present embodiment is：Temperature：25℃；Relative air humidity：80%；Light control is 10
Hour illumination, intensity of illumination 2000Lx.
Culture medium prescription is：MS+1.0mg/L 6-BA+1.0mg/L 2,4-D
Comparative example 2
A kind of abductive approach of the callus of the aseptic seedling by the preparation of amomum viosum rhizomes bud, includes the following steps：It will
The tip of a root of amomum viosum aseptic seedling is cut into the segment of 1cm, and by its tip of a root after sterile working, disinfection, traverse, which is inoculated in, have been prepared
And by being cultivated in the good culture medium of sterilization, 5 bottles are inoculated with, observed, recorded within every 3 days.Pass through culture screening life
Long good callus, and squamous subculture is carried out to the callus successfully induced, subculture is primary within 30 days.
Wherein the culture environment condition of the present embodiment is：Temperature：25℃；Relative air humidity：80%；Light control is 10
Hour illumination, intensity of illumination 2000Lx.
The culture medium prescription of the present embodiment is：MS+1.0mg/L 6-BA+1.0mg/L NAA
Above-described embodiment 1-6 and the inducing effect of comparative example 1,2 are compared, culture medium prescription and environmental condition are such as
Shown in table 2：
Table 2：The culture medium prescription and environmental condition of embodiment 1-6 and comparative example 1,2
The results are shown in Table 2 for above-described embodiment 1-6 and the inducing effect of comparative example 1,2：
Table 3：The inducing effect result of embodiment 1-6 and comparative example 1,2
| Group | Embodiment 1 | Embodiment 2 | Embodiment 3 | Embodiment 4 | Embodiment 5 | Embodiment 6 | Comparative example 1 | Comparative example 2 |
| Inductivity | 40% | 45% | 60% | 64% | 68% | 76% | 0 | 0 |
As shown in Table 3, the abductive approach of 1-6 of the embodiment of the present invention can induce out amomum viosum rhizomes bud aseptic seedling stem and
The callus of the tip of a root, and the inductivity highest of the callus of the abductive approach induction aseptic seedling stem of embodiment 3, and embodiment
The inductivity highest of the callus of the 6 abductive approach induction aseptic seedling tip of a root, therefore, embodiment 3 is induction nothing of the invention
The most preferred embodiment of the callus of vaccine stem, embodiment 6 are the best of the callus of the induction aseptic seedling tip of a root of the invention
Embodiment.And lack certain ingredients in induced medium of the present invention in the callus inducing medium of comparative example 1,2,
Callus cannot be induced, illustrates that abductive approach of the invention is obtained not by the simple combination of each component, then into
One step illustrates there is synergistic effect between Fiber differentiation based formulas each group conjunction of the present invention and the environmental condition of Fiber differentiation.
The induced medium of the callus of amomum viosum rhizomes bud of the invention and other induced mediums 1,2 are carried out
Compare that (environment condition of culture is identical：25 DEG C of temperature；Light application time 10h；Intensity of illumination：2000Lx；Air humidity：80%),
Formula see the table below：
Table 4：The formula and each component content of the embodiment of the present invention 3,6 induced mediums and other induced mediums 1,2
The embodiment of the present invention 3, the induction result of 6 induced mediums and other induced mediums 1,2 are as follows：
Table 5：The induction result of the embodiment of the present invention 3,6 induced mediums and other induced mediums 1,2
| Group | Embodiment 5 | Embodiment 6 | Induced medium 1 | Induced medium 2 |
| Inductivity | 60% | 78% | 5% | 15% |
As shown in Table 5, the inducing effect of induced medium of the invention is preferable, inductivity highest, and induced medium 1 is to use
In induction seed callus, induced medium 2 is the callus for inducing the aseptic seedling sprouted by seed, due to this
It is different from the above two to invent the explant used, the explant needed nutrient matter and inductive condition of plant different parts and organ
It is different, it is therefore desirable to different abductive approach to be developed, to improve inductivity.
The effect for going out callus of the stem of amomum viosum rhizomes bud aseptic seedling and the tip of a root is compared, in its culture item
Under part same case, comparing its time for going out callus, the quantity for going out callus and inductivity, (culture medium prescription is：
MS+1.5mg/L 6-BA+1.0mg/L2,4-D+0.5mg/LNAA, culture environment are：25 DEG C of temperature；Light application time 10h；Illumination
Intensity：2000Lx；Air humidity：80%) it, as a result see the table below.
Table 6：The stem of amomum viosum rhizomes bud aseptic seedling and the callus result out of the tip of a root
As shown in Table 6, the aseptic seedling stem sections of amomum viosum rhizomes bud, the callus time out of the tip of a root are shorter.Compare two kinds
Explant, under same culture conditions, the tip of a root is higher than the inductivity of stem section, the time of generation callus is shorter, and the tip of a root is more
It is easy to produce callus.This may it is more flourishing with the separate living tissue of aseptic seedling root, meristematic capacity is stronger related.Therefore, spring
The tip of a root of sand aseptic seedling is more suitable for the explant of callus induction.
The embodiments described above only express several embodiments of the present invention, and the description thereof is more specific and detailed, but simultaneously
It cannot therefore be construed as limiting the scope of the patent.It should be pointed out that coming for those of ordinary skill in the art
It says, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to protection of the invention
Range.Therefore, the scope of protection of the patent of the invention shall be subject to the appended claims.
Claims (10)
1. a kind of abductive approach of the callus of the aseptic seedling by the preparation of amomum viosum rhizomes bud, which is characterized in that with spring
The aseptic seedling of sand rhizomes bud obtains amomum viosum rhizomes bud aseptic seedling by sterilizing, inoculation and Fiber differentiation as explant
Callus；The induced medium that the Fiber differentiation uses is MS+1.0~2.5mg/L 6-BA+0.5~1.5mg/L 2,
4-D+0.5~1.5mg/L NAA.
2. special as described in claim 1 by the abductive approach of the callus of the aseptic seedling of amomum viosum rhizomes bud preparation
Sign is that the explant is the stem of the aseptic seedling of amomum viosum rhizomes bud.
3. special as claimed in claim 2 by the abductive approach of the callus of the aseptic seedling of amomum viosum rhizomes bud preparation
Sign is, the stem of the aseptic seedling for using amomum viosum rhizomes bud as explant induced medium for：MS+1.5mg/L
6-BA+1.0mg/L 2,4-D+0.5mg/L NAA。
4. the abductive approach of the callus of the aseptic seedling prepared as claimed in claim 2 or claim 3 by amomum viosum rhizomes bud,
It is characterized in that, the stem of the aseptic seedling of the amomum viosum rhizomes bud is prepared using following methods：First by amomum viosum rhizomes bud into
Row disinfection, obtains aseptic seedling through in vitro culture, then the stem of the aseptic seedling is cut into the segment of 2~3cm, stem section is placed vertically
It is inoculated in and has prepared and by being cultivated in the culture medium of sterilization.
5. special as described in claim 1 by the abductive approach of the callus of the aseptic seedling of amomum viosum rhizomes bud preparation
Sign is, the tip of a root of the aseptic seedling for amomum viosum rhizomes bud.
6. special as claimed in claim 5 by the abductive approach of the callus of the aseptic seedling of amomum viosum rhizomes bud preparation
Sign is, the tip of a root of the aseptic seedling for using amomum viosum rhizomes bud as explant induced medium for：MS+2.0mg/
L 6-BA+1.0mg/L 2,4-D+1.0mg/L NAA。
7. the abductive approach of the callus such as the aseptic seedling described in claim 5 or 6 by the preparation of amomum viosum rhizomes bud,
It is characterized in that, the tip of a root of the aseptic seedling of the amomum viosum rhizomes bud is prepared using following methods：First by amomum viosum rhizomes bud
It carries out disinfection, aseptic seedling is obtained through in vitro culture, then the tip of a root of the aseptic seedling is cut into the segment of 1~2cm, by tip of a root traverse
It is inoculated in and has prepared and by being cultivated in the culture medium of sterilization respectively.
8. special as described in claim 1 by the abductive approach of the callus of the aseptic seedling of amomum viosum rhizomes bud preparation
Sign is that the temperature of the Fiber differentiation is 22 DEG C~25 DEG C；Relative air humidity is 50~80%；Light control be 10~
20h；Intensity of illumination is 1500~2000Lx.
9. special as described in claim 1 by the abductive approach of the callus of the aseptic seedling of amomum viosum rhizomes bud preparation
Sign is that the aseptic seedling by the preparation of amomum viosum rhizomes bud is 30~50 days as the Fiber differentiation period of explant.
10. special as claimed in claim 9 by the abductive approach of the callus of the aseptic seedling of amomum viosum rhizomes bud preparation
Sign is that the Fiber differentiation period is 30 days.
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| CN110915442A (en) * | 2019-11-26 | 2020-03-27 | 广州市阔途生物科技有限公司 | Method for cutting and propagating root tips of rhizomes of amomum villosum |
| CN110915441A (en) * | 2019-11-26 | 2020-03-27 | 广州市阔途生物科技有限公司 | Cutting propagation method for rhizomes of amomum villosum |
| CN113317205A (en) * | 2021-07-15 | 2021-08-31 | 云南中医药大学 | Efficient artificial seedling culture method for culturing amomum villosum by utilizing basal stem cluster buds and node propagation |
| CN116711641A (en) * | 2023-07-12 | 2023-09-08 | 北京市农林科学院 | Method for rapid propagation of tea chrysanthemum tissue culture seedlings |
| CN117243126A (en) * | 2023-11-20 | 2023-12-19 | 云南省农业科学院药用植物研究所 | In-vitro preservation method and application of tsaoko amomum fruit germplasm resources |
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| CN110915442A (en) * | 2019-11-26 | 2020-03-27 | 广州市阔途生物科技有限公司 | Method for cutting and propagating root tips of rhizomes of amomum villosum |
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| CN110915442B (en) * | 2019-11-26 | 2021-10-08 | 广州市阔途生物科技有限公司 | Method for cutting and propagating root tips of rhizomes of amomum villosum |
| CN110915441B (en) * | 2019-11-26 | 2021-10-08 | 广州市阔途生物科技有限公司 | Cutting propagation method for rhizomes of amomum villosum |
| CN113317205A (en) * | 2021-07-15 | 2021-08-31 | 云南中医药大学 | Efficient artificial seedling culture method for culturing amomum villosum by utilizing basal stem cluster buds and node propagation |
| CN113317205B (en) * | 2021-07-15 | 2022-10-18 | 云南中医药大学 | Efficient artificial seedling culture method for culturing amomum villosum by utilizing basal stem cluster buds and node propagation |
| CN116711641A (en) * | 2023-07-12 | 2023-09-08 | 北京市农林科学院 | Method for rapid propagation of tea chrysanthemum tissue culture seedlings |
| CN116711641B (en) * | 2023-07-12 | 2024-04-26 | 北京市农林科学院 | Method for rapid propagation of tea chrysanthemum tissue culture seedlings |
| CN117243126A (en) * | 2023-11-20 | 2023-12-19 | 云南省农业科学院药用植物研究所 | In-vitro preservation method and application of tsaoko amomum fruit germplasm resources |
| CN117243126B (en) * | 2023-11-20 | 2024-01-23 | 云南省农业科学院药用植物研究所 | In-vitro preservation method and application of tsaoko amomum fruit germplasm resources |
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