CN106718934A - A kind of utilization plumular axis and radicle directly break up the bighead atractylodes rhizome regenerating system of adventitious bud - Google Patents
A kind of utilization plumular axis and radicle directly break up the bighead atractylodes rhizome regenerating system of adventitious bud Download PDFInfo
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- 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 the bighead atractylodes rhizome regenerating system that a kind of utilization plumular axis and radicle directly break up adventitious bud, including：Atractylodes macrocephala seed is cleaned, is sterilized, the Atractylodes macrocephala seed after sterilization is inoculated in MS culture mediums, cultivated to seed in dark place and sprouted, continuation is cultivated under being transferred to light；Radicle and plumular axis are cut, is cut into small pieces as explant, be seeded to Fiber differentiation on bud inducement cultivation base, its differentiation is grown green bud；Cut during green bud is forwarded to root media and cultivate, root induction；By the Transplantation of Regenerated Plantlets taken root, bighead atractylodes rhizome plant is obtained final product.The regenerating system that the present invention is provided substantially increases the proliferation frequency of the bighead atractylodes rhizome, has saved production cost, reproductive efficiency is improve, for the large-scale production of the medicinal bighead atractylodes rhizome provides technical support；And because the organ without callus occurs, the genetic stability of its plant is higher, this provides quality guarantee to improve bighead atractylodes rhizome quality, industrial seedling rearing and virus-free using plant gene engineering technology.
Description
Technical field
The present invention relates to the reproduction technique of medicinal material, specifically a kind of utilization plumular axis and radicle directly break up adventitious bud
Bighead atractylodes rhizome regenerating system.
Background technology
The bighead atractylodes rhizome ( Atractylodes macrocephalaKoidz.) belong to composite family [WTBX herbaceos perennial, do
Dry rhizome, gas delicate fragrance, sweet-bitter flavor is warm in nature, mainly originates in the ground such as the Zhejiang, Hunan, Hubei and Anhui of China（Chinese Pharmacopoeia
2010）, performance optimal is produced in latent, spy claims " in art ".《Sheng Nong's herbal classic》Middle first time is loaded with the medical value of the bighead atractylodes rhizome, is arranged
It is top grade.The bighead atractylodes rhizome is one of traditional Chinese medicine of China, and its active chemical includes volatile oil and polysaccharide etc..It is main in volatile oil
Contain atractylol, atractylone, sesquiterpenoids etc..The compounds such as volatile oil, lactone have stronger bioactivity.The bighead atractylodes rhizome
With antitumor, promote gastrointestinal motility, adjust the effect of gastrointestinal dysfunction（Chen Xiaoping etc., 2011）, with strengthening the spleen and replenishing qi,
Anti-senile dementia anti-aging is anti-oxidant, enhance immunity, the anticoagulant effect of anti-inflammatory diuresis（Wu Leqin etc., 2011）, it is " strong
The main component of the Chinese patent drug such as spleen ball ", " shenling baizhu powder ", " stomach pill of aucklandia and amomum fruit ".Additionally, the bighead atractylodes rhizome is still containing perhaps various other
There is the compound of pharmacological activity（Long Quanjiang etc., 2004;Chen Bingbing, 2012）.The bighead atractylodes rhizome is good Qi-tonifying drug simultaneously（Poplar
Pretty young woman etc., 2012）.
At present, Wild Plant Atractylodes macrocephala Koidz resource is endangered, the raw medicinal herbs bighead atractylodes rhizome of the major part from artificial growth used by the market,
And the artificial cultivation bighead atractylodes rhizome need to be by cumbersome, prolonged seedling raising process（Wang Liuzhu, 2009）.In recent years, people start to make
Tissue culture rapid propagating technology is used, this can within a short period of time obtain a large amount of bighead atractylodes rhizome test tube seedlings to meet the standardized production kind of the bighead atractylodes rhizome
Plant.The domestic report on the in vitro plant regeneration of the bighead atractylodes rhizome is less, mainly uses bighead atractylodes rhizome lateral bud（Shen Yinzhu etc., 1988;Peng Fei
Deng 2001）, blade（Chen Juan etc., 2006）, stem apex or root-like stock（Liang little Min etc., 2009）Sprout young shoot, young shoot conduct
Explant carries out its in vitro breeding.The technology greatly accelerates the artificial propagation speed of the bighead atractylodes rhizome compared with traditional seedling-raising technique,
But its proliferation frequency is relatively low, and organ by callus makes it be also easy to produce body cell hereditary variation, so as to influence
The stay in grade of regeneration plant.
The content of the invention
It is an object of the invention to provide the bighead atractylodes rhizome regenerating system that a kind of utilization plumular axis and radicle directly break up adventitious bud, to solve
The certainly existing bighead atractylodes rhizome artificial propagation time is long, and frequency is low, the poor problem of the quality stability of regeneration plant.
The purpose of the present invention is achieved through the following technical solutions：One kind directly breaks up adventitious bud using plumular axis and radicle
Bighead atractylodes rhizome regenerating system, comprise the following steps：
（a）Atractylodes macrocephala seed is cleaned, is sterilized, the Atractylodes macrocephala seed after sterilization is inoculated in MS culture mediums, at 25 ± 2 DEG C of dark
Culture to seed is sprouted, and continuation is cultivated under being transferred to light；
（b）Culture is taken out to during plant height 2-3cm, cuts radicle and plumular axis, the fritter that length is 0.3-0.5 cm is cut into, as outer
Implant, is seeded on bud inducement cultivation base, temperature be 25 ± 2 DEG C, light application time be 14h/d, intensity of illumination be 3000-4000
Lx Fiber differentiations, make to grow 2-3cm green buds long on its differentiation to explant；The bud inducement cultivation base refers to the MS in 1L
The culture medium that the agar of sucrose and 8g that TDZ, 0-0.5mgNAA, 30g of 0.5-1.5 mg are with the addition of in culture medium is obtained, it is described
The pH value of bud inducement cultivation base is 5.8-6.2；
（c）The green bud is cut to be forwarded in the blake bottle for filling root media, temperature be 25 ± 2 DEG C, the h/d of illumination 14,
Intensity of illumination 3000-4000 lx continue to cultivate, and root induction obtains regeneration plant；The root media refers in 1/2MS
The culture medium that the agar of the NAA of 0.5mg or the sucrose of IBA, 30g and 8g is obtained, the root media are with the addition of in culture medium
PH value be 5.8-6.2；
（d）The regeneration plant taken root and growth conditions are good is taken out, is transplanted to by vermiculite: sand: the mass ratio of Nutrition Soil
To continue to cultivate in the sterilization matrix of 1: 1: 1 composition, bighead atractylodes rhizome plant is obtained final product.
Step（a）Continue the cultivation temperature of culture under described light for 25 ± 2 DEG C, light application time are 14h/d, intensity of illumination
It is 3000-4000lx.
Step（a）Described sterilization refers to that the Atractylodes macrocephala seed after cleaning is 0.1% with mass concentration ratio before removal kind skin
The mercuric chloride immersion min of seed 10, aseptic water washing 3-5 times after 6 h are soaked in sterilized water, peels off kind of a skin；Mass ratio is used again
Concentration is 1.5% NaClO 10 min of sterilization, aseptic water washing 3-5 times.
Preferably, step（b）Described in bud inducement cultivation base refer to that with the addition of 1.5 mg in the MS culture mediums of 1L
The culture medium that the sucrose of TDZ, 0.2mgNAA, 30g and the agar of 8g are obtained.
Step（a）And step（b）Described MS culture mediums are to be prepared by the following method to form：By MS a great number of elements, MS
Trace element, MS organic matters, 30g sucrose, 8g agar are dissolved in 1L distilled water, and regulation pH value is 5.8-6.2,121 DEG C of sterilizing 15-
20min is obtained final product.
Step（c）Described 1/2MS culture mediums are to be prepared by the following method to form：1/2MS a great number of elements, MS is micro
Element, MS organic matters, 30g sucrose, 8g agar are dissolved in 1L distilled water, and regulation pH value is 5.8-6.2,121 DEG C of sterilizing 15-
20min is obtained final product.
The present invention using the bighead atractylodes rhizome different explants carry out adventitious bud induction differentiation test, establish the bighead atractylodes rhizome it is efficient from
Body rapid propagation method, aseptic seedling is obtained using sterilization method, plumular axis and radicle both specific explants is chosen, by spy
Fixed bud inducement cultivation base and root media carry out directly induction differentiation, establish the efficient rapid regeneration breeding system of the bighead atractylodes rhizome.
The regenerating system that the present invention is provided can be used for the genetic transformation of the bighead atractylodes rhizome and quick in vitro breeding, substantially increase the propagation of the bighead atractylodes rhizome frequently
Rate, shortens repoductive time, has saved production cost, improves production efficiency, is large-scale production and the quality of the medicinal bighead atractylodes rhizome
Improvement provides technical support；And because the organ without callus occurs, the genetic stability of its plant is higher, this
To be provided using plant gene engineering technology improvement bighead atractylodes rhizome quality, using Vitro Quick Reproduction technology industrial seedling rearing and virus-free
Quality guarantee.
Brief description of the drawings
The step of Fig. 1 is embodiment 1（2）The growth conditions figure of middle radicle.
The step of Fig. 2 is embodiment 1（2）The growth conditions figure of mesocotyl.
The step of Fig. 3 is embodiment 1（3）The growth conditions figure of regeneration plant.
The step of Fig. 4 is embodiment 1（4）By the growth conditions figure after Transplantation of Regenerated Plantlets to Nutrition Soil matrix.
Specific embodiment
Example below is used to further describe the present invention, but embodiment does not do any type of limit to the present invention
It is fixed.Unless stated otherwise, the reagent for using of the invention, method and apparatus are the art conventional reagent, method and apparatus.But
The invention is not limited in any way.
The preparation of MS culture mediums：MS a great number of elements, MS trace elements, MS organic matters, 30g sucrose, 8g agar are dissolved in 1L
In distilled water, regulation pH value is 5.8-6.2, and 121 DEG C of sterilizing 15-20min are obtained final product.
The preparation of 1/2 MS culture mediums：By 1/2MS a great number of elements, MS trace elements, MS organic matters, 30g sucrose, 8g agar
It is dissolved in 1L distilled water, regulation pH value is 5.8-6.2,121 DEG C of sterilizing 15-20min are obtained final product.
Embodiment 1
（1）The acquisition of aseptic seedling.Big, the full Atractylodes macrocephala seed of grain is chosen, is rinsed well with flowing water, be 75% with mass concentration ratio
Alcohol-pickled 1 min after, carry out disinfection.Sterilized using two-step method：With the mercuric chloride that mass concentration ratio is 0.1% before removal kind skin
The immersion min of seed 10, aseptic water washing 3-5 times after 6 h are soaked in sterilized water, peels off kind of a skin；It is with mass concentration ratio again
1.5% NaClO 10 min of sterilization, aseptic water washing 3-5 times.Seed after sterilization, is inoculated in and is not added with any growth regulator
MS culture mediums in, after 25 ± 2 DEG C of dark place's cultures are sprouted to seed, be transferred under light and continue culture, cultivation temperature：25±
2 DEG C, light application time：14h/d, intensity of illumination is：3500 lx.
（2）After continuing to cultivate 1 week after Atractylodes macrocephala seed sprouting, taken out when plant height is 2-3cm, cut radicle and plumular axis, made
It is explant, is cut into the fritter of 0.3-0.5 cm, is seeded to the differentiation of evoking adventive bud on bud inducement cultivation base, cultivation temperature：
25 ± 2 DEG C, light application time：14h/ days, intensity of illumination：3000-4000lx；Culture 30 days, has differentiated green bud, counts each of which
Explant Bud Differentiation number is 13.13 ± 2.75（Plumular axis）With 7.95 ± 1.72（Radicle）, Differentiation ration of adventitious buds is respectively
91.18%（Plumular axis）With 68.30%（Radicle）.Wherein bud inducement cultivation base is to the addition of 1.5 mg in the MS culture mediums of 1L
The agar mixing of the sucrose and 8g of TDZ, 0.2mgNAA, 30g, regulation pH value is the training after 5.8,121 DEG C of min of autoclaving 15
Support base.
（3）The 2-3 cm of robust growth on explant green buds long are cut, the culture for filling root media is forwarded to
In bottle, vegetable material illumination cultivation condition is 25 ± 2 DEG C, the h/d of illumination 14, the lx of intensity of illumination 3500, and root induction must regenerate
Plant；Wherein root media refers to the sucrose and the fine jade of 8g of NAA, 30g that 0.5mg is with the addition of in the 1/2MS culture mediums of 1L
Fat mixes, and regulation pH value is the culture medium after 5.8,121 DEG C of min of autoclaving 15.
（4）The regeneration plant that will have been taken root and growth conditions are good, opens culture bottle cap, middle hardening 2-3 days between culture
It is careful afterwards to take out plant, culture medium is cleaned, transplant to vermiculite, sand and the Nutrition Soil mixing sterilization matrix for being in mass ratio 1: 1: 1
In, cultivate in small alms bowl, pour permeable, film covering moisturizing notes divulging information and keeps 25 DEG C or so of environment temperature to be continued
Cultivate, obtain bighead atractylodes rhizome plant.
Embodiment 2
Method only varies by step with embodiment 1（2）Middle bud inducement cultivation base：1.0 mg are with the addition of in the MS culture mediums of 1L
TDZ, 30g sucrose and 8g agar mixing, regulation pH value be 5.8,121 DEG C of min of autoclaving 15, the training being prepared from
Support base.
Embodiment 3
Method only varies by step with embodiment 1（2）Middle bud inducement cultivation base：With the addition of 0.5mg's in the MS culture mediums of 1L
The agar mixing of the sucrose of TDZ, 0.2mgNAA, 30g and 8g, regulation pH value be 5.8,121 DEG C of min of autoclaving 15, preparation and
Into culture medium.
Embodiment 4
Method only varies by step with embodiment 1（2）Middle bud inducement cultivation base：1.0 mg are with the addition of in the MS culture mediums of 1L
TDZ, 0.2mgNAA, 30g sucrose and 8g agar mixing, regulation pH value be 5.8,121 DEG C of min of autoclaving 15, preparation
Culture medium.
Embodiment 5
Method only varies by step with embodiment 1（2）Middle bud inducement cultivation base：1.5 mg are with the addition of in the MS culture mediums of 1L
TDZ, 0.5mgNAA, 30g sucrose and 8g agar mixing, regulation pH value be 5.8,121 DEG C of min of autoclaving 15, preparation
Culture medium.
Embodiment 6
Method only varies by step with embodiment 1（2）Middle bud inducement cultivation base：1.5 mg are with the addition of in the MS culture mediums of 1L
TDZ, 30g sucrose and 8g agar mixing, regulation pH value be 5.8,121 DEG C of min of autoclaving 15, the training being prepared from
Support base.
Comparative example 1
Method only varies by step with embodiment 1（2）Middle bud inducement cultivation base：With the addition of 1.0mg's in the MS culture mediums of 1L
The agar mixing of the sucrose and 8g of 6-BA, 30g, regulation pH value is 5.8,121 DEG C of min of autoclaving 15, the culture being prepared from
Base.
Comparative example 2
Method with embodiment 1, by step（2）In " cut radicle and plumular axis " and be changed to " cutting true leaf and cotyledon "；Its Fiber differentiation
The formula of base is with embodiment 2.
Comparative example 3
Method with embodiment 1, by step（2）In " cut radicle and plumular axis bud " and be changed to " cutting true leaf and cotyledon "；Its induction training
The formula of base is supported with comparative example 1.
Comparative example 4
Method only varies by step with embodiment 1（2）Middle bud inducement cultivation base：With the addition of 2.0mg's in the MS culture mediums of 1L
The agar mixing of the sucrose of TDZ, 0.2mgNAA, 30g and 8g, regulation pH value be 5.8,121 DEG C of min of autoclaving 15, preparation and
Into culture medium.
Comparative example 5
Method only varies by step with embodiment 1（2）Middle bud inducement cultivation base：With the addition of 1.5mg's in the MS culture mediums of 1L
The agar mixing of the sucrose of TDZ, 1.2mgNAA, 30g and 8g, regulation pH value be 5.8,121 DEG C of min of autoclaving 15, preparation and
Into culture medium.
Embodiment 7
Under identical breeding condition and time, step in Statistics Implementation example and comparative example（2）Inductivity/% and each explant
The average bud number of body.Statistics is shown in Tables 1 and 2.
The statistics of the embodiment of table 1 and comparative example differentiation adventitious bud and callus induction rate
* different lowercase letter indication differences significantly (P<0.01)
2 comparative example of table 2 and 3 breaks up the statistics of adventitious bud and callus induction rate
Influence of the different bud inducement cultivation bases to Differentiation ration of adventitious buds can be seen that by the statistics of Tables 1 and 2 very big,
Wherein adventitious buds differentiation number of the radicle in different culture mediums has extremely significant difference（P<0.01）；With in bud inducement cultivation
6-BA is added in base to compare, either plumular axis explant or radicle explant, add bud induction in the bud inducement cultivation base of TDZ
Rate is higher, produce adventitious bud number more；Show that TDZ induction bighead atractylodes rhizome explants produce the effect of adventitious bud more preferably, but TDZ is to true
But effect is excessively poor for leaf and cotyledon, it is seen that TDZ has suitability only for differentiation plumular axis and radicle.Additionally, we are from embodiment
The data counted with comparative example compare discovery, and radicle and Epicotyl Explants of Phaseolus Differentiation ration of adventitious buds and adventitious bud number are subject to not
With significantly affecting for TDZ concentration（P <0.01）；When two kinds of explant Differentiation ration of adventitious buds are 1.5 mgL in TDZ concentration-1
When highest, and work as TDZ concentration for 2.0mgL-1When but show very undesirable；Under identical TDZ concentration conditions, epicotyl explant
The Differentiation ration of adventitious buds of body and each explant average division bud number are above radicle explant；And can see NAA concentration
When higher, the differentiation of bighead atractylodes rhizome radicle and Epicotyl Explants of Phaseolus adventitious bud can be suppressed.
From from the aspect of callus induction rate, radicle and plumular axis explant are in the step of embodiment 1（2）Middle induction is indefinite
Bud, cultivates during to 4 weeks, and radicle and plumular axis explant have directly differentiated adventitious bud from explant, is produced without callus
Raw, the growth conditions of its radicle are as shown in figure 1, the growth conditions of plumular axis are as shown in Figure 2；Cut green bud（Adventitious bud）Turn to plant to life
Root induction in root culture medium, cultivates 20 days or so, root restriction occurs in the part of the base in contact culture medium of bud, and 25 days start
Take root, root is long up to 2-3 cm after 40 d of culture, and its growth conditions is as shown in Figure 3；Treat that bighead atractylodes rhizome regeneration plant root is long to 2-3 cm
During left and right, after hardening 2-3 days, transplant into soil matrix, its growth conditions is as shown in Figure 4.
Additionally, in embodiment 1（4）" NAA " in the root media of step can use " IBA " replace, its actual effect with
Embodiment 1 is suitable.
Embodiment 8
Comparative example 6 is that Tao Yuan scapes in 2010 etc. exist《Medical biotechnology》" the bighead atractylodes rhizome stem tip tissue culture and fast delivered on periodical
Propagating technology optimizing research ".
Comparative example 7 is that Chen Juan in 2006 etc. exists《Chinese agronomy circular》Delivered on periodical " bighead atractylodes rhizome leaf regeneration plant and
The fast numerous research of Multiple Buds ".
Compared with documents 6 and documents 7, its experiment effect is shown in Table 3 to the method that the present invention is provided.
The present invention of table 3 and documents reproductive frequency and the comparing result of repoductive time
The propagation method of more existing some explants of propagation method of the invention is can be seen that from the data of table 3, its breeding frequency
Rate faster, repoductive time it is shorter, without callus, the quality of product is more stable.
Above-described embodiment is the present invention preferably implementation method, but embodiments of the present invention are not by the embodiment
Limitation, it is other it is any without departing from Spirit Essence of the invention and the change, modification, replacement made under principle, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Claims (4)
1. a kind of utilization plumular axis and radicle directly break up the bighead atractylodes rhizome regenerating system of adventitious bud, it is characterised in that comprise the following steps：
（a）Atractylodes macrocephala seed is cleaned, is sterilized, the Atractylodes macrocephala seed after sterilization is inoculated in MS culture mediums, at 25 ± 2 DEG C of dark
Culture to seed is sprouted, and continuation is cultivated under being transferred to light；
（b）Culture is taken out when being 2-3cm to plant height, cuts radicle and plumular axis, is cut into the fritter of a length of 0.3-0.5 cm as outer
Implant, is seeded on bud inducement cultivation base, temperature be 25 ± 2 DEG C, light application time be 14h/d, intensity of illumination be 3000-
4000lx Fiber differentiations, make to grow 2-3cm green buds long on its differentiation to explant；The bud inducement cultivation base refers in 1L
MS culture mediums in the addition of the culture medium that the sucrose of TDZ, 0-0.5mgNAA, 30g and the agar of 8g of 0.5-1.5 mg are obtained；
（c）The green bud is cut, is forwarded in the blake bottle for filling root media, be 25 ± 2 DEG C, the h/ of illumination 14 in temperature
D, intensity of illumination 3000-4000 lx continue to cultivate, and root induction obtains regeneration plant；The root media refers in 1/2MS
The culture medium that the agar of the NAA of 0.5 mg or the sucrose of IBA, 30g and 8g is obtained is with the addition of in culture medium；
（d）The regeneration plant taken root and growth conditions are good is taken out, is transplanted to by vermiculite: sand: the mass ratio of Nutrition Soil
To continue to cultivate in the sterilization matrix of 1: 1: 1 composition, bighead atractylodes rhizome plant is obtained final product.
2. utilization plumular axis according to claim 1 and radicle directly break up the bighead atractylodes rhizome regenerating system of adventitious bud, and its feature exists
In step（a）The cultivation temperature for continuing to cultivate under described light is 25 ± 2 DEG C, light application time is 14h/d, intensity of illumination is
3000-4000lx。
3. utilization plumular axis according to claim 1 and radicle directly break up the bighead atractylodes rhizome regenerating system of adventitious bud, and its feature exists
In step（a）And step（b）Described MS culture mediums are prepared by the following method and form：MS a great number of elements, MS is micro-
Secondary element, MS organic matters, 30g sucrose, 8g agar are dissolved in 1L distilled water, adjust pH value to 5.8-6.2,121 DEG C of sterilizing 15-
20min is obtained final product.
4. utilization plumular axis according to claim 1 and radicle directly break up the bighead atractylodes rhizome regenerating system of adventitious bud, and its feature exists
In step（c）1/2 described MS culture mediums are prepared by the following method and form：By 1/2MS a great number of elements, the micro units of MS
Element, MS organic matters, 30g sucrose, 8g agar are dissolved in 1L distilled water, adjust pH value to 5.8-6.2,121 DEG C of sterilizing 15-20min
Obtain final product.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110250009A (en) * | 2019-07-25 | 2019-09-20 | 恩施土家族苗族自治州农业科学院(恩施土家族苗族自治州硒应用技术与产品开发研究院) | A kind of method and its application expanding numerous Rhizoma Atractylodis Macrocephalae young plant seedling using Rhizoma Atractylodis Macrocephalae Multiple Buds |
| CN113383707A (en) * | 2021-06-24 | 2021-09-14 | 中国科学院合肥物质科学研究院 | Method for establishing high-efficiency in-vitro regeneration system of Qishu mature embryos |
| CN113475395A (en) * | 2021-07-06 | 2021-10-08 | 中国科学院合肥物质科学研究院 | Method for direct regeneration and in-vitro rooting of hypocotyls in Qishu |
| CN115843682A (en) * | 2022-10-28 | 2023-03-28 | 华中农业大学 | Method for inducing and regenerating tulip embryonic axis callus |
| CN117016394A (en) * | 2023-09-19 | 2023-11-10 | 中国中医科学院中药研究所 | Test-tube plantlet of bighead atractylodes rhizome, culture method thereof and method for culturing seedlings to be transplanted of bighead atractylodes rhizome |
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| CN113475395B (en) * | 2021-07-06 | 2022-05-03 | 中国科学院合肥物质科学研究院 | Method for direct regeneration and in-vitro rooting of hypocotyls in Qishu |
| CN115843682A (en) * | 2022-10-28 | 2023-03-28 | 华中农业大学 | Method for inducing and regenerating tulip embryonic axis callus |
| CN115843682B (en) * | 2022-10-28 | 2024-02-27 | 华中农业大学 | Method for inducing and regenerating tulip hypocotyl callus |
| CN117016394A (en) * | 2023-09-19 | 2023-11-10 | 中国中医科学院中药研究所 | Test-tube plantlet of bighead atractylodes rhizome, culture method thereof and method for culturing seedlings to be transplanted of bighead atractylodes rhizome |
| CN117016394B (en) * | 2023-09-19 | 2024-04-26 | 中国中医科学院中药研究所 | Test-tube plantlet of bighead atractylodes rhizome, culture method thereof and method for culturing seedlings to be transplanted of bighead atractylodes rhizome |
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