CN111543317A - Method for culturing and inducing callus by using platycodon grandiflorum anther - Google Patents
Method for culturing and inducing callus by using platycodon grandiflorum anther Download PDFInfo
- Publication number
- CN111543317A CN111543317A CN201910111696.8A CN201910111696A CN111543317A CN 111543317 A CN111543317 A CN 111543317A CN 201910111696 A CN201910111696 A CN 201910111696A CN 111543317 A CN111543317 A CN 111543317A
- Authority
- CN
- China
- Prior art keywords
- anther
- callus
- platycodon grandiflorum
- culture
- culturing
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Images
Classifications
-
- 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
-
- 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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Developmental Biology & Embryology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention provides a method for culturing and inducing callus by using platycodon grandiflorum anther, which comprises the following steps: collecting unopened buds, pretreating at low temperature for 24-48h, sterilizing, and inoculating on culture medium; the culture medium is MS culture medium, and sucrose, agar and hormone are added; dark culture was performed during anther callus induction. The method of the invention has the advantages of early callus starting, high induction rate, large callus volume and compact texture. The invention provides a basis for platycodon grandiflorum anther culture, haploid breeding, mutation breeding and the like.
Description
Technical Field
The invention belongs to the field of botany, and particularly relates to a method for culturing and inducing callus by using platycodon grandiflorum anther.
Background
Root of balloonflowerPlatycodon grandiflorum(Jacq.) A. DC. belongs to Campanulaceae, perennial dicotyledonous herbaceous plants, is a large amount of medicinal materials in China, has large demand, and is a plant for medicine, food and appreciation. The root of platycodon grandiflorum is used as a medicine, the main active ingredient of the root of platycodon grandiflorum is saponin, and the root of platycodon grandiflorum has wide pharmacological activities of relieving cough, resisting inflammation, reducing blood pressure and blood sugar, losing weight, resisting tumors, improving human immunity and the like.
The platycodon grandiflorum is mainly propagated by adopting seeds or asexual cuttage, but the seed propagation has the defects of long production period, low yield, low seed germination rate and low survival rate of cuttage branches, and the development of production is seriously limited.
The tissue culture technology is widely applied to various aspects such as platycodon grandiflorum variety improvement, germplasm preservation, rapid propagation, gene transfer and the like. Under the condition of in vitro culture of the platycodon grandiflorum tissues, the chromosomes of somatic cells are easy to generate various variations, and the frequency of gene mutation is greatly improved, so that the platycodon grandiflorum genetic resources are greatly enriched, and a new way is opened for improving medicinal plants. The tissue culture and rapid propagation technology can be used for providing sufficient high-quality seedlings for platycodon grandiflorum cultivation; in addition, the research on the tissue culture technology of the platycodon grandiflorum is carried out, and various materials meeting the requirements can be provided for the research on other aspects of the platycodon grandiflorum. The stem tip, stem segment and leaf in vitro material of the platycodon grandiflorum can be induced to differentiate into regenerated plants.
The study on the callus induction and plant regeneration process of the roots, stems and leaves of different explants of platycodon grandiflorum is carried out by utilizing a tissue culture technology, so that the platycodon grandiflorum is successfully induced to generate callus and differentiated seedlings are obtained. Finding out different callus inducing frequencies of the root, stem and leaf of the explant, and screening out the optimal callus inducing culture medium and seedling differentiation culture medium. The research of the vitro culture and plant regeneration test of the stem tip, stem segment and leaf of the platycodon grandiflorum by the cinnabar and the like. Experimental results of the comfortable radix and alpine forest show that the platycodon grandiflorum test-tube plantlet can induce the callus more easily, and the callus keeps stronger redifferentiation capability. The study of tissue culture of platycodon grandiflorum was conducted using epicotyl as explant, such as Yangliang paper and Qianzhigang. Research on the induction of platycodon grandiflorum callus and the formation of adventitious buds is carried out on dunn by taking ovules, stem tips, stem segments and root tips as explants. The result shows that on the culture medium of MS + 2.4-D3.0 mg/L + 6-BA 0.2mg/L, hypocotyl callus can form cluster buds, and the differentiation capability of adventitious buds is stronger; the differentiation ability of the root tip callus bud is inferior; the callus of the stem tip and the stem segment has no bud differentiation.
The platycodon grandiflorum flowers belong to amphoteric flowers, and according to literature reports and bagging selfing tests, the selfing and fruiting rate of the platycodon grandiflorum in a natural state is low, and fruiting mainly depends on cross pollination. The genetic breeding research of the platycodon grandiflorum is slow in progress, haploid plants are obtained through anther culture, homozygote is obtained through chromosome doubling, excellent pure line progeny selected from the homozygote is not separated, the performance is uniform, and the breeding period can be shortened. At higher induction frequency, the haplotype can show the recombined gamete type on the plant more fully, and new genetic resources and selection materials can be provided.
The callus is an important vector for plant rapid propagation, protoplast preparation and transgenic research, can provide good test materials for ploidy breeding, tissue culture rapid propagation, somatic cell hybridization, clone variation and genetic transformation of the callus, and finally provides an important basis for genetic improvement of the platycodon grandiflorum. In the past, callus induction research is carried out on roots, stems and leaves of different explants of platycodon grandiflorum. The platycodon grandiflorum has large anther, large amount and long flowering period, provides sufficient materials for anther culture, but does not see relevant reports for anther culture by using the platycodon grandiflorum at home and abroad.
Disclosure of Invention
In order to solve the defects of the prior art, the invention provides a method for culturing and inducing callus by using platycodon grandiflorum anther.
The invention adopts the following technical scheme:
the invention provides a method for culturing and inducing callus by using platycodon grandiflorum anther, which comprises the following steps: collecting unopened buds after noon in sunny days, wrapping with wet gauze, pre-treating in a sealed bag at 4 deg.C for 24-48h in dark environment, washing the buds with tap water and Tween 20 for 20 min, soaking in 75% ethanol for 30 s, sterilizing with 1% mercuric chloride for 8-10 min, washing with sterile water under aseptic condition of superclean bench for 3-5 times, taking out anther under dissecting mirror, and inoculating on culture medium; the culture medium is an MS culture medium, 30 g/L sucrose, 6 g/L agar and hormone are added, the pH value is 5.8, and sterilization is carried out for 20 min at the temperature of 120-; the hormone is 6-BA and/or 2,4-D, NAA, and the concentration of 6-BA is 0mg.L-1-2.0 mg.L-1(ii) a The concentration of the 2,4-D is 0mg.L-1-2.0 mg.L-1(ii) a The NAA concentration is 0mg.L-1-0.2 mg.L-1(ii) a Dark culture is carried out during the induction period of the anther callus, and the temperature is 25 +/-1 ℃.
The method provided by the invention can be applied to anther culture of white flower platycodon grandiflorum.
When the method provided by the invention is applied to anther culture of platycodon grandiflorum, the hormone is 1.0mg.L-16-BA and 2.0 mg.L-12,4-D 。
When the method provided by the invention is applied to anther culture of platycodon grandiflorum, the low-temperature pretreatment time is 48 hours.
The method provided by the invention can be applied to the cultivation of the anther of purple-flower platycodon grandiflorum.
When the method provided by the invention is applied to the cultivation of the anther of purple-flowered platycodon, the hormone is 1.0mg.L-16-BA 、1.0 mg.L-12,4-D and 0.2 mg.L-1NAA 。
When the method provided by the invention is applied to cultivation of the anther of purple-flower platycodon, the low-temperature pretreatment time is 24-48 h.
The invention has the beneficial effects that: the invention provides a method for culturing and inducing callus by utilizing platycodon grandiflorum anther. The invention provides a basis for platycodon grandiflorum anther culture, haploid breeding, mutation breeding and the like.
Drawings
FIG. 1 is a graph illustrating the effect of different times of low temperature pretreatment on callus induction.
Detailed Description
Example 1 experiment of culture of Platycodon grandiflorum anther for inducing callus
No.1 material and method
No.1.1 test Material
The test material varieties are white flower platycodon grandiflorum (BHJG) and purple flower platycodon grandiflorum (ZHJG), and are from the medicinal plant garden of Shandong agricultural university.
Test method No.1.2
Collecting unopened buds after noon in sunny days, wrapping with wet gauze, placing in a sealed bag, storing at 4 deg.C for 1 d (except for temperature pretreatment material) in dark environment, washing the buds with tap water and Tween 20 for 20 min, soaking in 75% ethanol for 30 s, sterilizing with 1% mercuric chloride for 8-10 min, washing with sterile water under aseptic condition of superclean bench for 3-5 times, taking out anther under dissecting mirror, and inoculating on culture medium. The inoculation density was 15 anthers/vial, three replicates per treatment, three replicates per vial.
MS culture medium is taken as an induction culture medium, 30 g/L of sucrose and 6 g/L of agar are added, the pH value is 5.8, and the mixture is sterilized at the temperature of 120 ℃ and 125 ℃ for 20 min. Dark culture is carried out during the induction period of the anther callus, and the temperature is 25 +/-1 ℃. After 30d, the induction rate of the callus and the size and the quality of the tissue block are investigated, and the growth condition of the callus is observed once a week. Callus induction rate (%) = (number of calli formed/number of anthers inoculated) × 100%.
No.1.2.1 Effect of different hormone ratios and concentrations on callus induction
The test materials of purple flower platycodon root and white flower platycodon root are respectively taken, anthers in the mononuclear border period are taken as explants, and the explants are respectively inoculated on callus induction culture media containing 6-BA and 2,4-D, NAA in different concentration combinations. And (5) counting the induction rate of the callus of each treatment at 30d after inoculation. Three levels of three hormones were established, respectively: 6-BA: 0 mg.L-1, 1.0 mg.L-1, 2.0 mg.L-1; 2, 4-D: 0 mg.L-1, 1.0 mg.L-1, 2.0 mg.L-1; NAA: 0 mg.L-1, 0.1 mg.L-1, 0.2 mg.L-1. The orthogonal experimental design (tables 1 and 2) of L9 (33) was used.
Influence of different temperature treatment on callus Induction No.1.2.2
White flower and purple flower platycodon grandiflorum are used as test materials, collected flower buds are wrapped by wet gauze, and are treated at low temperature of 4 ℃ for 1 d, 2d and 3 d, and the induction rate of each genotype is higher or lower after 30 d.
No.2 results and analysis
No.2.1 Effect of different hormone treatments on callus formation in anther culture of Platycodon grandiflorum
The anther in the uninucleate border period is used as an explant, and the 6-BA and 2,4-D, NAA which are commonly used in anther culture are used as exogenous hormones, and different levels are set for testing. As shown in Table 1, the healing rate induced by the treatment 1 without any exogenous hormone is 0, while the healing rate can be remarkably improved by adding the exogenous hormone into the culture medium, and the influence of the hormones on different genotype materials is found to be different through the range comparison of the healing rates of the three exogenous hormones. The effect of the 6-BA on the platycodon grandiflorum is the largest, the effect of the 2,4-D is the second, and the effect of the NAA is the smallest, which shows that the 6-BA has obvious effect on the anther culture of the platycodon grandiflorum. Different hormone combinations and concentration ratios have different induction effects on the platycodon grandiflorum callus.
After the inoculated anthers are cultured for 20 days under the condition of natural illumination at 25 ℃, light yellow callus begins to appear around some anthers, the growth of the callus is slow at the beginning, but the growth is gradually accelerated later, and after 45 days, the size of the medium callus is equal to that of the match heads.
Investigation after inoculation for 45 days shows that in the culture medium added with 1.0 mg.L-16-BA + 2.0 mg.L-12, 4-D, namely the treatment 6, the white flower platycodon grandiflorum callus induction rate is highest and reaches 88.9%, meanwhile, the white flower platycodon grandiflorum callus in the culture medium of the treatment 6 starts at the earliest time, the anther is enlarged and has light yellow color, and then white spots similar to matcha appear at the two ends, the texture is compact and the volume is largest, so that the white flower platycodon grandiflorum is the best hormone combination. Under the combined treatment of other hormones, the induced callus is yellow or yellow brown in color, loose in texture and small in volume, and has more white snowflake-shaped calluses.
TABLE 1 Effect of different hormone treatments on calli production by anther culture of Platycodon grandiflorum
No.2.2 influence of different hormones on callus generated by culturing anther of Philippine violet flower
As seen from Table 2, treatment 1 without any exogenous hormone induced a healing rate of 0, whereas the addition of exogenous hormone to the medium significantly increased the healing rate. The extremely poor comparison of the healing rates of the three exogenous hormones shows that the effect on 2,4-D of the purple-flowered platycodon root is the greatest, and the effect on NAA, 6-BA is the least.
After the inoculated anthers are cultured for 20 days under the condition of natural illumination at 25 ℃, faint yellow callus begins to appear around some anthers, the growth of the callus is slow at the beginning, but the growth is gradually accelerated later, and after 45 days, the size of the medium callus is equal to that of the match heads.
45 days after inoculation, the addition of 1.0mg.L was found-16-BA + 1.0 mg.L-1 2,4-D + 0.2 mg.L-1NAA is the culture medium of the treatment 5, the induction rate of calli of the Philippine violet root is the highest and reaches 97.8%, and meanwhile, the Philippine violet root is the culture medium of the treatment 5The medium callus has early starting, compact texture and largest volume, and is the best hormone combination of purple flower and platycodon grandiflorum.
TABLE 2 Effect of different hormone treatments on the callus formation in anther culture of Philippine Violet flower
The hormone plays a key role in the induction of the callus of the platycodon grandiflorum anther, and 1.0mg.L of hormone with different combinations is added in the hormone set in the experiment-16-BA + 2.0 mg.L-1The white flower platycodon grandiflorum callus on the culture medium of 2,4-D has the highest induction rate and the best quality, and the highest callus rate is 88.9%; adding 1.0mg.L-16-BA + 1.0 mg.L-1 2,4-D + 0.2 mg.L-1The induction rate of the calli of the purple flower platycodon grandiflorum on the NAA culture medium is the highest, the quality is the best, and the highest healing rate is 97.8%.
No.2.3 Effect of different genotypes on anther culture to generate callus
The callus is difficult to induce by anther culture, and different genotypes have great difference. The two genotype materials of the experiment can induce callus when the anther of which the microspore development is in the mononuclear border stage is subjected to callus induction (table 3), but the callus induction rate difference is large, which indicates that the genotype has great influence on the generation of anther callus in the anther culture. Among two tested materials with different genotypes, the induction rate of the purple-flowered platycodon root is higher, and the purple-flowered platycodon root belongs to the genotype which is easier to induce callus. Most of the callus that grows out is light yellow in color and compact in texture. The white platycodon grandiflorum callus has low induction rate and obvious difference with the purple platycodon grandiflorum, and the induced callus has loose texture and more white snowflake shapes.
TABLE 3 Effect of different genotypes on callus induction
Influence of different pretreatment temperatures on callus induction rates of No.2.4
The low-temperature pretreatment mainly has the function of providing a more suitable environment for inducing callus, and the culture effect of the anther of many plants after the low-temperature pretreatment is better than that without the low-temperature pretreatment. As can be seen from figure 1, the buds in the uninucleate side-by-side period of microspore development can be subjected to low-temperature (4 ℃) pretreatment for 24h, 48h and 72 h to induce callus. The highest induction rate is achieved by low-temperature treatment for two days, and the difference with that without low-temperature treatment is obvious. The suitable time for low temperature pretreatment of anthers indicating that microspore development is in the uninucleate edgewise stage is 48 h. In addition, the cure rate of purple-flower platycodon grandiflorum is less sensitive to low-temperature treatment time than that of white-flower platycodon grandiflorum.
Claims (7)
1. A method for culturing and inducing callus by using platycodon grandiflorum anther comprises the following steps: collecting unopened buds after noon in sunny days, wrapping with wet gauze, pre-treating in a sealed bag at 4 deg.C for 24-48h in dark environment, washing the buds with tap water and Tween 20 for 20 min, soaking in 75% ethanol for 30 s, sterilizing with 1% mercuric chloride for 8-10 min, washing with sterile water under aseptic condition of superclean bench for 3-5 times, taking out anther under dissecting mirror, and inoculating on culture medium; the culture medium is an MS culture medium, 30 g/L sucrose, 6 g/L agar and hormone are added, the pH value is 5.8, and sterilization is carried out for 20 min at the temperature of 120-; the hormone is 6-BA and/or 2,4-D, NAA, and the concentration of 6-BA is 0mg.L-1-2.0 mg.L-1(ii) a The concentration of the 2,4-D is 0mg.L-1-2.0 mg.L-1(ii) a The NAA concentration is 0mg.L-1-0.2 mg.L-1(ii) a Dark culture is carried out during the induction period of the anther callus, and the temperature is 25 +/-1 ℃.
2. The method for inducing callus by using anther culture of platycodon grandiflorum according to claim 1, wherein the callus is obtained by culturing platycodon grandiflorum anther: the method is applied to anther culture of white flower platycodon grandiflorum.
3. The method for inducing callus by using anther culture of platycodon grandiflorum according to claim 2, wherein:the hormone is 1.0mg.L-16-BA and 2.0 mg.L-12,4-D 。
4. The method for culturing and inducing callus by using anther of platycodon grandiflorum according to claim 3, wherein the callus is obtained by culturing and inducing anther of platycodon grandiflorum, comprising the following steps: the time of the low-temperature pretreatment is 48 hours.
5. The method for inducing callus by using anther culture of platycodon grandiflorum according to claim 1, wherein the callus is obtained by culturing platycodon grandiflorum anther: the method is applied to the cultivation of the anther of purple-flower platycodon grandiflorum.
6. The method for inducing callus by using anther culture of platycodon grandiflorum according to claim 5, wherein: the hormone is 1.0mg.L-16-BA 、 1.0 mg.L-12,4-D and 0.2 mg.L-1NAA 。
7. The method for inducing callus by using anther culture of platycodon grandiflorum according to claim 6, wherein: the low-temperature pretreatment time is 24-48 h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910111696.8A CN111543317A (en) | 2019-02-12 | 2019-02-12 | Method for culturing and inducing callus by using platycodon grandiflorum anther |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910111696.8A CN111543317A (en) | 2019-02-12 | 2019-02-12 | Method for culturing and inducing callus by using platycodon grandiflorum anther |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111543317A true CN111543317A (en) | 2020-08-18 |
Family
ID=71996196
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910111696.8A Withdrawn CN111543317A (en) | 2019-02-12 | 2019-02-12 | Method for culturing and inducing callus by using platycodon grandiflorum anther |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111543317A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115486369A (en) * | 2022-09-27 | 2022-12-20 | 广东海洋大学 | Method for improving induction rate of embryonic callus of myrtle anther |
| CN116098059A (en) * | 2023-02-09 | 2023-05-12 | 泉脉农业科技有限公司 | Method for inducing callus by co-culture of eustoma grandiflorum free microspores and anther |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105409770A (en) * | 2015-11-30 | 2016-03-23 | 浙江拓普药业股份有限公司 | Platycodon grandiflorum anther callus inducing method |
-
2019
- 2019-02-12 CN CN201910111696.8A patent/CN111543317A/en not_active Withdrawn
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105409770A (en) * | 2015-11-30 | 2016-03-23 | 浙江拓普药业股份有限公司 | Platycodon grandiflorum anther callus inducing method |
Non-Patent Citations (7)
| Title |
|---|
| 고정애等: "도라지Platycodongrandiflorum약배양에서저온처리가화분2형현상및배형성에미치는영향", 《KOREAN JOURNAL OF PLANT TISSUE CULTURE 》 * |
| 고정애等: "물레나물약배양에의한부정신초및식물체재분화", 《KOREAN JOURNAL OF PLANT RESOURCES》 * |
| 吴京姬等: "桔梗花药培养初报 ", 《植物研究》 * |
| 杨丽: "桔梗花药组织培养研究", 《中国优秀硕士学位论文全文数据库农业科技辑》 * |
| 杨丽等: "接种密度和蔗糖浓度对桔梗花药诱导率的影响 ", 《北方园艺》 * |
| 赵丽莉: "桔梗花药的发育时期及组织培养研究", 《中国优秀硕士学位论文全文数据库农业科技辑》 * |
| 赵丽莉等: "桔梗花药愈伤组织诱导培养基筛选", 《北方园艺》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115486369A (en) * | 2022-09-27 | 2022-12-20 | 广东海洋大学 | Method for improving induction rate of embryonic callus of myrtle anther |
| CN116098059A (en) * | 2023-02-09 | 2023-05-12 | 泉脉农业科技有限公司 | Method for inducing callus by co-culture of eustoma grandiflorum free microspores and anther |
| CN116098059B (en) * | 2023-02-09 | 2024-01-26 | 泉脉农业科技有限公司 | Method for inducing callus by co-culture of eustoma grandiflorum free microspores and anther |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN100586275C (en) | Fast replication method for dendrobium | |
| Moradi et al. | Micropropagation of strawberry by multiple shoots regeneration tissue cultures | |
| CN104273028B (en) | Method for rapid in-vitro propagation of Crassulaceae plant | |
| CN103430844B (en) | Method for gardenia tissue culture | |
| JP2022160084A (en) | Rapid propagation method of polygonatum cyrtonema hua | |
| CN102228005A (en) | Pinellia ternate tissue culture one-step speciation method | |
| Shahin | Isolation and culture of protoplasts: tomato | |
| Das et al. | Micropropagation of sweet orange, Citrus sinensis Osbeck. for the development of nucellar seedlings | |
| CN111543317A (en) | Method for culturing and inducing callus by using platycodon grandiflorum anther | |
| CN100361569C (en) | Method for preparing plant haploid embryo and haploid plant | |
| CN114342804A (en) | Method for promoting regeneration of camellia oleifera bud stem plant through light control | |
| CN112715367A (en) | Method for carrying out tissue culture subculture propagation on cornus walteri by utilizing lanthanum nitrate | |
| CN115885855B (en) | Method for establishing regeneration system by taking hypocotyl of tea tree kui as explant | |
| CN107410029A (en) | A kind of culture medium for improving long-grained nonglutinous rice and Xian round-grained rice friendship Anther Culture Efficiency and application | |
| CN100442971C (en) | Induction of peony embryoid | |
| CN114600772B (en) | Tissue culture method and rapid propagation method of michelia figo in remote mountains | |
| CN114424749A (en) | Liriope spicata in-vitro rapid propagation method | |
| CN105359961B (en) | It is a kind of to pass through the rescue isolated method for obtaining Hipeastrum vittalum (L Her.) Herb.- Amaryllisvittata Ait ' apple ' cenospecies of rataria | |
| Huang et al. | Chinese gooseberry, kiwifruit (Actinidia spp.) | |
| CN108293879B (en) | Rapid propagation method of euphorbia kansui | |
| CN108719046B (en) | Method for induced cultivation of hybrid liquidambar formosana tetraploid | |
| Zhai et al. | Shoot multiplication and plant regeneration in Caragana fruticosa (Pall.) Besser | |
| CN111557243A (en) | Tissue culture method of Dracocephalum rupestre and application thereof | |
| CN117044627B (en) | Tissue culture rapid propagation and in-vitro preservation method for alpine plant taraxacum | |
| CN107114245A (en) | One grows flax tissue culture method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20200818 |