CN108094203B - Preparation method of pseudo-ginseng seeds - Google Patents
Preparation method of pseudo-ginseng seeds Download PDFInfo
- Publication number
- CN108094203B CN108094203B CN201711402380.1A CN201711402380A CN108094203B CN 108094203 B CN108094203 B CN 108094203B CN 201711402380 A CN201711402380 A CN 201711402380A CN 108094203 B CN108094203 B CN 108094203B
- Authority
- CN
- China
- Prior art keywords
- pseudo
- ginseng
- seeds
- artificial
- culture
- 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.)
- Active
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
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
- A01H4/006—Encapsulated embryos for plant reproduction, e.g. artificial seeds
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention provides a preparation method of pseudo-ginseng seeds, which comprises the following steps: differentiating and culturing the pseudo-ginseng embryonic callus to obtain a somatic embryo, wherein the culture medium comprises the following components: SH +0.2-0.4 mg/L2, 4-D +28-32g/L sucrose +5-8g/L agar; continuously carrying out differentiation culture on the somatic embryos to obtain mature cotyledon-shaped embryos, wherein the culture medium comprises the following components: SH +28-32g/L sucrose +5-7g/L agar; taking the cotyledon-shaped embryo as an embedded propagule, taking SH +1-2 wt% of sucrose as artificial endosperm, taking 3-5 wt% of sodium alginate as artificial seed coat base, and preparing artificial seeds by adopting a dripping method. According to the preparation method of the pseudo-ginseng seed, the situation of seedling shortage in large-scale artificial cultivation of pseudo-ginseng is solved by using the modern biotechnology, and the pseudo-ginseng embryonic cells have the advantages of high growth speed and stable properties.
Description
Technical Field
The invention relates to the field of pseudo-ginseng cultivation, and in particular relates to a preparation method of pseudo-ginseng seeds.
Background
Notoginseng, also known as Tianqi, is an Araliaceae plant of Umbelliferae and is mainly distributed in Yunnan, Guangxi, Jiangxi, Sichuan and other places. The root of the Chinese medicinal material pseudo-ginseng is used as a medicinal part, and has the effects of removing blood stasis, stopping bleeding, reducing swelling and relieving pain. Can be used for treating hemoptysis, hematemesis, epistaxis, hematochezia, metrorrhagia, traumatic hemorrhage, thoracico-abdominal pain, and traumatic swelling and pain.
Pseudo-ginseng is a traditional and rare Chinese medicinal material in China, one of forty large medicinal material varieties, has an application history of over 400 years to date, and is famous at home and abroad, and the pseudo-ginseng has a name of a hemostatic and psychotropic medicine from ancient times. Due to the unique and obvious effects of pseudo-ginseng, the application is continuously expanded, the dosage of health care, chemical industry and medical treatment is continuously increased, particularly, related research and development products are in endless and huge demand when China enters the high-incidence period of cardiovascular and cerebrovascular diseases, and enterprises such as Tianqi toothpaste of Guangxi Wuzhou, Yunnan white drug group, Tianjin Shili, Guangzhou white Yunshan, Yuxi Hewei, Zhenbaoma pharmacy, Shanghai Huayu and the like are large user representatives of pseudo-ginseng. Annual demand has approached 8000 tons and the usage is still on the rise.
With the increasing market demand, the breeding problem becomes the focus of research. The traditional propagation mode of rhizome division and root induction seedling has low propagation rate and high production cost, and is not beneficial to resource protection. Although the problem of rapid propagation can be solved to a certain extent by using a tissue culture rapid propagation mode, the bottleneck problems of variation and germplasm degradation of callus after subculture for multiple generations, low transplanting survival rate of callus differentiated seedlings, high production cost and the like exist at the same time, and the large-scale cultivation production of panax notoginseng is limited to a great extent.
Artificial seeds are in fact closely related to the practical application of somatic embryos. The somatic embryo has the advantages of fast proliferation, high yield and stable heredity, and more importantly, the somatic embryo also has a bipolar structure, so that a complete plant can be formed in one step without inducing the formation of buds and roots step by step according to the conventional method. Therefore, the artificial seeds have wide application prospect. At present, no relevant report on a comparison system of pseudo-ginseng artificial seeds exists at home and abroad, and the pseudo-ginseng artificial seeds are still in a technological blank stage.
In view of the above, the present invention is particularly proposed.
Disclosure of Invention
The invention aims to provide a preparation method of pseudo-ginseng seeds, which solves the problem of seedling shortage in large-scale artificial cultivation of pseudo-ginseng by using a modern biotechnology, overcomes the defects of easy browning, variation, long production period, high cost and the like in conventional tissue culture of pseudo-ginseng, solves the problems of low pseudo-ginseng seed quantity and low reproduction rate, opens up a new way for cultivation of pseudo-ginseng seeds, realizes proliferation and differentiation of pseudo-ginseng somatic embryos, and ensures that the germination rate of the finally prepared pseudo-ginseng artificial seeds can reach more than 95 percent.
In order to achieve the above purpose of the present invention, the following technical solutions are adopted:
the invention provides a preparation method of pseudo-ginseng seeds, which comprises the following steps:
(A) differentiating and culturing the pseudo-ginseng embryonic callus to obtain a somatic embryo, wherein the culture medium comprises the following components: SH +0.2-0.4 mg/L2, 4-D +28-32g/L sucrose +5-8g/L agar;
(B) continuously carrying out differentiation culture on the somatic embryos to obtain mature cotyledon-shaped embryos, wherein the culture medium comprises the following components: SH +28-32g/L sucrose +5-7g/L agar;
(C) taking the cotyledon-shaped embryo as an embedded propagule, taking SH +1-2 wt% of sucrose as artificial endosperm, taking 3-5 wt% of sodium alginate as artificial seed coat base, and preparing artificial seeds by adopting a dripping method.
In the prior art, pseudo-ginseng is one of the traditional rare Chinese medicinal materials in China, forty large medicinal material varieties exist, the application history is over 400 years from now on, and pseudo-ginseng is famous at home and abroad and has the name of a hemostatic magical medicine from ancient times. Due to the unique and obvious effects of pseudo-ginseng, the application is continuously expanded, the dosage of health care, chemical industry and medical treatment is continuously increased, particularly, related research and development products are in endless and huge demand when China enters the high-incidence period of cardiovascular and cerebrovascular diseases, and enterprises such as Tianqi toothpaste of Guangxi Wuzhou, Yunnan white drug group, Tianjin Shili, Guangzhou white Yunshan, Yuxi Hewei, Zhenbaoma pharmacy, Shanghai Huayu and the like are large user representatives of pseudo-ginseng. Annual demand has approached 8000 tons and the usage is still on the rise.
With the increasing market demand, the breeding problem becomes the focus of research. The traditional propagation mode of rhizome division and root induction seedling has low propagation rate and high production cost, and is not beneficial to resource protection. Although the problem of rapid propagation can be solved to a certain extent by using a tissue culture rapid propagation mode, the bottleneck problems of variation and germplasm degradation of callus after subculture for multiple generations, low transplanting survival rate of callus differentiated seedlings, high production cost and the like exist at the same time, and the large-scale cultivation production of panax notoginseng is limited to a great extent.
Artificial seeds are in fact closely related to the practical application of somatic embryos. The somatic embryo has the advantages of fast proliferation, high yield and stable heredity, and more importantly, the somatic embryo also has a bipolar structure, so that a complete plant can be formed in one step without inducing the formation of buds and roots step by step according to the conventional method. Therefore, the artificial seeds have wide application prospect. At present, no relevant report on a comparison system of pseudo-ginseng artificial seeds exists at home and abroad, and the pseudo-ginseng artificial seeds are still in a technological blank stage.
The invention aims to solve the technical problems and provides a preparation method of pseudo-ginseng seeds, the pseudo-ginseng seeds are prepared by performing proliferation culture on pseudo-ginseng embryonic callus, then performing differentiation culture, and finally adopting a drip method under specific operating conditions to prepare artificial seeds.
Preferably, in step (A) of the present invention, the culture conditions are preferably dark culture at a temperature of 20 to 24 ℃ and are subcultured on the differentiation medium every 4 to 6 weeks, more preferably every 5 weeks.
The somatic embryos in this step are preferably those for 3-4 subcultures.
Preferably, in step (B) of the present invention, the culture conditions are also preferably performed under dark culture conditions, and the culture temperature is controlled to be between 30-35 ℃, so that the cultured somatic embryos can be more easily performed in the subsequent steps by controlling the appropriate culture conditions.
Preferably, the culture time is 50-60d, preferably 55d, and the length of the cotyledonary embryo obtained by the differentiation culture is preferably controlled to be 4-6mm, more preferably 5 mm.
In step (C) of the present invention, the dropping method is preferably carried out as follows: dripping semi-gel state sodium alginate containing cotyledon-shaped embryo into 5-6 wt% CaCl with suction tube2Performing ion exchange in the solution for 8-9min, and cleaning.
Wherein, in order to eliminate the drying of the impurities, the cleaning is preferably carried out by using distilled water, and the cleaning frequency is controlled between 2 and 3 times.
Preferably, the cotyledonary embryo is mixed with the sodium alginate in a ratio of 1-2: 100 mass ratio. And the above-mentioned semi-gel state is formed by mixing both, preferably heating and stirring, and the pH value is preferably controlled to be 6 to 7 by measuring the pH.
The pseudo-ginseng seeds prepared by the preparation method are placed in a solid culture medium of 1/2SH +10g/L sucrose +6g/L agar, the culture temperature is 25 ℃, the illumination intensity is 2000lx, the illumination is 16h/d, and the germination rate is more than 95%.
Compared with the prior art, the invention has the beneficial effects that:
(1) according to the preparation method of the pseudo-ginseng seeds, the situation of seedling shortage in large-scale artificial cultivation of pseudo-ginseng is solved by utilizing the modern biotechnology, the pseudo-ginseng embryonic cells have the advantages of high growth speed and stable properties, and the defects of easy browning, variation, long production period, high cost and the like in conventional tissue culture of pseudo-ginseng are overcome;
(2) the preparation method of the pseudo-ginseng seed also solves the problems of small amount of pseudo-ginseng seed and low reproduction rate, opens up a new way for cultivating the pseudo-ginseng seed, realizes the proliferation and differentiation of the pseudo-ginseng somatic embryo, and ensures that the germination rate of the finally prepared pseudo-ginseng artificial seed can reach more than 95 percent.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below.
Fig. 1 is a schematic diagram showing the detailed appearance of the seeds of panax notoginseng obtained by the manufacturing method of the embodiment 1 of the invention;
FIG. 2 is a schematic diagram of the appearance of the seeds of Panax notoginseng obtained by the method of example 2 of the present invention after 20 days of growth;
FIG. 3 is a detailed view of the seed coat broken by the root and the stem after the pseudo-ginseng seed grows for 20 days, which is obtained by the manufacturing method of the embodiment 2 of the invention;
FIG. 4 is a schematic diagram of the appearance of the seeds of Panax notoginseng obtained by the method of example 3 of the present invention after growing for 30 days;
FIG. 5 is a detailed view of the beginning of stem growth and leaf growth of the seeds of Panax notoginseng obtained by the method of example 3 of the present invention after 30 days;
fig. 6 is a schematic diagram of the shape of the pseudo-ginseng seed obtained by the manufacturing method of embodiment 4 of the present invention after growing for 60 d.
Detailed Description
Embodiments of the present invention will be described in detail below with reference to examples, but it will be understood by those skilled in the art that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
Example 1
The preparation method of the pseudo-ginseng artificial seeds comprises the following steps:
(1) proliferation of somatic embryos: inoculating the embryogenic callus to a proliferation medium consisting of: SH +0.2 mg/L2, 4-D +28g/L sucrose +5g/L agar, and subcultured on the same medium every 4 weeks for 1 time in the dark at a culture temperature of 24 ℃. Maintaining the differentiation capacity of the embryonic callus;
(2) differentiation of somatic embryos: inoculating the embryonic callus to a somatic embryo differentiation culture medium, wherein the culture medium comprises the following components: SH +32g/L sucrose +7g/L agar, dark culture, the culture temperature is 30 ℃. Obtaining mature cotyledonary shaped embryos (about 4mm in length) after 50 days;
(3) preparing pseudo-ginseng artificial seeds: pseudo-ginseng somatic embryos of about 4mm are used as embedded propagules, SH + 1% sucrose is used as artificial endosperm, and 3 wt% sodium alginate is used as an artificial seed coat matrix. Adopting a dripping method to prepare artificial seeds: on a clean bench, the somatic embryo is immersed into artificial endosperm containing sodium alginate, and the semi-gel state sodium alginate coated with the somatic embryo is dripped into 5 wt% CaCl by using a suction pipe2In the solution, each artificial seed only wraps one somatic embryo, and the ratio of cotyledonary embryo to sodium alginate is 1: 100 mass ratio. In CaCl2Ion exchange in the solution for 8min, solidifying into balls, taking out, and washing with distilled water twice to obtain pseudo-ginseng artificial seeds. Placing in 1/2SH +10g/L sucrose +6g/L agar solid culture medium, culturing at 25 deg.C under illumination of 2000lx under illumination of 16h/d, the germination rate is more than 95%, and the appearance of the pseudo-ginseng seeds prepared by manual work is shown in figure 1.
Example 2
The preparation method of the pseudo-ginseng artificial seeds comprises the following steps:
(1) proliferation of somatic embryos: inoculating the embryogenic callus to a proliferation medium consisting of: SH +0.4 mg/L2, 4-D +32g/L sucrose +8g/L agar, and subcultured on the same medium every 6 weeks for 1 time in the dark at a temperature of 20 ℃. Maintaining the differentiation capacity of the embryonic callus;
(2) differentiation of somatic embryos: inoculating the embryonic callus to a somatic embryo differentiation culture medium, wherein the culture medium comprises the following components: SH +28g/L sucrose +5g/L agar, dark culture is carried out, and the culture temperature is 35 ℃. Obtaining mature cotyledonary shaped embryos (about 6mm in length) after 50 days;
(3) preparing pseudo-ginseng artificial seeds: pseudo-ginseng somatic embryos of about 6mm are used as embedded propagules, SH + 1% sucrose is used as artificial endosperm, and 3 wt% sodium alginate is used as an artificial seed coat matrix. Adopting a dripping method to prepare artificial seeds: on a clean bench, the somatic embryo is immersed into artificial endosperm containing sodium alginate, and the semi-gel state sodium alginate coated with the somatic embryo is dripped into 6 wt% CaCl by using a suction pipe2In the solution, each artificial seed only wraps one somatic embryo, and the cotyledon-shaped embryo and sodium alginate are mixed in a ratio of 2: 100 mass ratio. In CaCl2Ion exchange in the solution for 9min, solidifying into balls, taking out, and washing with distilled water for three times to obtain the pseudo-ginseng artificial seeds. Placing in 1/2SH +10g/L sucrose +6g/L agar solid culture medium, culturing at 25 deg.C under illumination intensity of 2000lx for 16h/d, and allowing germination rate to reach above 95%, wherein the growth state of Notoginseng radix seed is shown in FIGS. 2-3.
Example 3
The preparation method of the pseudo-ginseng artificial seeds comprises the following steps:
(1) proliferation of somatic embryos: inoculating the embryogenic callus to a proliferation medium consisting of: SH +0.3 mg/L2, 4-D +30g/L sucrose +7g/L agar, and subcultured on the same medium every 5 weeks for 1 time in the dark at 22 ℃. Maintaining the differentiation capacity of the embryonic callus;
(2) differentiation of somatic embryos: inoculating the embryonic callus to a somatic embryo differentiation culture medium, wherein the culture medium comprises the following components: SH +30g/L sucrose +6g/L agar, dark culture, the culture temperature is 32 ℃. Mature cotyledonary embryos (approximately 5mm in length) were obtained after 55 d;
(3) preparing pseudo-ginseng artificial seeds: pseudo-ginseng somatic embryos of about 5mm are used as embedded propagules, SH + 1% sucrose is used as artificial endosperm, and 3 wt% sodium alginate is used as an artificial seed coat matrix. Adopting a dripping method to prepare artificial seeds: on a clean bench, the somatic embryo is immersed into artificial endosperm containing sodium alginate, and the semi-gel state sodium alginate coated with the somatic embryo is dripped into 6 wt% CaCl by using a suction pipe2In the solution, each artificial seed only wraps one somatic embryo, and the cotyledon-shaped embryo and sodium alginate are mixed in a proportion of 1.5: 100 mass ratio. In CaCl2Ion exchange in the solution for 9min, solidifying into balls, taking out, and washing with distilled water for three times to obtain the pseudo-ginseng artificial seeds. Placing in 1/2SH +10g/L sucrose +6g/L agar solid culture medium, culturing at 25 deg.C under illumination intensity of 2000lx for 16h/d, and allowing germination rate to reach above 95%, wherein the growth state of Notoginseng radix seed is shown in FIGS. 4-5.
Example 4
The preparation method of the pseudo-ginseng artificial seeds comprises the following steps:
(1) proliferation of somatic embryos: inoculating the embryogenic callus to a proliferation medium consisting of: SH +0.3 mg/L2, 4-D +31g/L sucrose +6g/L agar, and subcultured on the same medium every 5 weeks for 1 time in the dark at 23 ℃. Maintaining the differentiation capacity of the embryonic callus;
(2) differentiation of somatic embryos: inoculating the embryonic callus to a somatic embryo differentiation culture medium, wherein the culture medium comprises the following components: SH +30g/L sucrose +6g/L agar, dark culture, the culture temperature is 33 ℃. Mature cotyledonary embryos (approximately 5mm in length) were obtained after 60 days;
(3) preparing pseudo-ginseng artificial seeds: pseudo-ginseng somatic embryos of about 5mm are used as embedded propagules, SH + 1% sucrose is used as artificial endosperm, and 3 wt% sodium alginate is used as an artificial seed coat matrix. Adopting a dripping method to prepare artificial seeds: immersing the embryo in a person with sodium alginate on a clean benchDripping the semi-gel sodium alginate coated with the somatic embryo into 6 wt% CaCl in the endosperm2In the solution, each artificial seed only wraps one somatic embryo, and the cotyledon-shaped embryo and sodium alginate are mixed in a proportion of 1.5: 100 mass ratio. In CaCl2Ion exchange in the solution for 9min, solidifying into balls, taking out, and washing with distilled water for three times to obtain the pseudo-ginseng artificial seeds. Placing in 1/2SH +10g/L sucrose +6g/L agar solid culture medium, culturing at 25 deg.C under illumination intensity of 2000lx for 16h/d, and allowing germination rate to reach above 95%, wherein the growth state of Notoginseng radix seed is shown in FIG. 6.
Comparative example 1
The specific procedure was the same as in example 4 except that the steps (1) to (2) were omitted and the somatic embryo was directly dropped into the artificial endosperm using a pipette.
Comparative example 2
The specific procedure was the same as in example 4 except that in step (1), the medium composition was: SH +0.6 mg/L2, 4-D +20g/L sucrose +2g/L agar.
Comparative example 3
The specific procedure was the same as in example 4 except that in step (2), the medium composition was: SH +35g/L sucrose +8g/L agar.
Example 5
Spraying pullulan solution on the surface of the pseudo-ginseng artificial seeds, drying, putting into a sterile triangular flask, sealing with a sealing film, and storing in a refrigerator at 4 ℃, wherein 50 seeds are treated correspondingly in each example.
After 30 days, the artificial seeds of each example were inoculated into a solid medium of 1/2SH +10g/L sucrose +6g/L agar at a density of 1 pellet/cm2The depth is 0.5cm, and the culture conditions are as follows: the culture temperature is 25 ℃, the illumination intensity is 2000lx, the illumination is 16h/d, the humidity is 85%, the transformation condition of the seeds is observed after 30 days, and the germination rate is counted. The results are shown in Table 1.
The statistical basis of the forwarding rate is as follows: inoculating the pseudo-ginseng artificial seeds into humus, culturing at constant temperature for 1 month, counting the forwarding rate, and taking cotyledons and roots generated by somatic embryos as the standards for successful transformation.
Specific results are shown in table 1 below.
TABLE 1 statistics of results
As can be seen from table 1 above, the preparation method of pseudo-ginseng seeds according to the embodiment of the present invention can significantly improve the germination rate, and is worth of wide popularization and application.
Compared with the prior art, the invention has the beneficial effects that:
(1) according to the preparation method of the pseudo-ginseng seeds, the situation of seedling shortage in large-scale artificial cultivation of pseudo-ginseng is solved by utilizing the modern biotechnology, the pseudo-ginseng embryonic cells have the advantages of high growth speed and stable properties, and the defects of easy browning, variation, long production period, high cost and the like in conventional tissue culture of pseudo-ginseng are overcome;
(2) the preparation method of the pseudo-ginseng seed also solves the problems of small amount of pseudo-ginseng seed and low reproduction rate, opens up a new way for cultivating the pseudo-ginseng seed, realizes the proliferation and differentiation of the pseudo-ginseng somatic embryo, and ensures that the germination rate of the finally prepared pseudo-ginseng artificial seed can reach more than 95 percent.
While particular embodiments of the present invention have been illustrated and described, it would be obvious that various other changes and modifications can be made without departing from the spirit and scope of the invention. It is therefore intended to cover in the appended claims all such changes and modifications that are within the scope of this invention.
Claims (10)
1. A preparation method of pseudo-ginseng seeds is characterized by comprising the following steps:
(A) differentiating and culturing the pseudo-ginseng embryonic callus to obtain a somatic embryo, wherein the culture medium comprises the following components: SH +0.2-0.4 mg/L2, 4-D +29-32g/L sucrose +5-8g/L agar;
(B) continuously carrying out differentiation culture on the somatic embryos to obtain mature cotyledon-shaped embryos, wherein the culture medium comprises the following components: SH +28-32g/L sucrose +5-7g/L agar;
(C) taking the cotyledon-shaped embryo as an embedded propagule, taking SH +1 wt% of sucrose as artificial endosperm, taking 3 wt% of sodium alginate as artificial seed skin base, and preparing artificial seeds by adopting a dripping method;
in the step (B), the culture condition is dark culture, and the culture temperature is 30-35 ℃; the culture time is 50-60 days.
2. The method according to claim 1, wherein the culturing conditions in step (A) are dark culturing at 20-24 ℃ and subculture every 4-6 weeks on the differentiation medium.
3. The method of claim 2, wherein the differentiation medium is subcultured every 5 weeks in the step (A).
4. The method for preparing seeds of panax notoginseng as claimed in claim 1, wherein the somatic embryos obtained in step (a) are subcultured for 3-4 passages.
5. The method for preparing seeds of panax notoginseng according to claim 1, wherein the cultivation time in step (B) is 55 d.
6. The method for preparing seeds of Panax notoginseng as claimed in any one of claims 1 to 4, wherein the cotyledonary embryos have a length of 4 to 6mm in step (B).
7. The method for preparing seeds of Panax notoginseng as claimed in any one of claims 1 to 4, wherein the cotyledonary embryos have a length of 5mm in the step (B).
8. The method for preparing seeds of panax notoginseng according to any one of claims 1 to 4, wherein the dropping method in the step (C) comprises: dripping semi-gel state sodium alginate containing cotyledon-shaped embryo into 5-6 wt% CaCl with suction tube2Performing ion exchange in the solution for 8-9min, and cleaning.
9. The method of claim 8, wherein the washing in step (C) is performed with distilled water for 2-3 times.
10. The method for preparing seeds of panax notoginseng according to claim 8, wherein the ratio of the cotyledonary embryo to the sodium alginate in step (C) is 1-2: 100 mass ratio.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711402380.1A CN108094203B (en) | 2017-12-22 | 2017-12-22 | Preparation method of pseudo-ginseng seeds |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711402380.1A CN108094203B (en) | 2017-12-22 | 2017-12-22 | Preparation method of pseudo-ginseng seeds |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108094203A CN108094203A (en) | 2018-06-01 |
| CN108094203B true CN108094203B (en) | 2020-11-24 |
Family
ID=62212087
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711402380.1A Active CN108094203B (en) | 2017-12-22 | 2017-12-22 | Preparation method of pseudo-ginseng seeds |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108094203B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110199883B (en) * | 2019-07-11 | 2022-08-30 | 云南维和药业股份有限公司 | Cultivation method of panax notoginseng tissue culture seedlings |
| CN110771508B (en) * | 2019-11-26 | 2022-07-05 | 大连大学 | Preparation method of artificial blueberry seeds |
| CN111869565B (en) * | 2020-07-16 | 2023-04-25 | 云南农业大学 | Culture method for expanding propagation of pseudo-ginseng green embryogenic callus |
| CN113317203A (en) * | 2021-06-28 | 2021-08-31 | 大连大学 | Pogostemon cablin artificial seed culture method |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102907318B (en) * | 2011-08-01 | 2015-09-09 | 东北林业大学 | A kind of method utilizing the fast numerous pseudo-ginseng regeneration plant of bioreactor culture somatic embryo |
| CN104686340A (en) * | 2015-02-22 | 2015-06-10 | 杨业容 | Method for establishing cell suspension culture system of gynura divaricate (L.) DC. |
| CN106069791B (en) * | 2016-08-25 | 2018-04-24 | 文山苗乡三七科技有限公司 | A kind of pseudo-ginseng embryonic callus induction cultural method |
-
2017
- 2017-12-22 CN CN201711402380.1A patent/CN108094203B/en active Active
Non-Patent Citations (1)
| Title |
|---|
| 人参属代表植物人工种子包埋技术研究;郑惠元;《中国优秀硕士学位论文全文数据库 农业科技辑》;20190115(第12期);第8、16页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108094203A (en) | 2018-06-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108094203B (en) | Preparation method of pseudo-ginseng seeds | |
| JP7274233B2 (en) | Rapid breeding method of Huangjing | |
| CN105706900B (en) | Sterile seeding and seedling raising method for hybrid orchid and Tibet cymbidium hybrid seeds | |
| CN111226792B (en) | High-throughput breeding method for leaf seedlings of cymbidium sinense | |
| CN105010147A (en) | Special culture medium for improving tissue culture propagation speed of haworthia succulent plants and tissue culture method | |
| CN104160956A (en) | High efficient and fast culture method of anoectochilus roxburghii tissue cultured seedling | |
| CN109220790B (en) | In vitro propagation method of rhododendron simsii | |
| CN103548696B (en) | A kind of method of breeding blueberry seedling | |
| CN111183902B (en) | Tissue culture method for polygonatum sibiricum | |
| CN108077071B (en) | Culture medium for culturing vitex agnus-castus tissue and rapid propagation method | |
| CN114946657A (en) | Hispid fig tissue culture method | |
| CN104813938A (en) | Panax notoginseng tissue culture seedling raising method | |
| CN115581202B (en) | Method for regenerating new variety of Polygonatum cyrtonema Fabricius and in vitro seedling | |
| CN110741937A (en) | Rapid propagation method of polygonatum sibiricum | |
| CN105850737B (en) | A kind of alpine rose " young beauty " chromosome doubling method | |
| CN107593448A (en) | A kind of method that rapid propagation in vitro is carried out using eight treasures (choice ingredients of certain special dishes) red-spotted stonecrop aseptic seedling stem sections | |
| CN103621401B (en) | The method for building up of Tulipa edulis Clonal regeneration system | |
| CN108782244B (en) | Tissue culture method for longzhuguo | |
| CN107616093B (en) | Tissue culture rapid propagation method of Ardisia macrophylla | |
| CN111919751A (en) | Tissue culture method for murraya paniculata seeds | |
| CN111528098A (en) | Angelica sinensis tissue culture seedling rooting culture method | |
| CN115088617B (en) | Culture medium and method for polyploid dandelion plant tissue culture | |
| CN117136842B (en) | Micro-tissue culture and rapid propagation method of cudrania tricuspidata | |
| CN112753579B (en) | In-vitro culture and plant regeneration method for leaves of Chinese medicinal herb chlorophytum comosum | |
| CN110278872B (en) | Walnut somatic embryo rooting and seedling exercising transplanting 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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20180601 Assignee: Guangzhou Fenghua Shubao Biotechnology Co.,Ltd. Assignor: INSTITUTE OF SPECIAL ANIMAL AND PLANT SCIENCES OF CAAS Contract record no.: X2023980032410 Denomination of invention: A preparation method of Panax notoginseng seeds Granted publication date: 20201124 License type: Common License Record date: 20230222 |