Method for producing notoginsenoside and ginsenoside by utilizing panax notoginseng cotyledon embryo

Abstract

The invention relates to a method for producing notoginsenoside and ginsenoside by utilizing panax notoginseng cotyledon embryos, and belongs to the technical field of plant adventitious root tissue culture. The method comprises the following steps: inducing embryogenic calluses by utilizing immature zygotic embryos of panax notoginseng; proliferating the panax notoginseng embryogenic callus on a plate culture medium; s2, putting the pseudo-ginseng embryogenic callus obtained in the step S2 into a liquid culture medium, carrying out cell dispersion, and sieving by using a sieve with the aperture of 24-30 meshes; a mechanical stirring type bioreactor is filled with a liquid culture medium, and the globular embryo and the heart-shaped embryo are placed in the mechanical stirring type bioreactor to be cultured; s3, recovering the filtrate in the step S3, filtering by using a screen with the aperture of 30-80 meshes and 80-150 meshes to obtain uniform panax notoginseng embryogenic cells, and differentiating the panax notoginseng embryogenic cells in a triangular flask for the step S4; and drying the cotyledon embryo obtained in the step S4 at 45 DEG C, and grinding into powder. According to the method, large-scale generation of cotyledon embryos can be completed only in 4-6 weeks, the production process is remarkably accelerated, the method is suitable for industrial batch production requirements, and the production efficiency is high.

Description

Technical Field

[0001] The invention relates to the technical field of plant adventitious root tissue culture technology, and in particular to a method for producing notoginseng saponins and ginsenosides by utilizing cotyledon embryos of notoginseng. Background Art

[0002] Panax notoginseng, also known as Tianqi, belongs to the genus Panax in the Araliaceae family, along with ginseng, American ginseng, and bamboo ginseng. It is a perennial herbaceous plant with perennial roots. It is mainly distributed in Yunnan and Guangxi in China, growing under forests or on hillsides at an altitude of 400-1800 meters. Panax notoginseng usually takes more than three years to be harvested, and the harvest period is from October to December. The main medicinal parts are roots and rhizomes. "Supplement to Compendium of Materia Medica" records that "ginseng is the best for replenishing qi, and Panax notoginseng is the best for replenishing blood." Modern pharmacological studies have shown that Panax notoginseng has anti-inflammatory, lipid-regulating, anti-platelet aggregation, anti-tumor, anti-thrombotic, prevention and treatment of atherosclerosis and fatty liver, prevention of myocardial infarction, and promotion of liver regeneration and angiogenesis.

[0003] As the medicinal effects of Panax notoginseng are gradually discovered, wild Panax notoginseng has become extinct due to the excavation of the dry pond. Panax notoginseng cultivated in the field has serious problems such as continuous cropping obstacles, diseases, heavy metal residues in the soil and long growth cycle, which affect the quality of Panax notoginseng and restrict the development and application of Panax notoginseng products. As the country's requirements for the quality of medicinal plant products increase and the application of Panax notoginseng is gradually expanded to the fields of skin care products and cosmetics, Panax notoginseng cultivated in the field is difficult to meet the requirements. Since the cultivation process of Panax notoginseng cotyledon embryos is completely separated from the soil, does not involve pesticides, has no heavy metal pollution, and has a short cycle and is not affected by the external climate environment, it overcomes the above-mentioned problems of field cultivation and provides a new way for the production of Panax notoginseng saponin R1, ginsenoside Rg1 and ginsenoside Rb1. However, there are few reports on the method of producing Panax notoginseng saponin R1, ginsenoside Rg1 and ginsenoside Rb1 using Panax notoginseng cotyledon embryos.

[0004] In the related art, one scheme for producing Panax notoginseng cotyledon embryos is to first induce Panax notoginseng embryonic callus through different explants such as Panax notoginseng embryos, stem segments, young shoots, adventitious roots, etc. under different concentrations of hormones (2,4-D and 6-BA), and then use the embryonic callus to differentiate into somatic embryos on a hormone-free medium, and then use different concentrations of hormone GA to induce somatic embryos. 3Act on the somatic embryo to develop into a cotyledon embryo. The disadvantage of this scheme is that the entire culture cycle starting from the induction of embryonic callus is time-consuming, and the complexity and high cost of the process limit its application in industrial production. Another scheme is to induce the embryonic callus of Panax notoginseng through different explants such as Panax notoginseng seed embryos, stem segments, tender shoots, adventitious roots, etc. on different concentrations of hormones (2,4-D and 6-BA). The embryonic callus differentiates for 6 weeks on a low concentration of 2,4-D culture medium to form a spherical embryo, and the spherical embryo differentiates for 4 weeks in a bioreactor to form a cotyledon embryo containing a radicle morphology. The disadvantage of this scheme is that a large number of secondary embryos will be produced, that is, multiple embryos are combined together, affecting subsequent normal development. Secondly, deformed embryos may appear during the culture process in the reactor. In addition, the culture cycle of this scheme is relatively long, and the whole process needs to last for more than 10 weeks. At present, the production of saponins using bioreactors generally uses Panax notoginseng callus, but the saponin content of callus is low and the production efficiency is low. Therefore, it is necessary to provide a method for producing notoginsenosides and ginsenosides using notoginseng cotyledon embryos, which is suitable for industrial mass production needs and has high production efficiency. Summary of the invention

[0005] In order to overcome the problems existing in the background technology, the present invention provides a method for producing notoginsenosides and ginsenosides using Panax notoginseng cotyledon embryos. Large-scale production of cotyledon embryos can be completed in only 4-6 weeks, which significantly accelerates the production process, is suitable for industrial mass production needs, and has high production efficiency.

[0006] To achieve the above object, the present invention is implemented through the following technical solutions:

[0007] The present invention provides a method for producing notoginsenoside and ginsenoside by using notoginseng cotyledon embryos, comprising the following steps:

[0008] S1, using immature zygotic embryos of Panax notoginseng to induce embryonic callus;

[0009] S2, proliferating embryonic callus of Panax notoginseng on plate culture medium;

[0010] S3, placing the embryonic callus of Panax notoginseng obtained in step S2 into a liquid culture medium and performing cell dispersion, and sieving using a sieve with a 24-30 mesh aperture to obtain spherical embryos and heart-shaped embryos;

[0011] S4, a mechanical stirring bioreactor is filled with liquid culture medium, the spherical embryos and the heart-shaped embryos are placed in the mechanical stirring bioreactor for culture, and cotyledon embryos are obtained after 4-5 weeks;

[0012] S5, recovering the filtrate of step S3, filtering it with a sieve with an aperture of 30-80 or 80-150 meshes to obtain uniform embryonic cells of Panax notoginseng, placing the cells in a conical flask for differentiation, placing the conical flask on a shaker at a rotation speed of 100-120 rpm, and culturing in the dark to obtain spherical embryos and heart-shaped embryos, which are used in step S4;

[0013] S6. Dry the cotyledon embryos obtained in step S4 at 45° C. and grind the powder. Pass the powder through a No. 3 sieve to extract notoginsenoside and ginsenoside.

[0014] In the above technical solution, the plate culture medium in step S2 is WPM culture medium with a pH of 5.8 and 30 g / L sucrose, 2 mg / L 2,4-D, 1 g / L CH, 1 g / L PVP, 1 g / L activated carbon and 3 g / L Phytagel added.

[0015] In the above technical solution, the embryonic callus tissue described in step S3 is placed in a conical flask containing 30-50 mL WPM culture solution supplemented with 30 g / L sucrose, and a magnetic stirring rotor is placed in the flask to stir for 10 minutes to disperse the cells.

[0016] In the above technical solution, the mechanical stirring bioreactor in step S4 is a 7L mechanical stirring bioreactor.

[0017] In the above technical solution, the input parameters of the mechanical stirring bioreactor described in step S4 are pH 5.6-5.7, temperature 23-25°C, rotation speed 150-250rpm, and ventilation volume 1-3L / min.

[0018] In the above technical solution, the liquid culture medium described in step S4 is a WPM liquid culture medium with a pH of 5.8 and 50-70 g / L sucrose, 0.5-2 g / L glutamine, and 2-4 g / L MES added thereto.

[0019] In the above technical solution, the conical flask in step S5 contains 30-100 mL of liquid culture medium.

[0020] In the above technical solution, the liquid culture medium in step S5 is a WPM liquid culture medium supplemented with 50-80 g / L sucrose / maltose, 4-16.6 g / L MES, and 0.5-1 g / L glutamine.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] 1. The present invention proposes a new method for obtaining notoginseng saponins and ginsenosides (mainly notoginseng saponin R1, ginsenoside Rg1 and ginsenoside Rb1) of the medicinal ingredients of Panax notoginseng without destroying natural resources and relying on land. This method breaks through the limitation of relying on land resources in the traditional planting process, does not need to destroy the natural environment, ensures the green production of Panax notoginseng medicinal materials, and has no chemical residues, thereby realizing a pure and safe raw material supply. In addition, this method has a short culture cycle, and only 4-6 weeks are required to complete the large-scale generation of cotyledon embryos, which significantly accelerates the production process and is suitable for industrial mass production needs.

[0023] 2. The present invention produces notoginsenoside R1, ginsenoside Rg1 and ginsenoside Rb1 by culturing cotyledon embryos. The total saponin content in the cotyledon embryos obtained by the present invention is much higher than the saponin content in the callus tissue, which can improve the production efficiency of saponins and enhance product quality.

[0024] 3. The present invention has an efficient circular production, which obtains spherical embryos and heart-shaped embryos by directly utilizing the differentiated Panax notoginseng embryonic cells in the liquid culture medium, thereby eliminating the step of starting over from seed embryos. These Panax notoginseng embryonic cells can rapidly differentiate into a large number of spherical embryos and heart-shaped embryos in just 3 to 5 weeks, which can then be efficiently used for the large-scale production of cotyledon embryos, thus constructing a closed production loop.

[0025] 4. In order to meet the specific needs of different culture stages, the present invention adopts a strategy of step-by-step expansion of culture and combines a variety of culture methods and containers: embryonic callus induction stage - solid culture medium; globular embryo heart-shaped embryo differentiation stage - triangular flask liquid culture; cotyledon embryo differentiation stage - liquid fermentation tank culture. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the structures shown in these drawings without paying any creative work.

[0027] Figure 1 It is the effect diagram of each stage of forming cotyledon embryo of the present invention;

[0028] Figure 2 It is a composition diagram of embryonic callus 24-30 objects of the present invention;

[0029] Figure 3 It is a composition diagram of embryonic callus 30-80 mesh of the present invention;

[0030] Figure 4It is a composition diagram of embryonic callus 80-150 mesh of the present invention;

[0031] Figure 5 The invention discloses a spherical embryo and a heart-shaped embryo formed by the 80-150 mesh cells in a triangular flask. DETAILED DESCRIPTION

[0032] In order to make the objectives, technical solutions and advantages of the present invention more clear, the following part will further describe the embodiments of the present application in detail in conjunction with the accompanying drawings.

[0033] When the following description refers to the drawings, unless otherwise indicated, the same numbers in different drawings represent the same or similar elements. The embodiments described in the following exemplary embodiments do not represent all embodiments consistent with the present invention. Instead, they are only examples of devices and methods consistent with some aspects of the present invention as detailed in the attached claims.

[0034] In the description of the present invention, it should be understood that the terms "first", "second", etc. are only used for descriptive purposes and cannot be understood as indicating or implying relative importance. For those of ordinary skill in the art, the specific meanings of the above terms in the present invention can be understood according to specific circumstances. In addition, in the description of the present invention, unless otherwise specified, "plurality" refers to two or more. "And / or" describes the association relationship of associated objects, indicating that three relationships may exist. For example, A and / or B can represent: A exists alone, A and B exist at the same time, and B exists alone. The character " / " generally indicates that the previously associated objects are in an "or" relationship.

[0035] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in this specification are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more related listed items.

[0036] Example 1

[0037] A method for producing notoginsenosides and ginsenosides by using cotyledon embryos of Panax notoginseng, specifically comprising the following steps:

[0038] S1. Induce embryonic callus using immature zygotic embryos of Panax notoginseng: woody plant medium WPM, 0.5 mg / L 2,4-D, 1 g / L PVP, 1 g / L CH, 30 g / L sucrose, 3 g / L Phytagel (plant gel), pH 5.8.

[0039] S2. Embryogenic callus proliferated on a plate medium supplemented with 30 g / L sucrose, 2 mg / L 2,4-D, 1 g / L CH, 1 g / L PVP, 1 g / L activated carbon and 3 g / L Phytagel at a pH of 5.8.

[0040] S3. In a clean bench, use sterilized forceps to pick up embryonic callus tissue and place it in a flask containing 50 mL WPM culture medium supplemented with 30 g / L sucrose. Place a magnetic stirring rotor in the flask and stir for 10 minutes to disperse the cells. Sieve with a 24-30 mesh sieve to obtain spherical embryos and heart-shaped embryos.

[0041] S4 and 7L mechanical stirring bioreactors were filled with WPM liquid culture medium with pH 5.8 supplemented with 50g / L sucrose, 0.5g / L glutamine, and 2g / LMES. Spherical embryos and heart-shaped embryos were cultured in the 7L mechanical stirring bioreactor. The input parameters were pH 5.6, temperature 23°C, rotation speed 150rpm, ventilation volume 1L / min, and cotyledonary embryos were obtained after 4-5 weeks.

[0042] S5. Recover the filtrate of step S3, filter it with a sieve with an aperture of 30-80 or 80-150 mesh to obtain uniform embryonic cells of Panax notoginseng, and place it in a conical flask filled with 30 mL of liquid culture medium for differentiation. The liquid culture medium is WPM liquid culture medium supplemented with 50 g / L sucrose, 4 g / L MES, and 0.5 g / L glutamine. The conical flask is placed on a shaker with a rotation speed of 100-120 rpm and cultured in the dark to obtain spherical embryos and heart-shaped embryos, which are used in step S4 to obtain a large number of cotyledonary embryos with good morphology.

[0043] S6. Dry and grind the cotyledon embryos obtained in step S4 at 45°C, and pass the powder through a No. 3 (355μm) sieve for later use. The extraction and content determination of notoginsenoside R1, ginsenoside Rg1 and ginsenoside Rb1 refer to the extraction method in the 2020 edition of the "Pharmacopoeia of the People's Republic of China", and change the heating reflux extraction method to the ultrasonic extraction method. Preparation of the test solution: Take about 1g of the powder of this product, accurately weigh it, accurately add 10mL of ethanol, and ultrasonically extract for 0.5h. Preparation of the reference solution: Take an appropriate amount of notoginsenoside R1, ginsenoside Rg1 and ginsenoside Rb1 reference substances, accurately weigh them, and add methanol to make a mixed solution containing 0.4mg of each per 1mL. Chromatographic conditions: Chromatographic column: COSMOSIL Cholester (4.6IDx10mm); Detection wavelength: 203nm; Mobile phase: acetonitrile-water; Determination method: Accurately aspirate 20μL of reference solution and test solution respectively, inject into high performance liquid chromatography, and determine the content of saponins in cotyledon embryo, Panax notoginseng saponin R1 is 0.17mg / g, ginsenoside Rg1 is 0.33mg / g, ginsenoside Re is 0.43mg / g, ginsenoside Rb1 is 0.38mg / g, and total saponins are 1.31mg / g.

[0044] Example 2

[0045] A method for producing notoginsenosides and ginsenosides by using cotyledon embryos of Panax notoginseng, specifically comprising the following steps:

[0046] S1. Induce embryonic callus using immature zygotic embryos of Panax notoginseng: woody plant medium WPM, 0.5 mg / L 2,4-D, 1 g / L PVP, 1 g / L CH, 30 g / L sucrose, 3 g / L Phytagel (plant gel), pH 5.8.

[0047] S2. Embryogenic callus proliferated on a plate medium supplemented with 30 g / L sucrose, 2 mg / L 2,4-D, 1 g / L CH, 1 g / L PVP, 1 g / L activated carbon and 3 g / L Phytagel at a pH of 5.8.

[0048] S3. In a clean bench, use sterilized tweezers to pick up embryonic callus tissue and place it in a triangular flask containing 40 mL WPM culture medium supplemented with 30 g / L sucrose. Place a magnetic stirring rotor in the triangular flask and stir for 10 minutes to disperse the cells. Sieve with a 24-30 mesh sieve to obtain spherical embryos and heart-shaped embryos.

[0049] S4 and 7L mechanical stirring bioreactors were filled with WPM liquid culture medium with pH 5.8 supplemented with 60g / L sucrose, 1g / L glutamine, and 3g / LMES. Spherical embryos and heart-shaped embryos were cultured in the 7L mechanical stirring bioreactor. The input parameters were pH 5.6, temperature 24°C, rotation speed 200rpm, ventilation volume 2L / min, and cotyledonary embryos were obtained after 4-5 weeks.

[0050] S5. Recover the filtrate of step S3, filter it with a sieve with an aperture of 30-80 or 80-150 mesh to obtain uniform embryonic cells of Panax notoginseng, and place it in a conical flask filled with 60 mL of liquid culture medium for differentiation. The liquid culture medium is WPM liquid culture medium supplemented with 65 g / L maltose, 10 g / L MES, and 0.8 g / L glutamine. The conical flask is placed on a shaker with a rotation speed of 100-120 rpm and cultured in the dark to obtain spherical embryos and heart-shaped embryos, which are used in step S4 to obtain a large number of cotyledonary embryos with good morphology.

[0051] S6. Dry and grind the cotyledon embryos obtained in step S4 at 45°C, and pass the powder through a No. 3 (355μm) sieve for later use. The extraction and content determination of notoginsenoside R1, ginsenoside Rg1 and ginsenoside Rb1 refer to the extraction method in the 2020 edition of the "Pharmacopoeia of the People's Republic of China", and change the heating reflux extraction method to the ultrasonic extraction method. Preparation of the test solution: Take about 1g of the powder of this product, accurately weigh it, accurately add 10mL of ethanol, and ultrasonically extract for 0.5h. Preparation of the reference solution: Take an appropriate amount of notoginsenoside R1, ginsenoside Rg1 and ginsenoside Rb1 reference substances, accurately weigh them, and add methanol to make a mixed solution containing 0.4mg of each per 1mL. Chromatographic conditions: Chromatographic column: COSMOSIL Cholester (4.6IDx10mm); Detection wavelength: 203nm; Mobile phase: acetonitrile-water; Determination method: Accurately aspirate 20μL of reference solution and test solution respectively, inject into high performance liquid chromatography, and determine the content of saponins in cotyledon embryo, Panax notoginseng saponin R1 is 0.24mg / g, ginsenoside Rg1 is 0.36mg / g, ginsenoside Re is 0.37mg / g, ginsenoside Rb1 is 0.47mg / g, and total saponins are 1.44mg / g.

[0052] Example 3

[0053] A method for producing notoginsenosides and ginsenosides by using cotyledon embryos of Panax notoginseng, specifically comprising the following steps:

[0054] S1. Induce embryonic callus using immature zygotic embryos of Panax notoginseng: woody plant medium WPM, 0.5 mg / L 2,4-D, 1 g / L PVP, 1 g / L CH, 30 g / L sucrose, 3 g / L Phytagel (plant gel), pH 5.8.

[0055] S2. Embryogenic callus proliferated on a plate medium supplemented with 30 g / L sucrose, 2 mg / L 2,4-D, 1 g / L CH, 1 g / L PVP, 1 g / L activated carbon and 3 g / L Phytagel at a pH of 5.8.

[0056] S3. In a clean bench, use sterilized forceps to pick up embryonic callus tissue and place it in a triangular flask containing 30 mL WPM culture medium supplemented with 30 g / L sucrose. Place a magnetic stirring rotor in the triangular flask and stir for 10 minutes to disperse the cells. Sieve with a 24-30 mesh sieve to obtain spherical embryos and heart-shaped embryos.

[0057] S4 and 7L mechanical stirring bioreactors were filled with WPM liquid culture medium with pH 5.8 supplemented with 70g / L sucrose, 2g / L glutamine, and 4g / LMES. Spherical embryos and heart-shaped embryos were cultured in the 7L mechanical stirring bioreactor. The input parameters were pH 5.7, temperature 25°C, rotation speed 250rpm, ventilation volume 3L / min, and cotyledonary embryos were obtained after 4-5 weeks.

[0058] S5. Recover the filtrate of step S3, filter it with a sieve with an aperture of 30-80 or 80-150 mesh to obtain uniform embryonic cells of Panax notoginseng, and place it in a conical flask filled with 100 mL of liquid culture medium for differentiation. The liquid culture medium is WPM liquid culture medium supplemented with 80 g / L sucrose, 16.6 g / L MES, and 1 g / L glutamine. The conical flask is placed on a shaker with a rotation speed of 100-120 rpm and cultured in the dark to obtain spherical embryos and heart-shaped embryos, which are used in step S4 to obtain a large number of cotyledonary embryos with good morphology.

[0059] S6. Dry and grind the cotyledon embryos obtained in step S4 at 45°C, and pass the powder through a No. 3 (355μm) sieve for later use. The extraction and content determination of notoginsenoside R1, ginsenoside Rg1 and ginsenoside Rb1 refer to the extraction method in the 2020 edition of the "Pharmacopoeia of the People's Republic of China", and change the heating reflux extraction method to the ultrasonic extraction method. Preparation of the test solution: Take about 1g of the powder of this product, accurately weigh it, accurately add 10mL of ethanol, and ultrasonically extract for 0.5h. Preparation of the reference solution: Take an appropriate amount of notoginsenoside R1, ginsenoside Rg1 and ginsenoside Rb1 reference substances, accurately weigh them, and add methanol to make a mixed solution containing 0.4mg of each per 1mL. Chromatographic conditions: Chromatographic column: COSMOSIL Cholester (4.6IDx10mm); Detection wavelength: 203nm; Mobile phase: acetonitrile-water; Determination method: Accurately aspirate 20μL of reference solution and test solution respectively, inject into high performance liquid chromatography, and determine the content of saponins in cotyledon embryo, Panax notoginseng saponin R1 is 0.23mg / g, ginsenoside Rg1 is 0.36mg / g, ginsenoside Re is 0.39mg / g, ginsenoside Rb1 is 0.48mg / g, and total saponins are 1.46mg / g.

[0060] Saponin content in callus and cotyledon embryos

[0061]

[0062] As can be seen from the above data, the cotyledon embryos obtained by the method of the present invention have shown significant advantages in terms of total saponin content, and their content far exceeds the total saponin content in the callus cultured in the traditional way. Specifically, the total saponin content in the cotyledon embryos cultivated by the method of the present invention has reached 7-8 times the total saponin content in the callus. Not only has the production efficiency of saponins been greatly improved, more saponin products can be obtained at a lower cost in a shorter time, and the quality of the products has also been greatly improved. Due to the significant improvement of the total saponin content in the cotyledon embryos, the resulting saponin products have higher purity and stronger activity, thereby being able to better meet the needs of various fields such as medicine, health products, and cosmetics for high-quality saponin raw materials.

[0063] Finally, it should be noted that the above preferred embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail through the above preferred embodiments, those skilled in the art should understand that various changes can be made in form and details without departing from the scope defined by the claims of the present invention.

Claims

1. A method for producing notoginsenosides and ginsenosides using Panax notoginseng cotyledon embryos, characterized in that: The specific steps include: S1, using immature zygotic embryos of Panax notoginseng to induce embryonic callus; S2, proliferating embryonic callus of Panax notoginseng on plate culture medium; S3, placing the embryonic callus of Panax notoginseng obtained in step S2 into a liquid culture medium and performing cell dispersion, and sieving using a sieve with a 24-30 mesh aperture to obtain spherical embryos and heart-shaped embryos; S4, a liquid culture medium is placed in a mechanical stirring bioreactor, and the spherical embryos and the heart-shaped embryos are cultured in the mechanical stirring bioreactor to obtain cotyledon embryos after 4-5 weeks; S5, recovering the filtrate of step S3, filtering it with a sieve with an aperture of 30-80 or 80-150 meshes to obtain uniform embryonic cells of Panax notoginseng, placing the cells in a conical flask for differentiation, placing the conical flask on a shaker at a rotation speed of 100-120 rpm, and culturing in the dark to obtain spherical embryos and heart-shaped embryos, which are used in step S4; S6. Dry the cotyledon embryos obtained in step S4 at 45° C. and grind the powder. Pass the powder through a No. 3 sieve to extract notoginsenoside and ginsenoside.

2. The method for producing notoginsenosides and ginsenosides by using cotyledon embryos of Panax notoginseng according to claim 1, characterized in that: The plate culture medium in step S2 is WPM culture medium with a pH of 5.8 and 30 g / L sucrose, 2 mg / L 2,4-D, 1 g / L CH, 1 g / L PVP, 1 g / L activated carbon and 3 g / L Phytagel added.

3. The method for producing notoginsenosides and ginsenosides by using cotyledon embryos of Panax notoginseng according to claim 1, characterized in that: The embryonic callus tissue described in step S3 was placed in a conical flask containing 30-50 mL WPM culture solution supplemented with 30 g / L sucrose, and a magnetic stirring rotor was placed in the flask to stir for 10 minutes to disperse the cells.

4. The method for producing notoginsenosides and ginsenosides by using cotyledon embryos of Panax notoginseng according to claim 1, characterized in that: The mechanically stirred bioreactor in step S4 is a 7L mechanically stirred bioreactor.

5. The method for producing notoginsenosides and ginsenosides by using cotyledon embryos of Panax notoginseng according to claim 4, characterized in that: The input parameters of the mechanical stirring bioreactor described in step S4 are pH 5.6-5.7, temperature 23-25° C., rotation speed 150-250 rpm, and ventilation volume 1-3 L / min.

6. The method for producing notoginsenosides and ginsenosides by using cotyledon embryos of Panax notoginseng according to claim 1, characterized in that: The liquid culture medium described in step S4 is a WPM liquid culture medium supplemented with 50-70 g / L sucrose, 0.5-2 g / L glutamine, 2-4 g / L MES and a pH of 5.

8.

7. The method for producing notoginsenosides and ginsenosides by using cotyledon embryos of Panax notoginseng according to claim 1, characterized in that: In step S5, the conical flask is filled with 30-100 mL of liquid culture medium.

8. The method for producing notoginsenosides and ginsenosides by using cotyledon embryos of Panax notoginseng according to claim 7, characterized in that: The liquid culture medium is a WPM liquid culture medium supplemented with 50-80 g / L sucrose / maltose, 4-16.6 g / L MES, and 0.5-1 g / L glutamine.

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