A culture medium for growing ginger seedlings and a method for growing ginger

Abstract

The present invention belongs to the field of tissue culture technology and specifically relates to a culture medium for growing ginger seedlings and a method for growing ginger. Each 1000 mL of the culture medium contains: 20-25 g sucrose, 10-13 g agar, 0.16-0.19 g Hericium erinaceus fermentation product, 0.10-0.14 g plant fermentation product, 120-150 mg inositol, 580-600 mg KNO3, 130-150 mg NH4H2PO4, 240-270 mg MgSO4·7H2O, 20-24 mg 6-BA, 3-6 mg thiamine hydrochloride, 150-180 mg CaCl2, etc. The culture medium of the present invention can improve the survival rate of ginger seedlings, significantly increase the proliferation coefficient, and have a high rooting rate.

Description

Technical Field

[0001] The invention belongs to the technical field of plant tissue culture, and particularly relates to a culture medium for growing ginger seedlings and a method for culturing ginger. Background Art

[0002] Since ginger rarely reproduces by seed, relying primarily on asexual reproduction, this method, while maintaining the variety's desirable characteristics, also presents several challenges. Long-term asexual reproduction can easily lead to the accumulation and spread of viruses, bacteria, and other pathogens. These issues not only lead to a significant decrease in ginger yield, but also deteriorate in quality and weaken its disease resistance, impacting the economic value of ginger and the sustainability of agricultural production.

[0003] Tissue culture of ginger is a modern biotechnology method used to rapidly propagate high-quality ginger seedlings. This method not only effectively avoids diseases that may be carried by traditional propagation methods, but also allows for the mass production of genetically consistent plants in a relatively short period of time. Although ginger shoot tip tissue culture technology has obvious advantages in terms of rapid propagation and virus removal, it also faces some challenges and technical problems in actual operation: (1) Contamination problem: Incomplete disinfection of explants during tissue culture may lead to the invasion of contaminants such as bacteria and fungi, affecting the success rate of culture. (2) Difficulty in callus formation: Not all explants can be successfully induced to form callus, which may be related to factors such as the selection of explants, culture medium formulation, and the type and concentration of plant growth regulators. (3) Low differentiation rate: Even if callus is formed, its efficiency in differentiating into buds or roots may not be high, affecting subsequent plant regeneration. (4) Variation phenomenon: Changes in chromosome number or gene mutations may occur during tissue culture, resulting in trait variation in regenerated plants. This not only affects the quality of ginger, but also may reduce its commercial value. (5) Difficulty in large-scale production: Although regenerated plants can be successfully obtained under laboratory conditions, how to achieve large-scale production and commercial application remains a challenge. It is necessary to consider how to improve production efficiency, reduce production costs, and ensure the quality of plants.

[0004] Optimizing culture medium formulations to improve the success rate, differentiation rate, and plant quality of ginger stem tip tissue culture is a common method. MS culture medium is widely used for tissue culture of various plants, and its effectiveness can be further optimized by adding vitamins, organic acids, and other ingredients. However, existing ginger seedling culture media cannot meet the exogenous substance requirements of ginger at different growth stages, resulting in the optimized formula being unable to meet the needs of the entire growth cycle.

[0005] Therefore, there is an urgent need for a culture medium for growing ginger seedlings and a method for growing ginger. Summary of the Invention

[0006] The present invention aims to provide a culture medium for growing ginger seedlings and a method for growing ginger.

[0007] In order to achieve the above object, the present invention provides the following technical solutions:

[0008] A culture medium for growing ginger seedlings, comprising: per 1000 mL of the culture medium: 20-25 g sucrose, 10-13 g agar, 0.16-0.19 g Hericium erinaceus fermentation product, 0.10-0.14 g plant fermentation product, 120-150 mg inositol, 580-600 mg KNO3, 130-150 mg NH4H2PO4, 240-270 mg MgSO4·7H2O, 20-24 mg 6-BA, 3-6 mg thiamine hydrochloride, 150-180 mg CaCl2, 15-20 mg MnSO4·H2O, 0.07-0.12 mg CuSO4·5H2O, 0.2-0.5 mg 6-benzylaminopurine, and 0.2-0.6 mg indolebutyric acid; and a pH value of 5.7-6.0.

[0009] The preparation method of the Hericium erinaceus fermentation product comprises the following steps:

[0010] (1) Inoculate Hericium erinaceus strains onto a slant culture medium, place in a constant temperature incubator, and culture at 25-30°C for 8-10 days. 2 The mycelium is strong, white, and covers the entire slope, which means the activated mycelium is obtained;

[0011] (2) inoculating the activated bacterial strain into the seed culture medium, culturing at a speed of 150-170 r / min and 25-30° C. for 6-8 days to obtain a seed liquid;

[0012] (3) pouring the seed liquid into a fermentation medium for fermentation culture to obtain a fermentation material; crushing the fermentation material using a high-speed tissue crusher at a speed of 9000-10000 rpm for 10-15 minutes to homogenize and obtain a fermentation raw material;

[0013] (4) adjusting the pH of the fermentation raw material to 5-6 using 1 mol / L food-grade hydrochloric acid solution, adding the composite bacteria, and fermenting at 35-38°C for 25-30 hours to obtain a fermentation product;

[0014] (5) The fermentation product was filtered using a 0.22 μm filter membrane, concentrated, and spray-dried to obtain the Hericium erinaceus fermentation product.

[0015] Furthermore, the fermentation conditions in step (3) are: aerobic fermentation, inoculation amount of seed liquid accounting for 8-10% of the mass of the fermentation medium, culture temperature of 25-30°C, rotation speed of 160-180r / min, and culture time of 6-8 days.

[0016] Furthermore, the composite bacteria include Lactobacillus brevis, Lactobacillus plantarum and Bifidobacterium animalis.

[0017] Furthermore, the dosage of Lactobacillus brevis is 10 8 -10 9 CFU / mL fermentation raw material; the dosage of Lactobacillus plantarum is 10 7 -10 8 CFU / mL fermentation raw material; the dosage of animal Bifidobacterium is 10 9 -10 10 CFU / mL fermentation raw material.

[0018] The culture medium currently used for ginger seedling cultivation on the market mainly adds nutrients and growth regulators to MS culture medium, and different components need to be added at different stages, which is cumbersome and difficult to implement. The culture medium of the present invention is compounded in a specific proportion by multiple components, which work synergistically. At the same time, by adding Hericium erinaceus fermentation products to the system, the rooting rate of ginger seedlings can be improved. The fermentation broth of Hericium erinaceus is aerobically fermented and then fermented with composite bacteria, which contains a variety of plant hormones that can stimulate the formation and growth of roots. It also contains a variety of bioactive substances, such as polysaccharides, polyphenols and amino acids, which can promote cell division and differentiation and enhance the growth vitality of plants.

[0019] Furthermore, the method for preparing the plant fermentation product comprises the following steps:

[0020] (1) Mixing dried Rhodymenia palmata, astragalus, and green tea, and crushing the mixture to a size of less than 200 mesh to obtain a mixture;

[0021] (2) mixing a mixture and water in a weight ratio of 1:(8-10) to obtain a fermentation mixture; adding Lactobacillus casei to the fermentation mixture, performing anaerobic fermentation at a fermentation temperature of 35-38°C and a fermentation time of 16-20 hours, sterilizing, continuing to add Bifidobacterium bifidum, performing anaerobic fermentation at a fermentation temperature of 35-38°C and a fermentation time of 12-15 hours, sterilizing, and finally adding Bifidobacterium longum, performing anaerobic fermentation at a fermentation temperature of 35-38°C and a fermentation time of 7-10 hours, sterilizing after the fermentation is completed, and obtaining a fermentation mixture product;

[0022] (3) The fermentation mixture was filtered using a 0.22 μm filter membrane, and the filtrate was subjected to reduced pressure rotary evaporation and spray drying to obtain the plant fermentation product.

[0023] Furthermore, the weight ratio of palmate dulse, astragalus and green tea is 1:(1.2-1.4):(0.5-0.7).

[0024] Further, the amount of Lactobacillus casei is 10 8 -10 9 CFU / mL fermentation mixture; the amount of Bifidobacterium bifidum is 10 9 -10 10 CFU / mL fermentation mixture; the dosage of Bifidobacterium longum is 10 9 -10 10 CFU / mL fermentation mix.

[0025] Contamination and the appearance of abnormal strains are common problems in plant tissue culture, which can seriously affect the success rate of culture and the quality of the plants. Since ginger explants carry a large amount of bacteria, sterilization is relatively difficult. In the prior art, multiple sterilizations using ethanol can affect the rooting rate of ginger. The present invention reduces the number of contaminated explants by adding plant fermentation products to the culture medium and performing only one ethanol sterilization. Because the mixed fermentation product of palmate red algae, astragalus and green tea is added to the culture medium, the active components in the fermentation product can effectively inhibit the growth of bacteria and fungi through various mechanisms such as antibacterial, antifungal, antioxidant and immune enhancement, thereby improving the success rate of culture and the quality of the plants. At the same time, under these conditions, abnormal strains are reduced.

[0026] The present invention also provides a method for culturing ginger using the culture medium for ginger seedling cultivation prepared according to claims 1 to 9, comprising the following steps:

[0027] (1) Taking a ginger stem tip with a length of 0.5-0.7 cm, soaking it in 70% ethanol by volume for 30-40 seconds, then washing it with sterile water, and heat-treating it at 50°C for 5-8 minutes to obtain a pretreated ginger stem tip;

[0028] (2) The pretreated ginger stem tip is inoculated into a culture medium for ginger seedling cultivation for primary culture; the culture is carried out in a dark room at 27-29° C. and a relative humidity of 60-70% until white and green callus tissue and sprouts grow. When the sprouts grow to 4-5 cm, the upper leaves and leaf sheaths of the sprouts are cut off and the sprouts are placed back into the culture medium, maintaining the same temperature and humidity. The ginger sprouts are first cultured under a light intensity of 1700-1900 Lux for 24 hours and then cultured in a lightless environment for 24 hours in an alternating culture mode to obtain proliferating rooted ginger seedlings.

[0029] Compared with the prior art, the advantages and beneficial effects of the present invention are:

[0030] 1. The culture medium of the present invention is compounded in a specific proportion by multiple components to work synergistically. At the same time, by adding Hericium erinaceus fermentation products into the system, the rooting rate of ginger seedlings can be improved.

[0031] 2. The present invention reduces the number of contaminated explants by adding plant fermentation products to the culture medium and performing ethanol sterilization once. Furthermore, under this condition, abnormal strains are reduced. DETAILED DESCRIPTION

[0032] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0033] The raw materials used in the following examples of the present invention are all commercially available commodities:

[0034] Lactobacillus brevis, number: SHBCCD14346, was purchased from Shanghai Microbiological Collection Center.

[0035] Lactobacillus plantarum, number: CCTCCAB2010210, was purchased from China Center for Type Culture Collection.

[0036] Bifidobacterium animalis, accession number: SHBCCD24407AS1.1852, was purchased from China Center for Type Culture Collection.

[0037] Lactobacillus casei, number SHBCCD24737, was purchased from Shanghai Microbiological Collection Center.

[0038] Bifidobacterium bifidum, accession number: SHBCCD24408ATCC35914, was purchased from Shanghai Collection of Microorganisms.

[0039] Bifidobacterium longum was purchased from Shanghai Collection of Microorganisms, with the number SHBCCD24310.

[0040] Slant culture medium was purchased from Shanghai Yuanye Biotechnology Co., Ltd., product number: R22702 slant culture medium (containing agar).

[0041] The fermentation medium includes the following components: 9 g / L glucose, 4 g / L peptone, 3 g / L ammonium sulfate, 2 g / L magnesium sulfate heptahydrate, 0.7 g / L potassium dihydrogen phosphate, 0.6 g / L dipotassium hydrogen phosphate, 1.3 g / L sodium chloride and 2 g / L sodium selenite, pH = 7.1.

[0042] Hericium erinaceus strain, Beijing Huayan Shijia Quality Inspection Technology Co., Ltd., product number CICC50081.

[0043] Seed culture medium, Shandong Top Bioengineering Co., Ltd., product number MD032.

[0044] Example 1

[0045] This embodiment provides a culture medium for growing ginger seedlings, wherein each 1000 mL of the culture medium contains: 22 g sucrose, 11 g agar, 0.17 g Hericium erinaceus fermentation product, 0.13 g plant fermentation product, 140 mg inositol, 590 mg KNO3, 140 mg NH4H2PO4, 260 mg MgSO4·7H2O, 22 mg 6-BA, 4 mg thiamine hydrochloride, 160 mg CaCl2, 17 mg MnSO4·H2O, 0.1 mg CuSO4·5H2O, 0.3 mg 6-benzylaminopurine, and 0.5 mg indolebutyric acid, with a pH of 5.8;

[0046] The preparation method of the Hericium erinaceus fermentation product comprises the following steps:

[0047] (1) Inoculate Hericium erinaceus strains onto a slant culture medium, place it in a constant temperature incubator, and culture it at 27°C for 9 days. 2 The mycelium is strong, white, and covers the entire slope, which means the activated mycelium is obtained;

[0048] (2) inoculating the activated bacterial strain into the seed culture medium, culturing at a speed of 160 r / min and 27°C for 7 days to obtain the seed liquid;

[0049] (3) pouring the seed liquid into the fermentation medium for fermentation culture to obtain a fermentation material; crushing the fermentation material with a high-speed tissue crusher at a speed of 9500 rpm for 12 minutes, and homogenizing to obtain a fermentation raw material; the fermentation conditions are: aerobic fermentation, an inoculation amount of the seed liquid accounting for 9% of the mass of the fermentation medium, a culture temperature of 27°C, a rotation speed of 170 r / min, and a culture time of 7 days.

[0050] (4) The fermentation raw material was adjusted to pH 5.5 using 1 mol / L food-grade hydrochloric acid solution, the composite bacteria were added, and the fermentation was carried out at 36°C for 27 h to obtain a fermentation product;

[0051] (5) The fermentation product was filtered using a 0.22 μm filter membrane, concentrated, and spray-dried to obtain the Hericium erinaceus fermentation product.

[0052] The compound bacteria include Lactobacillus brevis, Lactobacillus plantarum and Bifidobacterium animalis. The dosage of Lactobacillus brevis is 10 9 CFU / mL fermentation raw material; the dosage of Lactobacillus plantarum is 10 7 CFU / mL fermentation raw material; the dosage of animal Bifidobacterium is 10 10 CFU / mL fermentation raw material.

[0053] The method for preparing the plant fermentation product comprises the following steps:

[0054] (1) Mixing dried dulse, astragalus and green tea in a weight ratio of 1:1.3:0.6, and crushing the mixture to less than 200 meshes to obtain a mixture;

[0055] (2) mixing the mixture and water in a weight ratio of 1:9 to obtain a fermentation mixture; adding Lactobacillus casei to the fermentation mixture, performing anaerobic fermentation at a fermentation temperature of 37° C. and a fermentation time of 18 h, sterilizing, continuing to add Bifidobacterium bifidum, performing anaerobic fermentation at a fermentation temperature of 37° C. and a fermentation time of 13 h, sterilizing, and finally adding Bifidobacterium longum, performing anaerobic fermentation at a fermentation temperature of 37° C. and a fermentation time of 8 h, sterilizing after the fermentation is completed, and obtaining a fermentation mixture product;

[0056] (3) The fermentation mixture was filtered using a 0.22 μm filter membrane, and the filtrate was subjected to reduced pressure rotary evaporation and spray drying to obtain the plant fermentation product.

[0057] The dosage of Lactobacillus casei is 10 9 CFU / mL fermentation mixture; the amount of Bifidobacterium bifidum is 10 9 CFU / mL fermentation mixture; the dosage of Bifidobacterium longum is 10 10 CFU / mL fermentation mix.

[0058] The method for cultivating ginger using a culture medium for ginger seedling cultivation comprises the following steps:

[0059] (1) A ginger stem tip with a length of 0.6 cm was taken, soaked in 70% ethanol by volume for 30 seconds, then washed with sterile water, and heat-treated at 50°C for 7 minutes to obtain a pretreated ginger stem tip;

[0060] (2) The pretreated ginger stem tip is inoculated into a culture medium for ginger seedling cultivation for primary culture; the culture is carried out in a dark room at 28°C and a relative humidity of 65% until white and green callus tissue and sprouts grow. When the sprouts grow to 4.5 cm, the upper leaves and leaf sheaths of the sprouts are cut off and the sprouts are placed back into the culture medium, maintaining the same temperature and humidity. The sprouts are first cultured under a light intensity of 1800 Lux for 24 hours and then cultured in a lightless environment for 24 hours in an alternating culture mode to obtain proliferating rooted ginger seedlings.

[0061] Example 2

[0062] This embodiment provides a culture medium for growing ginger seedlings, wherein each 1000 mL of the culture medium contains: 25 g sucrose, 10 g agar, 0.19 g Hericium erinaceus fermentation product, 0.10 g plant fermentation product, 150 mg inositol, 580 mg KNO3, 150 mg NH4H2PO4, 270 mg MgSO4·7H2O, 20 mg 6-BA, 6 mg thiamine hydrochloride, 150 mg CaCl2, 20 mg MnSO4·H2O, 0.07 mg CuSO4·5H2O, 0.5 mg 6-benzylaminopurine, and 0.6 mg indolebutyric acid, with a pH of 5.9;

[0063] The preparation method of the Hericium erinaceus fermentation product comprises the following steps:

[0064] (1) Inoculate Hericium erinaceus strains onto a slant culture medium, place it in a constant temperature incubator, and culture it at 30°C for 8 days. 2 The mycelium is strong, white, and covers the entire slope, which means the activated mycelium is obtained;

[0065] (2) inoculating the activated bacterial strain into the seed culture medium, culturing at a speed of 170 r / min and 30°C for 6 days to obtain the seed liquid;

[0066] (3) pouring the seed liquid into the fermentation medium for fermentation culture to obtain a fermentation material; crushing the fermentation material with a high-speed tissue crusher at a speed of 10,000 rpm for 15 minutes, and homogenizing to obtain a fermentation raw material; the fermentation conditions are: aerobic fermentation, an inoculation amount of 10% of the mass of the seed liquid in the fermentation medium, a culture temperature of 25°C, a rotation speed of 180 r / min, and a culture time of 6 days.

[0067] (4) The fermentation raw material was adjusted to pH 6 using 1 mol / L food-grade hydrochloric acid solution, the composite bacteria were added, and the mixture was fermented at 35°C for 30 h to obtain a fermentation product;

[0068] (5) The fermentation product was filtered using a 0.22 μm filter membrane, concentrated, and spray-dried to obtain the Hericium erinaceus fermentation product.

[0069] The compound bacteria include Lactobacillus brevis, Lactobacillus plantarum and Bifidobacterium animalis. The dosage of Lactobacillus brevis is 10 8 CFU / mL fermentation raw material; the dosage of Lactobacillus plantarum is 10 7 CFU / mL fermentation raw material; the dosage of animal Bifidobacterium is 10 9 CFU / mL fermentation raw material.

[0070] The method for preparing the plant fermentation product comprises the following steps:

[0071] (1) Mixing dried dulse, astragalus and green tea in a weight ratio of 1:1.2:0.7, and crushing the mixture to less than 200 meshes to obtain a mixture;

[0072] (2) mixing the mixture and water in a weight ratio of 1:10 to obtain a fermentation mixture; adding Lactobacillus casei to the fermentation mixture, performing anaerobic fermentation at a fermentation temperature of 38° C. and a fermentation time of 16 h, sterilizing, continuing to add Bifidobacterium bifidum, performing anaerobic fermentation at a fermentation temperature of 38° C. and a fermentation time of 15 h, sterilizing, and finally adding Bifidobacterium longum, performing anaerobic fermentation at a fermentation temperature of 38° C. and a fermentation time of 7 h, sterilizing after the fermentation is completed, and obtaining a fermentation mixture product;

[0073] (3) The fermentation mixture was filtered using a 0.22 μm filter membrane, and the filtrate was subjected to reduced pressure rotary evaporation and spray drying to obtain the plant fermentation product.

[0074] The dosage of Lactobacillus casei is 10 9 CFU / mL fermentation mixture; the amount of Bifidobacterium bifidum is 10 10 CFU / mL fermentation mixture; the dosage of Bifidobacterium longum is 10 10 CFU / mL fermentation mix.

[0075] The method for cultivating ginger using a culture medium for ginger seedling cultivation comprises the following steps:

[0076] (1) A ginger stem tip with a length of 0.5 cm was taken, soaked in 70% ethanol by volume for 40 seconds, then washed with sterile water, and heat-treated at 50°C for 8 minutes to obtain a pretreated ginger stem tip;

[0077] (2) The pretreated ginger stem tip is inoculated into a culture medium for ginger seedling cultivation for primary culture; the culture is carried out in a dark room at 29°C and a relative humidity of 70% until white and green callus tissue and sprouts grow. When the sprouts grow to 5 cm, the upper leaves and leaf sheaths of the sprouts are cut off and the sprouts are placed back into the culture medium, maintaining the same temperature and humidity. The ginger sprouts are first cultured under a light intensity of 1700 Lux for 24 hours and then cultured in a lightless environment for 24 hours in an alternating culture mode to obtain proliferating rooted ginger seedlings.

[0078] Comparative Example 1

[0079] The difference between this comparative example and Example 1 is: a culture medium for growing ginger seedlings, wherein each 1000 mL of the culture medium contains: 15 g sucrose, 18 g agar, 0.13 g Hericium erinaceus fermentation product, 0.17 g plant fermentation product, 240 mg inositol, 490 mg KNO3, 240 mg NH4H2PO4, 160 mg MgSO4·7H2O, 12 mg 6-BA, 14 mg thiamine hydrochloride, 60 mg CaCl2, 117 mg MnSO4·H2O, 0.3 mg CuSO4·5H2O, 0.1 mg 6-benzylaminopurine, 0.5 mg indolebutyric acid, and a pH of 6.0.

[0080] Comparative Example 2

[0081] The difference between this comparative example and Example 1 is that the composite bacteria include Lactobacillus casei, Bifidobacterium bifidum and Bifidobacterium longum. The dosage of Lactobacillus casei is 10 9 CFU / mL fermentation raw material; the dosage of Bifidobacterium bifidum is 10 7 CFU / mL fermentation raw material; the dosage of Bifidobacterium longum is 10 10 CFU / mL fermentation raw material.

[0082] Comparative Example 3

[0083] The difference between this comparative example and Example 1 is that the preparation method of the Hericium erinaceus fermentation product comprises the following steps:

[0084] (1) Inoculate Hericium erinaceus strains onto a slant culture medium, place it in a constant temperature incubator, and culture it at 27°C for 9 days. 2 The mycelium is strong, white, and covers the entire slope, which means the activated mycelium is obtained;

[0085] (2) inoculating the activated bacterial strain into the seed culture medium, culturing at a speed of 160 r / min and 27°C for 7 days to obtain the seed liquid;

[0086] (3) pouring the seed liquid into the fermentation medium for fermentation culture to obtain a fermentation material; crushing the fermentation material with a high-speed tissue crusher at a speed of 9500 rpm for 12 minutes, and homogenizing to obtain a fermentation raw material; the fermentation conditions are: aerobic fermentation, inoculation amount 9%, culture temperature 27°C, rotation speed 170 r / min, and culture time 7 days.

[0087] (4) The fermentation raw material was adjusted to pH 4.5 using 1 mol / L food-grade hydrochloric acid solution, 0.01% cellulase by weight of the fermentation raw material was added, and enzymatic hydrolysis was carried out at 65°C for 2 h. The pH was then adjusted to 7.0 using 1 mol / L food-grade sodium hydroxide solution, 0.01% protease by weight of the fermentation raw material was added, and enzymatic hydrolysis was carried out at 65°C for 3 h. The temperature was raised to 100°C and maintained for 15 min to obtain an enzymatic hydrolyzate;

[0088] (5) The enzymatic hydrolysate was filtered using a 0.22 μm filter membrane, concentrated, and spray-dried to obtain the Hericium erinaceus fermentation product.

[0089] Comparative Example 4

[0090] The difference between this comparative example and Example 1 is that the dry dulse, astragalus and green tea in a weight ratio of 1:1:1 are mixed and then crushed to less than 200 meshes to obtain a mixture.

[0091] Comparative Example 5

[0092] The difference between this comparative example and Example 1 is that the method for preparing the plant fermentation product comprises the following steps:

[0093] (1) Mixing dried dulse, astragalus and green tea in a weight ratio of 1:1.3:0.6, and crushing the mixture to less than 200 meshes to obtain a mixture;

[0094] (2) mixing a mixture and water in a weight ratio of 1:9 to obtain a fermentation mixture; adding Lactobacillus brevis to the fermentation mixture, performing anaerobic fermentation at a fermentation temperature of 37° C. and a fermentation time of 18 h, sterilizing, continuing to add Lactobacillus plantarum, performing anaerobic fermentation at a fermentation temperature of 37° C. and a fermentation time of 13 h, sterilizing, and finally adding Bifidobacterium animalis, performing anaerobic fermentation at a fermentation temperature of 37° C. and a fermentation time of 8 h, sterilizing after the fermentation is completed, and obtaining a fermentation mixture product;

[0095] (3) The fermentation mixture was filtered using a 0.22 μm filter membrane, and the filtrate was subjected to reduced pressure rotary evaporation and spray drying to obtain the plant fermentation product.

[0096] The dosage of Lactobacillus brevis is 10 9 CFU / mL fermentation mixture; the amount of Lactobacillus plantarum is 10 9 CFU / mL fermentation mixture; the dosage of animal Bifidobacterium is 10 10 CFU / mL fermentation mix.

[0097] Comparative Example 6

[0098] The difference between this comparative example and Example 1 is that the method for preparing the plant fermentation product comprises the following steps:

[0099] (1) Mixing dried dulse, astragalus and green tea in a weight ratio of 1:1.2:0.7, and crushing the mixture to less than 200 meshes to obtain a mixture;

[0100] (2) mixing the mixture and water in a weight ratio of 1:10 to obtain a fermentation mixture; adding mixed bacteria to the fermentation mixture, the mixed bacteria including Lactobacillus casei, Bifidobacterium bifidum and Bifidobacterium longum, and performing anaerobic fermentation at a fermentation temperature of 38° C. and a fermentation time of 38 h; sterilizing after the fermentation is completed to obtain a fermentation mixture product;

[0101] (3) The fermentation mixture was filtered using a 0.22 μm filter membrane, and the filtrate was subjected to reduced pressure rotary evaporation and spray drying to obtain the plant fermentation product.

[0102] The dosage of Lactobacillus casei is 10 9 CFU / mL fermentation mixture; the amount of Bifidobacterium bifidum is 10 10 CFU / mL fermentation mixture; the dosage of Bifidobacterium longum is 10 10 CFU / mL fermentation mix.

[0103] Performance Testing

[0104] Cultivation was performed using the methods of Examples 1-2 and Comparative Examples 1-6, with 100 seedlings inoculated. During the tissue culture process, observations were made every three days from the start of inoculation for 25 days. The number of contaminated explants, the number of abnormal plants, the proliferation coefficient, and the rooting rate were observed and calculated over the 25 days. The proliferation coefficient = (number of effective seedlings generated / number of seedlings inoculated), which indicates the ability of the inoculated tissue to differentiate into multiple seedlings during the planting process; a larger number indicates better results. The rooting rate (%) = (number of rooted seedlings / number of seedlings inoculated) × 100%.

[0105] The results are shown in Table 1.

[0106] Table 1 Test results

[0107]

[0108] From the above performance test results, it can be seen that the culture media of Examples 1-2 are excellent in culturing ginger, especially the comprehensive performance of Example 1 is the most outstanding, which is mainly due to the synergistic effect of the culture media components.

[0109] However, the comparative examples did not adopt the necessary technical solutions, resulting in significantly worse performance tests than the examples. In comparative example 1, the ratio of the culture medium components was changed, and it can be seen that the ginger seedling cultivation effect was poor. In comparative example 2, the strain of the composite bacteria was changed, and enzymatic hydrolysis was used in comparative example 3. The results show that the rooting rate decreased. In comparative example 4, the ratio of palmate red algae, astragalus, and green tea was changed, the strain was changed in comparative example 5, and mixed fermentation was used in comparative example 6. All of these led to an increase in the number of contaminated explants and abnormal strains to varying degrees. The above experimental results further demonstrate the importance of the technical solution defined in the present invention for its technical effects.

[0110] The above is a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A culture medium for growing ginger seedlings, characterized in that: The components of each 1000 mL culture medium are: 22 g sucrose, 11 g agar, 0.17 g Hericium erinaceus fermentation product, 0.13 g plant fermentation product, 140 mg inositol, 590 mg KNO3, 140 mg NH4H2PO4, 260 mg MgSO4·7H2O, 22.3 mg 6-BA, 4 mg thiamine hydrochloride, 160 mg CaCl2, 17 mg MnSO4·H2O, 0.1 mg CuSO4·5H2O, 0.5 mg indolebutyric acid, pH 5.8; The preparation method of Hericium erinaceus fermentation product comprises: (1) inoculating Hericium erinaceus strains onto a slant culture medium, placing the culture medium in a constant temperature incubator, culturing at 27°C for 9 days, and picking 1 cm 2 The mycelium is strong, white and covers the slope, and the activated strain is obtained; (2) the activated strain is inoculated into the seed culture medium, the speed is 160r / min, and the culture is carried out at 27℃ for 7 days to obtain the seed liquid; (3) the seed liquid is poured into the fermentation culture medium, fermented and cultured to obtain the fermentation material; the fermentation material is crushed by a high-speed tissue crusher, the speed is 9500rpm, the time is 12min, and the slurry is obtained to obtain the fermentation raw material; the fermentation conditions are: aerobic fermentation, the inoculation amount is 9% of the mass of the seed liquid in the fermentation culture medium, the culture temperature is 27℃, the speed is 170r / min, and the culture time is 7 days; (4) the fermentation raw material is adjusted to pH 5.5 with 1mol / L food-grade hydrochloric acid solution, the composite bacteria are added, and the fermentation is carried out at 36℃ for 27h to obtain the fermentation product; (5) the fermentation product is filtered with a 0.22μm filter membrane, concentrated, and spray-dried to obtain the Hericium erinaceus fermentation product; the composite bacteria are Lactobacillus brevis, Lactobacillus plantarum and Bifidobacterium animalis; the amount of Lactobacillus brevis is 10 9 CFU / mL fermentation raw material; the dosage of Lactobacillus plantarum is 10 7 CFU / mL fermentation raw material; the dosage of animal Bifidobacterium is 10 10 CFU / mL fermentation raw material; The preparation method of plant fermentation products includes: (1) mixing dry red algae, astragalus and green tea in a weight ratio of 1:1.3:0.6, crushing to less than 200 mesh, and obtaining a mixture; (2) mixing the mixture in a weight ratio of 1:9 with water to obtain a fermentation mixture; adding Lactobacillus casei to the fermentation mixture, performing anaerobic fermentation at a fermentation temperature of 37°C and a fermentation time of 18 hours, sterilizing, continuing to add Bifidobacterium bifidum, performing anaerobic fermentation at a fermentation temperature of 37°C and a fermentation time of 13 hours, sterilizing, and finally adding Bifidobacterium longum, performing anaerobic fermentation at a fermentation temperature of 37°C and a fermentation time of 8 hours, sterilizing after the fermentation is completed, and obtaining a fermentation mixture product; (3) filtering the fermentation mixture product using a 0.22 μm filter membrane, performing reduced pressure rotary evaporation on the filtrate, and spray drying to obtain a plant fermentation product; the amount of Lactobacillus casei is 10 9 CFU / mL fermentation mixture; the amount of Bifidobacterium bifidum is 10 9 CFU / mL fermentation mixture; the dosage of Bifidobacterium longum is 10 10 CFU / mL fermentation mix.

2. The method for culturing ginger using the culture medium according to claim 1, characterized in that: The following steps are involved: (1) Take a ginger stem tip with a length of 0.6 cm, soak it in 70% ethanol by volume for 30 seconds, then wash it with sterile water, and heat-treat it at 50°C for 7 minutes to obtain the pretreated ginger stem tip; (2) Inoculate the pretreated ginger stem tip into the culture medium for ginger seedling cultivation for primary culture; culture it in a dark room at 28°C and a relative humidity of 65% until white and green callus tissue and sprouts grow. When the sprouts grow to 4.5 cm, cut off the upper leaves and leaf sheaths of the sprouts, and place them back in the culture medium, maintain the same temperature and humidity, and culture them under a light intensity of 1800 Lux for 24 hours, and then culture them in a lightless environment for 24 hours in an alternating culture method to obtain ginger proliferating rooting seedlings.

3. A culture medium for growing ginger seedlings, characterized in that: The components of each 1000 mL culture medium are: 25 g sucrose, 10 g agar, 0.19 g Hericium erinaceus fermentation product, 0.10 g plant fermentation product, 150 mg inositol, 580 mg KNO3, 150 mg NH4H2PO4, 270 mg MgSO4·7H2O, 20.5 mg 6-BA, 6 mg thiamine hydrochloride, 150 mg CaCl2, 20 mg MnSO4·H2O, 0.07 mg CuSO4·5H2O, 0.6 mg indolebutyric acid, pH 5.9; The preparation method of Hericium erinaceus fermentation product comprises: (1) inoculating Hericium erinaceus strains onto a slant culture medium, placing the culture medium in a constant temperature incubator, culturing at 30°C for 8 days, and picking 1 cm 2 The mycelium is strong, white and covers the slope, and the activated strain is obtained; (2) the activated strain is inoculated into the seed culture medium, the speed is 170r / min, and the culture is carried out at 30℃ for 6 days to obtain seed liquid; (3) the seed liquid is poured into the fermentation culture medium for fermentation culture to obtain fermentation material; the fermentation material is crushed by a high-speed tissue crusher at a speed of 10000rpm for 15min, and homogenized to obtain fermentation raw material; the fermentation conditions are: aerobic fermentation, the inoculation amount is 10% of the mass of the seed liquid in the fermentation culture medium, the culture temperature is 25℃, the speed is 180r / min, and the culture time is 6 days; (4) the fermentation raw material is adjusted to pH 6 using 1mol / L food-grade hydrochloric acid solution, the composite bacteria are added, and the fermentation is carried out at 35℃ for 30h to obtain the fermentation product; (5) the fermentation product is filtered using a 0.22μm filter membrane, concentrated, and spray-dried to obtain Hericium erinaceus fermentation product; the composite bacteria are Lactobacillus brevis, Lactobacillus plantarum and Bifidobacterium animalis; the amount of Lactobacillus brevis is 10 8 CFU / mL fermentation raw material; the dosage of Lactobacillus plantarum is 10 7 CFU / mL fermentation raw material; the dosage of animal Bifidobacterium is 10 9 CFU / mL fermentation raw material; The preparation method of plant fermentation products includes: (1) mixing dry dulse, astragalus and green tea in a weight ratio of 1:1.2:0.7, crushing to less than 200 mesh, and obtaining a mixture; (2) mixing the mixture in a weight ratio of 1:10 with water to obtain a fermentation mixture; adding Lactobacillus casei to the fermentation mixture, performing anaerobic fermentation at a fermentation temperature of 38°C and a fermentation time of 16 hours, sterilizing, continuing to add Bifidobacterium bifidum, performing anaerobic fermentation at a fermentation temperature of 38°C and a fermentation time of 15 hours, sterilizing, and finally adding Bifidobacterium longum, performing anaerobic fermentation at a fermentation temperature of 38°C and a fermentation time of 7 hours, sterilizing after the fermentation is completed, and obtaining a fermentation mixture product; (3) filtering the fermentation mixture product using a 0.22 μm filter membrane, performing reduced pressure rotary evaporation on the filtrate, and spray drying to obtain a plant fermentation product; the amount of Lactobacillus casei is 10 9 CFU / mL fermentation mixture; the amount of Bifidobacterium bifidum is 10 10 CFU / mL fermentation mixture; the dosage of Bifidobacterium longum is 10 10 CFU / mL fermentation mix.

4. The method for culturing ginger using the culture medium according to claim 3, characterized in that: The following steps are involved: (1) Take a ginger stem tip with a length of 0.5 cm, soak it in 70% ethanol by volume for 40 seconds, then wash it with sterile water, and heat-treat it at 50°C for 8 minutes to obtain the pretreated ginger stem tip; (2) Inoculate the pretreated ginger stem tip into the culture medium for ginger seedling cultivation for primary culture; culture it in a dark room at 29°C and a relative humidity of 70% until white and green callus tissue and sprouts grow. When the sprouts grow to 5 cm, cut off the upper leaves and leaf sheaths of the sprouts, and place them back in the culture medium, maintain the same temperature and humidity, and culture them under a light intensity of 1700 Lux for 24 hours, and then culture them in a lightless environment for 24 hours in an alternating culture method to obtain ginger proliferating rooting seedlings.

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