A method for preserving and detecting the vitality of Gastrodia elata seeds

Abstract

The present invention provides a method for preserving and testing the vitality of Gastrodia elata seeds, belonging to the field of biotechnology. This method involves vacuum freeze-drying Gastrodia elata capsules harvested on the day of harvest to a moisture content of 1.5-2.5%, and then refrigerating the dried capsules. The Gastrodia elata seeds preserved using this method have an effective storage period of up to two years. The present invention also provides a method for testing the vitality of Gastrodia elata seeds using a water agar medium, which can be used in conjunction with Gastrodia elata seed preservation methods, laying a solid foundation for the conservation and innovative utilization of Gastrodia elata germplasm resources.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a method for preserving and detecting the vitality of Gastrodia elata seeds. Background Art

[0002] Gastrodia elata (Gastrodia elata Bl), also known as red arrow grass, mother's vine, and heli, is a perennial heterotrophic herb in the Orchidaceae family. Gastrodia elata is a dried tuber of the Orchidaceae family. Its primary active ingredient is gastrodin. It also contains various functional ingredients, such as p-hydroxybenzyl alcohol, p-hydroxybenzaldehyde, and polysaccharides. It has been traditionally used to treat headaches, dizziness, epilepsy, and amnesia. As a food and medicine, it possesses high health and edible value and is used in a variety of culinary dishes, including famous dishes like Gastrodia elata stewed chicken. As a heterotrophic herb, Gastrodia elata has no roots or leaves, small seeds and no endosperm. It can only complete the process of flowering and fruiting through sexual reproduction. During sexual reproduction, it needs germination fungi to promote the germination of Gastrodia elata seeds. During asexual reproduction, the germinated rice hemp and white hemp seeds need to symbiotically grow with honey fungus to become arrow hemp. Arrow hemp then sprouts, blooms, and bears fruit until the seeds mature into Gastrodia elata. The entire life cycle lasts two and a half years across three years.

[0003] Gastrodia elata seeds are small and simple, with a relatively large surface area and strong hygroscopicity. The embryo accounts for a large proportion, which promotes respiration and is more specific than other seeds. They are difficult to maintain under normal conditions. Furthermore, mature Gastrodia elata capsules easily crack, causing the seeds to scatter, resulting in low viability and difficulty in preservation. Gastrodia elata's long life cycle, the tendency for agronomic traits to deteriorate under vegetative cultivation, and the difficulty in seed preservation restrict the preservation and innovative utilization of Gastrodia elata germplasm resources, significantly impacting the selection and breeding of superior Gastrodia elata varieties and the protection of new varieties. Therefore, there is an urgent need to explore preservation methods that are compatible with the characteristics of Gastrodia elata seeds. While existing methods for preserving Gastrodia elata seeds exist, their effective shelf life remains relatively short. For example, CN106900506A discloses low-temperature refrigeration at 0°C to 6°C for six months. Furthermore, in previous research on the effects of different treatments on the viability of Gastrodia elata seeds during storage, the inventors' team found that vacuum freeze-drying Gastrodia elata seeds followed by vitrification and embedding can effectively preserve them for one year. Therefore, in the protection and innovative utilization of Gastrodia elata germplasm resources, exploring effective medium- and long-term preservation methods for Gastrodia elata seeds remains one of the technical problems that need to be urgently solved in this field. Summary of the Invention

[0004] In view of this, the object of the present invention is to provide a method for preserving Gastrodia elata seeds, wherein the Gastrodia elata seeds preserved by the method can have a storage period of up to 2 years and the vitality of the Gastrodia elata seeds can reach 37.64%.

[0005] The present invention also provides a method for detecting the vitality of Gastrodia elata seeds using water agar culture medium, which can be matched with the Gastrodia elata seed preservation method, and lays a solid foundation for the protection and innovative utilization of Gastrodia elata germplasm resources.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The invention provides a method for preserving gastrodia elata seeds. The method comprises: performing vacuum freeze-drying treatment on picked gastrodia elata capsules until the water content is 1.5-2.5%, and refrigerating and preserving the vacuum freeze-dried gastrodia elata capsules.

[0008] Preferably, the Gastrodia elata capsule is a capsule weighing 180 g to 220 g that matures after artificial pollination of Gastrodia elata when it bolts and blooms.

[0009] Preferably, the Gastrodia elata capsules are Gastrodia elata capsules picked on the day.

[0010] Preferably, the dried Gastrodia elata capsules are packaged with dry absorbent paper, the outer layer is then wrapped with absorbent cotton and placed in a 50 mL centrifuge tube, and color-changing silica gel is added to the centrifuge tube.

[0011] Preferably, the refrigerated storage temperature is 3°C to 6°C.

[0012] The present invention also provides a method for detecting the vitality of Gastrodia elata seeds, which comprises: evenly shaking the Gastrodia elata seeds preserved by the above method into a plate of water agar culture medium with germination bacteria, dark culturing at 20°C to 25°C, regularly observing the germination of the Gastrodia elata seeds, distinguishing the germination of the Gastrodia elata seeds with the naked eye or counting the number of Gastrodia elata protocorms.

[0013] Preferably, the main components of the water agar culture medium are water and agar, and the amount of agar added to the water agar culture medium is 6 g / L to 8 g / L.

[0014] Preferably, the germination fungus is Dendrobium officinale.

[0015] Beneficial effects of the present invention:

[0016] (1) The present invention vacuum freeze-dries mature Gastrodia elata capsules to a moisture content of 1.5-2.5%, and then stores them at 3°C-6°C. The seeds have an effective storage period of up to 2 years, which is relatively long. Moreover, the vitality of the Gastrodia elata seeds after 2 years of storage can reach 37.64%.

[0017] (2) The preservation method of the present invention uses a preservation tool that is low-cost and simple, and the preservation tube is small in size, requiring less storage space and is easy to manage.

[0018] (3) The present invention establishes for the first time a Gastrodia elata seed preservation and its supporting vitality detection system, which can effectively detect the vitality of Gastrodia elata seeds, laying a solid foundation for the protection of Gastrodia elata germplasm resources. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 : Flowchart of Gastrodia elata seed preservation.

[0020] Figure 2 : Discoloration of color-changing silica gel during the storage of Gastrodia elata seeds: a, A represents good preservation; b, B represents poor preservation.

[0021] Figure 3 : Changes in moisture content of Gastrodia elata seeds after vacuum freeze-drying for different treatment times.

[0022] Figure 4 : Germination of Gastrodia elata seeds stored at 4℃ for two years after vacuum freeze-drying on water agar medium.

[0023] Figure 5 : Effect of 4℃ storage on the vitality of Gastrodia elata seeds with different moisture contents: a, b, c, d, e are the TTC staining results of vacuum freeze-dried Gastrodia elata seeds with moisture contents of 5.8%, 3.9%, 3.0%, 2.1%, and 1.6% after storage at 4℃ for two years; A, B, C, D, E are the harvest results of Gastrodia elata seeds with corresponding moisture contents after being cultivated with fungi for half a year. DETAILED DESCRIPTION

[0024] The present invention provides a method for preserving Gastrodia elata seeds, comprising: subjecting harvested Gastrodia elata capsules to a vacuum freeze-drying process until the moisture content is 1.5-2.5%, and then refrigerating the freeze-dried Gastrodia elata capsules. The moisture content is preferably 1.6-2.1%. The Gastrodia elata seeds are located within the Gastrodia elata capsules, and the Gastrodia elata seeds are effectively preserved by preserving the capsules.

[0025] The Gastrodia elata capsules of the present invention are capsules of 180g to 220g that mature after artificial pollination of Gastrodia elata and arrowhead. Preferably, 180g to 220g of Gastrodia elata and arrowhead are selected for planting, and artificial pollination is performed when the arrowhead bolts and blooms. After the Gastrodia elata capsules mature, the same batch of Gastrodia elata capsules are picked at the same time. More preferably, 180g to 220g of Gastrodia elata and arrowhead with clear provenance information, no pests and diseases, no damage, and uniform size are selected and planted with fine river sand. After the arrowhead bolts and blooms and artificial pollination is performed, the Gastrodia elata capsules are picked at the same time after they crack and mature when gently pinched by hand. The Gastrodia elata and arrowhead of the present invention include red Gastrodia elata, yellow Gastrodia elata, green Gastrodia elata, black Gastrodia elata, and loose Gastrodia elata. The water content of the river sand of the present invention is preferably 39% to 41%, and the thickness of the river sand covering is preferably 2.4cm to 2.6cm.

[0026] When the weight of Gastrodia elata is between 180g and 220g, the fresh weight of its fruit is high and the number of fruits is also large. In the present invention, the capsules of Gastrodia elata weighing 180g to 220g are selected for artificial pollination and matured after artificial pollination, which is more economical than the capsules of Gastrodia elata weighing other weights. More preferably, the Gastrodia elata of the present invention is red Gastrodia elata weighing 180g to 220g.

[0027] The Gastrodia elata capsules described in the present invention are capsules harvested on the day of harvest. After harvesting, as the Gastrodia elata capsules are stored for an increasing amount of time, the energy stored in the seeds in the capsules is released through respiration, and the seed organic matter content and seed vigor also change to a certain extent. The present invention utilizes capsules harvested on the day of harvest for vacuum freeze-drying and refrigerated storage, which is more conducive to maintaining seed vigor after long-term storage.

[0028] The gastrodia elata capsules after vacuum freeze-drying of the present invention are packaged with dry absorbent paper, the outer layer is then wrapped with absorbent cotton and placed in a 50mL centrifuge tube, and then color-changing silica gel is added to the centrifuge tube. Preferably, the amount of color-changing silica gel added of the present invention is 2 / 10 of the centrifuge tube. By adding color-changing silica gel, the present invention can not only maintain the dryness of the storage environment, but also judge the airtightness of the storage environment according to the fading of the color-changing silica gel, thereby intuitively judging whether the gastrodia elata seeds have been effectively preserved. If the color-changing silica gel does not change color, it proves that the preservation effect is good. If the color-changing silica gel changes color, it proves that the preservation effect is poor. The seeds may have absorbed moisture, which will have a certain impact on the vitality of the seeds later. It is possible to intervene in advance to ensure the seed preservation effect ( Figure 2 ).

[0029] The present invention refrigerates the centrifuge tube containing the vacuum freeze-dried gastrodia capsule at a temperature of 3° C. to 5° C. More preferably, the refrigerated storage temperature is 4° C.

[0030] The effective storage period of the Gastrodia elata seeds preserved by the method of the present invention can reach 2 years, and the vitality of the Gastrodia elata seeds after being preserved for 2 years can reach 37.64%.

[0031] The present invention also provides a method for detecting the vitality of Gastrodia elata seeds, comprising: evenly dropping the Gastrodia elata seeds preserved by the above method onto a plate containing a water agar culture medium containing germination bacteria, incubating in the dark at 20°C to 25°C, regularly observing the germination of the Gastrodia elata seeds, visually distinguishing germination of the Gastrodia elata seeds, or counting the number of Gastrodia elata protocorms. Preferably, during the periodic observation of Gastrodia elata seed germination, the germination of the Gastrodia elata seeds is observed under a stereomicroscope every five days. The dark incubation temperature of the present invention is more preferably 22°C.

[0032] As an optional embodiment, the present invention takes out the preserved Gastrodia elata capsules from a sterile clean bench, wipes the capsule surface with alcohol, quickly sterilizes it by fire, and places it in a sterile petri dish for later use, cuts open the Gastrodia elata capsule pods, collects the Gastrodia elata seeds and mixes them, evenly shakes the Gastrodia elata seeds into a petri dish containing a culture medium with germination bacteria, and cultures them in the dark at 20°C to 25°C.

[0033] The water agar medium of the present invention comprises water and agar, with the amount of agar added being 6 g / L to 8 g / L, preferably 7 g / L. As an optional embodiment, the present invention mixes 7 g of agar with 1000 mL of tap water, and sterilizes the mixture at natural pH at 121° C. for 30 min to obtain the water agar medium.

[0034] The germination fungus of the present invention is Dendrobium officinale. The cultivation method of the Dendrobium officinale comprises the following steps: transferring a Dendrobium officinale fungus seed of about 1 square centimeter the size of a fingernail under sterile conditions to a water agar culture medium, culturing in the dark at 25°C until mycelium grows to 2 / 3 of the entire plate, and obtaining a water agar culture medium with germination fungi.

[0035] In the present invention, unless otherwise specified, all raw material components are commercially available products well known to those skilled in the art.

[0036] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions of the present invention. Obviously, the embodiments described are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0037] In a specific embodiment of the present invention, the TTC staining method for the vitality of Gastrodia elata seeds is:

[0038] Add an appropriate amount of seeds to the 0.25mL mark in a 1.5mL centrifuge tube. Use a pipette to draw up 1mL of 1% TTC solution. After standing in the dark at 30°C for 48 hours, pipette the seeds to mix thoroughly. After mixing, pipette the seed mixture onto the center of a glass slide. Remove the stain with absorbent paper. Under a stereomicroscope, randomly select three fields of view and observe the seed staining. Viable seed embryos will stain orange or pink, while inactive seed embryos will remain unstained or stain yellow. Count the total number of seeds and the total number of viable seeds in each field of view and calculate seed viability. Seed viability = (total number of viable seeds / total number of seeds counted) × 100%. Three replicates were performed for each sample.

[0039] In a specific embodiment of the present invention, the vacuum freeze dryer model used is: (Thermo Scientific) HetoPowerDry LL1500.

[0040] Example 1

[0041] Materials: The capsules and seeds of Gastrodia elata were provided by Guizhou Institute of Crop Varieties and Resources (Modern Chinese Medicinal Materials Research Institute). The Gastrodia elata was identified as Gastrodia elata Bl.f.elata, a protomorph of Gastrodia elata in the Orchidaceae family.

[0042] Step 1: Selection of capsules and seeds of arrowroot and gastrodia elata

[0043] Gastrodia elata (Gastrodia elata) with clear provenance, pest and disease-free, undamaged, and uniform size (120g, 150g, 180g, 200g, and 220g) were selected and covered with fine river sand (approximately 40% moisture) to a thickness of 2.5cm. After the Gastrodia elata bolted and flowered, artificial pollination was performed. When the Gastrodia elata capsules ripened and cracked with a gentle squeeze of the pectin, capsules from the same batch were harvested at the same time. Approximately 100 Gastrodia elata plants of each weight were planted repeatedly. The fresh weight and number of fruit from the bolting Gastrodia elata of different weights were calculated. The results are shown in Table 1.

[0044] Table 1 Effects of different weights of ramie on fresh fruit weight and fruit quantity

[0045]

[0046] It can be seen from Table 1 that when the weight of red Gastrodia elata is 120g, the fresh weight of its fruit is 1.41g, and the number of fruits is 70.29; when it is 150g, the fresh weight of its fruit is 1.67g, and the number of fruits is 79.11; when it is 180g, the fresh weight of its fruit is 1.84g, and the number of fruits is 85.21; when it is 200g, the fresh weight of its fruit is 1.83g, and the number of fruits is 84.3; when it is 220g, the fresh weight of its fruit is 1.99g, and the number of fruits is 88.55. The above data show that when the weight of Gastrodia elata is between 180g and 220g, its fruit fresh weight is higher and the number of fruits is also larger; the reason why Gastrodia elata weighing more than 220g was not selected for seed preservation is that it has higher economic benefits and is more suitable for market transactions.

[0047] Step 2: Choosing the storage time of Gastrodia elata capsules after picking

[0048] Gastrodia elata capsules of uniform size and maturity were harvested at the same time and stored at room temperature for different storage times (0, 1, 2, and 3 days). The viability of the capsules was then tested with three replicates to investigate the effect of storage time after harvest on the viability of Gastrodia elata seeds. The effects of post-harvest storage time on seed viability (measured using the TTC method) are shown in Table 2.

[0049] Table 2 Effects of different storage days on the vitality of Gastrodia elata seeds

[0050]

[0051] As can be seen from Table 2, its vitality gradually decreases with the increase of time. On the day of picking (0d), its TTC vitality is 90.22%, on the 1st day, its TTC vitality is 88.25%, on the 2nd day, its TTC vitality is 83.84%, and on the 3rd day, its TTC vitality is 79.17%. Its vitality gradually decreases with the increase of time. Therefore, after picking the Gastrodia elata capsule, it is best to preserve it when its vitality is higher, that is, on the day of picking.

[0052] Step 3: Processing of Gastrodia elata capsules by vacuum freeze drying

[0053] The Gastrodia elata capsules on the day of picking were randomly divided into 5 groups and placed in glass dishes. The Gastrodia elata capsules just covered the surface of the dishes. Then, the Gastrodia elata capsules in each group were placed in a vacuum freeze dryer for vacuum freeze drying. The vacuum freeze drying time was 24h, 30h, 36h, 42h, and 48h. The water content of the seeds in the Gastrodia elata capsules after vacuum freeze drying was measured, and 3 samples were measured each time. The changes in water content of the seeds in the Gastrodia elata capsules after vacuum freeze drying for different treatment times are shown in Figure 2. Figure 3 .

[0054] Depend on Figure 3 It can be seen that within the first 24 hours of vacuum freeze-drying, the moisture content of Gastrodia elata seeds loses rapidly, and the moisture content drops to 5.4%, which reaches the critical state of ultra-dry seeds (5% moisture content); after drying for 30 hours, the moisture content of Gastrodia elata seeds can reach 3.9%; after drying for 36 hours, the moisture content of Gastrodia elata seeds can reach 3.0%; after drying for 42 hours, the moisture content of Gastrodia elata seeds can reach 2.1%; after drying for 48 hours, the moisture content of Gastrodia elata seeds can reach 1.6%.

[0055] Step 4: Store Gastrodia elata seeds with different water contents at 4°C for two years

[0056] First, prepare 280 mg to 300 mg of Gastrodia elata capsules with different freeze-drying times in step 3 for each centrifuge tube, use dry absorbent paper to divide the Gastrodia elata capsules treated with different freeze-drying times, wrap the outer layer with absorbent cotton and place it in a centrifuge tube, then add color-changing silica gel that accounts for 2 / 10 of the centrifuge tube. Do not add color-changing silica gel to the control group, tightly cover the centrifuge tube cap, and then store in a 4°C refrigerator.

[0057] Step 5: Pay attention to the color changes of the color-changing silica gel during storage.

[0058] After two years of storage, the Gastrodia elata seeds with different moisture contents were tested for seed viability using TTC staining. The results are shown in Table 3.

[0059] Table 3 Vitality of Gastrodia elata seeds after two years of storage in different groups

[0060]

[0061] Note: Different lowercase letters indicate significant differences at the 0.05 level.

[0062] Table 3 Combination Figure 3 It shows that when the moisture content of Gastrodia elata seeds is 1.6%, the TTC staining seed vigor is 37.64% after being stored at 4°C for 2 years. When the moisture content of Gastrodia elata seeds is 2.1%, the TTC staining seed vigor is 36.22% after being stored at 4°C for 2 years. Compared with seeds vacuum-freeze-dried to a moisture content of 3-5.4%, the seed vigor is significantly improved after being stored at 4°C for 2 years.

[0063] Example 2

[0064] The vitality of the Gastrodia elata seeds preserved in Example 1 was determined using a germination culture medium:

[0065] (1) PDA culture medium: 200 g potatoes (peeled), 20 g glucose, 7 g agar, 1000 mL tap water, natural pH, sterilized at 121°C for 30 min.

[0066] (2) PDA comprehensive culture medium: 200g potatoes (peeled), 20g glucose, 3g potassium dihydrogen phosphate, 1.5g magnesium sulfate, 7g agar, V B1 110 mg, 1000 mL of tap water, natural pH, sterilize at 121°C for 30 min.

[0067] (3) Enriched PDA medium: 200 g potatoes (peeled), 20 g glucose, 3 g potassium dihydrogen phosphate, 1.5 g magnesium sulfate, 7 g agar, 1 g peptone, V B1 110 mg, 1000 mL of tap water, natural pH, sterilize at 121°C for 30 min.

[0068] (4) Water agar medium: 7 g agar, 1000 mL tap water, natural pH, sterilize at 121°C for 30 min.

[0069] (5) Cyclobalanopsis glauca leaf culture medium: Spread the sterilized Cyclobalanopsis glauca leaf culture medium on a plate containing PDA comprehensive culture medium. The method of using the Cyclobalanopsis glauca leaf culture medium is to inoculate the germinated fungi under sterile conditions and culture them in the dark until the leaves are infected with the fungi. Then transfer them to a plate with a sterilized wet sponge on the bottom for culture. Keep the sponge moist during the culture process.

[0070] (6) Cultivation of Dendrobium officinale: transfer a piece of Dendrobium officinale mother mushroom about the size of a fingernail shell to the culture medium (1) to (5) under sterile conditions and culture in the dark at 25°C until the mycelium grows to 2 / 3 of the entire plate.

[0071] (7) Gastrodia elata seed viability test (germination method using germination culture medium): The Gastrodia elata capsules preserved in Example 1 were removed from a sterile laminar flow hood. The capsule surface was wiped with 75% alcohol, sterilized by fire, and placed in a sterile petri dish for later use. The pods of the Gastrodia elata capsules were cut open with a scalpel, and the Gastrodia elata seeds were collected with a pipette and mixed. Finally, 100 mg of Gastrodia elata seeds were aseptically taken and evenly distributed on the petri dishes containing the respective germination culture mediums in the laminar flow hood.

[0072] (8) Statistics of germination of Gastrodia elata seeds: After sowing, the culture medium was placed indoors and incubated in the dark at 22°C. The germination of the Gastrodia elata seeds was observed using a stereo microscope every 5 days, and finally the germination of the Gastrodia elata seeds was distinguished by naked eyes. The statistical results are shown in Tables 4 and 5:

[0073] Table 4 Germination time of two-year-old freeze-dried Gastrodia elata seeds on different culture media (unit: d)

[0074]

[0075]

[0076] Table 5 Number of seeds germinated after two-year storage of freeze-dried Gastrodia elata seeds on different types of culture media (unit: grains)

[0077]

[0078] Among them, the water agar medium containing Dendrobium officinale had the best effect on promoting the germination of red Gastrodia elata seeds with a water content of 1.6% ( Figure 4 ), seeds germinated relatively early, germinating as early as 30 days, with a maximum of 234 seeds germinated (Table 4). Water agar with a moisture content of 2.1% had the second-best germination-promoting effect on red Gastrodia elata seeds. This suggests that vacuum freeze-dried red Gastrodia elata seeds stored at 4°C for two years, when stimulated by Dendrobium officinale, exhibited the best germination-promoting effect on water agar when the moisture content was 1.6%. Based on the complexity of medium preparation and germination time, water agar is the most suitable medium for indoor germination of Gastrodia elata seeds.

[0079] Example 3

[0080] The Gastrodia elata seeds preserved in Example 1 were cultured with fungi to verify their vitality:

[0081] 100mg of freeze-dried red Gastrodia elata seeds, stored at 4°C for two years and varying in moisture content, were mixed with walnut-sized pieces of germination fungus (Dendrobium candidum) and placed in a ventilated plastic bag. The bags were then kept at 20°C until the pieces turned white. The bags were then planted in a pot with Armillaria mellea and covered with fine river sand to a thickness of 3cm. After six months of cultivation at room temperature, the yields of white and rice Gastrodia elata were measured to verify the viability of the Gastrodia elata seeds.

[0082] After half a year of mixed fungus cultivation, Figure 5 As shown, when the vacuum freeze-dried Gastrodia elata seeds have a low moisture content of 1.6%, they produce more white hemp and rice hemp.

[0083] Summarize:

[0084] When the weight of the Gastrodia elata fruit is between 180g and 220g, the fresh weight of the fruit is heavier and the number of the fruit is larger; at the same time, after the capsule is picked and placed for 0 days, that is, within the same day, its vitality is higher, and it is more suitable for vacuum freeze-drying on the same day;

[0085] Previous research by the present inventors showed that after one year of storage at 4°C, although the vitality of vacuum-freeze-dried Gastrodia elata seeds was significantly higher than that of fresh Gastrodia elata seeds stored at 4°C and room temperature, there was no significant difference in seed vitality between vacuum-freeze-dried Gastrodia elata seeds with different water contents (Effects of Different Treatments on the Vitality of Gastrodia elata Seeds During Storage [J]. Chinese Medicinal Materials, 2018, 41(10): 2261-2265.). The data in this study calculated that the preservation rate of seed vitality over one year was 34-37%. According to this rule, the seed vitality should be 11.5-14% after two years of storage. However, further research by the present inventors found that after vacuum freeze-drying to a moisture content of 1.5-2.5%, the seed vitality could reach 36-38% after two years of storage at 4°C, which far exceeded the theoretical deduction.

[0086] Furthermore, when the moisture content of the seeds is between 1.6% and 2.1%, the average number of germinations on the water agar medium is also higher. At the same time, after cultivation with mixed bacteria, the yield is also higher. This shows that reducing the moisture content of Gastrodia elata seeds is very important for maintaining the vitality of Gastrodia elata seeds during long-term storage.

[0087] In short, when preserving Gastrodia elata seeds in the present invention, 180g to 220g of the arrow hemp should be selected as the best. At the same time, the harvested seeds should be preserved when their vitality is highest, and then they should be vacuum freeze-dried in time to a water content of 1.5 to 2.5%. At the same time, during preservation, the effect of the Gastrodia elata seed preservation process is identified by adding medium color-changing silica gel. After the preservation of the Gastrodia elata seeds is completed, the vitality is directly verified by germinating on a water agar medium to check the number mean. The above shows that the vacuum drying preservation method for preserving Gastrodia elata seeds is an effective medium- and long-term preservation method.

[0088] The above is only 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 method for preserving Gastrodia elata seeds, characterized in that: The storage method comprises: performing vacuum freeze-drying treatment on the harvested Gastrodia elata capsules until the water content is 1.5-2.5%, and refrigerating and storing the vacuum freeze-dried Gastrodia elata capsules; The dried Gastrodia elata capsules were divided into portions with dry absorbent paper, wrapped with absorbent cotton and placed in 50 mL centrifuge tubes, and color-changing silica gel was added to the centrifuge tubes. The refrigerated storage temperature is 3°C to 6°C; The gastrodia elata capsule is a capsule weighing 180g to 220g that matures after artificial pollination when the gastrodia elata shoots and flowers.

2. The storage method according to claim 1, wherein The Gastrodia elata capsules are Gastrodia elata capsules picked on the day.

3. A method for detecting the vitality of Gastrodia elata seeds, characterized in that: The detection method comprises: evenly dropping the Gastrodia elata seeds preserved by the method according to any one of claims 1 to 2 onto a plate of water agar medium with germination bacteria, culturing in the dark at 20° C. to 25° C., regularly observing the germination of the Gastrodia elata seeds, visually distinguishing the germination of the Gastrodia elata seeds or counting the number of Gastrodia elata protocorms; The main components of the water agar culture medium are water and agar, and the amount of agar added to the water agar culture medium is 6g / L to 8g / L; the germination fungus is Dendrobium officinale.

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