A cultivation substrate for promoting the growth of Dendrobium officinale tissue culture seedlings and its application in this process.

Abstract

This invention belongs to the field of seedling propagation technology, specifically providing a cultivation substrate for seedling health enhancement, comprising pine infected wood, fresh pine bark, and oilseed cake in a volume ratio of 1:(1-2):1. The invention also provides a method for preparing this cultivation substrate, including the following steps: mixing pine infected wood, fresh pine bark, and oilseed cake in the volume ratio and fermenting for 6-10 months; rinsing the fermentation product with water until 70-80% black water flows out, then drying until 50%-60% moisture remains, thus obtaining the cultivation substrate. This cultivation substrate for seedling health enhancement incorporates the use of particulate matter formed after the harmless treatment of pine infected wood, achieving efficient utilization of the pine infected wood and significantly reducing the production cost of Dendrobium officinale tissue culture seedling health enhancement while ensuring the normal growth of healthy seedlings. The invention also provides a method for health enhancement of Dendrobium officinale tissue culture seedlings, employing a two-stage health enhancement method, shortening the health enhancement time, and effectively improving the survival rate and later seedling preservation rate of the healthy tissue culture seedlings.

Description

Technical Field

[0001] This invention belongs to the field of seedling propagation technology, specifically relating to a growth substrate for Dendrobium tissue culture seedlings and its application in the growth of Dendrobium tissue culture seedlings. Background Technology

[0002] Dendrobium is a general term for perennial herbaceous plants belonging to the genus Dendrobium in the Orchidaceae family, possessing both ornamental and medicinal value. There are approximately 1500 species in the genus Dendrobium, with 75 species and 2 varieties found in my country, of which over 50 have medicinal value. Medicinal Dendrobium uses its fresh or dried stems for medicinal purposes. Its active ingredients include polysaccharides, alkaloids, and amino acids, and it possesses effects such as nourishing yin and clearing heat, benefiting the stomach and promoting fluid production, and moistening the lungs and relieving cough. Modern pharmacological research has proven that medicinal Dendrobium has anti-cancer, cancer-preventive, anti-aging, immune-enhancing, vasodilating, anti-platelet aggregation, anti-radiation, and blood sugar-lowering effects. It also has good therapeutic effects on gastrointestinal diseases, cataracts, arthritis, thromboangiitis obliterans, and chronic pharyngitis. In today's fast-paced society, people are paying increasing attention to their health, creating a considerable market for powerful health products like medicinal Dendrobium, indicating a huge potential for future development in the medicinal Dendrobium industry. However, due to its narrow habitat, low reproductive capacity under natural conditions, and over-harvesting, Dendrobium officinale in my country has become an endangered species. For the industry to develop, seedlings must come first; the propagation of Dendrobium officinale seedlings is the foundation and key to its industrial development. Only by ensuring the proper propagation of Dendrobium officinale seedlings can the long-term, stable, healthy, and sustainable development of my country's Dendrobium officinale industry be achieved.

[0003] Currently, medicinal Dendrobium is mainly propagated through tissue culture using aseptic sowing. The physiological and anatomical characteristics of tissue-cultured seedlings necessitate hardening and conditioning to ensure they adapt well to the autotrophic growth and microbial environment of the field after transplanting. The hardening, conditioning, and transplanting processes of Dendrobium tissue-cultured seedlings are crucial for their suitability for later planting and sale. However, conventional hardening and conditioning processes often use fermented pine bark, resulting in poor cultivation outcomes. Furthermore, the conditioning techniques are demanding and complex; even slight errors can lead to a large number of seedling deaths, severely impacting the actual effectiveness of rapid propagation of medicinal Dendrobium through tissue culture.

[0004] Pine wilt disease is a globally recognized major plant disease, often referred to as "the cancer of pine trees," capable of destroying entire pine forests and even entire regions of forest. Currently, the only method for treating infected pine logs in China is burning, which is a huge waste of resources. A more effective method is to harmlessly process infected pine logs into small particles or fragments through rotary cutting or pulverization, which can then be used as a substrate for the healthy growth of Dendrobium officinale tissue culture seedlings. This process increases the added value of infected pine logs. Summary of the Invention

[0005] The purpose of this invention is to overcome the problems in the existing technology, such as the waste of pine tree diseased wood resources, the poor effect of Dendrobium tissue culture seedling hardening and cultivation substrate, and the high requirements, cumbersome steps, and poor results of the hardening technology.

[0006] Therefore, the present invention provides a health-promoting cultivation substrate, comprising pine wood in a volume ratio of 1:(1-2):1, fresh pine bark, and cake fertilizer.

[0007] The present invention also provides a method for preparing the above-mentioned health-promoting cultivation substrate, characterized by comprising the following steps: mixing infected pine wood, fresh pine bark and cake fertilizer in a volume ratio and fermenting for 6-10 months; rinsing the fermentation product with water until 70-80% black water flows out, and then drying it until 50%-60% moisture remains, thereby obtaining the cultivation substrate.

[0008] The cultivation substrate for promoting the growth of Dendrobium tissue culture seedlings provided by this invention can be used for this purpose.

[0009] Therefore, the present invention provides a method for the healthy growth of Dendrobium tissue culture seedlings, comprising the following steps:

[0010] (1) Open the bottle to harden the seedlings, and clean, disinfect and dry the seedlings;

[0011] (2) First-time conditioning:

[0012] Plant the dried tissue culture seedlings from step (1) into a seedling container filled with cultivation substrate, control the temperature and humidity, replenish water daily, and allow them to mature for 7-14 days.

[0013] (3) Secondary strengthening:

[0014] Take out the Dendrobium seedlings that have completed the first conditioning in step (2), transplant them into a container filled with the above-mentioned cultivation substrate, control the temperature and humidity, and replenish water daily. The second conditioning is completed when new roots grow from the roots of the Dendrobium seedlings.

[0015] (4) Plant the Dendrobium seedlings that have completed the second conditioning process.

[0016] Specifically, step (1) above is as follows: take out the Dendrobium tissue culture seedlings that have reached the growth standard, open the bottle cap or sealing film and place them at room temperature for 1-3 days to harden the seedlings; wash the culture medium in the roots and soak them in disinfectant solution for 30-40 minutes, rinse them clean, and place them in a cool place to dry until the roots of the seedlings turn white and there is no water on the leaves.

[0017] Specifically, the above disinfectant solution is prepared by mixing 5-8 ml of Basifkarun pyraclostrobin emulsion and 12,000-15,000 ml of water.

[0018] Specifically, step (2) above is as follows: arrange the stems, leaves and roots of the tissue culture seedlings that have been dried in step (1), and plant them into seedling containers filled with cultivation substrate at a planting density of 16-20 seedlings / clump. Then place the containers with the tissue culture seedlings on a tray. Spray 3‰ carbendazim until liquid flows out of the container. After tilting and pouring off the excess water on the tray, place the tray containing the seedling containers on the shelf of the conditioning room. Maintain the temperature and humidity in the conditioning room, and replenish the seedlings with water daily. Conditioning takes 7-14 days.

[0019] Specifically, in step (2) above, the cultivation substrate is fermented pine bark with a particle size of 9-12 mm.

[0020] Specifically, step (3) above is as follows: take out the Dendrobium seedlings that have completed the first conditioning in step (2), clean the substrate attached to the roots, and transplant the seedlings into the container according to the planting quantity of 3-4 clumps / group. The cultivation substrate in the container is pre-filled to a height of 6-8cm. After the Dendrobium seedlings are placed upright, the substrate is increased to a height of 8-10cm to ensure that the top of the planted seedlings does not protrude from the pot opening. Spray 3‰ carbendazim spray until liquid flows out of the container, place the container on the shelf of the conditioning room, maintain the temperature and humidity in the conditioning room, and water the seedlings daily. The second conditioning is completed when new roots grow from the roots of the Dendrobium seedlings.

[0021] Specifically, in steps (2) and (3) above, the temperature is controlled at 20-30℃ and the air humidity is controlled at 60-65%.

[0022] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0023] (1) The cultivation substrate for health preservation provided by this invention includes the use of particulate matter formed after the harmless treatment of pine infected wood, which not only realizes the efficient utilization of pine infected wood, but also greatly reduces the production cost of health preservation of medicinal Dendrobium tissue culture seedlings while ensuring the normal growth of healthy seedlings.

[0024] (2) The Dendrobium tissue culture seedling conditioning method provided by this invention is simple to transplant and easy to operate. It improves and perfects the seedling conditioning time, conditioning steps, and phased conditioning time in the Dendrobium tissue culture conditioning process. The two-stage conditioning method is adopted for seedling conditioning, which shortens the conditioning time and effectively improves the survival rate and later seedling preservation rate of the conditioning tissue culture seedlings. The produced seedlings are sterile seedlings of the medicinal Dendrobium species that represent the excellent traits of the parent plant. There is no phenomenon of mixed varieties, and the seedling cultivation is of high quality.

[0025] The present invention will now be described in further detail with reference to the accompanying drawings. Attached Figure Description

[0026] Figure 1This is a flowchart of the method for promoting the health of Dendrobium tissue culture seedlings provided by the present invention.

[0027] Figure 2 This is a schematic diagram of the container transplanting structure during the first stage of the Dendrobium tissue culture seedling conditioning method provided by the present invention.

[0028] Figure 3 This is a schematic diagram of the container transplanting structure during the secondary conditioning process of the Dendrobium tissue culture seedling conditioning method provided by the present invention.

[0029] Attached labels: 1. Tray; 2. Primary conditioning container; 3. Cultivation substrate A; 4. Conditioning rack; 5. Dendrobium tissue culture seedling; 6. Cultivation substrate B; 7. Secondary conditioning container. Detailed Implementation

[0030] The technical solutions of the present invention will be clearly and completely described below with reference to embodiments. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Although representative embodiments of the present invention have been described in detail, those skilled in the art will understand that various modifications and changes can be made to the present invention without departing from the scope of the present invention. Therefore, the scope of the present invention should not be limited to the embodiments, but should be defined by the appended claims and their equivalents.

[0031] This invention provides a health-promoting cultivation substrate, comprising pine infected wood, fresh pine bark, and oilseed cake in a volume ratio of 1:(1-2):1, prepared through the following steps: The pine infected wood, fresh pine bark, and oilseed cake are mixed according to the volume ratio and fermented for 6-10 months; the fermentation product is rinsed with water until 70-80% black liquid flows out, then dried until 50%-60% moisture content remains, thus obtaining the cultivation substrate. The pine infected wood is preferably composed of 3-6 mm particles.

[0032] This invention also provides a method for promoting the health of Dendrobium tissue culture seedlings, comprising the following steps:

[0033] (1) Take out Dendrobium tissue culture seedlings that have reached the height and root growth standard, open the bottle cap or sealing film and place them at room temperature for 1-3 days to harden off; wash the culture medium on the roots with running water and soak them in disinfectant solution for 30-40 minutes, rinse them with clean water, put the cleaned and disinfected tissue culture seedlings into a hollow plastic frame and place them in a cool place to dry until the roots of the seedlings turn white and there is no water on the leaves;

[0034] A qualified tissue culture seedling generally requires a height of 5-8cm and 4-5 roots with a length of more than 5cm.

[0035] The disinfectant solution is prepared by mixing 5-8 ml of Basifuron-methyl pyraclostrobin emulsion and 12,000-15,000 ml of water;

[0036] (2) First-time conditioning:

[0037] After the tissue culture seedlings in step (1) have been dried, arrange the stems, leaves and roots, and plant them into seedling containers filled with cultivation substrate at a planting density of 16-20 seedlings / clump. Then place the containers with the tissue culture seedlings on a tray. Spray 3‰ carbendazim until liquid flows out of the container. After tilting and pouring off the excess water on the tray, place the tray containing the seedling containers on the shelf of the conditioning room. The temperature in the conditioning room is controlled at 20-30℃, the air humidity is controlled at 60-65%, and normal diffused light is ensured. Pay attention to the daily watering of the seedlings. In this environment, condition for 7-14 days.

[0038] The preferred cultivation substrate for primary health care is fermented pine bark with a particle size of 9-12 mm that has undergone high-temperature and high-pressure sterilization;

[0039] The preferred seedling containers for primary health care are rectangular or cylindrical plastic cultivation containers with a bottom diameter of 5-7cm, a mouth diameter of 6-10cm, and a height of 6-10cm; the trays are open, hollow rectangular structures with a bottom.

[0040] (3) Secondary strengthening:

[0041] Take out the Dendrobium seedlings that have completed the first conditioning in step (2), clean the substrate attached to the roots, and transplant the seedlings into the container at a planting rate of 3-4 clumps / group. The cultivation substrate in the container is pre-filled to a height of 6-8cm. After placing the Dendrobium seedlings upright, increase the substrate to a height of 8-10cm to ensure that the top of the planted seedlings does not protrude from the pot opening. Spray 3‰ carbendazim until liquid flows out of the container. Place the container on the shelf in the conditioning room. The temperature in the conditioning room is controlled at 20-30℃, the air humidity is controlled at 60-65%, and normal diffused light is ensured. Water the seedlings daily. The second conditioning is completed when new roots grow from the roots of the Dendrobium seedlings.

[0042] The preferred container for secondary health care is a plastic flowerpot with a bottom diameter of 10-12cm, a mouth diameter of 14-16cm, and a height of 15-17cm.

[0043] The curing chamber shelves used for primary and secondary curing are multi-layered, with each layer measuring 140×50×45cm in length×width×height.

[0044] (4) Plant the Dendrobium seedlings that have completed the secondary conditioning process on trees, stone substrates, rocks, or directly in pots at room temperature or sell them according to cultivation needs.

[0045] The effects of the cultivation substrate for improving the health of Dendrobium officinale tissue culture seedlings and the method for improving the health of Dendrobium officinale tissue culture seedlings of the present invention will be studied through specific embodiments below.

[0046] The pine trees infected with the disease used in this invention have all undergone conventional harmless treatment, and the fermented pine bark comes from Hubei Guanggu Lanyan (Luotian) New Energy Co., Ltd., a designated processing enterprise for infected pine trees. Example 1

[0047] This embodiment provides a method for the healthy development of Dendrobium tissue culture seedlings, including the following steps:

[0048] (1) Take out Dendrobium officinale tissue culture seedlings with a height of 5-8cm and 4-5 roots with a length of more than 5cm. After opening the bottle cap or sealing film, place them at room temperature for 3 days to harden off. Wash the culture medium on the roots with running water and soak them in disinfectant solution for 35 minutes. Rinse them with clean water. Place the cleaned and disinfected tissue culture seedlings in a hollow plastic frame and place them in a cool place to dry until the roots turn white and there is no water on the leaves.

[0049] The disinfectant solution is prepared by mixing 5 ml of Basifuron-methyl pyraclostrobin emulsion and 12000 ml of water;

[0050] (2) First-time conditioning:

[0051] like Figure 2 As shown, the tissue culture seedlings dried in step (1) are arranged with their stems, leaves and roots sorted out, and planted into seedling containers containing cultivation substrate A at a planting density of 18 seedlings / clump. The containers containing the tissue culture seedlings are then placed on a tray. 3‰ carbendazim is sprayed until liquid flows out of the container. The excess water on the tray is tilted and poured off. The tray containing the seedling containers is then placed on the shelf of the conditioning room. The temperature in the conditioning room is controlled at 20-30℃, the air humidity is controlled at 60-65%, and normal diffused light is ensured. Attention is paid to the daily watering of the seedlings. The seedlings are conditioning in this environment for 10 days.

[0052] The primary health-promoting cultivation substrate A is fermented pine bark with a particle size of 9-12mm that has been sterilized by high temperature and high pressure;

[0053] The seedling container for primary health care is a rectangular plastic cultivation container with a bottom diameter of 6cm, a mouth diameter of 8cm, and a height of 8cm; the tray is a hollow rectangular structure without a lid and with a bottom.

[0054] (3) Secondary strengthening:

[0055] Remove the Dendrobium seedlings that have undergone one round of conditioning in step (2), clean the substrate attached to the roots, and transplant the seedlings into containers at a planting density of 3 clumps per group, referring to... Figure 3Pre-fill the container with cultivation substrate B to a height of 7cm; after placing the Dendrobium seedlings upright, increase the cultivation substrate B to a height of 10cm, ensuring that the top of the planted seedlings does not protrude from the rim of the pot; spray with 3‰ carbendazim until liquid flows out of the container, place the container on the shelf in the conditioning room, control the temperature in the conditioning room at 20-30℃, control the air humidity at 60-65%, ensure normal diffused light, and water the seedlings daily; the second conditioning is completed when new roots grow from the roots of the Dendrobium seedlings.

[0056] The preferred container for secondary health care is a plastic flowerpot with a bottom diameter of 11cm, a mouth diameter of 15cm, and a height of 17cm.

[0057] The secondary health-promoting cultivation substrate B consists of pine infected wood, fresh pine bark, and cake fertilizer in a volume ratio of 1:1:1. It is prepared by the following steps: pine infected wood particles with a particle size of 3-6 mm, fresh pine bark, and cake fertilizer are mixed in the volume ratio and fermented for 8 minutes. Then, it is rinsed with water until 75% black water flows out, and then dried until 60% moisture is retained to make cultivation substrate B.

[0058] The curing chamber shelves used for primary and secondary curing are multi-layered, with each layer measuring 140×50×45cm in length×width×height.

[0059] (4) Plant the Dendrobium seedlings that have completed the second conditioning process.

[0060] After conditioning using the method described in this embodiment, the survival rate of Dendrobium officinale tissue culture seedlings reached 96%. After 60 days of conditioning, the preservation rate reached 90%. Example 2

[0061] This embodiment provides a method for the health enhancement of Dendrobium tissue culture seedlings. The difference between this method and that of Embodiment 1 is that the secondary health enhancement substrate B in step (3) includes pine infected wood, fresh pine bark and cake fertilizer in a volume ratio of 1:1:1. The method is prepared by the following steps: pine infected wood particles with a particle size of 9-12mm, fresh pine bark and cake fertilizer are mixed in a volume ratio and fermented for 8 minutes. After that, the mixture is rinsed with water until 75% black water flows out. The mixture is then dried until 60% moisture remains to prepare the cultivation substrate B.

[0062] After conditioning using the method described in this embodiment, the survival rate of Dendrobium officinale tissue culture seedlings reached 94%. After 60 days of conditioning, the preservation rate reached 89%. Example 3

[0063] This embodiment provides a method for the health enhancement of Dendrobium tissue culture seedlings. The difference between this method and that of Embodiment 1 is that the secondary health enhancement substrate B in step (3) includes pine sawdust, fresh pine bark and cake fertilizer in a volume ratio of 1:1:1. The substrate is prepared by the following steps: pine sawdust, fresh pine bark and cake fertilizer are mixed and fermented in a volume ratio for 8 minutes, then rinsed with water until 75% black water flows out, and then dried until 60% moisture is retained to make the cultivation substrate B.

[0064] After conditioning using the method described in this embodiment, the survival rate of Dendrobium officinale tissue culture seedlings reached 90%. After 60 days of conditioning, the preservation rate reached 85%. Example 4

[0065] This embodiment provides a method for the health enhancement of Dendrobium tissue culture seedlings. The difference between this method and that of Embodiment 1 is that the secondary health enhancement substrate B in step (3) includes pine infected wood, fresh pine bark and cake fertilizer in a volume ratio of 1:2:1. The method is prepared by the following steps: pine infected wood particles with a particle size of 9-12mm, fresh pine bark and cake fertilizer are mixed in a volume ratio and fermented for 8 minutes. Then, the mixture is rinsed with water until 75% black water flows out and dried until 60% moisture remains to prepare the cultivation substrate B.

[0066] After conditioning using the method described in this embodiment, the survival rate of Dendrobium officinale tissue culture seedlings reached 92%. After 60 days of conditioning, the preservation rate reached 86%.

[0067] Comparative Example 1:

[0068] This comparative example provides a method for the conditioning of Dendrobium tissue culture seedlings. The difference between this method and Example 1 is that the secondary conditioning substrate B in step (3) includes fresh pine bark and cake fertilizer in a volume ratio of 2:1. The substrate is prepared by the following steps: fresh pine bark and cake fertilizer are mixed and fermented in a volume ratio for 8 minutes, then rinsed with water until 75% black water flows out, and then dried until 60% moisture is retained to make the substrate B.

[0069] After conditioning using the method described in this comparative example, the survival rate of Dendrobium officinale tissue culture seedlings reached 92%. After 60 days of conditioning, the survival rate was 85%.

[0070] Comparative Example 2:

[0071] This comparative example provides a method for the conditioning of Dendrobium tissue culture seedlings. The difference between this method and Example 1 is that the cultivation substrate used in the secondary conditioning in step (3) is fermented pine bark that has been sterilized by high temperature and high pressure.

[0072] After conditioning using the method described in this comparative example, the survival rate of Dendrobium officinale tissue culture seedlings reached 95%. After 60 days of conditioning, the survival rate was 85%.

[0073] Comparative Example 3:

[0074] This comparative example provides a method for the conditioning of Dendrobium tissue culture seedlings. The difference between this method and Example 1 is that a one-step conditioning method is used. After the conditioning time in step (2) is extended to 35 days, step (3) is omitted and the value is directly determined.

[0075] After conditioning using the method in this comparative example, the survival rate of Dendrobium officinale tissue culture seedlings was only 70%. After 60 days of conditioning, the survival rate was only 60%.

[0076] Comparative Example 4:

[0077] This comparative example provides a method for the healthy growth of Dendrobium tissue culture seedlings. The difference between this method and Example 1 is that step (3) is: leave space for root extension in the flowerpot, fill the cultivation substrate B to be level with the surface of the pot, and expose the planted Dendrobium seedlings at the pot opening with the root base level with the pot opening.

[0078] After conditioning using the method in this comparative example, the survival rate of Dendrobium officinale tissue culture seedlings was only 75%. After 60 days of conditioning, the survival rate was only 65%.

[0079] Comparative Example 5:

[0080] This comparative example provides a method for improving the health of Dendrobium tissue culture seedlings. The difference between this method and Example 1 is that the transplanting substrate B in step (3) is pine bark.

[0081] After conditioning using the method described in this comparative example, the survival rate of Dendrobium officinale tissue culture seedlings was 90%. After 60 days of conditioning, the survival rate was only 85%.

[0082] Comparative Example 6:

[0083] This comparative example provides a method for the conditioning of Dendrobium tissue culture seedlings. The difference between this method and Example 1 is that step (2) is omitted. In step (1), the dried tissue culture seedlings directly enter step (3) to condition until new roots grow from the roots of the seedlings.

[0084] After conditioning using the method described in this comparative example, the survival rate of Dendrobium officinale tissue culture seedlings was 90%. After 60 days of conditioning, the survival rate was 80%.

[0085] The results of Examples 1-4 provided by this invention show that by using a two-stage conditioning method, simultaneously applying pine wilt-infested wood particles or sawdust of different sizes and formulation ratios to the conditioning cultivation of Dendrobium, excellent cultivation results were achieved, with seedling survival and retention rates both exceeding 90%. Example 1 showed the best effect, reaching 96%. This method can replace commonly used cultivation substrates for Dendrobium cultivation in tissue culture-based industrialized seedling production and routine seedling cultivation of medicinal Dendrobium.

[0086] As can be seen from Comparative Examples 1-6, after changing the cultivation substrate and conditioning method, the survival rate and later preservation rate of Dendrobium tissue culture seedlings were between 70% and 80%, which were much lower than those of the embodiments of the present invention. It can be seen that the present invention can significantly improve the survival rate and later preservation rate of Dendrobium tissue culture seedlings by combining the cultivation substrate with the secondary conditioning method.

[0087] The above examples are merely illustrative of the present invention and do not constitute a limitation on the scope of protection of the present invention. All designs that are the same as or similar to the present invention are within the scope of protection of the present invention.

Claims

1. A method for promoting the healthy growth of Dendrobium tissue culture seedlings, characterized in that, Includes the following steps: (1) Open the bottle to harden the seedlings, and clean, disinfect and dry the seedlings; (2) First-time conditioning: Plant the dried tissue culture seedlings from step (1) into a seedling container filled with cultivation substrate, control the temperature and humidity, and replenish water daily; (3) Secondary strengthening: Take out the Dendrobium seedlings that have completed the first conditioning in step (2), transplant them into a container filled with conditioning substrate, control the temperature and humidity, and water them daily. The second conditioning is completed when new roots grow from the roots of the Dendrobium seedlings. The conditioning substrate includes pine wood, fresh pine bark and cake fertilizer in a volume ratio of 1:(1-2):

1. (4) Plant the Dendrobium seedlings that have completed the second conditioning process.

2. The method for promoting the health of Dendrobium tissue culture seedlings as described in claim 1, characterized in that: The preparation method of the health-promoting cultivation substrate includes the following steps: mixing infected pine wood, fresh pine bark and cake fertilizer according to the volume ratio and fermenting them for 6-10 months; rinsing the fermentation products with water until 70-80% black water flows out, and then drying them until 50%-60% moisture remains to obtain the cultivation substrate.

3. The method for promoting the health of Dendrobium tissue culture seedlings as described in claim 1, characterized in that, The specific steps (1) are as follows: take out the Dendrobium tissue culture seedlings that have reached the growth standard, open the bottle cap or sealing film and place them at room temperature for 1-3 days to harden the seedlings; wash the culture medium from the roots and soak them in disinfectant solution for 30-40 minutes, rinse them clean, and place them in a cool place to dry until the roots of the seedlings turn white and there is no water on the leaves.

4. The method for promoting the health of Dendrobium tissue culture seedlings as described in claim 3, characterized in that: The disinfectant solution is prepared by mixing 5-8 ml of Basifkarun pyraclostrobin emulsion and 12,000-15,000 ml of water.

5. The method for promoting the health of Dendrobium tissue culture seedlings as described in claim 1, characterized in that, The specific steps (2) are as follows: the tissue culture seedlings that have been dried in step (1) are arranged with their stems, leaves and roots, and planted into seedling containers containing cultivation substrate at a planting density of 16-20 seedlings / clump. The containers containing the tissue culture seedlings are then placed on a tray. 3‰ carbendazim is sprayed until liquid flows out of the container. The excess water on the tray is tilted and poured off. The tray containing the seedling containers is then placed on the shelf in the conditioning room. The temperature and humidity in the conditioning room are maintained. The seedlings are watered daily and the conditioning process lasts for 7-14 days.

6. The method for promoting the health of Dendrobium tissue culture seedlings as described in claim 1, characterized in that: In step (2), the cultivation substrate is fermented pine bark with a particle size of 9-12 mm.

7. The method for promoting the health of Dendrobium tissue culture seedlings as described in claim 1, characterized in that, The specific steps (3) are as follows: Take out the Dendrobium seedlings that have completed the first conditioning in step (2), clean the substrate attached to the roots, and transplant the seedlings into the container at a planting rate of 3-4 clumps / group. The cultivation substrate in the container is pre-filled to a height of 6-8cm. After the Dendrobium seedlings are placed upright, the substrate is increased to a height of 8-10cm to ensure that the top of the planted seedlings does not protrude from the pot opening. Spray 3‰ carbendazim until liquid flows out of the container. Place the container on the shelf of the conditioning room, maintain the temperature and humidity in the conditioning room, and water the seedlings daily. The second conditioning is completed when new roots grow from the roots of the Dendrobium seedlings.

8. The method for promoting the health of Dendrobium tissue culture seedlings as described in claim 1, characterized in that: In steps (2) and (3), the temperature is controlled at 20-30℃ and the air humidity is controlled at 60-65%.

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