Method for inducing intermediate propagules of rare cymbidium faberi
By combining the culture medium prepared with double disinfectants and four hormones with the dark-light culture method, we can induce the intermediate reproductive bodies of the precious Cymbidium orchid, which solves the problem of difficulty in breeding precious Cymbidium orchids and realizes an efficient and rapid breeding method to meet market demand.
Patent Information
- Application Number
- CN202510947959.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-07-10
- Publication Date
- 2025-09-23
AI Technical Summary
The precious Cymbidium orchid is difficult to propagate through seedlings and asexual reproduction. The traditional division reproduction cycle is long and the reproduction coefficient is low, resulting in a scarcity of seedlings. In addition, wild Cymbidium orchids are over-harvested, making it difficult to meet market demand.
A method for inducing intermediate propagules is provided by using a two-step disinfection method with double disinfectants and a culture medium prepared in synergistic manner with four hormones, combined with a dark-light two-stage culture method, including disinfection with alcohol and mercuric chloride solution, MS culture medium prepared with IBA, NAA, TDZ, and 6-BA hormones, and inducing intermediate propagules under dark-light culture conditions.
Significantly reduce explant contamination and browning rate, improve breeding efficiency, shorten the breeding cycle, solve the problem of seedling scarcity, and provide technical support for the large-scale and factory-based breeding of precious Cymbidium orchids.
Smart Images
Figure FT_1 
Figure FT_2
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of rapid propagation of orchid plants, in particular to a method for inducing intermediate propagules of precious Cymbidium faberi. Background Art
[0002] Cymbidium faberi is a species of plant in the Orchidaceae family, with a long history of cultivation in China. The rare and precious Line Art Cymbidium faberi is a variation of the common Cymbidium faberi, with leaves striped with yellow and white. It is extremely rare and has high ornamental and economic value.
[0003] Cymbidium orchids are difficult to propagate through seedlings and asexual reproduction. Their seeds are tiny and their embryos are underdeveloped. In their natural state, they rely on a symbiotic relationship with specific fungi, making seedling propagation difficult. Traditional propagation by division is slow, has a low reproduction rate, and accumulates viruses.
[0004] To obtain rare orchid varieties, wild orchids have been over-harvested and are now listed as a national Class II protected plant. To overcome the reproductive barriers and address the scarcity of rare orchid seedlings, a novel method for breeding rare orchids by inducing intermediate propagules has been proposed. This method is of great significance for achieving large-scale, factory-based rapid propagation of rare orchids. Summary of the Invention
[0005] The purpose of the present invention is to provide a method for inducing intermediate propagules of precious line-art Cymbidium, to innovate the technical path for aseptically obtaining intermediate propagules from seedlings, and to provide new technologies and new options for large-scale and factory-based breeding of precious line-art Cymbidium.
[0006] The purpose of the present invention is achieved through the following technical solutions.
[0007] A method for inducing intermediate propagules of precious Cymbidium lineatum, comprising the following steps:
[0008] (1) Use a double disinfectant two-step disinfection method to disinfect and sterilize the precious Cymbidium orchid explants to reduce mortality and obtain sterile materials;
[0009] (2) Use a culture medium prepared with four hormones of appropriate species and concentrations to induce intermediate propagules;
[0010] (3) Use a two-stage dark-light culture method to reduce the browning rate.
[0011] The double disinfectants in step (1) are disinfectant 1 (alcohol) and disinfectant 2 (mercuric chloride solution).
[0012] The two-step disinfection method in step (1) is to clean the surface dust of the precious orchid with a detergent, rinse it under running water, transfer it to a sterile workbench, shake and soak it with disinfectant 1, and then continue to disinfect it with disinfectant 2, then wash the residual solution on the surface with sterile water, and use sterile filter paper to absorb the residual moisture on the surface of the material to obtain a sterile explant.
[0013] The plant growth regulators of different types and concentrations added to the induction culture medium in the above step (2) with synergistic effects are specifically: IBA, NAA, TDZ, and 6-BA.
[0014] The main components of the induction culture medium in the above step (2) include: MS, plant growth regulator, sucrose, agar, activated carbon, and the pH is adjusted to 5.8-6.0.
[0015] In the above step (2), the four hormones synergistic induction is to remove the roots of the obtained sterile explants with sterilized tools to obtain stem segments, and then inoculate the materials into the induction medium for synergistic induction.
[0016] In the above step (3), the dark-light two-stage culture method is that the first stage is dark culture for a certain period to reduce the browning rate of the precious line art orchid seedlings, and the second stage is light culture to induce the intermediate propagules.
[0017] In the above step (3), the light culture conditions were 24±2°C and the light intensity was set to 45 μmol / m 2 / s, and the photoperiod is 14h / d.
[0018] The precious thread art orchid used in the method of the present invention is sourced from Southwest University of Science and Technology.
[0019] The above scheme has the following beneficial effects:
[0020] The present invention proposes a new method for breeding precious line-art Cymbidium orchids by inducing intermediate propagules, which significantly reduces the contamination rate and browning rate of explants; through this method, the problems of traditional non-symbiotic germination of Cymbidium orchid seeds, variation of excellent genetic traits, low efficiency of division reproduction, and virus transmission are avoided; it breaks through the technical route of traditional tissue culture rapid propagation of Cymbidium orchids, shortens the breeding cycle of precious line-art Cymbidium orchids, solves the problem of scarcity of precious varieties of seedlings, and provides effective technical support for large-scale and factory-based breeding of precious line-art Cymbidium orchids. BRIEF DESCRIPTION OF THE DRAWINGS
[0021] Figure 1 The precious thread art orchid is sterilized according to Example 1 of the present invention.
[0022] Figure 2 This is the intermediate propagule induced in Example 1 of the present invention. DETAILED DESCRIPTION
[0023] The present invention is further described in detail below through specific examples, but the examples are not intended to limit the technical solutions of the present invention.
[0024] The precious thread art orchid used in the embodiment is from Southwest University of Science and Technology.
[0025] Example 1:
[0026] Medium preparation: Prepare a mixture of MS, sucrose, agar, activated carbon, IBA, NAA, TDZ, and 6-BA. Adjust the pH to 5.8-6.0 and sterilize by autoclaving at 121°C, 101.5 kPa for 20 minutes.
[0027] Explant disinfection and sterilization: Use a two-disinfectant, two-stage method. Take a rare Cymbidium orchid seedling, wash it with detergent, rinse it with running water, and transfer it to a sterile workbench. Use Disinfectant 1 for a period of time, then continue shaking and allowing it to stand for disinfection with Disinfectant 2. Rinse off any remaining disinfectant with sterile water, and then use sterile filter paper to absorb any remaining moisture on the surface of the material to obtain a sterile material.
[0028] Induction culture of intermediate propagules: Under sterile conditions, use sterilized tools to remove the roots of the precious Cymbidium orchid to obtain stem segments; then the material is inoculated into the induction culture medium and induced by the coordination of four hormones with different concentrations.
[0029] A two-stage dark-light culture method was used. In the first stage, dark culture was performed for a certain period of time to reduce the browning rate. In the second stage, light culture was performed for induction. After 30 days of culture, intermediate propagules occurred. After 70 days, the occurrence rate of intermediate propagules reached 83.3%.
Claims
1. A method for inducing intermediate propagules of precious Cymbidium lineatum, comprising the steps of: (1) Use a two-step disinfection method with double disinfectants to disinfect and sterilize the precious Cymbidium orchid explants to reduce mortality; (2) Use appropriate species and concentrations of four hormones to synergistically prepare a culture medium to induce intermediate propagules; (3) Use a two-stage dark-light culture method to reduce the browning rate.
2. The method for inducing intermediate propagules of precious Cymbidium lineatum according to claim 1, characterized in that: The double disinfectants in step (1) are disinfectant 1 (alcohol) and disinfectant 2 (mercuric chloride solution).
3. The method for inducing intermediate propagules of precious Cymbidium lineatum according to claim 1, characterized in that: The two-step disinfection method described in step (1) is to clean the surface dust of the thread art orchid with a detergent, rinse it under clean running water, transfer it to a sterile workbench, shake and soak it with disinfectant 1, and then continue to shake it with disinfectant 2, and let it stand for disinfection for a certain period of time to eliminate surface bacteria and fungi, wash the residual solution on the surface with sterile water, and use sterile filter paper to absorb the residual moisture on the surface of the material to obtain a sterile explant.
4. The method for inducing intermediate propagules of the precious Cymbidium lineatum according to claim 1, characterized in that: The formula of the four-hormone synergistic induction culture medium of appropriate species and concentrations described in step (2) is MS+6-BA+IBA+NAA+TDZ.
5. The method for inducing intermediate propagules of the precious Cymbidium lineatum according to claim 1, characterized in that: The sterile explants obtained in step (2) are excised with a sterilized inoculation tool to obtain stem segments, and the materials are inoculated into an induction medium for synergistic induction with four hormones.
6. The method for inducing intermediate propagules of the precious Cymbidium line art as claimed in claim 1, characterized in that: The dark-light two-stage culture method described in step (3) is that the first stage is dark culture for a certain period to reduce the browning rate of the precious line art Cymbidium seedlings, and the second stage is light culture.
7. The method for inducing intermediate propagules of the precious Cymbidium lineatum according to claim 1, characterized in that: The light culture conditions in step (3) are: temperature of 24±2°C, light intensity of 45 μmol / m 2 / s, and the photoperiod is 14h / d.