Induction and efficient culture method of dendrobium officinale adventitious roots
Through the culture method of Dendrobium officinale indeterminate roots, the sterile Dendrobium officinale seedlings induce the indeterminate roots and perform expansion and culture, the problem of unstable secondary metabolites production in Dendrobium officinale cell culture is solved, and efficient supply of Dendrobium officinale raw materials is achieved, and industrial production is supported.
Patent Information
- Application Number
- CN202510910387.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2023-11-27
- Publication Date
- 2025-08-12
AI Technical Summary
In the prior art, the production of secondary metabolites in Dendrobium officinale cell culture is unstable, and the lack of an uncertain root culture system makes it difficult to achieve industrial production.
A method for cultivating Dendrobium officinale is provided, including germination culture, proliferation culture, rooting culture, callus induced culture and uncertain root induced culture. A variety of explants of Dendrobium officinale are used to induce intractable roots and expand and culture.
The induction rate of Dendrobium officinale indeterminate roots was achieved between 72.30% and 88.74%, laying the foundation for the industrial production of Dendrobium officinale and solving the contradiction between supply and demand of raw materials.
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Abstract
Description
[0001] This case is application number: 202311590952.9, application date: November 27, 2023, invention name:
[0002] A divisional application for a method for cultivating adventitious roots of Dendrobium officinale. Technical Field
[0003] The invention relates to the technical field of plant tissue culture, in particular to a method for culturing adventitious roots of Dendrobium officinale. Background Art
[0004] Adventitious roots are roots that grow on the stems or leaves of plants. In most cases, the occurrence of adventitious roots is due to injury to plant organs or stimulation by external factors such as hormones and pathogenic microorganisms, and therefore manifests as a regenerative response of the plant. The occurrence and formation of adventitious roots is an important way of asexual reproduction of plants. Adventitious root culture has the advantages of short growth cycle, controllable conditions with small errors, single source of materials, consistent genetic background, economy and convenience, strong repeatability, high efficiency and year-round testing or growth. The adventitious root culture of medicinal plants can overcome the shortcomings of low content of active ingredients and instability in cell suspension culture. Therefore, the adventitious root culture of medicinal plants is an effective way to develop Chinese medicine resources.
[0005] Dendrobium officinale (Kimura et Migo) is a perennial epiphytic herb in the genus Dendrobium of the Orchidaceae family. It is named "Dendrobium officinale" for its iron-green stems. The Taoist Canon lists Dendrobium officinale as the first of the Nine Immortal Herbs, and contemporary botanical researchers call it the "Giant Panda" of the plant world. Modern scientific research has revealed that Dendrobium officinale possesses a range of benefits, including anti-tumor, increased saliva secretion, immune enhancement, liver protection, blood sugar lowering, and fatigue relief. It has also been approved as a medicinal ingredient for use in health foods.
[0006] In its natural state, Dendrobium officinale is primarily found on cliffs between 1,000 and 3,400 meters tall, or on broad-leaved trees covered with thick mosses and wild vines. It prefers relatively cool, humid environments. Because its seeds lack fully developed embryos and endosperm, their germination rate is low and their natural reproductive capacity is weak. Due to overharvesting and damage to the natural ecosystem, wild Dendrobium officinale resources are critically endangered and designated a Class II endangered medicinal plant under key protection. Research on tissue culture techniques for Dendrobium officinale began in the mid-1970s, and complete regenerated plants were successfully obtained. In the 1980s, the Zhejiang Academy of Medical Sciences pioneered the use of Dendrobium officinale tissue culture technology for production. In the decades that followed, a wave of interest in Dendrobium officinale tissue culture among Chinese researchers has led to significant breakthroughs in this field.
[0007] Currently, the technical challenges of rapid tissue culture propagation of Dendrobium officinale have been largely addressed, but the production of medicinal ingredients from Dendrobium officinale using biotechnology has yet to be reported. Adventitious root culture, a newly developed technique, offers stable and rapid production of secondary metabolites, making it a primary pathway for producing secondary metabolites from rhizomes of medicinal plants. The present invention aims to address the issues of unstable secondary metabolite production using Dendrobium officinale cell culture and the lack of adventitious root culture systems. Based on these issues, a method for direct induction and proliferation of adventitious roots of Dendrobium officinale is proposed. Summary of the Invention
[0008] The purpose of the present invention is to provide a method for cultivating adventitious roots of Dendrobium officinale.
[0009] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:
[0010] The present invention provides a method for cultivating adventitious roots of Dendrobium officinale, comprising the following steps:
[0011] (1) inoculating the Dendrobium officinale seeds into a start-up culture medium, a proliferation culture medium, and a seedling rooting culture medium in sequence, and performing germination culture, proliferation culture, and rooting culture, respectively, to obtain sterile Dendrobium officinale seedlings;
[0012] (2) performing callus induction culture or adventitious root induction culture on the sterile seedlings of Dendrobium officinale obtained in step (1) to obtain adventitious roots of Dendrobium officinale;
[0013] The method of callus induction culture is:
[0014] a. Inoculating the leaves, petioles or stem segments of the sterile seedlings of Dendrobium officinale described in step (1) into callus induction medium and callus proliferation medium in sequence, and performing light culture and dark culture respectively to obtain Dendrobium officinale callus;
[0015] b. Cutting the Dendrobium officinale callus described in step a and inoculating it into an adventitious root induction medium for dark culture to obtain Dendrobium officinale adventitious roots;
[0016] The method for the adventitious root induction culture is:
[0017] The leaves, petioles or stem segments of the sterile Dendrobium officinale seedlings in step (1) are inoculated into an adventitious root culture medium to perform adventitious root culture to obtain Dendrobium officinale adventitious roots.
[0018] Preferably, the starting culture medium in step (1) is: MS+agar 7.0-7.5 g / L+sucrose 25-30 g / L, with a pH of 5.8-6.0.
[0019] The proliferation medium is MS+agar 7.0-7.5 g / L+6-BA 1.5-2.0 mg / L+NAA 0.1-0.2 mg / L+sucrose 25-30 g / L, with a pH of 5.8-6.0:
[0020] The seedling rooting culture medium comprises: 1 / 2MS+agar 7.0-7.5 g / L+IBA 0.2-0.5 mg / L+activated carbon 0.2-0.5 g / L+sucrose 25-30 g / L, and the pH value is 5.8-6.0.
[0021] Preferably, the germination culture time in step (1) is 10 to 16 days; the proliferation culture time is 20 to 25 days; and the rooting culture time is 15 to 25 days.
[0022] Preferably, the callus induction medium in step a is MS+2,4-D 2.0-3.0 mg / L+1.0-2.0 mg / L NAA+sucrose 30-40 g / L+agar 7.2-7.6 g / L, pH=5.8-6.0.
[0023] The callus proliferation medium comprises MS, 1.0-1.5 mg / L 6-BA, 0.1-0.2 mg / L 2,4-D, 30-40 g / L sucrose, and 7.2-7.6 g / L agar, with a pH of 5.8-6.0.
[0024] Preferably, the light culture time in step a is 10 to 15 days, and the light intensity is 2500 to 3000 lx; the light-avoidance culture time is 20 to 30 days.
[0025] Preferably, the adventitious root induction medium in step b is: MS + agar 7.2-7.6 g / L + 1.5-2.5 mg / L NAA + sucrose 25-30 g / L, pH = 5.8-6.0.
[0026] Preferably, the adventitious root induction culture time in step b is 10 to 18 days.
[0027] Preferably, the adventitious root culture medium is MS+agar 7.2-7.6 g / L+NAA 0.5-0.8 mg / L+banana juice 15%+sucrose 30-40 g / L, pH=5.8-6.0; the adventitious root culture is kept away from light for 30-40 days.
[0028] Preferably, after obtaining the adventitious roots of Dendrobium officinale, the method further comprises inoculating the adventitious roots of Dendrobium officinale into an adventitious root proliferation culture medium for adventitious root proliferation culture to obtain a large number of adventitious roots of Dendrobium officinale.
[0029] Preferably, the adventitious root proliferation culture method is to first culture in the dark for 30 to 40 days, and then culture under 2500 to 3000 lx light for 10 to 20 days; the adventitious root proliferation culture medium is MS + agar 7.2 to 7.6 g / L + NAA 0.5 to 0.8 mg / L + banana juice 15% + sucrose 30 to 40 g / L, pH = 5.8 to 6.0.
[0030] Compared with the prior art, the present invention has the following beneficial effects:
[0031] The invention utilizes a variety of explants (callus tissue, leaves, protocorms, and stem segments) of sterile Dendrobium officinale seedlings to induce adventitious roots and then performs propagation culture. The induction rate of the adventitious roots is between 72.30% and 88.74%, laying a foundation for the future industrial production of important active ingredients of Dendrobium officinale by utilizing the adventitious roots of Dendrobium officinale, and simultaneously solving the contradiction between supply and demand of Dendrobium officinale raw materials. BRIEF DESCRIPTION OF THE DRAWINGS
[0032] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are merely embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying any creative work.
[0033] Figure 1 These are sterile seedlings of Dendrobium officinale.
[0034] Figure 2 It is the adventitious root of Dendrobium officinale. DETAILED DESCRIPTION
[0035] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.
[0036] Example 1
[0037] The present invention provides a method for cultivating adventitious roots of Dendrobium officinale, comprising the following steps:
[0038] (1) Disinfect the seeds of Dendrobium officinale with 70% alcohol by volume for 30 seconds, rinse twice with water, and then disinfect with 0.08% mercuric chloride for 6 minutes, and rinse twice with water.
[0039] (2) inoculating the Dendrobium officinale seeds into a start-up culture medium, a proliferation culture medium, and a seedling rooting culture medium in sequence, and performing germination culture for 10 days, proliferation culture for 20 days, and rooting culture for 15 days, respectively, to obtain sterile Dendrobium officinale seedlings;
[0040] The starting culture medium is: MS+agar 7.0 g / L+sucrose 25 g / L, with a pH of 5.8.
[0041] The proliferation medium is MS+agar 7.0 g / L+6-BA 1.5 mg / L+NAA 0.1 mg / L+sucrose 25 g / L, pH 5.8:
[0042] The seedling rooting medium is: 1 / 2MS+agar 7.0g / L+IBA 0.2mg / L+activated carbon 0.2g / L+sucrose 25g / L, with a pH value of 5.8.
[0043] (3) The leaves of the sterile seedlings of Dendrobium officinale (0.4 cm × 0.4 cm in size) were inoculated into callus induction medium and callus proliferation medium in sequence, and cultured under 2500 lx light for 1 day and dark for 20 days, respectively, to obtain Dendrobium officinale callus;
[0044] The callus induction medium is MS+2,4-D 2.0 mg / L+1.0 mg / L NAA+sucrose 30 g / L+agar 7.2 g / L, pH=5.8.
[0045] The callus proliferation medium MS+6-BA 1.0 mg / L+2,4-D 0.1 mg / L+sucrose 30 g / L+agar 7.2 g / L, pH=5.8.
[0046] (4) cutting the Dendrobium officinale callus and inoculating it into an adventitious root induction medium for 10 days in the dark to obtain the adventitious roots of Dendrobium officinale;
[0047] The adventitious root induction medium is: MS+agar 7.2g / L+1.5mg / L NAA+sucrose 25g / L, pH=5.8;
[0048] (5) The adventitious roots of Dendrobium officinale were cut into 1 cm lengths and inoculated into adventitious root proliferation medium for adventitious root proliferation culture, first cultured in the dark for 30 days, and then cultured in 2500 lx light for 10 days to obtain a large number of Dendrobium officinale adventitious roots.
[0049] The adventitious root proliferation culture medium is MS+agar 7.2 g / L+NAA 0.5 mg / L+banana juice 15%+sucrose 30 g / L, with a pH of 5.8.
[0050] Example 2
[0051] The present invention provides a method for cultivating adventitious roots of Dendrobium officinale, comprising the following steps:
[0052] (1) Disinfect the seeds of Dendrobium officinale with 80% alcohol by volume for 40 seconds, rinse with water three times, and then disinfect with 0.12% mercuric chloride for 8 minutes, and rinse with water three times.
[0053] (2) inoculating the Dendrobium officinale seeds into a start-up culture medium, a proliferation culture medium, and a seedling rooting culture medium in sequence, and performing germination culture for 16 days, proliferation culture for 25 days, and rooting culture for 25 days, respectively, to obtain sterile Dendrobium officinale seedlings;
[0054] The starting culture medium is: MS+agar 7.5 g / L+sucrose 30 g / L, with a pH of 6.0.
[0055] The proliferation medium is MS+agar 7.5 g / L+6-BA 2.0 mg / L+NAA 0.15 mg / L+sucrose 30 g / L, pH 6.0:
[0056] The seedling rooting medium is: 1 / 2MS+agar 7.5g / L+IBA 0.5mg / L+activated carbon 0.5g / L+sucrose 30g / L, with a pH value of 6.0.
[0057] (3) The petioles (0.5 cm segments) of the sterile seedlings of Dendrobium officinale were inoculated into callus induction medium and callus proliferation medium in sequence, and cultured under 3000 lx light for 15 days and in the dark for 25 days, respectively, to obtain Dendrobium officinale callus;
[0058] The callus induction medium is MS+2,4-D 3.0 mg / L+1.5 mg / L NAA+sucrose 40 g / L+agar 7.6 g / L, pH=6.0.
[0059] The callus proliferation medium MS+6-BA 1.5 mg / L+2,4-D 0.2 mg / L+sucrose 40 g / L+agar 7.6 g / L, pH=6.0.
[0060] (4) cutting the Dendrobium officinale callus and inoculating it into an adventitious root induction medium for 18 days in the dark to obtain the adventitious roots of Dendrobium officinale;
[0061] The adventitious root induction medium is: MS+agar 7.6 g / L+2.5 mg / L NAA+sucrose 30 g / L, pH=6.0;
[0062] (5) The adventitious roots of Dendrobium officinale were cut into 1.5 cm lengths and inoculated into adventitious root proliferation medium for adventitious root proliferation culture, first cultured in the dark for 40 days, and then cultured at 3000 lx light for 20 days to obtain a large number of Dendrobium officinale adventitious roots.
[0063] The adventitious root proliferation culture medium is MS+agar 7.6 g / L+NAA 0.8 mg / L+banana juice 15%+sucrose 40 g / L, with a pH of 6.0.
[0064] Example 3
[0065] The present invention provides a method for cultivating adventitious roots of Dendrobium officinale, comprising the following steps:
[0066] (1) Disinfect the seeds of Dendrobium officinale with 75% alcohol by volume for 35 seconds, rinse with water three times, and then disinfect with 0.1% mercuric chloride for 7 minutes and rinse with water three times.
[0067] (2) inoculating the Dendrobium officinale seeds into a start-up culture medium, a proliferation culture medium, and a seedling rooting culture medium in sequence, and performing germination culture for 13 days, proliferation culture for 23 days, and rooting culture for 20 days, respectively, to obtain sterile Dendrobium officinale seedlings;
[0068] The starting culture medium is: MS+agar 7.3 g / L+sucrose 28 g / L, with a pH of 5.9.
[0069] The proliferation medium is MS+agar 7.3g / L+6-BA 1.8mg / L+NAA 0.15mg / L+sucrose 28g / L, pH 5.9:
[0070] The seedling rooting medium is: 1 / 2MS+agar 7.3g / L+IBA 0.3mg / L+activated carbon 0.4g / L+sucrose 28g / L, with a pH value of 5.9.
[0071] (3) The stem segments (0.7 cm segments) of the sterile seedlings of Dendrobium officinale were inoculated into callus induction medium and callus proliferation medium in sequence, and cultured under 2800 lx light for 13 days and dark culture for 25 days, respectively, to obtain Dendrobium officinale callus;
[0072] The callus induction medium is MS+2,4-D 2.5 mg / L+1.5 mg / L NAA+sucrose 35 g / L+agar 7.4 g / L, pH=5.9.
[0073] The callus proliferation medium MS+6-BA 1.3 mg / L+2,4-D 0.15 mg / L+sucrose 35 g / L+agar 7.4 g / L, pH=5.9.
[0074] (4) cutting the Dendrobium officinale callus and inoculating it into an adventitious root induction medium for 14 days in the dark to obtain the Dendrobium officinale adventitious roots;
[0075] The adventitious root induction medium is: MS+agar 7.4 g / L+2 mg / L NAA+sucrose 28 g / L, pH=5.9;
[0076] (5) The adventitious roots of Dendrobium officinale were cut into 1.3 cm lengths and inoculated into adventitious root proliferation medium for adventitious root proliferation culture, first cultured in the dark for 35 days, and then cultured in 2800 lx light for 15 days to obtain a large number of Dendrobium officinale adventitious roots.
[0077] The adventitious root proliferation culture medium is MS+agar 7.4 g / L+NAA 0.7 mg / L+banana juice 15%+sucrose 35 g / L, with a pH of 5.9.
[0078] Example 4
[0079] The present invention provides a method for cultivating adventitious roots of Dendrobium officinale, comprising the following steps:
[0080] (1) Disinfect the seeds of Dendrobium officinale with 70% alcohol by volume for 30 seconds, rinse twice with water, and then disinfect with 0.08% mercuric chloride for 6 minutes, and rinse twice with water.
[0081] (2) inoculating the Dendrobium officinale seeds into a start-up culture medium, a proliferation culture medium, and a seedling rooting culture medium in sequence, and performing germination culture for 10 days, proliferation culture for 20 days, and rooting culture for 15 days, respectively, to obtain sterile Dendrobium officinale seedlings;
[0082] The starting culture medium is: MS+agar 7.0 g / L+sucrose 25 g / L, with a pH of 5.8.
[0083] The proliferation medium is MS+agar 7.0 g / L+6-BA 1.5 mg / L+NAA 0.1 mg / L+sucrose 25 g / L, pH 5.8:
[0084] The seedling rooting medium is: 1 / 2MS+agar 7.0g / L+IBA 0.2mg / L+activated carbon 0.2g / L+sucrose 25g / L, with a pH value of 5.8.
[0085] (3) inoculating the leaves of the sterile seedlings of Dendrobium officinale (0.4 cm×0.4 cm in size) into an adventitious root culture medium to obtain adventitious roots of Dendrobium officinale;
[0086] The adventitious root culture medium is MS+agar 7.2 g / L+NAA 0.5 mg / L+banana juice 15%+sucrose 30 g / L, pH=5.8; the adventitious root culture is carried out in the dark for 30 days.
[0087] (4) The adventitious roots of Dendrobium officinale were cut into 1 cm lengths and inoculated into adventitious root proliferation medium for adventitious root proliferation culture, first cultured in the dark for 30 days, and then cultured at 2500 lx light for 10 days to obtain a large number of Dendrobium officinale adventitious roots.
[0088] The adventitious root proliferation culture medium is MS+agar 7.2 g / L+NAA 0.5 mg / L+banana juice 15%+sucrose 30 g / L, with a pH of 5.8.
[0089] Example 5
[0090] The present invention provides a method for cultivating adventitious roots of Dendrobium officinale, comprising the following steps:
[0091] (1) Disinfect the seeds of Dendrobium officinale with 80% alcohol by volume for 40 seconds, rinse with water three times, and then disinfect with 0.12% mercuric chloride for 8 minutes, and rinse with water three times.
[0092] (2) inoculating the Dendrobium officinale seeds into a start-up culture medium, a proliferation culture medium, and a seedling rooting culture medium in sequence, and performing germination culture for 16 days, proliferation culture for 25 days, and rooting culture for 25 days, respectively, to obtain sterile Dendrobium officinale seedlings;
[0093] The starting culture medium is: MS+agar 7.5 g / L+sucrose 30 g / L, with a pH of 6.0.
[0094] The proliferation medium is MS+agar 7.5 g / L+6-BA 2.0 mg / L+NAA 0.15 mg / L+sucrose 30 g / L, pH 6.0:
[0095] The seedling rooting medium is: 1 / 2MS+agar 7.5g / L+IBA 0.5mg / L+activated carbon 0.5g / L+sucrose 30g / L, with a pH value of 6.0.
[0096] (3) inoculating the petioles (0.5 cm segments) of the sterile Dendrobium officinale seedlings into an adventitious root culture medium to obtain adventitious roots of Dendrobium officinale;
[0097] The adventitious root culture medium is MS+agar 7.6 g / L+NAA 0.8 mg / L+banana juice 15%+sucrose 40 g / L, pH=6.0; the adventitious root culture is carried out in the dark for 40 days.
[0098] (4) The adventitious roots of Dendrobium officinale were cut into 1.5 cm lengths and inoculated into adventitious root proliferation medium for adventitious root proliferation culture, first cultured in the dark for 40 days, and then cultured at 3000 lx light for 20 days to obtain a large number of Dendrobium officinale adventitious roots.
[0099] The adventitious root proliferation culture medium is MS+agar 7.6 g / L+NAA 0.8 mg / L+banana juice 15%+sucrose 40 g / L, with a pH of 6.0.
[0100] Example 6
[0101] The present invention provides a method for cultivating adventitious roots of Dendrobium officinale, comprising the following steps:
[0102] (1) Disinfect the seeds of Dendrobium officinale with 75% alcohol by volume for 35 seconds, rinse with water three times, and then disinfect with 0.1% mercuric chloride for 7 minutes and rinse with water three times.
[0103] (2) inoculating the Dendrobium officinale seeds into a start-up culture medium, a proliferation culture medium, and a seedling rooting culture medium in sequence, and performing germination culture for 13 days, proliferation culture for 23 days, and rooting culture for 20 days, respectively, to obtain sterile Dendrobium officinale seedlings;
[0104] The starting culture medium is: MS+agar 7.3 g / L+sucrose 28 g / L, with a pH of 5.9.
[0105] The proliferation medium is MS+agar 7.3g / L+6-BA 1.8mg / L+NAA 0.15mg / L+sucrose 28g / L, pH 5.9:
[0106] The seedling rooting medium is: 1 / 2MS+agar 7.3g / L+IBA 0.3mg / L+activated carbon 0.4g / L+sucrose 28g / L, with a pH value of 5.9.
[0107] (3) inoculating the stem segments (0.7 cm segments) of the sterile seedlings of Dendrobium officinale into an adventitious root culture medium to perform adventitious root culture to obtain adventitious roots of Dendrobium officinale;
[0108] The adventitious root culture medium is MS+agar 7.4 g / L+NAA 0.7 mg / L+banana juice 15%+sucrose 35 g / L, pH=5.9; the adventitious root culture is carried out in the dark for 35 days.
[0109] (4) The adventitious roots of Dendrobium officinale were cut into 1.3 cm in length and inoculated into adventitious root proliferation medium for adventitious root proliferation culture, first cultured in the dark for 35 days, and then cultured in 2800 lx light for 15 days to obtain a large number of Dendrobium officinale adventitious roots.
[0110] The adventitious root proliferation culture medium is MS+agar 7.4 g / L+NAA 0.7 mg / L+banana juice 15%+sucrose 35 g / L, with a pH of 5.9.
[0111] 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 cultivating adventitious roots of Dendrobium officinale, characterized in that: The steps include: (1) inoculating the Dendrobium officinale seeds into a start-up culture medium, a proliferation culture medium, and a seedling rooting culture medium in sequence, and performing germination culture, proliferation culture, and rooting culture, respectively, to obtain sterile Dendrobium officinale seedlings; (2) performing adventitious root induction culture on the sterile seedlings of Dendrobium officinale obtained in step (1) to obtain adventitious roots of Dendrobium officinale; The method for the adventitious root induction culture is: The leaves of the sterile Dendrobium officinale seedlings in step (1) are inoculated into an adventitious root culture medium to perform adventitious root culture to obtain Dendrobium officinale adventitious roots.
2. The method for cultivating adventitious roots of Dendrobium officinale according to claim 1, wherein: The starting medium in step (1) is: MS + agar 7.0-7.5 g / L + sucrose 25-30 g / L, pH 5.8-6.0; The proliferation medium is MS+agar 7.0-7.5 g / L+6-BA 1.5-2.0 mg / L+NAA 0.1-0.2 mg / L+sucrose 25-30 g / L, with a pH of 5.8-6.0: The seedling rooting culture medium comprises: 1 / 2MS+agar 7.0-7.5 g / L+IBA 0.2-0.5 mg / L+activated carbon 0.2-0.5 g / L+sucrose 25-30 g / L, and the pH value is 5.8-6.
0.
3. The method for cultivating adventitious roots of Dendrobium officinale according to claim 2, wherein: The time for the germination culture in step (1) is 10 to 16 days; the time for the proliferation culture is 20 to 25 days; and the time for the rooting culture is 15 to 25 days.
4. The method for cultivating adventitious roots of Dendrobium officinale according to claim 1, wherein: The adventitious root culture medium comprises MS+agar 7.2-7.6 g / L+NAA 0.5-0.8 mg / L+banana juice 15%+sucrose 30-40 g / L, with a pH of 5.8-6.
0. The adventitious root culture is carried out in a dark environment for 30-40 days.
5. The method for cultivating adventitious roots of Dendrobium officinale according to claim 1, wherein: After obtaining the adventitious roots of Dendrobium officinale, the method further includes inoculating the adventitious roots of Dendrobium officinale into an adventitious root proliferation culture medium for adventitious root proliferation culture to obtain a large number of adventitious roots of Dendrobium officinale.
6. The method for cultivating adventitious roots of Dendrobium officinale according to claim 5, wherein: The adventitious root proliferation culture method comprises the following steps: firstly culturing in darkness for 30 to 40 days, and then culturing under 2500 to 3000 lx light for 10 to 20 days; the adventitious root proliferation culture medium comprises MS+agar 7.2 to 7.6 g / L+NAA 0.5 to 0.8 mg / L+banana juice 15%+sucrose 30 to 40 g / L, with a pH of 5.8 to 6.0.