Methods for rapid propagation of Cymbidium orchids

Abstract

The present invention discloses a method for rapid propagation of Chinese orchids, comprising selecting a new bud with unopened leaves as an explant mother plant, cutting it vertically close to the base of a pseudobulb, and sterilizing the pseudobulb; inoculating the sterilized pseudobulb into an axillary bud induction medium, culturing in a dark environment, and inducing buds; excising the induced buds, inoculating the pseudobulbs after excision into a rhizome induction medium, culturing in a dark environment, and inducing rhizomes; proliferating and culturing the rhizomes, allowing them to differentiate and sprout, continuing to cultivate until the small buds grow into seedlings, and then performing rooting culture on the strong seedlings. The method for rapid propagation of Chinese orchids of the present invention avoids cutting the pseudobulbs in the early stage of sterilization, significantly reduces the mortality rate and browning rate, improves the induction success rate of Chinese orchid cut buds, and improves the rhizome proliferation rate and budding rate, thereby achieving rapid propagation of Chinese orchids and meeting the needs of commercial production of Chinese orchids.

Description

Technical Field

[0001] The invention belongs to the technical field of plant cultivation, and particularly relates to a method for rapidly propagating Cymbidium orchids by utilizing pseudobulbs to induce rhizomes. Background Art

[0002] Orchids, also known as Chinese orchids and known as orchids in ancient times, have a long history of cultivation in my country. Ancient people used the phrase "orchid chapter" to describe the beauty of poetry and prose, and "orchid jiao" to symbolize the true nature of friendship, and numerous poems and lyrics extol orchids. Orchids belong to the genus Orchid, a monocotyledonous perennial plant that grows as epiphytes or terrestrial herbs. They include seven species: Spring Orchid (C. goeringii), Hui Orchid (C. faberi), Jian Orchid (C. ensifolum), Black Orchid (C. sinense), Cold Orchid (C. kanran), Lotus Orchid (C. tortisepalum), and Spring Sword Orchid (C. tortisepalum var. longibracteatum). They have a unique fragrance and elegant color, making them highly ornamental.

[0003] The embryo of Chinese orchid seeds is incompletely developed, lacking endosperm and cotyledons. This makes germination difficult under natural conditions. Conventional propagation often relies on division, which is slow. Furthermore, long-term division can easily lead to the spread of viruses, which can degrade the characteristics of the species. Consequently, despite my country's long history of orchid cultivation and the domestication of many valuable orchids, achieving a critical mass and promoting their widespread availability has long limited the utilization of my country's high-quality orchid resources.

[0004] Tissue culture technology can rapidly produce and propagate superior orchid varieties, saving rare and endangered species and meeting the demands of large-scale commercial production. Cymbidium tissue culture is primarily divided into seed propagation and bud cutting. Seed propagation offers the advantage of easy pod disinfection, but it also suffers from a long propagation cycle and high rates of offspring variation. Bud cutting, on the other hand, produces seedlings with less variation and consistent growth, ensuring genetic stability, making it crucial for the propagation of superior varieties and the selection of new ones.

[0005] Existing methods for cutting orchid buds primarily use new buds as explants. The parent plant is first pre-treated with a disinfectant (sodium hypochlorite) (watered for about a week), then the new buds are cut and disinfected. After disinfection, the bud tip or stem tip growth point is directly cut to induce rhizome regeneration. Because the bud tissue is relatively tender, the explants experience high rates of browning and mortality after disinfection and cutting, resulting in a low success rate for cutting buds. Summary of the Invention

[0006] Based on this, one of the purposes of the present invention is to provide a method for rapid propagation of Cymbidium orbiculare, which can improve the success rate of inducing rhizomes from Cymbidium orbiculare explants and achieve rapid propagation of Cymbidium orbiculare.

[0007] The specific technical solutions for achieving the above-mentioned invention objectives include the following:

[0008] A method for rapid propagation of Cymbidium orchids, comprising the following steps:

[0009] (1) Select a new bud with unopened leaves as the explant mother plant, cut it vertically close to the base of the pseudobulb, and disinfect the pseudobulb;

[0010] (2) inoculating the disinfected pseudobulbs into axillary bud induction medium and culturing in a dark environment to induce budding;

[0011] (3) removing the buds induced in step (2), inoculating the pseudobulbs after the buds are removed into a rhizome induction medium, and inducing rhizomes in a dark environment;

[0012] (4) After the rhizomes are proliferated and cultured, they differentiate into buds, and continue to be cultured until the buds grow into seedlings, and then culture them to strengthen the seedlings and allow them to take root.

[0013] In some embodiments, the height of the new shoots in step (1) is 1 / 4 to 1 / 3 of the height of the mature plant.

[0014] In some embodiments, the method for disinfecting the pseudobulb described in step (1) comprises the following steps: removing the fleshy roots of the pseudobulb and the dark brown old tissue at the base, and peeling off 2 to 3 scaly bracts on the outside of the bud; after rinsing with running water, disinfecting with 75%±5% alcohol by shaking for 30±5 seconds, and washing with sterile water by shaking for 2 to 3 times, each time for 5±1 minutes; then disinfecting with 15%±3% bleach by shaking for 15±3 minutes; then peeling off the remaining bracts, and disinfecting with 7%±1% bleach by shaking for 5±1 minutes; washing with sterile water by shaking for 2 to 3 times, each time for 5±1 minutes; and finally placing the rinsed pseudobulb on sterile filter paper and air-drying.

[0015] In some embodiments, the culture temperature in step (2) is 25±2° C., and the culture time is 21 to 23 days.

[0016] In some embodiments, the rhizome induction medium in step (3) comprises: MS, 2.0 mg / L to 2.1 mg / L NAA, 25 mg / L to 35 mg / L sucrose, 0.45 mg / L to 0.55 mg / L activated carbon, 190 ml / L to 210 L coconut milk, and 7 mg / L to 9 mg / L agar.

[0017] In some embodiments, the culture temperature in step (3) is 25±2° C., and the culture time is 1 to 3 months.

[0018] In some embodiments, the culture medium for the proliferation culture in step (4) includes: MS, 1.45 mg / L to 1.55 mg / L NAA, 0.45 mg / L to 0.55 mg / L 6-BA, 1.55 to 2.05 mg / L peptone, 25 mg / L to 35 mg / L sucrose, 0.45 mg / L to 0.55 mg / L activated carbon, and 7 mg / L to 9 mg / L agar; and / or, the conditions for the proliferation culture are: culture temperature 25±2°C, and culture time 1 to 2 months.

[0019] In some embodiments, the culture medium for differentiation and budding in step (4) comprises: MS, 0.75 mg / L to 0.85 mg / L NAA, 3.95 mg / L to 4.05 mg / L 6-BA, 25 mg / L to 35 mg / L sucrose, 0.45 mg / L to 0.55 mg / L activated carbon, 190 ml / L to 210 ml / L coconut milk, 320 mg / L to 340 mg / L KH2PO4, and 7 mg / L to 9 mg / L agar; and / or, the culture conditions for differentiation and budding are: culture temperature 25±2°C, light intensity 45±5 μmol / (m·s -2 ), photoperiod 12±2h / d, culture time 25-35 days.

[0020] In some embodiments, the culture medium for rooting the seedlings in step (4) comprises: Huabao No. 1, 0.95 mg / L to 1.05 mg / L NAA, 1.95 mg / L to 2.05 mg / L peptone, 25 mg / L to 35 mg / L sucrose, 100 mg / L to 150 mg / L banana, 0.45 mg / L to 0.55 mg / L activated carbon, and 7 mg / L to 9 mg / L agar; and / or, the culture conditions for rooting the seedlings are: culture temperature 25±2°C, light intensity 45±5 μmol / (m·s -2 ), photoperiod 12±2h / d, culture time 60 to 90 days.

[0021] In some embodiments, the Chinese orchid is Chunlan, Huilan, Jianlan, Molan, Hanlan, Lianbanlan or Chunjian.

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

[0023] The method for rapid propagation of Cymbidium orchids of the present invention comprises the following steps: selecting new buds as explant mother plants, cutting off pseudobulbs, inducing the pseudobulbs to sprout first, cutting off the buds, and then inducing rhizomes; and simultaneously coordinating the culture medium formulas at different stages of the regeneration process. Under this concept, cutting of the pseudobulbs in the early stage of disinfection is avoided, the mortality rate and browning rate are greatly reduced, the induction success rate of the Cymbidium orchid cut buds is improved, the rhizome proliferation rate and sprouting rate are improved, the rapid propagation of Cymbidium orchids is achieved, and the demand for commercial production of Cymbidium orchids is met. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 The present invention is the pseudobulb after disinfection in Example 1.

[0025] Figure 2 This is the bud induced from the pseudobulb in Example 1 of the present invention.

[0026] Figure 3 This is the rhizome induced in Example 1 of the present invention.

[0027] Figure 4 This is the proliferation of rhizomes in Example 1 of the present invention.

[0028] Figure 5 This is the sprouting of rhizome differentiation in Example 1 of the present invention.

[0029] Figure 6 The invention provides a method for strengthening the seedlings and taking root in Example 1. DETAILED DESCRIPTION

[0030] To facilitate understanding of the present invention, the present invention will be described more fully below. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the understanding of the present disclosure more thorough and comprehensive.

[0031] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as commonly understood by those skilled in the art to which this invention pertains. The terms used in this specification are for the purpose of describing specific embodiments only and are not intended to limit the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0032] The present invention provides a method for rapidly propagating Cymbidium orchids by inducing rhizomes from pseudobulbs, comprising the following steps:

[0033] (1) Selection and disinfection of explants

[0034] Select robust, disease-free orchids, and new buds with unopened leaves of moderate size as explants (the larger the size, the more difficult it is to disinfect and the higher the contamination rate. If the size is too small, it will be more affected by the damage of the disinfectant and the lower the survival rate of the explant. Take new buds with unopened leaves and a height of one-third of the size of the adult plant as explants, which can reduce the contamination rate and ensure the survival rate of the explants). Use a clean and sharp scalpel to close to the base of the pseudobulb, cut the bud vertically and ensure the integrity of the bud (compared with the existing orchid bud cutting technology, there is no need to pre-treat the explant mother plant, which reduces the damage caused by the pre-treatment to the mother plant). Remove the fleshy roots of the pseudobulb and the dark brown old tissue at the base, and peel off 2 to 3 scaly bracts on the outside of the bud; rinse with running water to remove dust attached to the bud; then move it into a clean bench, shake and disinfect it with 75% ± 5% alcohol for 30 ± 5 seconds, and shake and rinse it twice with sterile water for 5 ± 1 minutes each; then shake and disinfect it with 15% ± 3% bleach for 15 ± 3 minutes; then peel off the remaining bracts, shake and disinfect it with 7% ± 1% bleach for 5 ± 1 minute; shake and rinse it with sterile water 2 to 3 times for 5 ± 1 minute each; finally, place the rinsed pseudobulb on sterile filter paper and blow dry.

[0035] (2) Bud induction

[0036] The pseudobulbs are inoculated into an axillary bud induction medium and cultured in a dark environment at a temperature of 25±2°C. After one week of culture, multiple milky white buds are induced from the pseudobulbs. After 15±1 days of culture, buds 0.5-1 cm in height are formed. The axillary bud induction medium contains the following components: MS, 0.45mg / L-0.55mg / L NAA, 0.45mg / L-0.55mg / L 6-BA, 25mg / L-35mg / L sucrose, 0.45mg / L-0.55mg / L activated carbon, 190ml / L-210ml / L coconut milk, and 7-9mg / L agar (after preparation, adjust the pH to 5.4-5.6 and sterilize at 121±2°C, 15-20 psi for 15-25 minutes).

[0037] (3) Induction of rhizomes

[0038] After buds are induced, the induced buds are excised, and the pseudobulbs after bud excision are inoculated into a rhizome induction medium, cultured in a dark environment for 1 to 3 months, and rhizomes are induced from the incisions; the rhizome induction medium contains the following components: MS, 2.0 mg / L to 2.1 mg / L NAA, 25 mg / L to 35 mg / L sucrose, 0.45 mg / L to 0.55 mg / L activated carbon, 190 ml / L to 210 L coconut milk, and 7 mg / L to 9 mg / L agar (after preparation, the pH is adjusted to 5.4-5.6, and the mixture is sterilized at 121±2° C. and 15-20 pounds for 20±5 minutes).

[0039] (4) Rhizome proliferation

[0040] The induced rhizomes are transferred to a proliferation medium for proliferation culture. The components of the rhizome proliferation medium are: MS, 1.45mg / L-1.55mg / L NAA, 0.45mg / L-0.55mg / L 6-BA, 1.55-2.05mg / L peptone, 25mg / L-35mg / L sucrose, 0.45mg / L-0.55mg / L activated carbon, and 7mg / L-9mg / L agar (after preparation, adjust the pH to 5.4-5.6 and sterilize at 121±2°C, 15-20 psi for 20±5 minutes).

[0041] (5) Differentiation and budding of rhizomes

[0042] The proliferated rhizomes were transferred to differentiation medium at a temperature of 25 ± 2 °C and a light intensity of 45 ± 5 μmol / (m·s -2 ) with a photoperiod of 12 ± 2 h / d. After 25 to 35 days of culture, small green shoots will differentiate from the rhizomes. Continue culturing until the shoots are approximately 3 cm long. The shoot induction medium contains the following components: MS, 0.75 mg / L to 0.85 mg / L NAA, 3.95 mg / L to 4.05 mg / L 6-BA, 25 mg / L to 35 mg / L sucrose, 0.45 mg / L to 0.55 mg / L activated carbon, 190 ml / L to 210 ml / L coconut water, 320 mg / L to 340 mg / L KH2PO4, and 7 mg / L to 9 mg / L agar.

[0043] (6) Strong seedlings and rooting

[0044] The rootless seedlings with a height of 3 to 5 cm were inoculated into the seedling rooting medium for seedling rooting culture. The culture temperature was 25 ± 2 ° C and the light intensity was maintained at 45 ± 5 μmol / (m·s -2 ), with a photoperiod of 12 ± 2 h / d. The seedling medium components are: Huabao No. 1, 0.95 mg / L–1.05 mg / L NAA, 1.95 mg / L–2.05 mg / L peptone, 25 mg / L–35 mg / L sucrose, 100 mg / L–150 mg / L banana, 0.45 mg / L–0.55 mg / L activated carbon, and 7 mg / L–9 mg / L agar.

[0045] The method for rapidly propagating Chinese orchids by utilizing pseudobulbs to induce rhizomes is applicable to all orchid varieties such as Chunlan, Huilan, Jianlan, Molan, Hanlan, Lianbanlan and Chunjian.

[0046] The present invention will be described in detail below with reference to specific embodiments and accompanying drawings.

[0047] Example 1 A method for rapidly propagating Cymbidium orchids by inducing rhizomes using pseudobulbs

[0048] This embodiment uses black orchid as material and uses pseudobulbs to induce rhizomes to quickly propagate Chinese orchids, specifically including the following steps:

[0049] (1) Selection and disinfection of explants

[0050] Select a healthy, disease-free, and insect-free black orchid, take a new bud with unopened leaves and a height of one-third of the size of the adult plant as the explant mother plant, use a clean and sharp scalpel close to the base of the pseudobulb, cut the bud vertically and ensure the integrity of the bud, remove the pseudobulb fleshy root and the dark brown old tissue at the base, peel off 2 to 3 scale-like bracts outside the bud; rinse with running water to remove dust attached to the bud; then move it into a clean bench, shake and disinfect with 75% alcohol for 30 seconds, shake and rinse twice with sterile water for 5 minutes each; then shake and disinfect with 15% bleach for 15 minutes; then peel off the remaining bracts, shake and disinfect with 7% bleach for 5 minutes; shake and rinse with sterile water 2 to 3 times for 5 minutes each; finally, place the rinsed pseudobulb on sterile filter paper and blow dry ( Figure 1 for the sterilized pseudobulbs).

[0051] (2) Bud induction

[0052] The pseudobulbs were inoculated into axillary bud induction medium (MS + 0.5 mg / L NAA + 0.5 mg / L 6-BA + 30 mg / L sucrose + 0.5 mg / L activated carbon + 200 ml / L coconut milk + 8 mg / L agar) and cultured in a dark environment at 25°C. After 1 week of culture, multiple milky white buds were induced from the pseudobulbs. After 15 days, buds with a height of 0.5 to 1 cm were induced. Figure 2 ).

[0053] (3) Induction of rhizomes

[0054] After the buds were induced, the buds were cut off and the pseudobulbs were inoculated into rhizome induction medium (MS, 2 mg / L NAA, 30 mg / L sucrose, 0.5 mg / L activated carbon, 200 ml / L coconut milk, 8 mg / L agar) and cultured in the dark for 3 months. Rhizomes were induced from the incision ( Figure 3 ).

[0055] (4) Rhizome proliferation

[0056] The induced rhizomes were transferred to proliferation medium (MS+1.5mg / L NAA+0.5mg / L 6-BA+2mg / L peptone+30mg / L sucrose+0.5mg / L activated carbon+8mg / L agar) for proliferation culture ( Figure 4 ).

[0057] (5) Differentiation and budding of rhizomes

[0058] The proliferated rhizomes were transferred to differentiation budding medium (MS + 0.8 mg / L NAA + 4 mg / L 6-BA + 30 mg / L sucrose + 0.5 mg / L activated carbon + 200 ml / L coconut milk + 340 mg / L KH2PO4 + 8 mg / L agar), cultured at 25 °C and maintained at a light intensity of 45 μmol / (m·s -2 ) and the photoperiod is 12h / d. After one month of cultivation, the rhizomes differentiate into green buds. Continue to cultivate until the buds grow to about 3cm in length. Figure 5 ).

[0059] (6) Strong seedlings and rooting

[0060] Rootless seedlings with a height of 3-5 cm were inoculated into a seedling rooting medium (Huabao No. 1 + 1 mg / L NAA + 2 mg / L peptone + 30 mg / L sucrose + 100 mg / L banana + 0.5 mg / L activated carbon + 8 mg / L agar) for seedling rooting culture. The culture temperature was 25 ° C and the light intensity was maintained at 45 μmol / (m·s -2 ), the photoperiod is 12h / d.

[0061] After 45 days of cultivation, the rooting rate reaches 98%, the length can reach 3cm to 5cm, and the root system is strong ( Figure 6 ).

[0062] Experimental Example 1 Optimization of treatment scheme for rhizome induction

[0063] To investigate the optimal treatment regimen for rhizome induction, sterilized explants (new shoots) were subjected to different treatments for rhizome induction (Example 1, Step 3). After three months of treatment, the induction rate, mortality rate, and browning rate of the explants were statistically analyzed. Each treatment was repeated three times, with 50 explants treated each time. The results are shown in Table 1.

[0064] Table 1 Comparison of results of different rhizome induction methods

[0065]

[0066] As shown in Table 1, the explants after sterilization were directly transferred to the rhizome induction medium. The mortality rate and browning rate of the explants were reduced, but the rhizome induction rate was lower (processing 3). After sterilization, the explants were directly cut, which significantly increased the mortality rate and browning rate of the material. The larger the cutting wound area, the more serious the browning degree, and the induction success rate was 0 (processing 2). Cutting out the stem tip was material, which could induce rhizome, but the explant mortality rate was higher, and the induction success rate was low (processing 4).

[0067] The method of Example 1 of the present invention (i.e., treatment 1) was used to transplant the sterilized explants into an axillary bud induction medium. Due to damage to the explants during sterilization, some of the explants died, but the mortality rate was low. After culturing for 3 weeks, the explants adapted to the environment of the sterile culture medium, recovered their activity, and induced buds 0.5-1 cm high at their growing points. The induced buds were then excised, and the pseudobulbs after the buds were excised were inoculated into a rhizome induction medium and cultured in a dark environment for 1-3 months, whereby rhizomes were induced from the incisions.

[0068] In summary, the method of the present invention avoids cutting the pseudobulbs in the early stage of disinfection, greatly reduces the mortality rate and browning rate, and improves the success rate of bud cutting of Cymbidium by inducing buds first and then inducing rhizomes after removing the buds, thereby realizing the rapid reproduction of Cymbidium.

[0069] The technical features of the above-mentioned embodiments can be combined arbitrarily. In order to make the description concise, not all possible combinations of the technical features in the above-mentioned embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0070] The above-described embodiments merely illustrate several implementations of the present invention, and while their descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the patent. It should be noted that a person skilled in the art would be able to make numerous variations and improvements without departing from the spirit of the present invention, all of which fall within the scope of protection of the present invention. Therefore, the scope of protection of the patent for this invention shall be determined by the appended claims.

Claims

1. A method for rapid propagation of Cymbidium orchids, characterized in that: The following steps are involved: (1) Select a new bud with unopened leaves as the explant mother plant, cut it vertically close to the base of the pseudobulb, and disinfect the pseudobulb; (2) Inoculate the disinfected pseudobulbs into an axillary bud induction medium and culture in a dark environment to induce budding; the axillary bud induction medium comprises: MS, 0.45 mg / L to 0.55 mg / L NAA, 0.45 mg / L to 0.55 mg / L 6-BA, 25 mg / L to 35 mg / L sucrose, 0.45 mg / L to 0.55 mg / L activated carbon, 190 ml / L to 210 ml / L coconut milk, and 7 mg / L to 9 mg / L agar; (3) excising the buds induced in step (2), inoculating the pseudobulbs after excision of the buds into a rhizome induction medium, and culturing in a dark environment to induce rhizomes; the rhizome induction medium comprises: MS, 2.0 mg / L-2.1 mg / L NAA, 25 mg / L-35 mg / L sucrose, 0.45 mg / L-0.55 mg / L activated carbon, 190 ml / L-210 L coconut milk, and 7 mg / L-9 mg / L agar; (4) transferring the induced rhizomes to a proliferation medium for proliferation culture, and then transferring the proliferated rhizomes to a differentiation medium for bud differentiation. The buds are cultured continuously until they grow into seedlings, and then the rootless seedlings with a height of 3 to 5 cm are inoculated into a seedling rooting medium for seedling rooting culture; The proliferation medium comprises: MS, 1.45 mg / L-1.55 mg / L NAA, 0.45 mg / L-0.55 mg / L 6-BA, 1.55-2.05 mg / L peptone, 25 mg / L-35 mg / L sucrose, 0.45 mg / L-0.55 mg / L activated carbon, and 7 mg / L-9 mg / L agar. The differentiation medium comprises: MS, 0.75 mg / L-0.85 mg / L NAA, 3.95 mg / L-4.05 mg / L 6-BA, 25 mg / L-35 mg / L sucrose, 0.45 mg / L-0.55 mg / L activated carbon, 190 ml / L-210 ml / L coconut water, 320 mg / L-340 mg / L KH2PO4, and 7 mg / L-9 mg / L agar. The seedling rooting culture medium comprises: Huabao No. 1, 0.95 mg / L~1.05 mg / L NAA, 1.95 mg / L~2.05 mg / L peptone, 25 mg / L~35 mg / L sucrose, 100 mg / L~150 mg / L banana, 0.45 mg / L~0.55 mg / L activated carbon, and 7 mg / L~9 mg / L agar.

2. The method for rapid propagation of Cymbidium according to claim 1, wherein The height of the new sprout in step (1) is 1 / 4 to 1 / 3 of the adult plant.

3. The method for rapid propagation of Cymbidium according to claim 1, wherein: The disinfection method of the pseudobulb described in step (1) comprises the following steps: removing the fleshy roots of the pseudobulb and the dark brown old tissue at the base, and peeling off 2 to 3 scaly bracts outside the bud; after rinsing with running water, vibrating and disinfecting with 75%±5% alcohol for 30±5 seconds, and vibrating and washing with sterile water 2 to 3 times, each time for 5±1 minutes; then vibrating and disinfecting with 15%±3% bleach for 15±3 minutes; then peeling off the remaining bracts, and vibrating and disinfecting with 7%±1% bleach for 5±1 minutes; and vibrating and washing with sterile water 2 to 3 times, each time for 5±1 minutes; and finally placing the rinsed pseudobulb on sterile filter paper and air-drying.

4. The method for rapid propagation of Cymbidium according to claim 1, wherein: In step (2), the culture temperature is 25±2° C. and the culture time is 21 to 23 days.

5. The method for rapid propagation of Cymbidium according to claim 1, wherein: The culture conditions in step (3) are: culture temperature 25±2°C, and culture time 1 to 3 months.

6. The method for rapid propagation of Cymbidium according to claim 1, wherein: The conditions for the proliferation culture in step (4) are: culture temperature 25±2°C, and culture time 1 to 2 months.

7. The method for rapid propagation of Cymbidium according to claim 1, wherein: The culture conditions for the differentiation and budding in step (4) are: culture temperature 25±2°C, light intensity 45±5 umol / (m·s -2 ), photoperiod 12±2 h / d, culture time 25~35 days.

8. The method for rapid propagation of Cymbidium according to claim 1, wherein: The culture conditions for the rooting of the seedlings in step (4) are: culture temperature 25±2°C, light intensity 45±5 umol / (m·s -2 ), photoperiod 12±2 h / d, culture time 60-90 days.

9. The method for rapid propagation of Cymbidium according to any one of claims 1 to 8, characterized in that: The Chinese orchid is Chunlan, Huilan, Jianlan, Molan, Hanlan, Lianbanlan or Chunjian.

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