Preparation method for artificial seeds of dendrobium candidum

A technology of artificial seeds and dendrobium candidum, applied in gardening methods, botanical equipment and methods, gardening tools/equipment, etc., can solve the problems of artificial seed germination rate, seedling rate and storage conditions, and achieve convenient storage and transportation , the effect of increasing the germination rate

Inactive Publication Date: 2012-05-02
澄思源生物科技(上海)有限公司
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AI-Extracted Technical Summary

Problems solved by technology

Modern research has made a lot of explorations in artificial endosperm, embedding methods, artificial seed coats, storage methods, etc...
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Method used

Although domestic relevant Dendrobium officinale artificial seed makes the report, only succeeds in germination on the substratum of the artificial seed of making, does not realize the germination on planting substrate and outdoor seedbed. The invention uses the artificial endosperm and artificial seed coat made of modern nano-materials, which can not only improve the air permeability, water retention and water absorption of the sodium alginate embedding system, shorten the germination time...
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Abstract

The invention discloses a preparation method for artificial seeds of dendrobium candidum. The method comprises: inducing dendrobium candidum stems in a culture medium so as to obtain protocorms, which are then subjected to proliferation culture, and transferring the protocorms into an MS culture solution with 0.5mg/L of ABA (abscisic acid) for shaking culture, changing the culture solution every 10d, after 30d, selecting a protocorm as an embedding propagule, making artificial seeds by an instillation method, taking the MS culture medium, 4mg/L of 6-BA, 0.4mg/L of NAA, 2% of sodium alginate and 0.2% of carbendazim to prepare basic artificial endosperm, adding 2% of chitosan and 5.0-10% of nanometer TiO2 into sodium alginate so as to form an artificial seed coat matrix, and carrying out research on germination and storage of artificial seeds. The preparation method of the invention can enhance seed germination rate and make the seeds convenient for storage and transportation. Adding of a special material into artificial endosperm or an artificial seed coat in the invention can promote germination of the artificial seeds or prolong storage of the artificial seeds.

Application Domain

Plant tissue cultureHorticulture methods

Technology Topic

GerminationDendrobium candidum +10

Examples

  • Experimental program(1)

Example Embodiment

[0019] The technical features of the present invention will be further described below in conjunction with specific embodiments.
[0020] (1) Induction of protocorm
[0021] Choose healthy plants of Dendrobium officinale with vigorous growth and disease-free, put every two sections into a 500ml beaker, add a small amount of washing powder, tie the cup with a single layer of gauze, and then tie it tightly with engineering thread and place it under the tap Rinse for 30 minutes, and adjust the amount of flushing water to make the stems of Dendrobium officinale float in the water. The main function is to wash away the external dust, large particle colonies, hyphae, etc. Bring the finished materials to the clean bench in the sterile room for secondary disinfection and sterilization: remove the bracts of the axillary buds of the stems, quickly soak them in 70% alcohol for 25s, and then use 0.1% mercury Soak the solution for 10 minutes, shake the beaker continuously during the sterilization process to make the sterilization more thorough, and then rinse with sterile water 5 times to remove the surface drug residue.
[0022] Cut the above-mentioned material into one section per section, remove the part of the stem that is soaked with the drug, inoculate it into a pre-made differentiation and proliferation medium tissue culture bottle, and place it on the medium for cultivation. Culture conditions: temperature 26℃, light time 12h, light intensity 2000Lx, 30 days later, protocorm was induced. The differentiation and proliferation medium is a mixture of MS medium, 4 mg/L 6-BA (6-benzylaminopurine) and 0.4 mg/L NAA (naphthalene acetic acid).
[0023] (2) Production of artificial seeds of Dendrobium officinale
[0024] The protocorm was transferred into MS culture medium containing 0.5mg/L ABA and cultured with shaking (100r/min). The culture medium was changed every 10 days. After 30 days, the length and width were about 0.5-1.5mm×2.0-3.4mm. The bulb serves as an embedded propagule. Take MS+4mg/L 6-BA (6-benzylaminopurine)+0.4mg/L NAA (naphthalene acetic acid)+2% sodium alginate+0.2% carbendazim as the basic artificial endosperm, in order to increase ventilation and maintain Water is easy to absorb. On this basis, 1.5-2.0% starch, 1.0-2.0% activated carbon, 1.0-2.0% water retaining agent and 1.0-3.0% nano-TiO are added 2 , Add 2% chitosan and 5.0%-10% nano-TiO in sodium alginate 2 It is an artificial seed coat substrate.
[0025] On an ultra-clean workbench, use tweezers to immerse the protocorm in the embedding solution of MS medium, 4mg/L 6-BA, 0.4mg/L NAA, 2% sodium alginate and 0.2% carbendazim (I.e. artificial endosperm), stir evenly with a glass rod to make the protocorm and embedding agent fully mixed evenly. After 5-10 minutes, use a dropper to drop the semi-gel state sodium alginate coated with protocorm into the 2% CaCl of the artificial seed coat matrix 2 The solution is solidified, and each drop is controlled to contain a protocorm. After 30 minutes, it is rinsed with culture solution for 15 minutes to obtain artificial seeds.
[0026] Germination experiment of artificial seeds:
[0027] The germination experiments were carried out on different germination substrates. The germination substrates were MS medium, planting substrate, and seedbed. The germination rate of artificial seeds in the seed MS medium within 30 days is 98.0%, the germination rate in the planting substrate can reach 75.0%, and the germination rate in the outdoor seedling bed can still reach 70%.
[0028] The germination rate of Dendrobium candidum seeds under natural conditions is only 5%, and the germination rate of artificial seeds on the market is only 80%. Our products have greatly improved the germination rate of Dendrobium candidum artificial seeds.
[0029] Storage experiment of artificial seeds:
[0030] Put the prepared artificial seeds into a sterilized and dried Erlenmeyer flask, seal them with a sealing film, and store them at 4°C for 30d and 90d, respectively.
[0031] After storing the prepared artificial seeds for 30 days, the germination rate on the MS medium is 95.0%, the germination rate after 60 days can still reach 90.0%, and the seedling rate can reach 85.0%.
[0032] There are still big problems with the storage of artificial seeds on the market. With the extension of storage time, the germination rate of artificial seeds will decrease significantly. The artificial seeds we prepare can still maintain a high germination rate after storage.
[0033] Although there are reports on the production of artificial seeds of Dendrobium officinale in China, the artificial seeds produced successfully germinated only on the culture medium, and did not achieve germination on the planting substrate and outdoor seedbed. The artificial endosperm and artificial seed coat made of modern nano materials in the present invention can not only improve the air permeability, water retention and water absorption of the sodium alginate embedding system, shorten the germination time of artificial seeds, but also inhibit the reproduction and growth of microorganisms. Achieve antibacterial and antiseptic effects, and promote the growth of the protocorm in the artificial seed colloid, thereby increasing the germination rate and seedling rate of the artificial seed in the planting substrate and outdoor seedbed, providing technical support for the practical application of Dendrobium candidum Thus opened up a new way for the breeding of Dendrobium officinale.
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Description & Claims & Application Information

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