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Method for breeding anti-reversible tissue culture seedlings of anoectochilus roxburghii

A technique for tissue culture of seedlings and culture medium, applied in horticultural methods, botanical equipment and methods, horticulture, etc.

Inactive Publication Date: 2012-06-27
GUIZHOU NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the process of tissue culture, combined with induction treatment to cultivate stress-resistant seedlings, there is no report

Method used

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  • Method for breeding anti-reversible tissue culture seedlings of anoectochilus roxburghii
  • Method for breeding anti-reversible tissue culture seedlings of anoectochilus roxburghii
  • Method for breeding anti-reversible tissue culture seedlings of anoectochilus roxburghii

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] 1. The first generation of rhizome culture: Take the rhizome of golden clematis, wash it with water, soak it with washing powder for 1 hour, then rinse it with tap water for 2 hours, cut off the leaves (keep the leaf sheath), and then use it on the ultra-clean workbench. Soak in 75% alcohol for 30 seconds, rinse once with distilled water, then add mercury chloride (add a few drops of Tween-20) for 12 minutes, and keep stirring, wash with sterile water 5 times. On the ultra-clean workbench, use a scalpel to cut the sterilized stem segment into a small segment with a length of about 1.5 cm, and insert the stem segment upright into MS+BA1.5mg / L+KT0.5mg / Medium of L+NAA1.5mg / L+sucrose 30g / L+0.5-0.6mg / Lthiourea+0.5-0.6mg / LVc. Place it under the conditions of light 12 hours / day, light intensity 1500lx-2000lx, and temperature 23±2°C. Aseptic buds can be obtained after culturing for 60 days.

[0019] It should be noted that 0.5-0.6 mg / L thiourea and 0.5-0.6 mg / L Vc were added...

Embodiment 2

[0028] Comparison test of different media for the primary culture of rhizomes: inoculate the stem segments on medium 1 and medium 2, and count the induction rate after 30 days. The results are shown in Table 4:

[0029] Table 2 Primary culture verification test

[0030]

[0031]

Embodiment 3

[0033] Membrane permeability change experiments of conventional tissue cultured seedlings and stress-resistant tissue cultured seedlings under stress:

[0034] High-temperature stress treatment: get the conventional tissue culture seedlings and the stress-resistant tissue culture seedlings of the same transplanting time in 3 pots, place the SPX-150-GB type light incubator (in the light incubator, the light intensity is 2000lx, and the light time is 12h / In d), carry out high temperature stress treatment, the temperature is set to 28 ℃, 33 ℃, with the control placed at 23 ℃, take the mature leaves of the same part on the 0th, 1st, 2nd, 3rd, 4th day of treatment and carry out membrane permeation For the determination of sex, each index was repeated three times for each treatment.

[0035] Determination of the permeability of the plasma membrane: use the conductivity meter method, take 0.200g of the treated leaves, cut them off, wash them with tap water, and then rinse them twice...

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Abstract

The invention discloses a method for breeding anti-reversible tissue culture seedlings of anoectochilus roxburghii. The method comprises the following steps of: A1, primary culture; A2, multiplication culture; and A3, rooting culture and anti-reversible induction: taking rootless seedlings out of culture bottles on an ultra-clean workbench, and inoculating the rootless seedlings to a rooting culture medium in a direct inserting mode, wherein the rooting culture medium contains 1 / 2 MS, 3 mg / L of NAA and 20 g / L of sucrose; a self-made resistance inducer containing 1.3 mg / L of ABA, 2.5 mg / L of SA and 30 mg / L of CTS is also added in the rooting culture medium; the culture is performed under the conditions that the illumination is 12 hours every day, the intensity of light is 1,500 to 2,000lx and the temperature is 23 + / - 2 DEG C, and 2 to 3 roots per plant can be achieved in 60 days; and A4, seedling production and transplanting. By combining quick propagation and stress tolerance of the seedlings, the invention provides the method for breeding the anti-reversible tissue culture seedlings of the anoectochilus roxburghii.

Description

technical field [0001] The invention belongs to the technical field of group cultivation of seedlings, and in particular relates to a method for cultivating stress-resistant tissue cultured seedlings of orchid orchids. Background technique [0002] Anoectochilus roxburghii (Wall.) Lindl., also known as Anoectochilus roxburghii (Wall.) Lindl., is a plant of the genus Anoectochilus roxburghii. Known as "King of Medicine", "Tiger of Medicine", "Golden Grass", "Miraculous Medicine", "Black Ginseng" and other laudatory titles; in addition to medicinal purposes, it has the effects of lowering blood lipids and anti-oxidation, and has mild medicinal properties, It is neither hot nor dry. Residents in Taiwan use it as a precious health supplement, and have developed a series of products such as beverages and oral liquids. In addition, because of its beautiful leaves, it is hailed as the holy product among the orchids by orchid growers. However, due to the strict requirements on the ...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 龚宁李光李亚军
Owner GUIZHOU NORMAL UNIVERSITY
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