Tissue culture method of Gentiana dahurica Fisch for rapid in vitro reproduction

A technology of tissue culture and chinensis, which is applied in the field of rapid propagation in vitro by tissue culture of Xiaoqinjiao, can solve the problems such as the inability of resources to meet the market demand, the destructive destruction of Xiaoqinjiao population, the destruction of germplasm resources, etc., to achieve rapid cell division and expansion, The effect of promoting cell division and expansion, and promoting germination and growth

Inactive Publication Date: 2012-08-15
CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the demand for Gentiana miltiorrhiza is increasing year by year. The medicinal materials of Gentiana miltiorrhiza have long relied on wild excavation to meet the needs of domestic and foreign marke

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0026] Example 1 A method for rapid propagation in vitro in tissue culture of Gentiana chinensis includes the following steps:

[0027] ⑴ Selection and disinfection of explants: Take the stem tips of two-year-old Gentiana spp. seedlings in the greenhouse as explants, soak and sterilize them with a mass concentration of 0.1% mercury for 8 minutes on an ultra-clean workbench. After rinsing with ionized water for 4 to 6 times, the sterilized stem tip of the explant is obtained.

[0028] ⑵Inoculation of explants: inoculate the sterilized stem tips of explants on callus induction medium, and inoculate 1 sterilized explant stem tips in every 25ml callus induction medium; 25℃±1℃, light intensity 30~60 umol. M -2 . s -1 , Illumination time is 12~14 h.d -1 Cultivated in the light incubator for 20-25 days, light yellow dense callus formed at the base of the stem tip.

[0029] Callus induction medium is to add 100ml of macro element mother liquor, 10ml trace element mother liquor, 10ml mot...

Example Embodiment

[0037] Example 2 A method for rapid propagation in vitro in tissue culture of Gentiana chinensis includes the following steps:

[0038] ⑴ Selection and disinfection of explants: Take the stem tip of the two-year-old Gentiana seedlings in the greenhouse as explants, soak and sterilize them with 0.2% mercury liters on a clean bench for 5 minutes, and sterilize them. After rinsing with ionized water for 4 to 6 times, the sterilized stem tip of the explant is obtained.

[0039] ⑵Inoculation of explants: inoculate the sterilized stem tips of explants on callus induction medium, and inoculate 1 sterilized explant stem tips in every 25ml callus induction medium; 25℃±1℃, light intensity 30~60 umol. M -2 . s -1 , Illumination time is 12~14 h.d -1 Cultivated in the light incubator for 20-25 days, light yellow dense callus formed at the base of the stem tip.

[0040] Callus induction medium is to add 100ml of macro element mother liquor, 10ml trace element mother liquor, 10ml mother liquo...

Example Embodiment

[0048] Example 3 A method for rapid propagation in vitro in tissue culture of Gentiana chinensis includes the following steps:

[0049] ⑴Explant selection and disinfection treatment: Take the stem tip of the two-year-old Gentiana chinensis seedlings in the greenhouse as explants, soak and sterilize them with a mass concentration of 0.15% mercury on a clean bench for 6 minutes, and sterilize them. After rinsing with ionized water for 4 to 6 times, the sterilized stem tip of the explant is obtained.

[0050] ⑵Inoculation of explants: inoculate the sterilized stem tips of explants on callus induction medium, and inoculate 1 sterilized explant stem tips in every 25ml callus induction medium; 25℃±1℃, light intensity 30~60 umol. M -2 . s -1 , Illumination time is 12~14 h.d -1 Cultivated in the light incubator for 20-25 days, light yellow dense callus formed at the base of the stem tip.

[0051] Callus induction medium is to add 100ml of macro element mother liquor, 10ml trace element...

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Abstract

The invention relates to a tissue culture method of Gentiana dahurica Fisch for rapid in vitro reproduction. The method comprises the following steps: (1) selection and sterilization process of explant, comprising taking the stem tip of seedling of biennial Gentiana dahurica Fisch in a greenhouse as the explant, dipping for sterilization and washing so as to obtain the sterilized explant stem tip; (2) inoculation of explant, comprising inoculating the explant stem tip after sterilization at a callus tissue induction culture medium so as to form callus tissue; (3) proliferation and differentiation culture of the callus tissue, comprising inoculating the callus tissue to a callus tissue proliferation culture medium to culture so as to form single buds of Gentiana dahurica Fisch; (4) induction of root growth, comprising separating the single buds of Gentiana dahurica Fisch and afterwards transplanting into the root growing culture medium to culture so as to form complete regenerated plantlets; (5) sterilization of seedling medium; (6) transplanting of plant, comprising transplanting the complete regenerated plantlets into the seedling substrate subjected to sterilization directly, and acclimating the seedling so as to grow normal Gentiana dahurica Fisch plants. According to the method, the problems of overlong seedling period and low reproduction coefficient of Gentiana dahurica Fisch are overcome, and the method is easy to operate and can be used for rapid factory seedling culture.

Description

technical field [0001] The invention relates to tissue culture and in vitro rapid propagation of plants, in particular to a method for in vitro rapid propagation of Gentiana miltiorrhiza tissue culture. Background technique [0002] Gentiana is a perennial herbaceous plant belonging to the genus Gentianaceae (Gentianaceae). It is a famous traditional Chinese and Tibetan medicinal material in my country. It is recorded in the 2005 edition of "Chinese Pharmacopoeia" and Tibetan Medicine Chronicles. It is distributed in alpine grasses at an altitude of 800-4500m. Meadows, thickets, forest margins, hillside grasslands, ravines and river beaches. In my country, Gentiana miltiorrhiza is mainly distributed in northern Sichuan, northwest, northwest, North China, northeast and other regions. It is a commonly used traditional Chinese and Tibetan medicinal material. The dried root is used as medicine and can be used to treat tonsillitis, urticaria and rheumatoid arthritis. . At present...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 徐文华周国英
Owner CHINA ACAD OF SCI NORTHWEST HIGHLAND BIOLOGY INST
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