Rhodida plant callus and suspension cell granule culture method

A technology of callus and culture method, which is applied in the direction of plant cells, tissue culture, biochemical equipment and methods, etc., to achieve the effects of reducing production costs, good dispersion, and protecting the ecological environment

Inactive Publication Date: 2009-09-16
BEIJING FORESTRY UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Embodiment 1

[0033] 1. Take the young stems of Rhodiola grandiflora as explants, wash them with running water for 1 hour, rinse them with 70% ethanol on the ultra-clean workbench for 10 seconds, and then soak them in 0.5% effective chlorine disinfectant for 30 minutes. Replace the disinfectant solution once, then rinse with sterile water for 5 times, blot dry the water with sterile filter paper, inoculate under sterile conditions on the following medium without growth regulators for cultivation: MS basic medium + sucrose 30g / L + Agar 5.5g / L (the pH value of the medium is 5.8-6.0, and the medium is sterilized at 121°C for 15min). Due to the presence of endophytic fungi in Rhodiola grandiflora leaves, 1-1.5ml / L of PPM (plant preservative mixture, plant cell technology, Inc) needs to be added to the culture medium; the culture conditions described are temperature 25°C, light intensity 24 μmol m -2 the s -1 , the photoperiod is 16h / d;

[0034] 2. Inoculate the sterile young leaves obtained ...

Embodiment 2

[0038] 1. Use Rhodiola grandiflora seeds as explants, wash and soak in running water for 1 hour, rinse with 70% ethanol for 30 seconds on the ultra-clean workbench, rinse with sterilized deionized water for 5 times; sterilize with 1.5% NaClO for 7 -10min, then rinse with sterile water for 5 times, dry the water with sterile filter paper, inoculate under sterile conditions on the culture medium without growth regulator: MS basic medium + sucrose 30g / L + agar 5.5g / L (the pH value of the medium is 5.8-6.0, and the medium is sterilized at 121°C for 15 minutes); the culture conditions described are temperature 25°C, light intensity 24μmol m -2 the s -1 , the photoperiod is 16h / d; after inoculation, the embryos start to germinate, and become complete seedlings after one week, and the height of the seedlings can reach 3-4cm after 30 days;

[0039] 2. Cut off the radicle and cotyledons of the 1-month-old sterile seedlings obtained in step 1, cut the young stems into about 1 cm with ...

Embodiment 3

[0043] 1. Use Rhodiola grandiflora seeds as explants, wash and soak in running water for 1 hour, rinse with 70% ethanol for 30 seconds on the ultra-clean workbench, rinse with sterilized deionized water for 5 times; sterilize with 1.5% NaClO for 7 -10min, then rinsed with sterile water 5 times, blotted dry with sterile filter paper, inoculated under aseptic conditions on the following medium for cultivation: MS basic medium+sucrose 30g / L+agar 5.5g / L (medium The pH value is 5.8-6.0, and the culture medium is sterilized at 121°C for 15 minutes); the culture conditions described are temperature 25°C, light intensity 24μmol m -2 the s -1 , the photoperiod is 16h / d; after inoculation, the embryos start to germinate, and become complete seedlings after one week, and the height of the seedlings can reach 3-4cm after 30 days;

[0044] 2. Cut off the tender stems of the 1-month-old sterile seedlings obtained in step 1, and inoculate them into the following callus induction medium for ...

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Abstract

The invention discloses a Rhodida plant callus and suspension cell granule production method, comprising: (1) constructing a Rhodida plant aseptic culture system to culture and obtain aseptic Rhodida plants with roots, stems and leaves; (2) inoculating the roots, stems or leaves of the Rhodida plants into a callus induction culture medium to carry out callus induction culture and successive transfer culture to obtain good suspension cell granules originated from different organs of the Rhodida plants. The good suspension cell line originated from different organs of the Rhodida plants has the characteristics of fast growing speed, good dispersibility, high content of secondary metabolites (wherein the content scope of Salisorosides Rosavin and tyrosol are respectively 1-3% and 0.5-0.7% of the dry weight of the suspension cells) and the like, is very stable in successive transfer culture and degradation phenomenon does not exist.

Description

technical field [0001] The invention relates to a plant tissue or cell culture method, in particular to a method for large-scale production of callus and excellent suspended cell particles from different organs of plants of the genus Rhodiola (Rhodiola L.), belonging to the field of plant tissue or cell culture . Background technique [0002] Rhodiola is a perennial herb or subshrub plant of the genus Rhodiola (Rhodiola L.) in the family Crassulaceae. It is a traditional Tibetan medicine and a traditional Chinese medicinal plant. It is mainly used for the treatment of pneumonia, infectious diseases and clearing vascular fever. , has the effects of clearing heat, detoxifying and drying dampness, and enjoys the reputation of "plateau ginseng". For decades, the research on Rhodiola plants at home and abroad has been in the ascendant. Rhodiola not only has the functions of anti-oxidation, anti-fatigue, anti-hypoxia, anti-cold, anti-aging, anti-microwave radiation and other simi...

Claims

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Application Information

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IPC IPC(8): C12N5/04C12N5/02
Inventor 史玲玲王莉戴向辰王彩云刘玉军
Owner BEIJING FORESTRY UNIVERSITY
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