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Culture method for rosmarinic acid high-yielding suspension cells of purple perilla

A technology of rosmarinic acid and suspended cells, which is applied to plant cells and other directions, can solve the problems of non-green chemical synthesis of rosmarinic acid, low yield of rosmarinic acid and high cost, and achieves the promotion of cell division and enlargement and yield. Increase and increase the effect of production

Inactive Publication Date: 2018-11-16
ZHONGBEI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The invention provides a method for producing high-yield rosmarinic acid by suspending perilla cells to solve the problem that the yield of natural plant secondary metabolite rosmarinic acid is low, but chemically synthesized rosmarinic acid is not green and the cost is high , this method is not limited by seasons and natural ecological conditions, and can stably, efficiently and quickly obtain a large amount of perilla callus with high content, providing theoretical basis and technical reference for large-scale culture of perilla cells with rosmarinic acid

Method used

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Examples

Experimental program
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Effect test

Embodiment

[0031] (1) Acquisition of perilla sterilized vaccine

[0032] The perilla seeds were rinsed twice with tap water, sterilized with 70% (V / V) ethanol for 30 seconds, washed with sterile water for 3 times, then soaked in 0.1% mercuric solution for 10 minutes, and then washed with sterile water for 3 minutes. -5 times, not less than 5 minutes each time, to completely remove the highly toxic HgCl 2 . The sterilized seeds were inoculated into tissue culture flasks containing 30 mL of hormone-free MS basic medium (adding 3% sucrose, agar, pH 5.8, sterilized at 121° C. for 20 min) to germinate. At 25 ± 1 ° C, 16 hours of light per day. 13-day-old aseptic seedlings, hypocotyls grow to 2 to 3 cm, and cotyledons can be used as explants when they unfold.

[0033] (2) Induction of callus

[0034] Under sterile conditions, the hypocotyls of the normal and robust perilla aseptic seedlings obtained in step (1) were cut into 0.5cm segments, and the cotyledons were cut into 0.2cm 2 Inocula...

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Abstract

The invention discloses a culture method for rosmarinic acid high-yielding suspension cells of purple perilla. The method comprises the specific steps: acquiring a sterile seedling of purple perilla;inducing calluses with excellent textures by means of an explant of the sterile seedling; by taking the calluses of purple perilla as a raw material, optimizing the optimum condition for producing rosmarinic acid produced by a fluid suspension culture by adopting a Box-Behnken central combinational design and a response surface method; and determining and verifying the optimum condition of suspension culture of the cells of purple perilla. The output of the rosmarinic acid is improved by nearly 10 times, and meanwhile, the calluses yielding the rosmarinic acid highly can be more quickly obtained, so that the cost is reduced, the indoor culture space is saved, the calluses can be produced all the year, the culture benefit is increased obviously, and the culture method is not limited by seasons and natural ecological conditions. A lot of calluses of purple perilla relatively high in content can be obtained stably, efficiently and quickly, and a theoretical basis and technical reference for culturing the rosmarinic acid by the cells of the purple perilla on a large scale are provided. The culture method has a wide application prospect and remarkable social and economical benefits.

Description

technical field [0001] The invention belongs to the technical field of plant cell culture and production of secondary metabolites, in particular to a method for culturing perilla suspension cells with high yield of rosmarinic acid. Background technique [0002] Basil (Perilla frutescens (L.) Britt.), also known as Eden, Baisu, Chisu, etc., is an annual herb of the Lamiaceae, native to China, mainly distributed in China, India, Japan, Korea and other countries. It is one of the first batch of 60 kinds of Chinese herbal medicines that are both food and medicine promulgated by the Ministry of Health of my country. Perilla leaves contain a variety of natural chemical components with physiological activities, such as anthocyanins, luteolin, apigenin, catechins, rosmarinic acid (RosA), etc., of which rosmarinic acid content is relatively high . Rosmarinic acid is a natural water-soluble antioxidant with a variety of biological activities, and has obvious anti-inflammatory, anti-o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04
CPCC12N5/04
Inventor 李会珍李娜张志军
Owner ZHONGBEI UNIV