Method for transferring Tibetan medicinal effective component into receptor cell using somatic cell cross breeding technology

A technology of medicinal ingredients and hybridization technology, applied in the field of plant cell engineering, can solve problems such as affecting curative effect, lack of wild resources, and unfavorable foreign trade export

Inactive Publication Date: 2007-05-09
SHANDONG UNIV
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Problems solved by technology

[0016] At present, due to the lack of wild resources of the above-mentioned authentic Tibetan medicinal materials, they cannot meet the needs of pharmaceutical production and foreign trade export. The main method is to use other plants of the same genus as substitutes for mixing, which will not only affect the curative effect, but also not conducive to foreign trade export.

Method used

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  • Method for transferring Tibetan medicinal effective component into receptor cell using somatic cell cross breeding technology
  • Method for transferring Tibetan medicinal effective component into receptor cell using somatic cell cross breeding technology
  • Method for transferring Tibetan medicinal effective component into receptor cell using somatic cell cross breeding technology

Examples

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Effect test

example 1

[0075] Example 1: Using asymmetric somatic cell hybridization to transfer the effective components of Swertia chinensis in Western Sichuan to Bupleurum chinensis

[0076] 1 Materials and methods

[0077] 1.1 The source of parental protoplasts Suspension cell lines were established from Bupleurum angustifolia callus. The suspension cell line subcultured for 3 days was used to separate and prepare protoplasts as recipients.

[0078] The protoplasts were isolated from the callus of Swertia chinensis in western Sichuan, and the method was the same as that of wheat. The protoplasts were subjected to 380μW / cm 2 After irradiating 1min and 2min of ultraviolet rays, they were used as fusion donors.

[0079] 2 Fusion and cultivation of protoplasts

[0080] The donor and acceptor were divided into 1×10 6 The density per ml was mixed at a ratio of 1:1, and fused by PEG method.

[0081] 3 Identification of hybrid properties of regenerated callus and regenerated plants

[0082] Inclu...

example 2

[0086] Example 2: Utilizing asymmetric somatic hybridization to transfer the active ingredients of Gentiana chinensis to Bupleurum chinensis

[0087] Method and steps are the same as in Example 1.

[0088] Results: The strength of Gentiana tuberosa protoplasts was 380μw / cm 2 After ultraviolet irradiation for 0 min, 1 min, and 2 min, respectively, they fused with Bupleurum protoplasts under the induction of PEG. The hybrid morphology (Fig. 6), chromosome, isozyme (Fig. 7) and RAPD (Fig. 8) analysis of 20 single-cell clones regenerated by fusion showed that 18 of them were somatic hybrid cell lines. Chromosomal observation showed (Fig. 2) that the number of chromosomes in the regenerated plants of the 11 clones was distributed in the range of 11-13, and the isoenzyme and RAPD analysis showed that they all had parental characteristic bands, confirming that they were interfamily somatic hybrids. The results of the determination of the content of the active ingredient (gentiopicr...

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Abstract

The process of transferring Tibetan medicinal effective component into receptor cell by means of somatic cell cross breeding technology includes the following main steps: 1) selecting parent material gene types; 2) dissociating the receptor protoplaster; 3) UV irradiating donor protoplaster; 4) fusing the donor protoplaster and the receptor protoplaster and culturing; 5) identifying hybrid callus and regenerated plant; and 6) detecting target characteristic in somatic cell hybrid. The present invention obtains cell line with high medicinal component content and fast growth by means of somatic cell cross breeding technology and may be used in obtaining famous and rare Tibetan medicinal effective component.

Description

technical field [0001] The invention relates to a method for transferring medicinal components of Tibetan medicinal materials into recipient cells by using somatic cell hybridization technology. It belongs to the field of plant cell engineering. Background technique [0002] Tibetan medicine is a traditional treasure of the Tibetan culture of the Chinese nation. It has a research history of thousands of years. Some unique Tibetan medicinal plants on the Qinghai-Tibet Plateau have attracted much attention at home and abroad because of their remarkable medicinal effects. [0003] Biotechnology is a technology that applies biological research results to increase the quantity and quality by engineering means, so as to meet the increasing human demand for biological products. Over the past 30 years, great progress has been made in many fields, but the application in traditional medicinal materials has just started. So far, my country has mainly carried out research on the follow...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/05C12N5/14A01H4/00
Inventor 向凤宁夏光敏蔡云飞李子东江莉
Owner SHANDONG UNIV
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