Method for obtaining callus and suspension cells under induction of Taxus chinensis test tube plantlet

Abstract

The invention discloses a method for obtaining callus and suspension cells under induction of Taxus chinensis test tube plantlet. The method comprises the following steps: (1) seed testa peeling: mechanical cracking is utilized and the testa is peeled manually; (2) explants disinfection: disinfection is carried out for 90 seconds with 70% alcohol and carried out for 12 minutes with 0.1% mercuric chloride, and washing is carried out by 5 times with sterile water for standby; (3) pretreatment of seed: seed soaking is carried out by 1-5 days at normal temperature hermetically with sterile water; (4) zygotic embryo separation; (5) induction of test tube plantlet: embryo is inoculated to a test tube plantlet induced culture substrate; (6) induction of callus; (7) subculture of callus; and (8) suspension culture of callus. The method has the characteristics of being capable of simply, conveniently and quickly obtaining Taxus chinensis callus and utilizing the callus to provide material for the suspension culture of cells of Taxus chinensis or further screen high-yield paclitaxel cell strains to provide high-quality screening material, thereby saving a great quantity of manpower and material resources and improving the labor efficiency.

Description

technical field [0001] The invention belongs to the fields of plant tissue culture and cell engineering, and in particular relates to a method for inducing callus and suspension cells from yew test-tube plantlets. Background technique [0002] Paclitaxel (trade name Taxol) is a natural tetracyclic diterpene alkaloid isolated from the bark of Taxus L. in the 1960s and 1970s. Its unique effect in tumor therapy has attracted widespread attention. According to relevant information, it is estimated that the demand for paclitaxel injections at home and abroad in the early 21st century is about 20 million sticks, that is, 600 kg of paclitaxel is needed, and the current market price is 1200-1800 yuan per stick. With the increasing demand for paclitaxel, the yield extracted from the yew tree can no longer meet the needs of the pharmaceutical market. In addition, the natural resources of yew are becoming increasingly exhausted due to the excessive exploitation, and the natural renewa...

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a set of method for inducing callus by using the test-tube plantlets of Taxus chinensis, and at the same time provide a method for using the callus to carry out suspension culture to obtain suspension cells, so as to overcome the high pollution of callus directly induced by stem segments and seed embryos High efficiency, low efficiency or defects in the method, in order to obtain suspension cells with good dispersion performance quickly, efficiently and on a large scale, to provide raw materials for the cell suspension culture of Taxus chinensis or to provide high-quality screening materials for further screening of high-yield paclitaxel cell lines

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