Method using hypocotyl of catharanthus roseus for cultivating regenerated plants

Abstract

The invention relates to the technical field of biology, in particular to a method using the hypocotyl of catharanthus roseus for cultivating regenerated plants. The method comprises the following steps: placing the seeds of catharanthus roseus on an MS (Murashige and Skoog) medium for culturing, and placing the hypocotyl thereof on a callus-inducing medium for culturing until the first pair of cotyledons branches out, so as to obtain callus; inoculating the callus on a differentiation medium for culturing, so as to obtain the green callus; inoculating the green callus on an adventitious bud inducing medium for culturing, so as to obtain the adventitious bud; placing the adventitious bud on a rooting medium for culturing, so as to obtain the test-tube plantlet; and rapidly propagating thetest-tube plantlet on a rapid-propagation medium, so as to obtain the regenerated plant. By using the method of the invention, the adventitious bud differentiation rate of the hypocotyl of catharanthus roseus reaches 81% and the adventitious bud rooting rate thereof reaches 100%.

Description

technical field [0001] The invention relates to a method for cultivating regenerated plants by utilizing periwinkle hypocotyls, and belongs to the field of biotechnology. Background technique [0002] Vinca (Catharanthus roseus) is a plant of the genus Vinca in the family Apocynaceae, which contains more than 100 biologically active terpenoid indole alkaloids (TIAs), such as Vinblastine and Vincristine ) has anti-cancer effects, while Ajmalicine and Serpentine can resist high blood pressure and have antibacterial and sedative effects. Among them, vinorelbine, a semi-synthetic derivative of vinblastine, has been widely used in clinical medicine and is currently the most widely used antineoplastic drug. Vinca has become the most economical and practical medicinal model plant. [0003] In actual production and application, due to the extremely low content of terpene indole alkaloids in Changchun flowers, especially the content of diterpenoid indole alkaloids, such as vinblast...

AI Technical Summary

Problems solved by technology

It has been reported that plants regenerated from hairy roots of Vinca are mainly used in the transformation system mediated by Agrobacterium rhizogenes, but the plants regenerated through hairy roots are deformed and sterile, which is not conducive to the implementation of genetic engineering strategies

Agrobacterium tumefaciens-mediated transformation is currently the most widely used plant transgenic method, which induces callus to regenerate normal complete plants by infecting explants (such as leaves, stems, etc.) other than hairy roots , so as to obtain transgenic strains. So far, direct transformation of Agrobacterium tumefaciens to obtain transgenic periwinkle plants has not been reported. The main reason is that it is impossible to establish an efficient and mature periwinkle explant regeneration system except for hairy roots.

Although there is a research report on using the embryogenesis of periwinkle somatic cells to regenerate plants, the regeneration cycle required by this method is long, the frequency of somatic embryogenesis is low, synchronization cannot be achieved, and deformed or rootless seedlings are often formed during development. Can establish a rapid and stable culture system using somatic embryo regeneration, which limits the transgenic engineering of periwinkle

[0004] Find through literature search to prior art, do not see and utilize periwinkle hypocotyl of the present invention to cultivate the relevant report of the method for regenerated plant

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