Taraxacum kok saghyz Rodin genetic transformation method

Abstract

The invention provides a Taraxacum kok saghyz Rodin genetic transformation method. The Taraxacum kok saghyz Rodin genetic transformation method includes the steps that (1) young and tender leaves of Xinjiang Taraxacum kok saghyz Rodin germplasm is taken and subjected to cutting processing, and put in a bud induction medium with MS+1.5-1.6mg / L6-BA+0.1mg / LNAA+20g / L sucrose and pH being 5.8 for culture; after 50 days of culture, the excellent Taraxacum kok saghyz Rodin germplasm suitable for tissue culture is selected; the excellent Taraxacum kok saghyz Rodin germplasm is transferred to a rootingmedium with 1 / 2MS+20g / L sucrose and pH being 5.8 to become a regenerated Taraxacum kok saghyz Rodin tissue culture seedling; (2) the seedling leaves are taken and cut, and pre-cultured on the bud induction medium for 2 days; and (3) an agrobacterium infection solution is added, dip-dyeing is conducted, the dip-dyed material is transferred into the bud induction medium, and co-culture is performedor 3 days in the dark; the co-cultured material is transferred to a bud induction medium containing 400 mg / L Car to be cultured for one week; the cultured material is transferred to a screening medium with MS+1.5-1.6mg / L 6-BA+0.1mg / L NAA+20g / L sucrose+400mg / L Car and 8-10mg / L Hyg and pH being 5.8 to induce regeneration of resistant buds; and the material with resistant buds is transferred to a new rooting medium with 1 / 2MS+20g / L sucrose+400mg / L Car and 8-10mg / L Hyg and pH being 5.8, so that a Taraxacum kok saghyz Rodin resistant regenerated plant is obtained. According to the Taraxacum kok saghyz Rodin genetic transformation method, the induction rate of resistant regenerated buds is greater than 30%, the rate of malformation of transgenic regenerated seedlings is low, and the positive detectable rate of transgenic Taraxacum kok saghyz Rodin is greater than 90%.

Description

technical field [0001] The invention relates to the field of plant biotechnology, in particular to a method for genetic transformation of rubber grass. Background technique [0002] Rubber grass, also known as Russian dandelion (Taraxacum kok saghyz Rodin), is a plant of the genus Dandelion in the Cichorium family of the Compositae family. It is native to Kazakhstan, Europe, and Xinjiang in my country. Because the root of rubber grass contains high-quality rubber, the content can reach more than 20%, and it gradually becomes the second natural rubber resource. [0003] In recent years, scholars at home and abroad have carried out some research on rubber grass tissue culture technology. For example, rubber grass leaves are used as explants to induce adventitious buds or callus. However, due to the self-incompatible propagation characteristics of rubber grass, the strains of rubber grass are seriously mixed. Due to the differences in tissue culture conditions and methods amo...

AI Technical Summary

Problems solved by technology

For example, rubber grass leaves are used as explants to induce adventitious buds or callus, but due to the self-incompatibility of rubber grass, the strains are seriously mixed.

Due to the differences in tissue culture conditions and methods among different strains, it is easy to cause problems such as low bud induction rate and low regenerative ability of regenerated seedlings, and it is difficult to be used for genetic transformation of rubber grass, and in the process of genetic transformation, resistance The induction rate of regenerated buds is extremely low, the genetic transformation cycle is long, and the deformity rate is high, so it is difficult to effectively obtain transgenic strains of rubber grass

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