A kind of genetic transformation method of rubber grass

Abstract

The invention provides a method for genetic transformation of rubber grass, comprising: (1) taking the young leaves of Xinjiang rubber grass germplasm, cutting them, and putting them in: MS+1.5~1.6mg / L6‑BA+0.1mg / LNAA+20g / L sucrose, cultivated in the bud induction medium of pH5.8; After 50 days, select the excellent germplasm of rubber grass suitable for tissue culture; transfer to: 1 / 2MS+20g / L sucrose, the rooting medium of pH5.8, become Tissue culture seedlings of regenerated rubber grass; (2) cut the leaves of the seedlings and pre-cultivate them on the bud induction medium for 2 days; (3) add Agrobacterium infection solution and transfer them to the bud induction medium for co-cultivation in the dark 3 days; transfer to bud induction medium containing 400mg / L Car for one week; 10mg / L Hyg, pH5.8 on the selection medium, induce the regeneration of resistant shoots; transfer to 1 / 2MS+20g / L sucrose+400mg / L Car and 8~10mg / L Hyg, pH5.8 new rooting culture Basically, regenerated plants with resistance to rubber grass are obtained; the genetic transformation method of rubber grass of the present invention has the induction rate of resistant regeneration buds > 30%, the plant deformity rate of transgenic regenerated seedlings is low, and the positive detection rate of transgenic rubber grass > 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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