Genetic transformation method of creeping type ground-cover chrysanthemum

Abstract

The invention relates to a method for genetic transformation of creeping ground-cover chrysanthemum, which belongs to the field of molecular breeding of chrysanthemum. RNA of a chrysanthemum variety zhongshanhongfeng is taken as a template to amplify to synthesize a DREBa gene, the gene is cloned to a T-vector, a target fragment of the DREBa gene obtained by double digestion is connected with pCAMBIA 1301, and then agrobacterium is transformed to obtain a bacterial liquid of the agrobacterium containing a recombinant plasmid pCAMBIA1301-DREBa; and a leaf disc of an aseptic seedling is infected by the bacterial liquid of the agrobacterium for 10min, and a rooted seedling with hygromycin resistance is obtained through the total culture for 3d, the debacteria culture for 3d and the screening culture. The PCR detection proves that a foreign gene has been transferred into DNA of a genome of the ground-cover chrysanthemum. The method aims at the creeping ground-cover chrysanthemum to establish a genetic transformation system of the creeping ground-cover chrysanthemum for the first time, provides a theoretical basis and an experimental evidence for molecular breeding work of the ground-cover chrysanthemum, and can effectively promote the progress of the molecular breeding of the ground-cover chrysanthemum.

Description

1. Technical field [0001] The invention relates to a genetic transformation method of creeping ground cover chrysanthemum, which is specially used for the molecular breeding of creeping ground cover chrysanthemum and belongs to the field of chrysanthemum molecular breeding. 2. Background technology [0002] Ground cover chrysanthemum is a kind of unique ground cover plant with comprehensive functions of greening, beautifying, coloring and aromatizing at the same time. department concerned. Therefore, screening and cultivating new varieties of creeping ground cover chrysanthemum with excellent characters has great practical value and economic significance. [0003] In recent years, with the development of plant genetic engineering, molecular breeding has overcome the limitations of conventional breeding, providing new means and ways for improving chrysanthemum varieties with new genetic resources. The Agrobacterium-mediated method has become the preferred method in the tran...

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to solve the problems of strong genotype dependence, low transformation frequency, difficult regeneration of explants after transformation, poor repeatability, and difficult seedling formation of transformed cells in the transgene of chrysanthemum mediated by Agrobacterium tumefaciens. The genetic transformation conditions of creeping ground cover chrysanthemum have been systematically and deeply studied, and a set of genetic transformation system suitable for creeping ground cover chrysanthemum has been established, laying the foundation for the molecular breeding of creeping ground cover chrysanthemum

Images