Method for efficiently and rapidly stabilizing gene transformation for tomatoes

Abstract

The invention discloses a method for efficiently and rapidly stabilizing gene transformation for tomatoes. The method comprises the steps that 1, activation of stains is conducted; 2, preparation of plant materials is conducted; 3, infection is conducted; 4, screening and culturing are conducted. According to the method, the tomato transgenic efficiency is increased, and the transformation rate can reach over 30 percent. The concentration of bacterial liquid is reduced, the infection time is prolonged, and on one hand, agrobacterium pollution is avoided; on the other hand, the infection efficiency is increased. The infection bacterial liquid is not added with drugs for assisting in infection, so that damage to tomato leaves is reduced, the survival rate and the regeneration rate of leaf culture are increased; in a traditional method, drugs such as acetosyringone which assists in infection are usually added, damage is caused to leaves, and the regeneration capacity of the leaves is reduced. By means of carriers carrying strong expressed reporter genes, transgenic plants can be obtained directly by screening through observation, and tedious work in traditional detection is avoided. The cycle of transgenes is shortened.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, in particular to a method for efficient, rapid and stable gene transformation of tomato. Background technique [0002] Tomato (Lycopersicone sculentum Mill.) is an important food crop in the world. It has many varieties, high yield, rich nutrition and wide application. Micro-Tom is a tomato mutant, which retains the basic characteristics of common tomato as a model plant, but the plant is short and has a shorter life cycle, which has the great advantage of saving research space and shortening research time. With the rapid development of molecular biology and genetic engineering in recent years, the research focus has turned to the deep mining of excellent genes in tomato germplasm resources, and the identification and determination of molecular markers and gene expression sequences. At the same time, research results on genetic improvement such as stress resistance, storage and tran...

AI Technical Summary

Problems solved by technology

Although the genetic transformation technology route of tomato is mature, the traditional method is laborious, time-consuming, cumbersome and inefficient, and too few transgenic plants are obtained, which brings obstacles to the later identification of transgenic plants

The operation process of the traditional Agrobacterium-mediated method is cumbersome, and many problems are prone to occur during the transgenic process, such as the selection of suitable vectors, the selection of suitable strains, the contamination of miscellaneous bacteria, the inability of Agrobacterium to be inhibited, and the concentration of bacteria solution is too high or too high. Low results in unsuccessful transgenics, control of the duration of infection, etc. These problems will affect the efficiency and speed of tomato transgenics. The transgenic efficiency of tomato plants obtained by traditional methods is less than 1%.