High-efficiency agrobacterium-mediated transformation method of soybean root hairs

Abstract

The invention discloses a high-efficiency agrobacterium-mediated transformation method of soybean root hairs. The method includes the following steps: 1) obtaining an agrobacterium rhizogenes lawn carrying a target gene, performing lawn suspension by utilizing a liquid co-culture medium, and adjusting an OD600 value to 0.6-1.0; 2) splitting soybean seeds germinated in a germination medium along the hilum so that half seeds can be obtained, putting the half seeds into the liquid co-culture medium containing the suspension lawn prepared by the step 1), and performing infection for 15-30 min; 3)distributed the half seeds infected by the step 2) in an empty culture container, and performing co-culture in the dark for 3-5 days with a temperature of 23-25 DEG C; and 4) cutting the roots of thehalf seed co-cultured by the step 3), distributing the half seeds in a culture medium, performing dark culture with the temperature of 23-25 DEG C, and selecting positive roots with lengths of 4-6 cmto cut for individual culturing after the roots are grown out. The method is short in operation time, high in rooting rate and positive incidence, and simple and efficient; and explants are not easilypolluted by agrobacterium and other competitors.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and in particular relates to a method for transforming soybean root hairs mediated by high-efficiency Agrobacterium. Background technique [0002] Agrobacteriam rhizogenes is a Gram-negative soil bacterium capable of infecting most dicotyledonous plants. Plants infected with Agrobacterium rhizogenes produce many rapidly growing adventitious roots called hairy or hairy roots [4] . Agrobacterium rhizogenes has important application value in plant transgenic research. It is infected through plant wounds, and its T-DNA is randomly inserted into the chromosome of the host cell, co-expressed with the gene of the host cell, and induces the plant to produce hairy roots [1] . [0003] As early as 1907, Smith and Townsend discovered that Agrobacterium rhizogenes could induce plants to form hairy roots. In 1934, Hildebrand explained this phenomenon again in his research on apple trees. [2] . Th...

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Problems solved by technology

This method is cumbersome to operate and is easily contaminated by Agrobacterium and other miscellaneous bacteria

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