The invention relates to an
Agrobacterium rhizogenes-mediated
transgene Stevia rebaudiana genetic transformation method. The method comprises: pre-cultured
Stevia rebaudiana stem apex explant is immersed into an
Agrobacterium rhizogenes solution of pRI101-ANDNA
plasmid carrying
target gene, treatments such as infection, co-culture and sterilization are performed, the hairy root is cultured in a
kanamycin-containing screening culture medium, the anti-
kanamycin hairy root is formed on the explant through induction, the hairy root is induced to form
callus, differentiation of adventitious
bud is performed, the adventitious
bud continuously grows to form the test-tube
plantlet, and a PCR amplification method is adopted to identify the
transgene plant carrying out the
target gene. With the technical scheme, the
target gene can be transformed into the
Agrobacterium rhizogenes, such that the steps of separation from the
protoplast and culture are eliminated compared with genetic transformation of
polyethylene glycol and other chemical methods; and compared with genetic transformation of
gene gun and other physical methods, the method of the present invention has characteristics of no requirement of expensive equipment, and simple operation technology.