High-frequency induction and genetic transformation method for pear hairy roots of Agrobacterium rhizogene-mediated balsam

Abstract

The invention relates to a high-frequency induction and genetic transformation method for pear hairy roots of Agrobacterium rhizogene-mediated balsam, comprising the following steps: selecting R1000 agrobacterium rhizogene strains to infect the stems or scutellum of seedlings in test tubes with the seedling age of 10-20 days, co-culturing in an MS culture medium containing IBA5.0 for 1-2 days to obtain co-cultured explants, and performing bacteria-free culture on the explants on a 1 / 2MS0 solid medium containing cephalosporin with the concentration of 500mg L<-1> for 8-10 days to obtain the balsam pear hairy roots. The balsam pear hairy roots acquired by the invention has the rooting frequency and rooting density, which respectively reaches 84.2% and 12.3%, and the total content of balsam pear protodioscin in the balsam is 33.43mg / g, which is respectively 69.1%, 48.6% and 49.8% higher than those in the root, stem and leaf of the seedling in a test tube. The method of the invention lays a foundation for breeding new fine-quality and high-yield balsam pear varieties and utilizing Ri plasmid as a vector to perform genetic transformation, and provides an effective path for adopting the balsam pear hairy roots to culture and biosynthesize medical components such as balsam pear saponin and the like.

Description

technical field

[0001] The invention belongs to the technical fields of plant genetic engineering and cell engineering. Specifically, it relates to a method for Agrobacterium rhizogenes mediating high-frequency induction and genetic transformation of bitter melon hairy roots. Background technique

[0002] Modern research has confirmed that balsam pear saponin is the main active ingredient in balsam pear, which has pharmacological effects such as hypoglycemic, anti-tumor, anti-AIDS, anti-oxidation, anti-fertility, antibacterial, anti-virus, and improving human immunity. Compared with cell culture and callus culture, Agrobacterium rhizogenes ( Agrobacterium rhizogenes ) Ri plasmid transformation induced hairy roots have the characteristics of rapid growth, hormone autotrophy, simple growth conditions, high content of secondary metabolites and stability, high degree of differentiation, and not easy to mutate. compound of. Therefore hairy root culture technology is considered...