Generation of Transgenic Soybean (Glycine max) via Particle Bombardment of Embryogenic Cultures

Abstract

This protocol describes one method for generating transgenic soybean (Glycine max) using particle bombardment of embryogenic cultures. Embryogenic cultures consist of proliferating masses of somatic embryos and are initially obtained from the cotyledons of immature seeds. Embryogenic cultures are bombarded with small DNA-coated particles and the cells that receive, incorporate and express the DNA are selected, based on a co-delivered antibiotic resistance gene. For recovery of whole plants from the antibiotic resistant embryogenic tissues, the tissues are first placed on a medium conducive to embryo development. Mature embryos are then dried for a short period and placed on a germination medium. Plantlets are then gradually exposed to ambient light and humidity conditions, prior to transfer to a greenhouse. Operators need to be constantly monitoring the protocol and observant of all outcomes. These procedures also are mostly descriptive and need to be precisely followed to be successful. © 2016 by John Wiley & Sons, Inc.