Transformation of nuclear DNA in meristematic and embryogenic tissues

Abstract

Particle bombardment/biolistic delivery is a very popular method of genetic transformation of diverse targets including cells and intact tissues. Delivery of DNA through particle bombardment is genotype and species independent, nevertheless, an efficient protocol for large-scale generation of transgenic plants through embryogenic tissues with a high (≥80%) shoot regeneration efficiency is a prerequisite. Young embryogenic tissues or multiple shoot buds in early stages of induction are the most suited target tissues for recovery of transgenic plants. We describe the protocol for delivery of foreign genes using particle delivery system (Biorad gene gun, PDS-1000/He) in to the meristematic tissues of embryonic axes derived from mature seeds of castor. With the optimized physical and biological parameters, putative transformants were obtained at a frequency of 1.4% through particle gun bombardment of castor embryo axes. Also, transformation of embryogenic calli of sorghum using particle inflow gun (PIG) is described. © 2013 Springer Science+Business Media, LLC.