7ElectroporationThe Electroporation of cells can be used to deliver DNA into plant cells andprotoplasts. The vectors used can be simple plasmids. The genes of interest require plantregulatory sequences, but no specific sequences are required for integration. Material isincubated in a buffer solution containing DNA and subjected to high-voltage electricalpulses. The DNA migrates then through high voltage induced pores in the plasmamembrane and integrates into the genome. Initially, protoplasts were used fortransformation, but one of the advantages of the system is that both intact cells andtissues (such as callus cultures, immature embryos and inflorescence material) can beused. However, the plant material used for Electroporation may require specifictreatments, such as pre- and post-electroporation incubations in high osmotic buffers. Theefficiency of electroporation is very dependent on the condition of the plant material usedand the electroporation and tissue treatment conditions chosen. The advantage of thissystem is all the electroporated cells are in the same physiological state.This method canbe used both for plant and animal transformation.MicroinjectionIn this method, which is mostly used for animal cells, DNA solution is directlyinjected into the nucleus of the cell. Typically, a microinjection assembly consists of alow power stereoscopic dissecting microscope and two micromanipulators, one for aglass micropipette to hold the nucleus by partial suction and the other for a glass injectionneedle to introduce the DNA into the nucleus. Generally, 2 picolitre (2 X 10-12litre) ofDNA solution is injected into the nucleus. The transgene integration occurs at randomsites in the genome.Other methods includes macroinjection, liposome mediated transformation, ultrasound mediated DNA transfer, DNA transfer via pollen etc is also used in planttransformation.
Read more here: Source link