DNA Protoplastos

A very high dosage can provoke the death of all the cells and a sub-dosage can lead to the appearance of escapes (plants not transformed) (BESPALHOK F., et al WAR; OLIVEIRA, 2000). The use of the biolstico process is sufficiently ample e, when compared with the majority of the methods right-handers of introduction of DNA in plants, the bombardeamento of particles presents some advantages. It is one versatile technique highly and of fciladaptao, being able to be applied the great variety of cells and fabrics, including suspensions (Klein et al., 1989; Fromm et al., 1990), meristemticos calluses (Vasil et al., 1985), fabrics (McCabe & Martinelli, 1993), immature embryos (Southgate et al., 1998) and somatic embryos (Finer & McMullen, 1991; Santarm & Blacksmith, 1997). This technique has allowed the regeneration of transgnicas plants in reproduzvel way and with little variability between experiments (Luthra et al., 1997). Eletroporao of Protoplastos the eletroporao of protoplastos is an used method to introduce macro-molecules in vegetal cells. Protoplastos is cells unprovided of cellular walls, and, theoretically, they can be isolated of any fabric vegetal. In well established conditions of fabric culture, the protoplastos reconstitute its walls, are divided, formed colonies, calluses and regenerate plants, for embriognese or organognese (ABDULLAH et al., 1985; PANIS et al., 1993).

The eletroporao consists of the application of a pulse of high voltage to a solution contends DNA and protoplastos in suspension, inducing reversible pores in the cellular membranes, resulting in flows of ons and molecules through the deformed membrane (CHANG, 1989). FROMM et al. (1985) they had used the technique of eletroporao in protoplastos of maize and tobacco, having thus offered an alternative of transformation of plants, since the Agrobacterium system was considered inefficient in monocotiledneas. POLIETILENOGLICOL (PEG) Polietilenoglicol (PEG), used in combinaocom Ca+2, alkaline Mg+2 and pH, promotes the linking of the exgeno DNA to the surface of the protoplastos. .