Electroporation of intact embryonic leaflets of peanut: Gene transfer and stimulation of regeneration capacity

Abstract

Direct gene transfer into peanut intact embryonic leaflets was performed through electroporation. In transient bglucuronidase expression assays, maximal expression was obtained by using pulses of 625 V cm 21 in EPRm (modified electroporation) buffer supplemented with 75 mM NaCl. Kanamycin-resistant plants were obtained, and the presence of the nptII gene was demonstrated by PCR analysis. The positive effect of electroporation on the efficiency of in vitro regeneration was demonstrated. Explants submitted to field strengths between 500 and 625 V cm 21 displayed a significantly increased number of shoots and originated faster growing calluses relative to control explants. Whereas in control explants callus formation occurred only at the petiolule, electroporated leaflets developed additional organogenic calluses on the foliar lamina.

AMELIORATION DES PLANTES Biotechnologies et arachide

“…Depuis, des arachides transgéniques ont été produites par bombardement de particules [16][17][18] ou via la bactérie Agrobacterium tumefaciens portant une résistance à un antibiotique [19][20][21]. Plus récemment, la mise au point d'une méthode d'électroporation a permis un transfert direct de gène dans des folioles embryonnaires (plus faciles à manipuler que les protoplastes) avec une bonne qualité de régénération [22].…”

Antifungal Activity in Transgenic Peanut (Arachis hypogaeaL.) Conferred by a Nonheme Chloroperoxidase Gene

“…Transformation of peanut has been accomplished by several different methods, including particle bombardment (Higgins et al 2004;Livingstone and Birch, 1999;Magbanua et al 2000;Ozias-Akins et al 1993;Yang et al 1998), Agrobacterium-mediated transformation (Cheng et al 1997;Rohini and Rao, 2001;Sharma and Anjaiah, 2000), and electroporation (De Padua et al 2000). Individual genes that confer agronomic traits have been integrated into the peanut genome such as bialophos resistance (bar) for herbicide tolerance (Brar et al 1994), Bacillus thuringiensis (Bt) toxin cryIA(c) for insect resistance (Singsit et al 1997), viral nucleocapsid or coat protein genes for virus resistance (Higgins et al 2004;Li et al 1997;Magbanua et al 2000;Sharma and Anjaiah, 2000;Yang et al 1998;Yang et al 2004), chitinase, glucanase, and oxalate oxidase to control fungal diseases (Chenault et al 2005;Chenault et al 2002;Livingstone et al 2005;Rohini and Rao, 2001).…”