Plant & Animal Genomes XV Conference

January 13-17, 2007
Town & Country Convention Center
San Diego, CA

Targeted Mutagenesis As A Tool For Tobacco Crop Improvement

Emilie Julio¹ , Frédéric Laporte² , Stéphanie Reis¹ , Christophe Rothan² , François Dorlhac de Borne¹

¹  Altadis - Institut du Tabac, Domaine de la Tour, 769 route de Sainte-Alvère, 24100 Bergerac, FRANCE

²  UMR Physiologie et Biotechnologie Végétales, IBVI-INRA Bordeaux, 33883 Villenave d'Ornon Cedex, FRANCE

Nicotiana tabacum is a model widely used in functional genomics with transgenesis; however, genetically modified organisms are not accepted by consumers in Europe. Targeted mutagenesis as a non transgenic approach was assessed on a demonstration gene involved in alkaloid metabolism. A population of 4.000 EMS-mutagenized M2 families was created. Single Strand Conformation Polymorphism (SSCP) was used to target mutations. Eleven putative mutants were identified by screening 1344 M2 families. Mutations identified in DNA pools were validated by sequencing. Individual plants carrying missense or truncation mutations were studied for their phenotype. Homozygous plants for one truncation were identified, and the expected phenotype was observed in the field. These plants have been used as genitors to introduce this mutation into elite lines. Backcrosses are being performed to recover the elite line background in combination with SSCP analysis to follow the mutation. The amphidiploid nature of tobacco avoids problems related to fertility. Efficiency of this method to create novel genetic variation and to develop cultivars has been demonstrated for the first time in tobacco.