CONICET | Buscador de Institutos y Recursos Humanos

A Quick and easy method for DNA extraction from apples infected with Botrytis cinerea Fernández G, Fernández Baldo M, Sansone G, Calvente V, Salinas E, Benuzzi D, Sanz Ferramola M I Tecnología Qca y Biotecnología. U.N.S.L. E-mail:jgfernandez@unsl.edu.ar Botrytis cinerea is an important phytopathogenic fungus responsible for grey mould in fruit apple. Several methods of fungi genomic DNA extractions have been used. B. cinerea have been molecularly characterized in transposa or vacuma distinguished by the presence of transposable elements. The objective of this work was obtaining an extraction method of DNA by B. cinerea from infected apple. Fruit were wounded with a punch, and inoculated with 20 ul of a conidial suspension. Apples were processed in a mortar in liquid nitrogen to homogenize. DNA quality was determined through the absorbance at 260/280 nm. DNA concentration was evaluated in agarose gel 1% by comparison with standard molecular weight marker. DNA quality was validated through its consistent amplification using PCR technique, with primers of ribosomal regions 18S (IGS spacer) and the transposable elements Flipper and Boty. Products were analyzed on agarose gel 2% by comparison with standard molecular weight marker. PCR amplificationindicated presence of transposable elements Flipper, Boty and ribosomal IGS spacer. Fungi genomic DNA was molecularly characterized in subpopulation transposa. This methodology for DNA extraction is fast, simple and renders genomic DNA of good quality, free of proteins and secondary compounds. Then, we obtained an extraction method of DNA by B. cinerea directly from homogenized infected apple. Supported by SeCyT-UNSL 22/Q941