Novel methodology for detection of Botrytis cinerea from apple symptomless

DELIA BENUZZI; GASTÓN FERNÁNDEZ; MARTÍN A. FERNÁNDEZ BALDO; GABRIELA SANSONE; VIVIANA CALVENTE; ELOY SALINAS; MARÍA ISABEL SANZ FERRAMOLA

San Juan

Congreso; 2da Reunión Conjunta de Sociedades de Biología de la República Argentina: XIII Jornada de la Sociedad Argentina de Biología, XXIX Reunión Anual de la Sociedad de Biología de Cuyo, XVIII Jornadas Científicas de la Sociedad de Biología de Córdoba; 2011

Universidad Nacional de San Juan - Sociedad Argentina de Biología - Sociedad de Biología de Cuyo

         NOVEL METHODOLOGY FOR         DETECTION OF Botrytis cinerea        FROM APPLE SYMPTOMLESS                      Benuzzi D, Fernández G, Fernández Baldo M, Sansone G,                            Calvente V, Salinas E, Sanz Ferramola M I.                            Tec. Qca. y Biotec.  UNSL. E-mail:jgfernandez@unsl.edu.ar   Gray mold caused by Botrytis cinerea, one of the major diseases of apple fruits in post-harvest. Their detection in disease early stages and symptomless, is essential to establish control strategies. This study developed a methodology for detection of B. cinerea from apple symptomless, through a protocol of extraction DNA and amplification for PCR the IGS ribosomal spacer and transposable elements (flipper and boty). Red Delicious apples were wounded with a punch, and inoculated with 20 µl of a conidial suspension (106 conidia/mL) of B. cinerea; after 3 days of inoculation and without symptoms of disease, the fruits were processed in a mortar in liquid nitrogen to homogenize the samples. Pure cultures of B. cinerea (3 days old) were used as positive control and healthy apples as negative control. The extraction method was easy and allowed obtain genomic DNA of B. cinerea from samples of apple symptomless, which is of sufficient quantity and quality for amplify by PCR the regions of interest. PCR amplification indicates the presence of both transposable elements and IGS spacer. Fungus genomic DNA was molecularly characterized in subpopulation transposa type. Pure cultures are also observed transposable elements and IGS spacer, while in healthy apples were not detected. In conclusion, the proposed methodology was able to detect the presence of B. cinerea in apple symptomless (gray mold) for its possible application in the preventive control of latent diseases.