First Report of Damping-off Caused by Pythium aphanidermatum on Leaf Beet (Beta vulgaris subsp. vulgaris) in Italy.

Abstract

During July 2010, symptoms of crown and root rot were observed on leaf beet (Beta vulgaris L. subsp. vulgaris) grown in a commercial field near Torino (northern Italy). The first symptoms developed 25 days after sowing with temperatures ranging from 25 to 30°C, and 20% of plants were affected. Affected plants were stunted and leaves showed chlorosis and suddenly wilted. The collar and young stems were affected first and appeared brown, water-soaked, and were characterized by a soft rot. Eventually, all affected plants collapsed. Thin aerial mycelia were visible on the surface of the infected plants if maintained at a high relative humidity. Tissue fragments of 1 mm2 were excised from the margins of the lesions, dipped in a solution containing 1% sodium hypochlorite, and plated on potato dextrose agar (PDA) and on a medium selective for oomycetes (2). Plates were incubated under constant fluorescent light at 22 ± 1°C for 5 days. Five isolates, grown on V8 medium (vegetable mix 300 g; agar 15 g; CaCO3 1.5 g; distilled water 1 liter) and observed under light microscope showed the morphological characters of Pythium aphanidermatum (3). This result was confirmed by PCR and sequence analysis. The internal transcribed spacer (ITS) region of rDNA of a single isolate (Py 7/10) was amplified using the primers ITS1/ITS4 and sequenced. BLAST analysis (1) of the 815 bp segment showed a 99% homology with the sequence of P. aphanidermatum (GenBank Accession JN695786). The nucleotide sequence has been assigned to the GenBank Accession JX462954. Pathogenicity tests were performed twice on B. vulgaris subsp. vulgaris grown in 2-liter pots, containing a steam disinfested organic peat substrate (70% black peat, 30% white peat, pH 5.5 to 6, N 110 to 190 mg L-1, P2O5 140 to 230 mg L-1, K2O 170 to 280 mg L-1), infested with wheat and hemp kernels colonized with a strain of P. aphanidermatum at a rate of 1 g L-1. Ten seeds per pot were sown in four pots filled with the infested medium, while the same number of seeds were sown in non-infested substrate. Plants were kept in two growth chambers, at 20 and 27°C. The first symptoms developed 7 days after the artificial inoculation. After 20 days, 70% of plants were infected at 27°C, while 10% were affected at 20°C. Control plants remained healthy at both temperatures. P. aphanidermatum was consistently reisolated from the lesions. To our knowledge, this is the first report of damping off of B. vulgaris subsp. vulgaris caused by P. aphanidermatum in Italy. The importance of the disease, at present limited, could increase in areas where leaf beet is intensively grown.