First Report of Sugar Beet Seedling Damping-Off Caused by Binucleate Rhizoctonia AG-A in China.

Abstract

Sugar beet (Beta vulgaris L.) is grown worldwide and produces one-third of the world's sugar supply. Sugar beet seedling Rhizoctonia damping-off is an important disease mainly caused by Rhizoctonia solani AG-2, AG-4, and AG-5 (2). In 2010, diseased sugar beet seedlings with about 20% incidence affected by damping-off, which showed dark brown lesions on the stems just below the soil surface and portions of the roots, were collected from nurseries in three locations in Heilongjiang province, northeast China. Root fragments taken from the margins of healthy tissues and lesions on roots were surface disinfected with 0.5% sodium hypochlorite for 2 min, rinsed with sterile water, then placed on potato dextrose agar (PDA) and incubated at 25°C in the dark. Three (designed HLJ-RAA1, HLJ-RAB1, HLJ-RAB2) of nine Rhizoctonia isolates were obtained from diseased tissues and preliminarily identified as binucleate Rhizoctonia (BNR) anamorph (teleomorph Ceratobasidium Rogers) species-like. Fungal colonies were white with large amounts of floccose, aerial hyphae. Hyphal cells were determined to be binucleate when stained with 4'-6-diamidino-2-phenylindole (DAPI) (1). No sclerotia were produced after 14 days on PDA. Average hyphal diameter of the three isolates were 4.2, 4.3, and 4.8 μm, respectively. Further, the internal transcribed spacer (ITS) region of rDNA was amplified from the genomic DNA extracted from hyphae by bead beating in 2% CTAB solution using stainless steel beads with primers ITS1 and ITS4. The ITS sequences (GenBank Accession Nos. JX073668, JX073669, and JX073670) were over 99% identical to those of more than 50 Ceratobasidium sp. AG-A isolates (e.g., GenBank Accession No. JQ688054.1; strain HY-15). Therefore, based on morphological and molecular characteristics, these isolates were identified to be BNR AG-A. To determine the pathogenicity of the isolates, sugar beet (cv. HI0305) seedlings were inoculated with wheat seeds colonized with each of the isolated Rhizoctonia strains (one seed per seedling), and grew in pots under greenhouse conditions (3). After 3 weeks, some inoculated plants showed damping-off as observed in the nurseries, whereas noninoculated control plants (sterile wheat seeds only) remained healthy. Disease incidence from the trials averaged 53.3%, 70%, and 53.3% for the isolates HLJ-RAA1, HLJ-RAB1, and HLJ-RAB2, respectively. The three BNR cultures of the pathogens were consistently reisolated from symptomatic roots, and their identities confirmed by morphological and molecular characteristics as described above, fulfilling Koch's postulates. BNR AG-A was previously reported to be pathogenic to soybean, pea, snap bean, and pak choy in China (4). However, to our knowledge, this is the first report of BNR AG-A causing sugar beet seedling damping-off in China. Sugar beet is often grown in crop rotation with soya bean and such a rotation could increase the risk of soilborne infection to either crop by BNR AG-A.