First Report of Rhizoctonia solani AG 4HG-III Causing Potato Stem Canker in South Africa.

Abstract

Black scurf and stem canker caused by Rhizoctonia solani Kühn (teleomorph: Thanathephorus cucumeris Frank Donk) are potato diseases of worldwide economic importance (4). R. solani consists of 13 anastomosis groups (AGs) of which AG 3-PT is considered the dominant causal agent of potato diseases globally (1,4). However, other AGs such as AG 2-1, 5, and 8 have been reported to cause potato diseases (1,4). In February 2013, potato stem samples (cv. Mondial) displaying dark brown lesions resembling those caused by Rhizoctonia stem canker were obtained from a commercial field in Limpopo Province, South Africa. Symptomatic tissue was disinfected with 1% NaOCl for 1 min, rinsed in sterile water, and 4-mm stem pieces excised from the margins of symptomatic tissues and plated on 2% water agar supplemented with 20 mg/l of chloramphenicol. Single hyphal tips taken from fungal isolates identified as R. solani based on morphological traits (3) were transferred to potato dextrose agar. DNA was isolated from the resulting cultures and ITS region of rDNA was sequenced as previously described (2). The resulting sequences of three of the isolates, Rh 81, Rh 82, and Rh 83 (KF712285, KF712286, and KF712287), were 99% similar to those of AG 4 HG-III found in GenBank (DQ102449 and AF354077). Therefore, based on molecular methods, these three isolates were identified as R. solani AG4 HG-III. To determine pathogenicity of the AG4 HG-III isolates, certified disease free mini-tubers (Generation 0, cv. Mondial, produced in tunnels) were used in pot trials. PDA plugs of each isolate were added to 10 g of barley grains, which had been sterilized by autoclaving for two consecutive days at 121°C for 30 min, and were incubated for 14 days until fully colonized. Ten colonized barley grains were placed 10 mm above each mini-tuber planted in 5l pots containing sterile potting mixture of sand:clay:pinebark (1:1:1). Ten tubers were inoculated with each isolate. Uninoculated, sterile barley grains were applied to the control treatment. Mini-tubers were grown in a greenhouse maintained at 22°C with light for a 12 h day. After 7 weeks, five plants for each isolate were destructively sampled and assessed for stem canker symptoms. At 120 days after sowing, the remaining five plants per treatment were assessed for blemishes on progeny tubers. The stem canker incidences of plants inoculated with Rh 81, Rh 82, and Rh 83 were 25, 25, and 50%, respectively, whereas no symptoms were observed in control plants. Sclerotia formation and blemishes were not observed on any of the progeny tubers, which might indicate that these strains are only able to infect stems, or that environmental conditions were not suitable for tuber blemish or black scurf development. R. solani AG4 HG-III was consistently re-isolated from symptomatic stems displaying brown lesions, and the identity of the re-isolates were confirmed by molecular tests as previously described, thereby fulfilling Koch's postulates. To our knowledge, this is the first report of R. solani AG4 HG-III causing stem canker on potato in South Africa and worldwide. Knowledge of which AGs are present in crop production systems is important when considering disease management strategies such as crop rotation and fungicide treatments (3).