Pythium species have a wide host range and are important pathogens of many agricultural crops. In Zimbabwe, 15 isolates of Pythium have been obtained from symptomatic tobacco (Nicotiana tabacum) in the new float seedling production system. This production system now accounts for 25 to 30% of the tobacco industry's annual requirement of 975 million seedlings. Disease symptoms are observed usually 5 to 6 weeks after sowing as wilting and yellowing of leaves followed by rotting of the roots, and in severe cases, seedling death. Up to 70% seedling loss has been reported in commercial seedbeds. In a study to fulfill Koch's postulates and to determine the susceptibility of 16 commercially grown tobacco cultivars, seedlings were produced in float trays and inoculated individually with 1 ml of Pythium (Isolate Py 19) spores (1 × 104 CFU/ml) and mycelium pipetted around the base of the stem of each seedling 9 weeks after sowing. First symptoms appeared 7 to 10 days after inoculation as yellowing and wilting of leaves. When seedlings were pulled, the lower portion of the stem and roots were brown and rotted. Seedling mortality averaged 29% and disease incidence was 96 to 100% among cultivars. All 16 tobacco cultivars were susceptible to Pythium root and stem rot and there were no significant (P > 0.05) differences in their susceptibility to the disease. The pathogen was reisolated from the inoculated seedlings. The representative isolate (Py 19) sent to Centraalbureau voor Schimmelcultures, the Netherlands was identified as Pythium myriotylum Drechsler (CBS Accession No. 125021) (2). P. aphanidermatum, causing black stem rot, and P. debaryanum and P. ultimum, responsible for damping-off (3), have been reported in the predominant conventional soil-based tobacco seedling production system, but do not cause economic losses. However, stem and root rot caused by P. myriotylum threaten the float seedling production system in Zimbabwe, although a chemical curative control of the disease has been recommended and is now widely practiced. P. myriotylum has previously been reported in the tobacco float seedling production system in South Carolina (1). To our knowledge, this constitutes the first published report of P. myriotylum on tobacco in Zimbabwe. References: (1) M. G. Anderson et al. Plant Dis. 81:227, 1997. (2) Centraalbureau voor Schimmelcultures. Retrieved from http://www.cbs.knaw.nl , 2010. (3) A. J. Masuka et al. List of Plant Diseases in Zimbabwe. Department of Research and Extension and Tobacco Research Board. Harare, Zimbabwe, 2003.

The study was undertaken to identify and characterize Pythium isolates associated with root rot disease of tobacco seedlings as a first step towards developing management strategies for the pathogen. A total of 85 Pythium isolates were collected from diseased tobacco seedlings during 2015-2016 tobacco growing season. The isolates were identified to species level using sequencing of the internal transcribed spacer region. Thereafter, a subset of the isolates was tested for sensitivity to the commonly used fungicides, metalaxyl, azoxystrobin and a combination of fenamidone/propamocarbby growing isolates on Potato Dextrose Agar plates amended with the fungicides. The sequence analysis of the ITS-rDNA identified Pythium myriotylum as the dominant Pythium species associated with the root rot of tobacco seedlings in Zimbabwe. Pythium aphanidermatum and P. insidiosum were also identified albeit at lower frequencies. Phylogenetic analyses of the ITS region of the P. myriotylum isolates showed little sequence diversity giving rise to one distinct clade. The fungicide sensitivity tests showed that metalaxyl provided the best control of P. myriotylum in vitro, as compared to other fungicides. To the best of our knowledge, this is the first comprehensive study to determine and characterize Pythium species associated with root rot of tobacco in the float seedling production system in Zimbabwe. K E Y W O R D Sfloat tray seedling production, fungicide sensitivity, ITSrDNA, Pythium spp., root rot, sequencing, tobacco