The incidence of Phoma ligulicola in Tasmanian pyrethrum seed and methods of managing seedborne mycoflora were determined. Fourteen different fungi were regularly isolated from seed, including Alternaria tenuissima, Stemphylium botryosum, and P. ligulicola, which have been documented as pathogens of pyrethrum. Comparisons between the incidence of these fungi from seed surface-sterilized with sodium hypochlorite and nontreated seed indicated they occurred both within and on the outside of the seed. A polymerase chain reaction (PCR) test for the detection of P. ligulicola was also developed, with a detection limit of 800 fg of fungal DNA. The assay detected infested seed lots down to an incidence of 0.5%. Reliable amplification of the target DNA was achieved with the addition of bovine serum albumin to reduce the influence of inhibitors from pyrethrum seed. Agar plate tests and PCR demonstrated variability with pyrethrum cultivars in the presence and viability of P. ligulicola in seed. Effective management strategies for the reduction of seedborne P. ligulicola included the regular use of fungicides for reducing foliar disease intensity in the seed fields prior to harvest. Seed treatments with fludioxonil and thiabendazole/thiram also significantly reduced the incidence of seedborne P. ligulicola and increased seed germination and seedling survival.

The isolation frequency of Microsphaeropsis sp. in spring in association with necrotic lesions on leaves in Tasmanian pyrethrum ( Tanacetum cinerariifolium ) fields has increased substantially since first identification in 2001. Examination of morphological features and sequencing of the internal transcribed spacer region (ITS) resulted in the identification of a new species, herein described as Microsphaeropsis tanaceti sp. nov. The pathogenicity of three M. tanaceti isolates to two pyrethrum cultivars was confirmed by inoculating glasshouse-grown plants in three experiments. No significant differences in the susceptibility of the two cultivars to infection by M. tanaceti were found. Symptoms were tan-coloured spots which coalesced around the margins of the leaves. Therefore, the name 'tan spot' is proposed for this new disease of pyrethrum. The sensitivity of seven M. tanaceti isolates to difenoconazole and azoxystrobin, commonly used fungicides for the management of foliar diseases in spring, was assessed under in vitro conditions. Sensitivity testing for difenoconazole was conducted using a mycelial growth assay on potato dextrose agar, whilst testing for sensitivity to azoxystrobin used a conidial germination assay on water agar. Microsphaeropsis tanaceti was found to be more sensitive to azoxystrobin than difenoconazole, with complete inhibition of conidial germination at concentrations above 0·625 μ g a.i. mL -1. By comparison, concentrations of 50 μ g a.i. difenoconazole mL -1 or greater were required for significant inhibition of mycelial growth. It therefore appears likely that there is currently some control of tan spot as a result of the use of azoxystrobin and to a lesser extent, difenoconazole, for the control of other diseases.

To improve sampling efficiency and precision in the assessment of white mould (caused by Sclerotinia sclerotiorum) disease incidence on bean (Phaseolus vulgaris), the spatial characteristics of epidemics were characterized in 54 linear transects in 18 bean fields during 2008-2010 in northern Tasmania, Australia. The incidence of diseased pods and plants was assessed prior to harvest. Distributional and correlation-based analyses indicated the incidence of diseased pods was characterized by a largely random pattern at the individual plant scale, with some patches of similar disease levels on pods occurring at a scale of 1AE5 m or greater. Collectively, these results suggested epidemics may be dominated by localized sources of inoculum. Sequential sampling approaches were developed to estimate or classify disease incidence above or below provisional thresholds of 3, 5 and 15% incidence on pods near harvest. Achieving prespecified levels of precision by sequential estimation was possible only when disease incidence on pods was greater than approximately 4% and sampling was relatively intense (i.e. 10 pods evaluated on each of at least 64 plants). Using sequential classification, correct decisions on disease status were made in at least 95% of independent validation datasets after assessment of only 10AE1-15 plants, depending on classification threshold and error rates. Outcomes of this research provide the basis for implementing more efficient sampling and management strategies for this disease in Australian fields.

Ray blight caused by Phoma ligulicola is an important disease of pyrethrum in Australia, and successful management relies upon the fungicides, azoxystrobin and difenoconazole. Azoxystrobin and difenoconazole were introduced into pyrethrum production in 2001. The sensitivity of P. ligulicola isolates collected in 2003 to azoxystrobin (n ¼ 56) and difenoconazole (n ¼ 61) was tested. Testing for sensitivity to azoxystrobin and difenoconazole used a conidial germination and mycelial growth assay respectively. For each fungicide, the effective dose required to reduce mycelial growth or conidial germination by 50% (EC 50 ) was determined by probit analysis. The EC 50 values ranged from 0.007 to 0.193 lg/ml for azoxystrobin and 0.04 to 13.8 lg/ml for difenoconazole. No evidence was found for cross-resistance between azoxystrobin and difenoconazole in this baseline population. This information serves as important baseline data for tracking future changes in sensitivities of P. ligulicola to these fungicides.

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Downy mildew is a serious threat to opium poppy production globally. In recent years, two pathogen species, Peronospora somniferi and Peronospora meconopsidis, which induce distinct symptoms, have been confirmed in Australia. In order to manage the spread of these pathogens, identifying the sources of inoculum is essential. In this study, we assessed pathogen presence associated with poppy seed. We developed PCR and qPCR assays targeting the coxI and coxII gene regions, for the detection, differentiation, and quantification of P. somniferi and P. meconopsidis in poppy seed. These results were complemented and compared with direct seed histological examination and a seed washing combined with viability staining for oospore detection. The majority of seed lots from all harvest years contained detectable P. meconopsidis, the earliest (1987) predating the first official record of the disease in Tasmania (1996). In contrast, only seed lots harvested in 2012 or later contained P. somniferi, evidence of its more recent introduction. P. meconopsidis contamination was estimated to be as high as 33.04 pg DNA/g of seed and P. somniferi as high as 35.17 pg DNA/g of seed. Incidence of pathogen contamination of seeds, estimated via a group testing protocol, ranged from 0 to 9% (P. meconopsidis) or 0 to 11% (P. somniferi). Mycelia were predominately found external to the seed coat. Seed washing and viability staining demonstrated that putatively viable oospores were present in the majority of seed lots. Transmission testing confirmed both pathogens can be successfully transmitted from infested seed to infected seedling. PCR and qPCR pathogen assays were found to be reliable and offer a routine test for determining pathogen inoculum in poppy seeds.

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