A new pathogen of pyrethrum (Tanacetum cinerariifolium) causing anthracnose was described as Colletotrichum tanaceti based on morphological characteristics and a four-gene phylogeny consisting of rDNA-ITS, b-tubulin (TUB2), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and actin (ACT) gene sequences. The fungus produced perithecia in culture, requiring an opposite mating type isolate in a heterothallic manner. The initial infection strategy on pyrethrum leaves involved the formation of appressoria followed by production of multilobed infection vesicles in the epidermal cells. Infection and colonization then proceeded through thinner secondary hyphae, which resulted in the initial production of water-soaked lesions followed by black necrotic lesions. The infection process was suggestive of a hemibiotrophic infection strategy. Moreover, phylogenetic analysis clearly showed that C. destructivum, C. higginsianum and C. panacicola were separate species that also had similar intracellular hemibiotrophic infection strategies as C. tanaceti, which all clustered in the C. destructivum complex. Colletotrichum spp. were detected at 1% incidence in seed of 1 of 19 seed lines, indicating the potential for seed as a source of inoculum into crops. Colletotrichum tanaceti was detected in leaf lesions from 11 of 24 pyrethrum fields surveyed between April and July 2012, at a frequency of 1Á3-25Á0% of lesions. Anthracnose probably contributes to the complex of foliar diseases reducing green leaf area in pyrethrum fields in Australia.
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.
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.
White mold (Sclerotinia sclerotiorum) of leguminous crops in New York is generally managed with preventive applications of fungicides. However, no research has been conducted during the last decade to assess the sensitivity of the S. sclerotiorum population to fungicides or compare their performance under field conditions. The sensitivity of S. sclerotiorum to boscalid, fluazinam, and thiophanate-methyl was assessed in 151 isolates from 15 fields across New York using an agar dilution method with discriminatory concentrations. In addition, the effective concentration at which mycelial growth is reduced by 50% (EC50) was estimated for one representative isolate from each field. The efficacy of commercial formulations of each fungicide on white mold incidence in plants and pods was also tested in two field trials (2015 and 2016). The EC50 values ranged from 0.068 to 0.219, 0.001 to 0.002, and 1.23 to 2.15 µg/ml for boscalid, fluazinam, and thiophanate-methyl, respectively. Evidence of resistance was not found using the discriminatory concentration tests. The mycelial growth inhibition relative to the control ranged from 56 to 83%, 66 to 84%, and 53 to 83% at discriminatory concentrations of boscalid (5 µg a.i./ml), fluazinam (0.05 µg a.i./ml), and thiophanate-methyl (5 µg a.i./ml), respectively. Fourteen isolates with mycelial growth inhibition lower than 60% at 5 µg/ml of thiophanate-methyl, did not exhibit point mutations within a partial sequence of the β-tubulin gene. In the field trials, fungicides effectively reduced white mold incidence on plants by 75% (2015) and 93% (2016) and on pods by 81% (2015) and 87% (2016), both relative to the nontreated plots. However, fungicide applications led to significant increases in pod yield, relative to the nontreated plots, only in 2015 when the incidence of white mold on plants and pods were higher (85 and 49.2%) than in 2016 (31.3 and 10.3%). Although fungicide resistance was not detected, and thus control failures reported by New York snap bean growers may be due to other factors, further monitoring of sensitivity within the S. sclerotiorum population is encouraged as well as the use of rational systems to base their judicious and economic use.
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