Neofabraea vagabunda is the prevalent cause of bull's eye rot, one of the main postharvest diseases of apple, in many producing areas, but its biology has not been studied in detail. The molecular identification, by DNA sequencing of the b-tubulin region, of 41 isolates collected from apples showing bull's eye rot in the Emilia-Romagna region confirmed N. vagabunda as the main species in Italy. A biological and morphological characterization of N. vagabunda isolates was performed in vitro. Assays at temperatures ranging from 0 to 30°C carried out on 10 isolates demonstrated: (i) a marked influence of temperature on colony morphology, conidial production, conidial size and mycelial growth, showing the cold-tolerant character of N. vagabunda; and (ii) that culture at 15°C on tomato agar (TA) for 14 days is a rapid and reliable method to favour pathogen conidial production. Trials performed on 38 isolates using these incubation conditions recorded the presence of two N. vagabunda morphotypes, differing for colony morphology, conidial size, conidiomata formation and temperature requirement. The alkalizing ability of the pathogen during growth on TA was also demonstrated for the first time. The pathogenicity of 25 N. vagabunda isolates was proved in vivo on artificially infected Cripps Pink apples. A pH increase was also recorded in apple tissue infected by N. vagabunda isolates (on average 0.2 and 0.3 units of pH after 60 and 120 days of incubation, respectively), suggesting that the N. vagabunda transition from quiescence to necrotrophic colonization in apples could involve the secretion of alkalizing compounds.

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The fungicide cyflufenamid (phenyl-acetamide, Fungicide Resistance Action Committee [FRAC] code U6) was approved for use in Italy in 2011 as Takumi (Certis Europe, Utrecht, The Netherlands) to control Podosphaera xanthii (Castagne) U. Braun. & N. Shishkoff, the main causal agent of cucurbit powdery mildew. Considering that strains of this pathogen have developed resistance to strobilurin (5) and demethylation inhibitor (DMI) (4) fungicides, cyflufenamid represented a viable alternative to control this disease. However, this fungicide is also prone to resistance development as illustrated by resistance of P. xanthii in Japan (3). In the 2012 and 2013 growing seasons, significant declines in cyflufenamid efficacy were observed in two experimental fields in the Apulia (AP) and Emilia-Romagna (ER) regions of Italy on Cucumis melo and Cucurbita pepo, respectively. Takumi had been applied four times at the recommended field rate of 0.15 liter/ha (15 μg/ml of active ingredient [a.i.]) each growing season since 2010 in each field. Powdery mildew-infected leaf samples were collected in 2012 from both fields (25 isolates from AP and 19 from ER), and from five gardens (one isolate per garden); while in 2013, samples were collected only from the ER field (two polyconidial isolates). Isolates were maintained on detached zucchini cotyledons (1). Sensitivity of the isolates to cyflufenamid was determined by leaf disk bioassays (4) using Takumi at 0.01, 0.1, 1, 10, 20, and 50 μg a.i./ml. The 50% effective concentration (EC50) and the minimum inhibitory concentration (MIC) values were calculated (2). Isolates collected in ER and the gardens in 2012 all had an EC50< 0.01 μg/ml, and the MIC ranged from <0.01 to <1 μg/ml. Isolates from AP in 2012 had elevated EC50 values, from 0.230 to >50 μg/ml, and MIC values from <10 to >50 μg/ml; by 2013, the EC50 values of ER isolates ranged from 3.35 to >50 μg/ml. Based on the mean EC50 value of 0.0019 μg/ml for sensitive isolates of P. xanthii in Japan (2), isolates from both the ER field and gardens in 2012 were considered sensitive to cyflufenamid. Additionally, EC50 values of AP isolates from 2012 and ER isolates from 2013 were greater than those of sensitive isolates, indicating a shift in sensitivity toward resistance to cyflufenamid (resistance factor >100 [2]). Consequently, poor control of powdery mildew with cyflufenamid applications in the AP and ER trials was most likely a result of fungicide resistance. Isolates from these fields were exposed to selection pressure for fungicide resistance because cyflufenamid was applied more times than permitted in the label instructions. However, control of powdery mildew in 2013 was not as effective as in previous years in commercial fields in AP (C. Dongiovanni, personal communication). This observation, combined with proof of reduced sensitivity of some P. xanthii strains in Italy to cyflufenamid, highlights the need for implementing resistance management strategies to minimize the risk of fungicide resistant strains developing in cucurbit fields. References: (1) B. Álvarez and J. A. Torés. Bol. San. Veg. Plagas 23:283, 1997. (2) M. Haramoto et al. J. Pest. Sci. 31:397, 2006. (3) H. Hosokawa et al. Jpn. J. Phytopathol. 72:260, 2006. (4) M. T. McGrath et al. Plant Dis. 80:697, 1996. (5) M. T. McGrath and N. Shishkoff. Plant. Dis. 87:1007, 2003.

In three closely related genera of the Proteaceae, namely, Lomatia R.Br., Stenocarpus R.Br. and Strangea Meisn. the pollen presenter and stigma at anthesis are covered by specialised cells. The cells are spiral-walled, contain polyphenolic material and are produced from the stigma, and in Lomatia from the pollen presenter. The spiral wall-thickenings are formed on the inside of the primary wall late in the development of the cells when they are budded off from the stigma or presenter surface. Pollen from the anthers is placed onto these cells from where it is taken to other flowers for cross-pollination. The spiral-walled cells prevent contact between self-pollen and the stigma and provide a method whereby cross-pollen can only reach the stigma if self-pollen has been removed. The form of the cells and the mechanism for preventing autogamy is not found in any other plant group.

We characterized Plasmopara viticola populations utilizing PCR-RFLP technique to detect a point mutation known to cause resistance to carboxylic acid amides (CAA) fungicides. Sensitivity of these samples to the mandipropamid fungicide was assayed by a leaf-disc method. In this work, we provide the first evidence about the presence of mandipropamid-resistant populations of P. viticola from commercial vineyards in Italy. Improving the knowledge about development of resistant populations could enhance the current grapevine downy mildew management strategies and minimize the risk of the spread of mandipropamid and other CAA-resistant populations.