Effects of a penthiopyrad and picoxystrobin fungicide mixtureon phoma stem canker (Leptosphaeria spp.) on UK winteroilseed rape

© Koninklijke Nederlandse Planteziektenkundige Vereniging 2016. This is a pre-copyedited, author-produced PDF of an article accepted for publication in European Journal of Plant Pathology following peer review. The final publication [Sewell, T.R., Moloney, S., Ashworth, M. et al., European Journal of Plant Pathology (2016) 145: 675-685, first published online April 5, 2016] is available at Springer via doi: http://dx.doi.org/10.1007/s10658-016-0916-8In the UK, fungicides are often used to controlphoma stem canker on winter oilseed rape. Field trialswere established near Boxworth, Cambridgeshire for fourcropping seasons (2011/2012, 2012/2013, 2013/2014 and2014/15) to test the efficacy of a new fungicide mixtureRefinzar® (penthiopyrad + picoxystrobin) by comparisonto an existing fungicide Proline 275® (prothioconazole)against phoma stem canker (Leptosphaeria spp.) andthe effect on winter oilseed rape (cv. Catana) yield. Ineach season, weather data were collected from a weatherstation at Boxworth and the release of ascospores wasmonitored using a nearby Burkard spore sampler. Thepatterns of ascospore release differed between seasonsand related to weather conditions. Fungicidespenthiopyrad + picoxystrobin and prothioconazole wereapplied in October/November when 10 % of plants hadphoma leaf spotting (T1, early), 4/8 weeks after T1 (T2,late) or at both T1 and T2 (combined). When phoma leafspot symptoms were assessed in autumn/winter,penthiopyrad + picoxystrobin and prothioconazole bothdecreased numbers of phoma leaf spots caused byL. maculans; there were few leaf spots caused byL. biglobosa. Penthiopyrad + picoxystrobin andprothioconazole both reduced phoma stem canker severitybefore harvest compared to the untreated control butdid not increase yield in these seasons when epidemicswere not severe. In 2013/2014, the presence ofL. maculans and L. biglobosa in upper stem lesions orstem base cankers was determined by species-specificPCR. The proportions of stems with L. maculans DNAwere much greater than those with L. biglobosa DNA forboth upper stem lesions and basal stem cankers. Theseresults suggest that both penthiopyrad + picoxystrobinand prothioconazole can decrease phoma stem cankerseverity on winter oilseed rape in severe disease seasons.Peer reviewe

Quantitative PCR analysis of abundance of airborne propagules of Leptosphaeria species in air samples from different regions of Poland

When airborne propagules of Leptosphaeria maculans and L. biglobosa were collected in Poland at three ecologically different sites from 1 September to 30 November in 2004 to 2008, using a Hirst-type seven-day volumetric spore trap, there were fluctuations in timing of ascospore release and diverse ratios between airborne propagules of both species depending on season, field location and weather conditions. The detection was done using the microscope as well as quantitative PCR with species-specific primers targeted against fragments of β-tubulin genes and quantified with a dual-labelled fluorescent probe approach. This detection chemistry is described for the first time for L. maculans and L. biglobosa. Its advantage over the previous ITS-based SYBR-Green chemistry resides in improved sensitivity and the virtual absence of false positives in the detection of these fungi. There were significant, positive correlations between data obtained using visual assessment of ascospore numbers and DNA concentrations that were measured by qPCR. Climatic differences between the oilseed rape growing regions could have significantly affected the biological processes of pseudothecial maturation and ascospore development of the pathogens. The data suggest that regular rain events of intermediate intensity recorded in the Maritime region favoured the maturation of the pathogen more than the drier weather recorded in the Silesia or Pomerania regions. It was observed that the number of rainy days was of greater importance than the cumulative rainfall to obtain the generative sporulation of the pathogen. Accurate detection of airborne inoculum of pathogenic Leptosphaeria spp. facilitates improved targeting of disease management decisions for oilseed rape protection against phoma stem canker and stem necrosis diseases.Peer reviewe

Assessing quantitative resistance against Leptosphaeria maculans (phoma stem canker) in Brassica napus (oilseed rapte) in young plants

Quantitative resistance against Leptosphaeria maculans in Brassica napus is difficult to assess in young plants due to the long period of symptomless growth of the pathogen from the appearance of leaf lesions to the appearance of canker symptoms on the stem. By using doubled haploid (DH) lines A30 (susceptible) and C119 (with quantitative resistance), quantitative resistance against L. maculans was assessed in young plants in controlled environments at two stages: stage 1, growth of the pathogen along leaf veins/petioles towards the stem by leaf lamina inoculation; stage 2, growth in stem tissues to produce stem canker symptoms by leaf petiole inoculation. Two types of inoculum (ascospores; conidia) and three assessment methods (extent of visible necrosis; symptomless pathogen growth visualised using the GFP reporter gene; amount of pathogen DNA quantified by PCR) were used. In stage 1 assessments, significant differences were observed between lines A30 and C119 in area of leaf lesions, distance grown along veins/petioles assessed by visible necrosis or by viewing GFP and amount of L. maculans DNA in leaf petioles. In stage 2 assessments, significant differences were observed between lines A30 and C119 in severity of stem canker and amount of L. maculans DNA in stem tissues. GFP-labelled L. maculans spread more quickly from the stem cortex to the stem pith in A30 than in C119. Stem canker symptoms were produced more rapidly by using ascospore inoculum than by using conidial inoculum. These results suggest that quantitative resistance against L. maculans in B. napus can be assessed in young plants in controlled conditions. Development of methods to phenotype quantitative resistance against plant pathogens in young plants in controlled environments will help identification of stable quantitative resistance for control of crop diseases