Attached and detached leaves of 60 potential host species were inoculated in the greenhouse and laboratory with aeciospores of Cronartium ribicola J.C. Fisch. from six Finnish locations and of Cronartium flaccidum (Alb. & Schw.) Wint. from 20 locations in Finland and Sweden in 2011. Candidate hosts represented 16 plant families: Solanaceae, Verbenaceae, Asclepiadaceae, Grossulariaceae, Paeoniaceae, Balsaminaceae, Gentianaceae, Scrophulariaceae, Loasaceae, Tropaeolaceae, Acanthaceae, Myricaceae, Phrymaceae, Plantaginaceae, Orobanchaceae, and Apocynaceae. Inoculations of C. flaccidum produced uredinia after 2 weeks and (or) telia after 4 weeks of incubation on 25 hosts. Inoculation trials identified several new hosts for C. flaccidum in Fennoscandia, namely Impatiens balsamina, Swertia fedtschenkoana, Loasa tricolor, Myrica gale, Verbena canadensis, Saxifraga spp., Paeonia obovata, and Veronica daurica. Myricaceae and Saxifragaceae represent new host families for these rusts. Cronartium ribicola formed uredinia or telia on 10 species: Ribes spp. (7 species/cultivars), Pedicularis palustris subsp. palustris, Bartsia alpina, and Loasa triphylla. Results suggest wider alternate host ranges for both C. flaccidum and C. ribicola than previously recognized. Spores were virulent regardless of their source location, suggesting a lack of host-specificity among Fennoscandian populations of Cronartium.Key words: alternate hosts, Scots pine blister rust, white pine blister rust.
Genetic mapping of quantitative trait loci (QTL) for resistance to Melampsora leaf rust was performed in two willow families: the progeny from a backcross between Salix viminalis and a hybrid S. viminalis×Salix schwerinii (population S 1 ), and the F 1 progeny of a cross between two S. viminalis (population S 3 ). Disease levels were scored in the field for three consecutive years. In the laboratory, five different rust strains were sprayed onto leaf disks and the following resistance components were scored: latent period, diameter and number of uredinia, and flecking. One major QTL and 14 smaller were identified in the S 1 host population. One rust strain, that represents a Melampsora form with limited incidence on S. viminalis, showed lower aggressiveness to the S 1 host population together with a different pattern in QTLs. In the S 3 host population, we detected 13 QTLs for rust resistance, of which two were located to the same genomic regions as those found for the S 1 population. We showed that the strongest QTL cosegregated with a gene homologous to a known Toll/ interleukin receptor-nucleotide binding site-leucine-rich repeat resistance gene in poplar. The importance of the identified QTLs is discussed in relation to breeding for durable resistance.
The disease development and population structure of Puccinia graminis f. sp. avenae, which causes stem rust on oats, were studied to investigate if sexual reproduction plays an important role in the epidemiology of the disease. The genetic population structure of P. graminis f. sp. avenae in Sweden was investigated by sampling 10 oat fields in July and August 2008 and seven fields during the same period in 2009. Nine single‐pustule isolates were first used to test simple sequence repeat (SSR) markers developed for P. graminis f. sp. tritici. Eleven of the 68 tested SSR markers were useful for genotyping P. graminis f. sp. avenae. For the main study, DNA from single uredinia was extracted and the SSR markers were used to genotype 472 samples. Both allelic and genotypic diversity were high in all fields, indicating that P. graminis f. sp. avenae undergoes regular sexual reproduction in Sweden. No significant relationship between genetic and geographic distances was found. Disease development was studied on two farms during 2008 and 2009. The apparent infection rates ranged between 0·17 and 0·55, indicating the potential for rapid disease development within fields. The incidence of oat stem rust has increased recently in Sweden. One possible explanation is a resurgence of its alternate host, barberry (Berberis spp.), after the repeal of the barberry eradication law in 1994. Barberry is present in several grain‐producing areas in Sweden, which supports the conclusion that P. graminis f. sp. avenae undergoes regular sexual reproduction there.
Demethylation inhibitor (DMI) and succinate dehydrogenase inhibitor (SDHI) fungicides are currently relied upon for the control of septoria tritici blotch (STB) in European wheat fields. However, multiple mutations have occurred over time in the genes encoding the targeted proteins that have led to a practical loss of fungicide efficacies. Among the different amino acid substitutions in Zymoseptoria tritici associated with resistance to these fungicides, S524T in CYP51 (DMI target) and H152R in SdhC (SDHI target) are regarded as conferring the highest resistance factors to DMI and SDHI, respectively. To facilitate further studies on the monitoring and selection of these substitutions in Z. tritici populations, a multiplex allele‐specific quantitative PCR (qPCR) assay allowing for estimation of both allele frequencies in bulk DNA matrices was developed. The assay was then used on complex DNA samples originating from a spore trap network set up in Belgium, Denmark, Sweden, and Ireland in 2017 and 2018, as well as on leaf samples with symptoms. The S524T allele was present in all field samples and its proportion was significantly higher in Ireland than in Belgium, whereas the proportion of H152R was only sporadically present in both countries. The frequency of S524T varied greatly in the airborne inoculum of all four countries; however, the H152R allele was never detected in the airborne inoculum. The method developed in this study can be readily adopted by other laboratories and used for multiple applications including resistance monitoring in field populations of Z. tritici.
BACKGROUND: Over the past decade, demethylation inhibitor (DMI) and succinate dehydrogenase inhibitor (SDHI) fungicides have been extensively used to control to septoria tritici blotch, caused by Zymoseptoria tritici on wheat. This has led to the development and selection of alterations in the target-site enzymes (CYP51 and SDH, respectively). RESULTS: Taking advantage of newly and previously developed qPCR assays, the frequency of key alterations associated with DMI (CYP51-S524T) and SDHI (SDHC-T79N/I, C-N86S and C-H152R) resistance was assessed in Z. tritici-infected wheat leaf samples collected from commercial crops (n = 140) across 14 European countries prior to fungicide application in the spring of 2019. This revealed the presence of a West to East gradient in the frequencies of the most common key alterations conferring azole (S524T) and SDHI resistance (T79N and N86S), with the highest frequencies measured in Ireland and Great Britain. These observations were corroborated by sequencing (CYP51 and SDH subunits) and sensitivity phenotyping (prothioconazole-desthio and fluxapyroxad) of Z. tritici isolates collected from a selection of field samples. Additional sampling made at the end of the 2019 season confirmed the continued increase in frequency of the targeted alterations. Investigations on historical leaf DNA samples originating from different European countries revealed that the frequency of all key alterations (except C-T79I) has been gradually increasing over the past decade. CONCLUSION: Whilst these alterations are quickly becoming dominant in Ireland and Great Britain, scope still exists to delay their selection throughout the wider European population, emphasizing the need for the implementation of fungicide antiresistance measures.
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