Alder decline caused by Phytophthora alni has been one of the most important diseases of natural ecosystems in Europe during the last 20 years. The emergence of P. alni subsp. alni —the pathogen responsible for the epidemic—is linked to an interspecific hybridization event between two parental species: P. alni subsp. multiformis and P. alni subsp. uniformis. One of the parental species, P. alni subsp. uniformis, has been isolated in several European countries and, recently, in North America. The objective of this work was to assess the level of genetic diversity, the population genetic structure, and the putative reproduction mode and mating system of P. alni subsp. uniformis. Five new polymorphic microsatellite markers were used to contrast both geographical populations. The study comprised 71 isolates of P. alni subsp. uniformis collected from eight European countries and 10 locations in North America. Our results revealed strong differences between continental populations (Fst = 0.88; Rst = 0.74), with no evidence for gene flow. European isolates showed extremely low genetic diversity compared with the North American collection. Selfing appears to be the predominant mating system in both continental collections. The results suggest that the European P. alni subsp. uniformis population is most likely alien and derives from the introduction of a few individuals, whereas the North American population probably is an indigenous population.

A simple and reliable method for preparation of whole nuclei of a common oomycete, Phytophthora infestans, is described for laser flow cytometry. The ease of preparation, the absence of detectable debris and aggregates, and the precision in determinations of DNA content per nucleus improve interpretation and understanding of the genetics of the organism. Phytophthora infestans is the pathogen that causes potato and tomato late blight. The genetic flexibility of P. infestans and other oomycete pathogens has complicated understanding of the mechanisms of variation contributing to shifts in race structure and virulence profiles on important agricultural crops. Significant phenotypic and genotypic changes are being reported in the apparent absence of sexual recombination in the field. Laser flow cytometry with propidium iodide is useful in investigating the nuclear condition of the somatic colony of field strains of P. infestans. The majority of the studied strains contain a single population of nuclei in nonreplicated diplophase. However, mean DNA content per nucleus varies considerably among isolates confirming the heterogeneity of the nuclear population in regard to C-value, for field isolates. Nuclear DNA content varies from 1.75× to 0.75× that of nuclei in a standard strain from central Mexico. Some strains contain two to three populations of nuclei with differing DNA contents in the mycelium and are heterokaryons. Such a range in DNA content suggests DNA-aneuploidy, but direct confirmation of aneuploidy will require microscopy of chromosomes. Heterokaryosis and populations of nuclei of differing DNA content necessarily confound standardized assays used worldwide in crop breeding programs for determination of race profiles and virulence phenotypes of this important pathogen. © 2010 International Society for Advancement of Cytometry

The gene Yr5 in wheat has generally been known to confer resistance to Puccinia striiformis f. sp. tritici races worldwide. However, a few reports indicated that some isolates of the stripe rust pathogen were virulent on the wheat genotypes with Yr5 in India, Australia, and China (Nagarajan 1986; Wellings and McIntosh 1990; Zhang et al. 2020). In April 2020, a stripe rust infection was observed on Triticum spelta 'Album' (TSA), the Yr5 donor, provided by Turkey Seed Gene Bank in a wheat breeding field at the experimental station of Akdeniz University in the Antalya province of Turkey. The sample was tested on the seedlings of the Yr single-gene lines in the Avocet S (AvS) background including Yr5 near-isogenic line (AvSYr5NIL), TSA, and susceptible bread wheat variety 'Morocco' according to the procedure reported by Wan et al. (2014), and found to be highly virulent on Morocco with infection type (IT) 9 and virulent on AvSYr5NIL and TSA with IT 7. To confirm this novel finding, single-pustule isolates derived from this sample were obtained from TSA and re-tested on the differentials and TSA. The Yr5-virulent isolate yielded similar infection types on these plants further verifying previous results. The single-pustule isolates were determined to be avirulent (Avr) (IT 0 to 6) on AvSYr1NIL, AvSYr8NIL, AvSYr9NIL, AvSYr10NIL, AvSYr32NIL, and AvSYr43NIL and virulent (Vr) (IT 7 to 9) on AvSYr5NIL, AvSYr6NIL, AvSYr7NIL, AvSYr17NIL, AvSYr24NIL, AvSYr27NIL, AvSYr44NIL, AvSYrSpNIL, and TSA. The isolates were identified as a new race and temporarily named TRVR20-5 based on its Avr/Vr pattern. Furthermore, to confirm the presence of Yr5, the genomic DNA of fresh leaves of AvSYr5NIL and TSA used in the virulence testing were isolated using NucleoSpin Plant II (Macherey-Nagel, Germany) according to the manufacturer's procedure. The genomic DNA samples were amplified with the primers STS-7 (5'-GTACAATTCACCTAGAGT-3') and STS-8 (5'-GCAAGTTTTCTCCCTATT-3') (Chen et al. 2003) using a thermal cycler (T-100, BioRad, USA), and the resulting PCR products were digested with DpnII and visualized on an agarose gel. AvSYr5NIL and TSA had the fragment linked to the resistance allele (308 bp) whereas AvS and Morocco had the fragment linked to the susceptible allele (181 bp). Thus, both wheat genotypes were confirmed to carry Yr5, and the new Pst race virulent to Yr5 was confirmed. To the best of our knowledge, this is the first report of a Pst race virulent to Yr5 in Turkey, where stripe rust epidemics have occured in 2 out of every 5 years with 1-5% crop losses in over 25% of the wheat growing areas (Chen 2020). In a nutshell, monitoring the Pst races virulent to Yr5 with a potential to cause devastating epidemics worldwide is crucial due to the fact that international wheat breeding programs have been using Yr5 to develop wheat cultivars with resistance to stripe rust.

Mushroom sciarid fly Lycoriella ingenua (Dufour, 1839) comb. nov., is one of the most common fly pests affecting the cultivation of Agaricus bisporus

Two fungi were isolated from grapevines in Michigan vineyards with Eutypa dieback symptoms: Eutypa lata and Eutypella vitis. These fungi are difficult to distinguish morphologically but are genetically distinct as determined by sequencing of the internal transcribed spacer (ITS) regions. The ITS regions of 25 Eutypa lata and 15 Eutypella vitis isolates were sequenced. Eutypa lata sequences were more variable than those of Eutypella vitis. Polymerase chain reaction (PCR) primers were designed for each species and evaluated against isolates of both fungi as well as 11 closely related Diatrypaceous fungi and 23 isolates of other fungi representing various pathogenic, saprophytic, and endophytic genera on grape and other small fruit crops. The primers were specific for their intended species. A nested multiplex PCR protocol was developed and used to successfully detect these fungi in wood samples from cankers with and without stromata from naturally infected vines as well as in artificially inoculated, potted canes. The primers developed in this study will assist in our abilities to diagnose and study the roles of Eutypa lata and Eutypella vitis in Eutypa dieback development.