A New Virulent Race of Wheat Stripe Rust Pathogen (Puccinia striiformis f. sp. tritici) on the Resistance Gene Yr5 in Turkey

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 stri