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First Report of Races TKTTP and TKKTP of Puccinia graminis f. sp. tritici with Virulence to Wheat Stem Rust Resistance Gene Sr24 in Turkey and Tunisia.

Severe wheat stem rust caused by Puccinia graminis Pers.:Pers. f. sp. tritici Erikss. (Pgt) can result in complete crop failure. In recent years, the increasing frequency and the early onset of stem rust in Central West Asia and North Africa (CWANA) has become a big concern. The Sr24 resistance gene, one of the most effective stem rust resistance genes effective against most P. graminis f. sp. tritici races worldwide, has been widely deployed. Until the recent establishment of virulence to Sr24 within the Ug99 lineage of the pathogen in Africa (Hei et al. 2020; Jin et al. 2008; Patpour et al. 2015), Iraq (Nazari et al., 2021), occasional detections of races virulent to Sr24 were reported in South Africa (Le Roux and Rijkenberg 1987), India (Bhardwaj et al. 1990), Germany (Olivera Firpo et al. 2017), Georgia (Olivera, et al. 2019), and Western Siberia (Skolotneva et al., 2020). During the rust surveys conducted in Sinops, Samsun, and Kastomonu in the Black Sea region in northern Turkey in 2018, 19 isolates were collected. Single pustule (SP) isolates were developed and used in race analysis in the Biosafety Rust Laboratory, Regional Cereal Rust Research Center (RCRRC), Izmir, Turkey. Sample recovery, experimental procedures for pre-inoculation, inoculation, incubation, and race typing were conducted as previously described (Nazari et al. 2021). Among the tested SP isolates, two isolates showed a high infection type (IT) of 33+ on the Sr24 tester line (Little Club/Agent) and a low infection type of 11+ for the source of Sr31 (Benno/6*LMPG-6). Eight SP isolates were further developed from the high IT 33+ pustules collected from the Sr24 tester line. After spore multiplications, they were used in inoculation of the 20 North American stem rust single-gene lines used to differentiate races of P. graminis f. sp. tritici, plus Trident (Sr38+), Siouxland (Sr24+Sr31), and Sisson (Sr31+Sr36). Five SP-derived isolates with IT 33+ on the Sr24 single-gene line collected from Samsun (Alacam - Etyemez; Location: N 41.61889 E 35.55722) and Sinop (Merkez-Sanlıoglu; Location: N 41.85556 E 35.04889) were identified as race TKKTP and the remaining three SP isolates as race TKTTP. In 2020, we detected two isolates of TKKTP among the stem rust samples from Tunisia submitted to RCRRC. These two isolates were collected from bread wheat cultivars Heydna and Tahmet at a trial site near Bou Salem in Western Tunisia (Location: N 36.5351 E 8.95486). Based on the negative results of the Stage 1 test using a suite of four real-time polymerase chain reaction assays diagnostic for the Ug99 race group developed by Szabo (2012), these two races should not belong to the Ug99 race group when compared to the reference Ug99 race TTKTT from Kenya. These races were virulent to Sr5, Sr21, Sr9e, Sr7b, Sr6, Sr8a, Sr9g, Sr9b, Sr30, Sr17, Sr9a, Sr9d, Sr10, SrTmp, Sr24, Sr38, and SrMcN. In addition to these genes, race TKTTP was virulent to Sr36. Both races were avirulent to Sr11 and Sr31. To our knowledge, this is the first report of P. graminis f. sp. tritici races with the Sr24 virulence in Turkey and Tunisia. The results reflect an increasing trend of virulence to Sr24 in the pathogen populations, and raise a great concern given the deployment of the Sr24 resistance gene in widely grown wheat cultivars worldwide.