Isolates of Puccinia graminis f. sp. tritici belonging to the Ug99 race group are virulent to a broad spectrum of resistance genes, rendering most of the world's wheat germplasm susceptible to stem rust (3). Following the initial detection of Ug99 (TTKSK, North American [NA] race notation) in Uganda, virulence to the widely used Sr31 resistance gene has been reported from Kenya, Ethiopia, Sudan, and Iran (2,3). In November 2009, a wheat genotype suspected to carry Sr31 showed a susceptible response to stem rust in a disease nursery (29°08′05.02′′S, 30°38′29.18′′E), inoculated with race TTKSP, near Greytown in KwaZulu-Natal, South Africa. Inoculation of urediniospores of the field collection (isolate UVPgt60) onto seedlings of line Federation4*/Kavkaz confirmed virulence for Sr31. In three independent, replicated, and comparative seedling tests, eight single-pustule isolates of UVPgt60 all typed to race PTKST following the NA race nomenclature. These isolates produced compatible infection types (ITs) (3+ to 4) on the Sr31 testers Gamtoos, Sr31/6*LMPG, Federation4*/Kavkaz, Kavkaz, and Clement, whereas isolate UVPgt59 (TTKSP) was avirulent (ITs ;1 to 1) on these genotypes. In addition to Sr31 virulence, the new race differed from TTKSP by producing a lower IT (2 to 2++) on Cns_T.mono_ deriv., the accepted entry for Sr21 in the NA differential set. The UVPgt60 isolates were clearly avirulent on Einkorn (Sr21) (IT ;1=), a response that also differed from those produced by BPGSC, TTKSF, and TTKSP (IT 2). With the exception of Sr21, UVPgt60 isolates had a virulence pattern similar to race TTKST (1), notably the virulence combination for Sr24 and Sr31. Isolate UVPgt60.6 was randomly selected for testing on additional Sr genes and South African wheat cultivars and breeding lines. Similar to the race identification experiments seedling tests were duplicated and compared with reactions produced by TTKSP and other races. Greenhouse temperatures for all seedling tests ranged between 18 and 25°C. On the basis of primary leaf responses, PTKST is avirulent (ITs 0; to 2++) for Sr13, 14, 21, 22, 25, 26, 27, 29, 32, 33, 35, 36, 37, 39, 42, 43, 44, Em, Tmp, and Satu and virulent (ITs 3 to 4) for Sr5, 6, 7b, 8a, 8b, 9a, 9b, 9d, 9e, 9g, 10, 11, 16, 17, 24, 30, 31, 34, 38, 41, and McN. From 103 South African wheat cultivars and lines tested as seedlings, 59 and 47 were susceptible (IT ≥ 3) to races PTKST and TTKSP, respectively. Simple-sequence repeat analysis (4) with selected primer pairs showed that PTKST clusters with isolates belonging to the Ug99 lineage. Subsequent to the collection made at Greytown, stem rust sampled in December 2009 from naturally infected breeders' lines at Cedara (29°32′19.59′′S, 30°16′03.50′′E), KwaZulu-Natal, revealed five isolates with a virulence profile similar to PTKST. On the basis of current evidence it appears that PTKST may be an introduction to South Africa rather than a single-step mutation from local stem rust races.
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Ug99 is a devastating race of Puccinia graminis f.sp. tritici possessing virulence against resistant genes Sr31 and Sr24. This race is highly adoptive and has spread quite rapidly with 13 known variants covering 13 different countries. For reducing the vulnerability of wheat in south Asia to the Ug99, breeding durable resistant varieties is important. India, second largest wheat producer, falls in the predicted pathway of Ug99. Most of the Indian germplasm possesses Sr31 and Sr24 in their background. HUW468, a well adopted variety of north eastern plains zone (NEPZ) of India, carries durable resistance gene Sr2. To strengthen it, a MABB program was initiated to introgressed two major genes (Sr50 and SrWeb) using a donor line PMBWIR4 from CIMMYT. The foreground selection was performed with Xgwm47 for SrWeb and IB267 for Sr50 followed by the background selection by using 128 polymorphic SSR markers covering all chromosomes. Backcross progenies of HUW468 were screened in the field condition by using of Pgt race 21A-2 at IARI, Regional Station, Indore located in the central India. Superior selected lines from BC2F4:5 generation was planted at three locations in India namely; Varanasi, Indore and Dharwad. HUW468-09-25-47-09 and HUW468-09-25-47-56 were selected from BC2F5 generation having Sr50 and SrWeb along with Sr2 gene, superior agronomic performance and with 93.5% and 92.7% genome recovery, respectively. These two lines also possess 6-10 % yield superiority over the recipient parent HUW468. These lines have been submitted for registration in NBPGR (National Bureau of Plant Genetic Resources), India.
The basidiomycetous fungus, Puccinia graminis f. sp. tritici (Pgt) causes stem rust disease as one of the most destructive wheat pathogens, worldwide. TTKSK and other Pgt races under Ug99 race group are considered as major threats to wheat production in east Africa and CWANA region by defeating the stem rust resistance gene Sr31, while its ineffectiveness was reported in Iran in 2007. Race TKTTF of Pgt caused a severe stem rust epidemic in southern Ethiopia in 2013, and was spread to Europe through 2016 Sicily outbreak. This research describes race identification of Iranian isolates collected during the widespread distribution of stem rust in 2014-16. Purified urediniospores of 123 Pgt isolates were separately inoculated on seedlings of 20 North American differential wheat cultivars carrying different Sr resistance gene/s. Infection types were recorded at 14 days post inoculation (dpi) using Stakman et al. 0-4 scale. Based on the letter code nomenclature, we identified the Pgt races TKTTF, TTTTF, TTKSK, TTKTK, PKTTF, TKSTF, PKSTF, PKTTC, PTRTF, PTTTF, PKSTC, TTRTF, TKSTC and PKRTF in Iran. TKTTF and TTTTF were determined as prevalent Iranian Pgt races. This is the first report of race TTKTK, a new variant of Ug99 race group with virulence on Sr31 and SrTmp resistance genes, in Iran. Since TTKTK primarily occurred in south west of Iran, the migration route for this new race seems to be similar to race TTKSK. The high race variation observed in this study could indicate a high genetic diversity among P. graminis f. sp. tritici populations in Iran, as a wheat center of origin.
Stem and leaf rusts affect the winter and spring wheat in the Novosibirsk region. During 2008-2017 leaf rust incidence was generally moderate, from 20 to 40%. A leaf rust outbreak occurred in 2015 when incidence increased up to 80%. Leaf rust severity on the 'Thatcher' NILs ranged from immune or resistant to highly susceptible host response with maximum severity of 90S. Lines carrying genes Lr17, Lr18, Lr24, Lr29, Lr35, Lr37, Lr44, and LrW remained almost free of infection for the whole time of inspection. Genes Lr12, Lr13, Lr28, Lr34, and Lr38 exhibited moderate resistance but they did not provide sufficient level of resistance in favorable conditions. Since race-specific genes Lr24 and Lr29 are still effective in the neighboring Novosibirsk and Omsk regions, they might be recommended for breeding purposes in Western Siberia.
In 2016 stem rust was more prevalent and widespread in the region than ever before. Disease incidence ranged between 4.5 - 60% with high severity up to 80S in six fields from seven observed locations. The 4th ISRTN and varieties carrying Sr31 of West Siberian germplasm were assessed in field trials to monitor the virulence of the local population. There was no virulence to Sr9b, Sr9e, Sr20, Sr28, Sr29, Sr33, Sr39, Sr40, SrWld, Sr2 complex. Possible virulence to Sr6, Sr11, Sr12, Sr13, Sr17, Sr24, Sr25, Sr30, Sr31, Sr35, Sr38, Sr44, Sr57 was observed with low frequency. Entries genotyped for gene Sr31 were scored as MS and S. However, follow up race analysis work is needed to determine the actual stem rust races present and confirm the suspected possible observed virulence on Sr31.
Wheat stripe (Puccinia striiformis f. sp. tritici,=Pst) and stem (Puccinia graminis f. sp. tritici =Pgt) rusts are the most important wheat disease in Egypt as well as present in all wheat growing areas. This study to evaluate a set of tester lines of wheat carrying stripe Yr's, stem Sr's rust genes and selected Egyptian varieties have been studied for their response to Pst and Pgt at adult plant stage under field conditions in Sakha Agriculture Research Station, during the 2011 to 2014 growing seasons. The results revealed that stripe rust, it has been observed that the new race Yr27-virulence to Pst. In addition pathotypes were virulent for Yr2, Yr6, Yr7, Yr8, Yr9, Yr27, while Yr18 showed moderate susceptibility. On the other hand, Yr1, Yr5, Yr10, Yr15, Yr17, Yr32 and YrSP exhibited high levels of resistance. Regarding, evaluation of resistance genes sources of stem rust on ICARDA, CIMMYT wheat germplasm, and Egyptian wheat varieties released i.e. Misr1 and Misr2 which having Ug99_resistance genes Sr2 and Sr25 were found susceptible to Pgt, also Sr31 recorded infection moderately susceptible to susceptible at adult stage. Genes Sr2 complex, Sr24, Sr26, Sr27, and Sr32 were resistant at adult plant stages. The combination of Sr26 with Sr2 and Sr25 provided stem rust resistance in some CIMMYT wheat germplasm. The objectives of this work are: race analysis of wheat stem and stripe rust disease, evaluation the level and distribution of wheat stripe and stem rust in Egypt, and identification the resistance genes in commercial varieties or new promising lines using standard and molecular genetic markers. Egyptian germplasm such as Misr1, and Misr2 and others tester lines of wheat carrying stem rust Sr's were evaluative under field condition at adult stage in Egypt during 2014 growing season, Egyptian cultivars Misr1 and Misr2 were susceptible rated 10S-20S and Sr31 rated MSS. that results clearly presence a new Sr31-virulence. On other hand, genes Sr2 complex, Sr24, Sr26, Sr27 and Sr32 were resistant and combination of Sr26 with (Sr2 and Sr25) produced stem rust resistance in some CIMMYT wheat germplasm. Shahin et al., 2015, in APS Annual Meeting, Aug. 1-5, Pasadena, CA, US, (In Press).
The stem rust resistance genes Sr31 and Sr50 in wheat were both derived from translocations of the short arm of chromosome 1 from rye and conferred resistance to all field isolates of Puccinia graminis f. sp tritici (Pgt) for many years, preventing their distinction as different resistance specificities. We now show that Sr50 confers resistance against the Ug99 strain that overcomes Sr31, whereas a mutant Pgt strain virulent towards Sr50 is avirulent towards Sr31. Because lack of recombination between wheat and rye chromosome arms precludes genetic mapping and so map-based cloning of Sr50, we used a combination deletion mutagenesis and large DNA fragment cloning in bacterial artificial chromosome (BAC) vectors to define this resistance locus. Sequence analysis of a BAC contig spanning the smallest deletion detected with DNA markers at the Sr50 locus identified six coiled coil nucleotide binding site leucine-rich repeat (CC-NB-LRR) genes and a chymotrypsin inhibitor gene closely related to genes at the orthologous barley powdery mildew resistance locus, Mla. Sequencing of these genes from two EMS-induced mutants that had lost no DNA markers revealed mutation in one of the CC-NB-LRR orthologs of Mla. Transgenic complementation tests in stem rust susceptible wheat proved this gene to be Sr50. A survey of a set of rye accessions identified several carrying the gene but occurring in different Mla gene haplotypes based on DNA gel blot patterns and copy number of Mla orthologs. Several different powdery mildew and rust resistance genes including TmMla1 from T. monococcum, 23 Mla alleles from barley and stem rust resistance genes Sr33 from Aegilops tauschii and Sr50 from rye are all members of the Mla clade of cereal R genes. The gene Sr50,was initially thought to be allelic to Sr31, however, appearance of Ug99 showed that this is a different gene and is rye ortholog of barley Mla powdery mildew resistance gene. The cloning of Sr50 gives us an opportunity to screen the rye germplasm for presence of Sr50 and allows us to now do functional analysis of the various domains and understand the mechanism of resistance. The cloning also helps to add very effective resistance to gene cassette. Sr50 is effective against all the stem rust isolates around the globe
Stem (black) rust is a potentially important disease in northern, western and southern Iran. A new Pgt race with virulence to gene Sr31 appeared in Iran in 2007. Similar races have spread in Africa and some CWANA countries. In 2014 stem rust was widespread in western, northern, northwestern and central Iran, but at low severities. Thirty-nine stem rust samples were collected for race analysis. After purification and increase each isolate was inoculated to a set of 20 North American differentials in the greenhouse. Infection types were recorded 12-14 days after inoculation using the scale described by McIntosh et al. (1995, Wheat Rusts: An Atlas of Resistance Genes, CSIRO, East Melbourne, Australia). Races TKSTC (59%), TKTTC (20%), TTTTC, KTTSK (virulent on plants with Sr31), TTSTC, PTTTF and TTTTF were detected. Race TKSTC was common in western, northwestern and central Iran. Except for avirulence to Sr17 this race is similar to the race (TKTT) that caused a stem rust epidemic in Ethiopia in 2013.
Frequent emergence of new variants in the Puccinia graminis f. sp. tritici Ug99 race group in Kenya has made pathogen survey a priority. We analyzed 140 isolates from 78 P. graminis f. sp. tritici samples collected in Kenya between 2008 and 2014 and identified six races, including three not detected prior to 2013. Genotypic analysis of 20 isolates from 2013 and 2014 collections showed that the new races TTHST, TTKTK, and TTKTT belong to the Ug99 race group. International advanced breeding lines were evaluated against an isolate of TTKTT (Sr31, Sr24, and SrTmp virulence) at the seedling stage. From 169 advanced lines from Kenya, 23% of lines with resistance to races TTKSK and TTKST were susceptible to TTKTT and, from two North American regional nurseries, 44 and 91% of resistant lines were susceptible. Three lines with combined resistance genes were developed to facilitate pathogen monitoring and race identification. These results indicate the increasing virulence and variability in the Kenyan P. graminis f. sp. tritici population and reveal vulnerabilities of elite germplasm to new races.
In much of the world, resistance to stem rust in wheat, caused by Puccinia graminis f. sp. tritici, is based at least in part on the gene Sr31. During February 1999, high levels of stem rust infection were observed on entries in wheat (Triticum aestivum) grown in a nursery at Kalengyere Research Station in Uganda. Because several of the rusted entries were known to carry the 1BL-1RS chromosome translocation containing the Sr31, Lr26, and Yr9 genes for rust resistance, virulence to Sr31 was suspected. Urediniospores, collected in bulk from rusted stems of seven entries containing Sr31, were suspended in light mineral oil and sprayed on primary leaves of 7-day-old seedlings of South African wheat cv. Gamtoos (=Veery #3, pedigree: Kvz/Buho‘S’//Kal/BB). Plants were kept overnight at 19 to 21°C in a dew chamber before placement in a greenhouse at 18 to 25°C. After ≈14 days, urediniospores were collected from large, susceptible-type stem rust pustules and subsequently increased on Gamtoos, which served as a selective host for the new rust culture, designated Pgt-Ug99. Pathogenicity of Pgt-Ug99 was studied in seedling tests of available wheats containing Sr31, as well as other stem rust differential lines. All seedling tests were conducted at least three times in independent inoculations. Isolate Pgt-Ug99 was not virulent to Avocet‘S’/Yr9 (Australian line containing Sr26) or Oom Charl (South African cultivar) but was virulent to the other Sr31 testers: Alondra ‘S’, Bobwhite, Chokka, Clement, Federation/Kavkaz, Gamtoos, Grebe, Kavkaz, Letaba, Line E/Kavkaz, RL6078, and Veery ‘S’. Virulence to Sr31 (infection types [ITs] 3-3 to 3++4) was clearly contrasted by the low reactions (ITs 0; to 1) produced by UVPgt53, a South African pathotype avirulent to Sr31. According to the reactions of the differential lines, Pgt-Ug99 is avirulent to Sr21, -22, -24, -25, -26, -27, -29, -32, -33, -34, -35, -36, -39, -40, -42, and -43, Agi, and Em and virulent to Sr5, -6, -7b, -8a, -8b, -9b, -9e, -9g, -11, -15, -17, -30, -31, and -38. Virulence to the T. ventricosum-derived gene Sr38, which is linked to Lr37 and Yr17 and occurs in cultivars from Australia, the United Kingdom, and the United States, was not known previously (1). Both Pgt-Ug99 and UVPgt53 produced a continuum of ITs (; to 2+3) on Petkus rye (obtained from the USDA-ARS National Small Grains Collection, Aberdeen, ID), the original Sr31 donor source. Pgt-Ug99 did not appear more virulent than UVPgt53 on Petkus. All triticales tested, as well as oat cv. Overberg, were highly resistant to Pgt-Ug99. According to McIntosh et al. (1), Huerta-Espino mentioned a Sr31-virulent culture from Turkey, but this could not be confirmed. Should the Sr31-virulent pathotype migrate out of Uganda, it poses a major threat to wheat production in countries where the leading cultivars have resistance based on this gene.
The appearance and spread of races of Puccinia graminis f. sp. tritici with virulence for the Sr31 resistance gene has renewed interest in breeding for durable resistance to stem rust of wheat. Since the occurrence of stem rust has been low in South Africa until the detection of race TTKSF in 2000, breeding for resistance to this disease has not been a primary objective. The aim of this study was to test bread wheat cultivars and lines at the seedling stage for their infection response to stem rust, thus determining their level of resistance or vulnerability. A collection of 65 bread wheat entries was tested with one USA race, two Eastern African races, and three South African races of P. graminis f. sp. tritici. The Eastern African and South African races all belong to the Ug99 lineage. The cultivars Duzi, Caledon, Elands, PAN 3364, PAN 3191, SST 047, SST 399, and Steenbras produced resistant infection types (IT <3) to all races. The molecular marker Sr24#50 indicated the presence of Sr24 in 12 entries. In cultivars resistant to TTTTF, TTKSF, and TTKSP but susceptible to TTKSK and PTKST, the iag95 DNA marker indicated the presence of Sr31 in five wheat lines. Using the cleaved amplified polymorphic sequence marker csSr2, Sr2 was detected in PAN 3377, Inia, and Steenbras. Few South African wheat cultivars appear to have a broad-based resistance to stem rust, as 88% of the entries were susceptible as seedlings to at least one of the races tested. Diversification of resistance sources and more directed breeding for stem rust resistance are needed in South Africa.