Endemic and panglobal genetic groups, and divergence of host-associated forms in worldwide collections of the wheat leaf rust fungus Puccinia triticina as determined by genotyping by sequencing.

The wheat leaf rust fungus, Puccinia triticina, is found in the major wheat growing regions of the world and is a leading cause of yield loss in wheat. Populations of P. triticina are highly variable for virulence to resistance genes in wheat and adapt quickly to resistance genes in wheat cultivars. The objectives of this study were to determine the genetic relatedness of worldwide collections of P. triticina using restriction site associated genotyping by sequencing. A total of 558 isolates of P. triticina from wheat producing regions in North America, South America, Europe, the Middle East, Ethiopia, Russia, Pakistan, Central Asia, China, New Zealand, and South Africa were characterized at 6745 single nucleotide loci. Isolates were also tested for virulence to 20 near-isogenic lines that differ for leaf rust resistance genes. Populations that were geographically proximal were also more closely related for genotypes. In addition, groups of isolates within regions that varied for genotype were similar to groups from other regions, which indicated past and recent migration across regions. Isolates from tetraploid durum wheat in five different regions were highly related with distinct genotypes compared to isolates from hexaploid common wheat. Based on a molecular clock, isolates from durum wheat found only in Ethiopia were the first to diverge from a common ancestor form of P. triticina that is found on the wild wheat relative Aegilops speltoides, followed by the divergence of isolates found worldwide that are virulent to durum wheat, and then by isolates found on common wheat.

Multilocus Genotypes of the Wheat Leaf Rust Fungus Puccinia triticina in Worldwide Regions Indicate Past and Current Long-Distance Migration.

Many plant pathogenic fungi have a global distribution across diverse ecological zones and agricultural production systems. Puccinia triticina, the wheat leaf rust fungus, is a major pathogen in many wheat production areas of the world. The objective of this research was to determine the genetic relatedness of P. triticina in different worldwide regions. A total of 831 single-uredinial isolates collected from 11 regions were characterized for multilocus genotype at 23 simple sequence repeat loci and for virulence to 20 lines of wheat with single genes for leaf rust resistance. A total of 424 multilocus genotypes and 497 virulence phenotypes were found. All populations had high heterozygosity and significant correlation between virulence and molecular variation, which indicated clonal reproduction. The populations from North America and South America, Central Asia and Russia, and the Middle East and Europe were closely related for multilocus genotypes and many individual isolates from other continental regions were closely related. Twenty-seven multilocus genotypes were found in more than one continental region, and 13 of these had isolates with identical virulence phenotypes. The wide geographic distribution of identical and highly related multilocus genotypes of P. triticina indicated past and more recent migration events facilitated by the spread of clonally produced urediniospores.

Vulnerability of Barley to African Pathotypes of Puccinia graminis f. sp. tritici and Sources of Resistance.

The emergence of widely virulent pathotypes (e.g., TTKSK in the Ug99 race group) of the stem rust pathogen (Puccinia graminis f. sp. tritici) in Africa threatens wheat production on a global scale. Although intensive research efforts have been advanced to address this threat in wheat, few studies have been conducted on barley, even though pathotypes such as TTKSK are known to attack the crop. The main objectives of this study were to assess the vulnerability of barley to pathotype TTKSK and identify possible sources of resistance. From seedling evaluations of more than 1,924 diverse cultivated barley accessions to pathotype TTKSK, more than 95% (1,844) were found susceptible. A similar high frequency (910 of 934 = 97.4%) of susceptibility was found for the wild progenitor (Hordeum vulgare subsp. spontaneum) of cultivated barley. Additionally, 55 barley lines with characterized or putative introgressions from various wild Hordeum spp. were also tested against pathotype TTKSK but none was found resistant. In total, more than 96% of the 2,913 Hordeum accessions tested were susceptible as seedlings, indicating the extreme vulnerability of the crop to the African pathotypes of P. graminis f. sp. tritici. In total, 32 (1.7% of accessions evaluated) and 13 (1.4%) cultivated and wild barley accessions, respectively, exhibited consistently highly resistant to moderately resistant reactions across all experiments. Molecular assays were conducted on these resistant accessions to determine whether they carried rpg4/Rpg5, the only gene complex known to be highly effective against pathotype TTKSK in barley. Twelve of the 32 (37.5%) resistant cultivated accessions and 11 of the 13 (84.6%) resistant wild barley accessions tested positive for a functional Rpg5 gene, highlighting the narrow genetic base of resistance in Hordeum spp. Other resistant accessions lacking the rpg4/Rpg5 complex were discovered in the evaluated germplasm and may possess useful resistance genes. Combining rpg4/Rpg5 with resistance genes from these other sources should provide more durable resistance against the array of different virulence types in the Ug99 race group.

Genetic mapping of resistance to the Ug99 race group of Puccinia graminis f. sp. tritici in a spring wheat landrace CItr 4311.

KEY MESSAGE: A gene for Ug99 resistance from wheat landrace CItr 4311 was detected on the long arm of chromosome 2B. Wheat landrace CItr 4311 has seedling resistance to stem rust caused by Puccinia graminis f. sp. tritici race TTKSK and field resistance to the Ug99 race group. Parents, F1 seedlings, 121 doubled haploid (DH) lines, and 124 recombinant inbred lines (RILs) developed from a cross between CItr 4311 and the susceptible line LMPG-6 were evaluated for seedling resistance to race TTKSK. Goodness-of-fit tests indicated that a single dominant gene in CItr 4311 conditioned the TTKSK resistance. The 90 K wheat iSelect SNP platform was used to genotype parents and the DH population. The seedling resistance locus was mapped to the chromosome arm 2BL. Parents and the DH population were evaluated for field resistance in Kenya. One major QTL for the field resistance was consistently detected in the same region on 2BL as the seedling resistance. Using KASP assays, five linked SNP markers were used to verify the result in the 124 RIL, 35 wheat accessions, 46 DH lines from the LMPG-6/PI 165194 cross and F1 seedlings, and susceptible bulks derived from crosses between six resistant landraces with LMPG-6. Race specificity, mapping results, and haplotype similarity with lines with Sr9h (Gabo 56, Timstein, and PI 670015), support the hypothesis that the Sr gene in CItr 4311 and the landraces is Sr9h. The KASP assays developed in this study will be useful for pyramiding the TTKSK resistance from CItr 4311 with other Sr genes effective against Ug99.

Development of a Greenhouse Screening Method for Adult Plant Response in Wheat to Stem Rust.

Screening for adult plant resistance in wheat to stem rust, caused by Puccinia graminis f. sp. tritici, is generally conducted in field plots. Although such evaluations are successful if managed properly, field ratings are time consuming, expensive, weather dependent, and open to inoculum of unwanted races or other confounding diseases. The objective of this study was to develop a dependable system of screening the response of adult plants to stem rust under greenhouse conditions. A comparison of inoculation methods and incubation environments showed that plants inoculated with urediniospores suspended in water, followed by a 24 h dew period in a plastic chamber constructed in a greenhouse, gave the most consistent results. Measurements of response type, stem rust severity, and frequency in follow-up experiments indicated that the most reliable infection was obtained when plants sprayed with 1.25 mg urediniospores per ml water were incubated in the plastic chamber. Using the optimized protocol, a Kariega × Avocet S doubled haploid population was inoculated with two P. graminis f. sp. tritici races. Depending on the race, composite interval mapping showed flag leaf infection type to be significantly influenced by regions on chromosomes 6A, 6D, and 7D. Stem rust severity and reaction type mapped to chromosomes 6D and/or 6A. The Lr34/Yr18/Sr57 gene derived from Kariega on chromosome 7D affected the rust response on flag leaves but not on stems of greenhouse-grown plants. This study showed that phenotyping and genetic analysis of especially major effect stem rust resistance in adult wheat plants is possible and reproducible under controlled conditions in a greenhouse.

Rapid Identification of Resistance Loci Effective Against Puccinia graminis f. sp. tritici Race TTKSK in 33 Spring Wheat Landraces.

Wheat breeders worldwide are seeking new sources of resistance to Puccinia graminis f. sp. tritici race TTKSK. To prioritize field-resistant landraces for follow-up genetic studies to test for the presence of new resistance genes, seedling response to P. graminis f. sp. tritici race TTKSK, molecular markers linked to specific Sr genes, segregation ratios among progeny from crosses, and bulked segregant analyses (BSA) were used. In total, 33 spring wheat landraces with seedling resistance to P. graminis f. sp. tritici race TTKSK were crossed to a susceptible genotype, LMPG-6. The segregation ratios of stem rust reactions in F2 seedlings fit a single dominant gene model in 31 populations and progeny from two crosses gave ambiguous results. Using the 90K wheat single-nucleotide polymorphism genotyping platform, BSA showed that the seedling resistance in 29 accessions is probably controlled by loci on chromosome 2BL. For the three remaining accessions, BSA revealed that the seedling resistance is most likely controlled by previously unreported genes. For confirmation, two populations were advanced to the F2:3 and screened against P. graminis f. sp. tritici race TTKSK. Segregation of the F2:3 families fit a 1:2:1 ratio for a single dominant gene. Using the F2:3 families, BSA located the TTKSK locus on chromosome 6DS to the same location as Sr42.

First Report of Puccinia triticina (Leaf Rust) Race FBPT on Wheat in South Africa.

Eleven isolates of Puccinia triticina Erikss. collected from bread wheat (Triticum aestivum L.) in the Western Cape during the 2010 annual rust survey were pathotyped to a race not previously recorded in South Africa. Replicated race analysis on seedlings of 16 Thatcher (Tc) near-isogenic differential lines at two rust laboratories confirmed avirulence (infection types [ITs] 0; to 2+) for lines with Lr1, Lr2a, Lr9, Lr11, Lr16, Lr24, and Lr26 and virulence (ITs 3 to 4) for lines with Lr2c, Lr3, Lr3ka, Lr10, Lr14a, Lr17, Lr18, Lr30, and LrB. Thatcher lines with LrB, Lr10, Lr14a, and Lr18 were added as the fourth set to the 12 original differential lines (1,2). This profile codes to race FBPT according to the North American system, and, based on these differentials, resembles a P. triticina isolate from Gwebi, Zimbabwe, in 2012 (Z. A. Pretorius, unpublished data). When additional single gene lines were tested with FBPT (race 3SA147 according to the ARC-Small Grain Institute rust notation procedure), lines with Lr2b, Lr15, Lr19, Lr20, Lr21, Lr25, Lr27+31, Lr28, Lr29, Lr32, Lr36, Lr38, Lr45, Lr47, Lr50, Lr51, and Lr52 were effective, whereas lines with Lr3bg, Lr23, Lr28, and Lr33 were ineffective. In adult plant tests in a greenhouse, Thatcher lines containing Lr12 (IT ;1c), Lr13 (IT ;1c), Lr22a (IT 1), Lr35 (IT ;1+), and Lr37 (IT ;1) were resistant, whereas Thatcher (Lr22b) was susceptible. Of 146 South African cultivars and lines infected as seedlings with 3SA147 (FBPT), 83% were resistant (IT ≤ 2) and 5% showed within-line variation. Entries showing compatible ITs with 3SA147 (FBPT) were also susceptible to either or both of 3SA133 (PDRS) and 3SA146 (MCDS). In addition, the new race was genotyped using 16 simple sequence repeat (SSR) primer-combinations (3). Three single pustule isolates of 3SA147 (FBPT) were identical and showed 82% and 76% similarity with the recently described races 3SA146 (MCDS) and 3SA145 (CCPS), respectively (4). Minimum spanning network analysis confirmed this close genetic relationship among the three races. However, since their virulence phenotypes differ, it is proposed that 3SA147 (FBPT) is not a stepwise mutation from either 3SA145 (CCPS) or 3SA146 (MCDS), but rather a foreign introduction into South Africa. As most current breeding lines and wheat cultivars are resistant, it is unlikely that race FBPT will threaten wheat production in South Africa, but its detection underlines the fact that new P. triticina variants have been occurring at regular intervals in the region. References: (1) J. A. Kolmer et al. Austr. J. Agric. Res. 58:631, 2007. (2) D. L. Long and J. A. Kolmer. Plant Dis. 79:525, 1989. (3) L. J. Szabo and J. A. Kolmer. Mol. Ecol. Notes 7:708, 2007. (4) T. Terefe et al. Plant Dis. 95:611, 2011.

Identification of Resistance to Races of Puccinia graminis f. sp. tritici with Broad Virulence in Triticale (×Triticosecale).

Triticale (×Triticosecale), an amphiploid of wheat (mainly Triticum turgidum) and cereal rye (Secale cereale), is an excellent source of resistance to wheat stem rust, caused by Puccinia graminis f. sp. tritici. A collection of 567 triticale accessions originating from 21 countries was evaluated at the seedling stage for reaction to races of P. graminis f. sp. tritici with broad virulence, including TTKSK, TRTTF, and TTTTF. A high frequency (78.4%) of accessions was resistant to race TTKSK, with low infection types ranging from 0; to X. A selection of 353 TTKSK-resistant accessions was evaluated for reaction to three South African isolates of P. graminis f. sp. tritici with single and/or combined virulences to stem rust resistance genes SrSatu, Sr27, and SrKw present in triticale. Genes SrSatu, Sr27, and SrKw were postulated to be present in 141 accessions and contributed to TTKSK resistance. The remaining 212 resistant accessions may possess uncharacterized genes or combinations of known genes that could not be determined with these isolates. These accessions were further evaluated for resistance to races TTKST, TPMKC, RKQQC, RCRSC, QTHJC, QCCSM, and MCCFC. Resistance remained effective across the entire set of races in the majority of the accessions (n = 200), suggesting that the resistances are effective against a broad spectrum of virulence. In all, 129 (79.6%) resistant accessions with noncharacterized genes were resistant to moderately resistant in field stem rust nurseries at Debre Zeit (Ethiopia) and St. Paul (Minnesota). Results from evaluating F2 populations derived from resistant-susceptible crosses revealed that resistance to TTKSK in triticale was conferred mostly by single genes with dominant effects.

Identification of adult plant resistance to stripe rust in the wheat cultivar Cappelle-Desprez.

Following the appearance of stripe rust in South Africa in 1996, efforts have been made to identify new sources of durable resistance. The French cultivar Cappelle-Desprez has long been considered a source of durable, adult plant resistance (APR) to stripe rust. As Cappelle-Desprez contains the seedling resistance genes Yr3a and Yr4a, wheat lines were developed from which Yr3a and Yr4a had been removed, while selecting for Cappelle-Desprez derived APR effective against South African pathotypes of the stripe rust fungus, Puccinia striiformis f. sp. tritici. Line Yr16DH70, adapted to South African wheat growing conditions, was selected and crossed to the stripe rust susceptible cultivar Palmiet to develop a segregating recombinant inbred line mapping population. A major effect QTL, QYr.ufs-2A was identified on the short arm of chromosome 2A derived from Cappelle-Desprez, along with three QTL of smaller effect, QYr.ufs-2D, QYr.ufs-5B and QYr.ufs-6D. QYr.ufs-2D was located within a region on the short arm of chromosome 2D believed to be the location of the stripe rust resistance gene Yr16. An additional minor effect QTL, QYr.ufs-4B, was identified in the cv. Palmiet. An examination of individual RILs carrying single or combinations of each QTL indicated significant resistance effects when QYr.ufs-2A was combined with the three minor QTL from Cappelle-Desprez, and between QYr.ufs-2D and QYr.ufs-5B.

First Report of a New TTKSF Race of Wheat Stem Rust (Puccinia graminis f. sp. tritici) in South Africa and Zimbabwe.

Seven races have been described in the Ug99 race group of Puccinia graminis f. sp. tritici (2). Ug99-related races previously recorded in South Africa are TTKSF, TTKSP, and PTKST (4). In December 2010, severe stem rust infection of the winter wheat cv. Matlabas was observed for the first time in South Africa. Race analysis using the 20 North American (NA) stem rust differential lines and letter code system classified the race as TTKSF. In comparative infection studies in a greenhouse, cv. Matlabas seedlings were susceptible (infection type [IT] 4) to isolate UVPgt61/1 (TTKSF+) collected from Afrikaskop in the eastern Free State, whereas the cultivar was resistant (IT 1 to 2) to stem rust isolates 2013 (TTKSF), UVPgt55 (TTKSF), UVPgt59 (TTKSP), and UVPgt60 (PTKST). Isolate 2013 represents the original collection of race TTKSF in South Africa (1). In addition to the NA differentials, no variation in the IT range of seedlings of lines with Sr7a, 8b, 12, 13, 14, 16, 18, 19, 22, 25, 26, 27, 28, 29, 32, 33, 34, 35, 39, 41, 42, 43, 44, Em, R, Tt2, and Satu was observed between UVPgt61/1 and UVPgt55. With the exception of cv. Matlabas, ITs of 106 South African cultivars likewise did not differentiate UVPgt61/1 and UVPgt55. Seedling IT studies were conducted at least twice. Microsatellite analysis (4) showed that all single pustule isolates established from the original Matlabas isolate formed part of the Ug99 group. When characterized with selected single nucleotide polymorphisms (SNPs), all single pustule isolates shared an identical genotype that differed from UVPgt55 (TTKSF), a foreign introduction into South Africa (1,3). SNP genotype analysis suggests that UVPgt61/1 is genetically dissimilar to UVPgt55, as is Zim1009, another TTKSF+ isolate that was collected from Birchenough in Zimbabwe. Studies are underway to determine the identity of the defeated Sr gene in Matlabas and the cultivar has been added to the South African stem rust differential set. TTKSF+ is the eighth race detected in the Ug99 group. Since no other cultivars or advanced lines were found to carry the Matlabas gene, it is unlikely that race TTKSF+ will threaten wheat production in South Africa. However, the occurrence of a new Ug99-related race emphasizes the variability within this internationally important group. References: (1) W. H. P. Boshoff et al. Plant Dis. 86:922, 2002. (2) R. F. Park et al. Euphytica 179:109, 2011. (3) B. Visser et al. Mol. Plant Pathol. 10:213, 2009. (4) B. Visser et al. Euphytica 179:119, 2011.

A multiple resistance locus on chromosome arm 3BS in wheat confers resistance to stem rust (Sr2), leaf rust (Lr27) and powdery mildew.

Sr2 is the only known durable, race non-specific adult plant stem rust resistance gene in wheat. The Sr2 gene was shown to be tightly linked to the leaf rust resistance gene Lr27 and to powdery mildew resistance. An analysis of recombinants and mutants suggests that a single gene on chromosome arm 3BS may be responsible for resistance to these three fungal pathogens. The resistance functions of the Sr2 locus are compared and contrasted with those of the adult plant resistance gene Lr34.

First Report of a New Wheat Leaf Rust (Puccinia triticina) Race with Virulence for Lr12, 13, and 37 in South Africa.

A new race of Puccinia triticina was collected from common wheat (Triticum aestivum) in the Eastern and Western Cape provinces during the annual rust survey in 2009. Six single-pustule isolates from a field collection, which were shown to be a new race in preliminary analyses, were inoculated onto seedlings of 16 Thatcher (Tc) near-isogenic differential lines (1) and other tester lines with known Lr genes. Standard procedures for inoculation, incubation, and rust evaluation were followed (4) and all infection studies were repeated. The low infection type of Lr18 was confirmed at 18°C. All six isolates were avirulent (infection types [ITs] 0; to 2) to Lr1, 2a, 2c, 9, 11, 16, 18, and 24 and virulent (ITs 3 to 4) to Lr3, 3ka, 10, 14a, 17, 26, 30, B, and Tc (control). The new race, named 3SA145 according to the ARC-Small Grain Institute notation, corresponds to race CCPS in the North American system (1). On the basis of seedling ITs of the extended Lr gene set, 3SA145 was avirulent (ITs 0; to 22+) to Lr2b, 19, 21, 23, 25, 28, 29, 32, 36 (E84081), 38, 45, 47 (KS90H450), 50 (KS96WGRC36), 51 (R05), and 52 and virulent to Lr3bg, 15, 20 (Thew), 27+31 (Gatcher), and 33. Lines containing the adult plant resistance (APR) genes Lr12 (RL6011, IT 3++), Lr13 (CT263, IT 3), Lr22b (Tc, IT 4), and Lr37 (RL6081, IT 3) were susceptible in the adult stage to 3SA145, whereas lines with the APR genes Lr22a (RL6044, IT ;1), Lr34 (RL6058, IT Z1), and Lr35 (RL6082, IT ;1) were resistant in controlled infection studies in a greenhouse. A control, the common race (3SA133), was virulent only on Tc adult plants. In seedlings, 3SA133 was avirulent to Lr15, 17, 26, and 27+31, but unlike 3SA145, it was virulent to Lr1, 2c, 11, 18, 24, and 28. Races 3SA133 and 3SA145 did not differ in their virulence to the remaining seedling genes. Virulence to Lr37 has been reported in several countries, including Australia, Canada, Uruguay, and the United States (1,2). Prior to the detection of 3SA145, adult plants of RL6081 were resistant to all wheat leaf rust races in South Africa. In 2009, however, RL6081 showed severity levels of up to 30S at certain Western Cape trap plot sites. Of 124 South African bread wheat cultivars and advanced breeding lines tested at the seedling stage, 3SA145 was virulent to 48, whereas 3SA133 was virulent to 36 entries. A further six entries were heterogeneous in their reaction to 3SA145. In adult plant infection studies of 48 South African spring wheats in a greenhouse, 19 were susceptible (flag leaf IT ≥3) and 22 were resistant to 3SA145. Seven entries showed a Z3 flag leaf IT indicating adult plant resistance. According to a simple sequence repeat (SSR) study using 17 primer-pair combinations described by Szabo and Kolmer (3), 3SA145 showed 30% homology with the dominant South African races. Although virulence to Lr12 and Lr13 has been known in different leaf rust races in South Africa, to our knowledge, this is the first report of combined virulence to Lr12, 13, and 37. The SSR data and unique avirulence/virulence profile suggest that 3SA145 may be an exotic introduction to South Africa. References: (1) J. A. Kolmer et al. Plant Dis. 89:1201, 2005. (2) B. McCallum and P. Seto-Goh. Can. J. Plant Pathol. 31:80, 2009. (3) L. Szabo and J. Kolmer. Mol. Ecol. Notes 7:708, 2007. (4) T. Terefe et al. S. Afr. J. Plant Soil 26:51, 2009.

Detection of Variants of Wheat Stem Rust Race Ug99 (Puccinia graminis f. sp. tritici) in Zimbabwe and Mozambique.

The migration of Ug99 variants of Puccinia graminis f. sp. tritici is of concern to global wheat production (1). Seven races have been characterized in the Ug99 lineage (3), three of which occur in South Africa (4). During surveys of wheat fields for Ug99 in Zimbabwe and Mozambique in August and September 2010, high stem rust severities were found at Chiredzi, Chisumbanje, and Birchenough in Zimbabwe and at Rotanda in Mozambique. Stem rust was widespread in the lowlands (<800 m above sea level) of Zimbabwe and trace amounts were present in the mid-altitude areas. In Mozambique, stem rust was only observed at Rotanda (sample Moz1001). Collections from Chiredzi (samples Zim1004 and Zim1005), Chisumbanje (Zim1006), and Birchenough (Zim1009 and Zim1010) yielded viable urediniospores for infection studies. According to race analysis conducted on seedlings of the North American stem rust differential set (2) in a greenhouse at 18 to 25°C, Zim1005 and Zim1006 were typed as PTKST and Zim1004 and Zim1009 as TTKSF. Both TTKSF and PTKST were detected in the Zim1010 sample. Race analysis experiments were conducted three times. Urediniospores of isolate Moz1001 were not viable in infection studies, but yielded fungal DNA for simple sequence repeat (SSR) analysis. Using eight selected SSR primer combinations (4), all six isolates clustered within the Ug99 lineage. Isolates Zim1005, Zim1006, Zim1009, Zim1010, and Moz1001 and the stem rust control races TTKSF, TTKSK, and PTKST grouped into two main clusters, with Zim1009 and Zim1010 clustering together and sharing 88% similarity with the rest of the isolates. Zim1005 and Zim1006 were identical to TTKSF and TTKSK, respectively. Zim1004 shared 96% genetic similarity with the TTKSP control, with these two sharing 74% genetic similarity with the remaining isolates. The SSR data correlated with the infection data, except for Zim1004, which was typed as TTKSF but clustered close to TTKSP. Wheat cvs. SC Nduna, SC Shine, SC Stallion, SC Smart, Kana, Insiza, and Dande are predominant in Zimbabwe. Cv. SC Stallion and other unidentified cultivars were susceptible to P. graminis f. sp. tritici in the field in Zimbabwe. In Mozambique, the tall, local cv. Sitsonko was susceptible to P. graminis f. sp. tritici but no infections were observed on SC Nduna or SC Shine. The similarity in P. graminis f. sp. tritici races in Zimbabwe, South Africa, and Mozambique suggests that inoculum is exchanged within the region and explains the detection of race PTKST in South Africa in 2009. Trajectory models showed winds originating at Birchenough in October 2009, where stem rust was observed, passing directly over KwaZulu-Natal, South Africa within 48 to 72 h. Race PTKST was confirmed from collections in KwaZulu-Natal in November 2009 (4). The confirmation of Sr31 virulence in race PTKST in Zimbabwe is important because it provides new geographical records for an Ug99-related race and puts Southern African cultivars with 1B.1R resistance at risk. References: (1) D. Hodson. Euphytica 179:93, 2011. (2) Y. Jin et al. Plant Dis. 92:923, 2008. (3) R. F. Park et al. Euphytica 179:109, 2011. (4) B. Visser et al. Euphytica 179:119, 2011.

First Report of Virulence for the Wheat Leaf Rust (Puccinia triticina) Resistance Gene Lr32 in South Africa.

The wheat leaf rust resistance gene Lr32 was transferred from Aegilops tauschii Coss. to bread wheat (Triticum aestivum L.) (1). Despite virulence for Lr32 in some isolates from Bulgaria, Israel, and Turkey, the gene has been reported to be effective in Australia, Mexico, the United States, and South Africa (1,2). A leaf rust isolate that differed in its avirulence/virulence profile from previously recorded races of Puccinia triticina Eriks. in South Africa was collected from triticale (× Triticosecale) in the Western Cape in 2005. According to the South African leaf rust differential set (3), this isolate (UVPt19) was avirulent for Lr3a, 3bg, 3ka, 10, 11, 16, 20, 26, and 30 and virulent for Lr1, 2a, 2b, 2c, 14a, 15, 17, 24, and Thatcher (Tc, control). Except for Lr20 in cv. Thew, all differentials are Tc near-isogenic lines. In comparison with known South African races (3), it differed from race 3SA132 at the Lr10 locus. Using standard rust pathology protocols (3), an expanded set of Lr gene lines (non Tc lines indicated) showed that UVPt19 is avirulent on wheat seedlings containing Lr9, 19, 21, 25, 27+31 (Gatcher), 29, 36 (ER84018), 37, 41 (KS91WGRC10), 44, 45, 47 (KS90H450), 50 (KS96WGRC36), 51 (R05), and 52, and virulent for Lr12, 22a, 23, 28, 32, 33, and 35. In the seedling stage, UVPt19 was virulent for the temperature sensitive genes Lr13, 18, and 34 at 25°C, but produced lower infection types (ITs) on Lr18 and 34 at 14 to 18°C. Seedlings of Pavon 76 (Lr46) were resistant (IT ;1=) to UVPt19. The susceptible response of lines carrying Lr32 was confirmed by high ITs (3++4) on RL5713/2*Mq, RL6086 (TcLr32), and RL5713/2*Mq//6*Palmiet. A control isolate (UVPt9) produced ITs ;1+, ;1+, and ;;1= on these lines, respectively. UVPt19 was virulent on line RL6092 (TcLr20) but avirulent on Thew. When tested on adult plants of lines RL6011 (TcLr12), CT263 (TcLr13), RL6044 (TcLr22a), RL6058 (TcLr34), RL6082 (TcLr35), RL6081 (TcLr37), and Tc (control), UVPt19 was only virulent (IT 3+) on CT263 and Tc. Flag leaves of RL6011 (IT ;1), RL6044 (IT 1), RL6058 (IT Z3-), RL6082 (IT 0;), and RL6081 (IT ;1) were resistant. UVPt19 was virulent on seedlings of 11 of 13 triticale cultivars and lines tested as opposed to UVPt9, which was virulent to only one entry. From a collection of 105 South African bread wheat cultivars and elite breeding lines, UVPt19 was virulent on 13 and five were mixed in their response to this isolate. All IT experiments were repeated. Although virulence has emerged for Lr32 in South Africa, the gene has not been used in local cultivars. Previously, McIntosh et al. (1) also reported that Lr32 has not been exploited in wheat production. On the basis of current evidence, UVPt19 appears to be potentially more damaging to triticale than bread wheat. Furthermore, the race seems rare because it was not collected in a recent wheat leaf rust survey in South Africa (3). References: (1) R. A. McIntosh et al. The Wheat Rusts: An Atlas of Resistance Genes, CSIRO-Kluwer, Dordrecht, the Netherlands, 1995. (2) Z. A. Pretorius. Phytophylactica 21:195, 1989. (3) T. Tarekegn et al. S. Afr. J. Plant Soil 26:51, 2009.

First Report of a Puccinia graminis f. sp. tritici Race Virulent to the Sr24 and Sr31 Wheat Stem Rust Resistance Genes in South Africa.

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. References: (1) Y. Jin et al. Plant Dis. 92:923, 2008. (2) K. Nazari et al. Plant Dis. 93:317, 2009. (3) R. P. Singh et al. Adv. Agron. 98:271, 2008. (4) B. Visser et al. Mol. Plant Pathol. 10:213, 2009.

Detection of Virulence to Resistance Gene Sr36 Within the TTKS Race Lineage of Puccinia graminis f. sp. tritici.

The stem rust resistance gene Sr36 confers a near-immune resistance reaction to many races of Puccinia graminis f. sp. tritici and is highly effective against race TTKSK (syn. Ug99), which possesses unusually broad virulence combinations. Because this gene is widely used in United States soft winter wheat germplasm and cultivars, it has been considered to be an important source of resistance to TTKSK. In 2007, moderately susceptible infection responses were observed on wheat lines and cultivars carrying Sr36 in a field screening nursery for stem rust at Njoro, Kenya. We derived 18 single-pustule isolates from stem rust samples collected from the 2007 Njoro nursery. The isolates were evaluated for virulence on 20 North American stem rust differential lines and on wheat lines and cultivars carrying Sr36, Sr31+Sr36, and Sr24+Sr31. Of the 18 isolates, 10 produced infection types 3+ to 4 on line W2691SrTt-1 (monogenic for Sr36) and other lines that carry Sr36 and belonged to a new virulence phenotype that was not detected in previous years. These isolates were identified as race TTTSK. The remaining eight isolates were identified as races TTKSK (five isolates) and TTKST (three isolates), with avirulence and virulence, respectively, to Sr24. Thirteen simple sequence repeat (SSR) markers were used to examine the genetic relationships among the three races in the TTKS lineage. All isolates in the lineage shared an identical SSR genotype and were clearly different from North American races. In all, 16 wheat cultivars and 60 elite breeding lines, postulated to possess Sr36, were susceptible to race TTTSK. The occurrence of race TTTSK with combined virulence on Sr31 and Sr36 has further broadened the virulence spectrum of the TTKS lineage and rendered an important source of resistance ineffective.

Detection of Virulence to Resistance Gene Sr24 Within Race TTKS of Puccinia graminis f. sp. tritici.

The stem rust resistance gene Sr24 is effective against most races of Puccinia graminis f. sp. tritici, including race TTKS (syn. Ug99), and is used widely in commercial wheat cultivars worldwide. In 2006, susceptible infection responses were observed on wheat lines and cultivars carrying Sr24 in a field stem rust screening nursery at Njoro, Kenya. We derived 28 single-pustule isolates from stem rust samples collected from the 2006 Njoro nursery. The isolates were evaluated for virulence on 16 North American stem rust differential lines; on wheat lines carrying Sr24, Sr31, Sr38, and SrMcN; and on a wheat cultivar with a combination of Sr24 and Sr31. All isolates were identified as race TTKS with additional virulence on Sr31 and Sr38. These isolates were divided into two groups: group A (seven isolates and the two control isolates), producing a low infection type, and group B (21 isolates), producing a high infection type on Sr24, respectively. Isolates of group B represented a new variant of race TTKS with virulence to Sr24. Eighteen simple sequence repeat (SSR) markers were used to examine the genetic relationship between these two groups of isolates in race TTKS and five North American races (MCCF, QCCQ, RCRS, RTHS, and TPMK) that are representative of distinct lineage groups. All isolates of race TTKS shared an identical SSR genotype and were clearly different from North American races. The virulence and SSR data indicated that the new variant of race TTKS with Sr24 virulence likely has arisen via mutation within the TTKS genetic lineage. We propose to revise the North American stem rust nomenclature system by the addition of four genes (Sr24, Sr31, Sr38, and SrMcN) as the fifth set. This revision recognizes the virulence on Sr31 and differentiates isolates within race TTKS into two separate races: TTKSK and TTKST, with avirulence and virulence on Sr24, respectively. The occurrence of race TTKST with combined virulence on Sr24 and Sr31 has substantially increased the vulnerability of wheat to stem rust worldwide.

First Report of Asian Soybean Rust Caused by Phakopsora pachyrhizi on Kudzu in South Africa.

Asian soybean rust was first reported on soybean in South Africa (SA) in 2001 (3). The disease has occurred in all ensuing seasons, particularly in the humid, eastern production regions, causing significant losses in soybean fields not protected by fungicides. In April 2005, rust-infected Pueraria lobata (kudzu) was detected near Nelspruit, Mpumalanga, SA. At this location (25°20'41″S, 30°43'30″E), kudzu plants occurred abundantly on road sides, edges of pine plantations, and in natural vegetation. Most vines were infected, with abaxial surfaces of older leaves often showing 100% severity. Following inoculation with rust spores collected from kudzu, soybean line PI200492 (Rpp1) produced tan lesions typical of a susceptible reaction for Asian soybean rust. PI230970 (Rpp2), PI462312 (Rpp3), and PI459025 (Rpp4) showed red-brown lesions typical of a resistant reaction. Using Ppm1/Ppa2 and Ppm1/Ppm2 primer combinations, the amplification profiles of the internal transcribed spacer region (1) of rust DNA extracted from primary leaves of line PI200492 infected with spores collected from kudzu positively identified the pathogen as Phakopsora pachyrhizi. The Ppm1/Pme2 primer combination specific for P. meibomiae (1) did not yield an amplification product. The Qualiplate ELISA test kit (EnviroLogix Inc., Portland, ME) verified the identification of P. pachyrhizi on an original kudzu sample as well as the leaf material used for DNA analysis. A survey of kudzu at the Nelspruit site during July 2005 confirmed the presence of the pathogen during the offseason for soybean. At that time, incidence of kudzu rust remained high, but few leaves showed high severity. The susceptibility of kudzu to Asian soybean rust has been reported in controlled infection studies in SA (2). To our knowledge, this is the first report of P. pachyrhizi causing rust on a large, naturally occurring kudzu population in SA. References: (1) R. D. Frederick et al. Phytopathology 92:217, 2002. (2) A. Nunkumar. M.Sc. thesis. University of KwaZulu-Natal, South Africa, 2006. (3) Z. A. Pretorius et al. Plant Dis. 85:1288, 2001.

Characterization of Seedling Infection Types and Adult Plant Infection Responses of Monogenic Sr Gene Lines to Race TTKS of Puccinia graminis f. sp. tritici.

Stem rust, caused by Puccinia graminis f. sp. tritici, historically was one of the most destructive diseases of wheat and barley. The disease has been under effective control worldwide through the widespread use of host resistance. A number of stem rust resistance genes in wheat have been characterized for their reactions to specific races of P. graminis f. sp. tritici. Adult plant responses to race TTKS (also known as Ug99) of monogenic lines for Sr genes, a direct measurement of the effectiveness for a given gene, have not been investigated to any extent. This report summarizes adult plant infection responses and seedling infection types for monogenic lines of designated Sr genes challenged with race TTKS. High infection types at the seedling stage and susceptible infection responses in adult plants were observed on monogenic lines carrying Sr5, 6, 7a, 7b, 8a, 8b, 9a, 9b, 9d, 9g, 10, 11, 12, 15, 16, 17, 18, 19, 20, 23, 30, 31, 34, 38, and Wld-1. Monogenic lines of resistance genes Sr13, 22, 24, 25, 26, 27, 28, 32, 33, 35, 36, 37, 39, 40, 44, Tmp, and Tt-3 were effective against TTKS both at the seedling and adult plant stages. The low infection types to race TTKS observed for these resistance genes corresponded to the expected low infections of these genes to other incompatible races of P. graminis f. sp. tritici. The level of resistance conferred by these genes at the adult plant stage varied between highly resistant to moderately susceptible. The results from this study were inconclusive for determining the effectiveness of resistance genes Sr9e, 14, 21, and 29 against race TTKS. The understanding of the effectiveness of individual Sr genes against race TTKS will facilitate the utilization of these genes in breeding for stem rust resistance in wheat.