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Wheat stem rust (SR), caused by Puccinia graminis f. sp. tritici, (Pgt) is considered one of the most destructive diseases of the wheat crop. As Sr24 and Sr31 are the most widely used resistance genes in the Southern Cone of America, wheat crops in this region is under threat of SR outbreaks posed by the potential migration of virulent Pgt Ug99-lineage races (Ug99+). Efforts have to be made to develop adapted lines resistant to Ug99+. Genes Sr26, Sr32 and Sr39 are effective to both Ug99+ and local races of the pathogen. This work is aimed to pyramid two and three of the resistance genes in two locally adapted wheat cultivars (G?nesis 2375 and G?nesis 6.87). Donor lines of Sr26, Sr32 and Sr39 (developed by I. Dundas, University of Adelaide, Australia) and molecular markers Sr26#43, csSr32#1 and Sr39#22r (developed by R. Mago et al., University of Adelaide) were used. Lines with two-gene combinations were developed in two steps. First, tree-way crosses were made by crossing heterozygous F1 plants (derived from crossings donor lines) to either one of the two adapted wheat cultivars. Subsequently, tree-way F1 plants were genotyped and only those with two-gene combinations were backcrossed (BC) twice to the adapted cultivars. Among three-way F1 plants, two-genes combinations were confirmed for Sr26+Sr32 (8 out of 31), Sr26+Sr39 (2 of 115) and Sr32+Sr39 (26 out of 103). In the BC1F1 generation, Sr26+Sr32, Sr26+Sr39 and Sr32+Sr39 combinations corresponded with 9, 9 and 45 out of 99, 27 and 241 plants, respectively. In 2017, 1345 BC2F1 plants are being grown to obtain BC2F2. We plan to intercross plants with two-gene combinations to obtain lines with the three genes which will be used as sources of resistance to develop cultivars with presumably longer lasting resistance to wheat SR.
In 2016 rust surveys were carried out in all the four key wheat growing regions: South Rift (June, July), Mount Kenya (July), North Rift (September) and Central Rift (part of August and September). A total of 304 farms were sampled. Stem rust was detected in 235 (78.3%), yellow rust in twenty-eight (9.3%) and leaf rust in fourteen (4.7%) of the farms. Stem and yellow rust were detected in all the wheat growing regions while leaf rust was detected in South, North and Central Rift. Stem rust infection ranged from TR to 90S with maximum infection in Central Rift (88.3%), Mt. Kenya region (80.3%); South Rift (76.5%) and North Rift (72.4%). Yellow rust infection ranged TR to 60S with maximum infection in Central Rift (16.7%); North Rift(13.3 %) and minimum infection in South Rift( 4.9%),) and Mt. Kenya region ( 1.7%). Leaf rust infection ranged from trace to 50S with maximum infection in North Rift (10.2%) minimum infection in Central Rift (3.3 %) and South Rift (1.2%). Fifty percent of the eight previously released wheat varieties are now susceptible to the Ug99 race. Race analysis results from AAFC Canada suggested the presence of TTKSK which was dominating in North Rift and TTKSK, TTKST and TTTTF were dominant in the screening nursery at Njoro. Yellow rust in the region has increased in the current year owing to the incursion of a probable new race AF2012 which has resulted in increased disease severity on varieties and materials tested in the International nurseries at KALRO, Njoro.
The rapid appearance of new races of rust pathogens with virulence for the major seedling resistance genes in wheat has intensified the focus to discover adult plant resistance (APR) genes in wheat and utilize them in breeding programs for sustainable wheat production. The experimental breeding line 'Copio' developed by the International Maize and Wheat Improvement Centre (CIMMYT) in Mexico has exhibited high levels of APR to all three rusts including the African stem rust Ug99 race group. To dissect the mechanism of APR in Copio it was crossed with APAV#1, which is susceptible to all three rusts and a population of 176 F4:F5 recombinant inbred lines (RILs) was developed at CIMMYT. Both parental lines were found to be susceptible (IT >3) at the seedling stage to races TTKSK and TKTTF, which ensures the field data from Africa will be applicable for APR mapping. Seedling tests were also conducted on the RIL population using the predominant Pakistani race RRTTF, and Chi-squared tests indicated segregation of two stem rust seedling genes (?2 test P value of 0.00002). Both parents were also tested for the known APR genes Lr34/Yr18/Sr57, Lr46/Yr29/Sr58, Lr67/Yr46/Sr55 and Sr2/Yr30 using molecular markers and results indicate that APAV#1 does not carry any known APR genes, while Copio might have Lr46 and Sr2. This population was tested in four field environments (US, Pakistan, Mexico, and Kenya) for leaf, stem and yellow rusts during 2015-16 and 2016-17. Disease severity distributions of all three rusts for the RILs across all environments were continuous, suggestive of quantitative and polygenic resistance.
We are using genotyping by sequencing (GBS) as a genotyping platform and anticipate having preliminary mapping results available by spring 2018.
Wheat plays a vital role in multifaceted farming system of Pakistan. Like other many other countries, Pakistan's sustainable wheat production is also continuously threatened by a number of biotic and abiotic stresses. Among the biotic stresses, three rust diseases of wheat have been the most devastating. Stem rust was effectively controlled with adoption of the semi-dwarf spring wheats of the Green Revolution. However, the threat of the evolution of Ug99 race of stem rust in East Africa and its migration to Iran cannot be neglected. The Chance of of Ug99 migrating from Iran into Pakistan, coupled with the presence of dangerous new races of stripe and leaf rusts invites enormous efforts for development of rust resistant varieties for sustainable production of the wheat in the country. In this regard the Wheat Program, NARC, Pakistan initiated an intensive breeding program with financial and technical support of USDA and CIMMYT. Diverse sources of resistance to the three rusts particularly to the stem rust race Ug99 were introduced from CIMMYT. Through the rigorous selection procedure, four rusts resistant wheat varieties (NARC 2011, Pakistan 2013, Zincol 2016 and Borlaug 2016) have been released. These varieties are also resistant to Ug99. The varieties i.e. NARC 2011, Borlaug 2016 and Zincol 2016 are performing well in irrigated areas whereas Pakistan 2013 is suitable for rainfed conditions. The variety Zincol 2016 has high Zn content (35 ppm) in grain as compared to national standard check variety (25 ppm). These varieties are not only higher yielding but also possess good grain quality and other desirable traits. A considerable quantity of seed of the varieties is already present in the national seed system and will reduce the risk of Ug99 threat.
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.
To reduce losses caused by rusts, regular and timely replacement of susceptible varieties with new high yielding, rust resistant varieties must occur. Data from a farmer survey carried out across Pakistan (Punjab, Sindh, KPK and Baluchistan) in 2014 enabled an analysis of the uptake of rust resistant variety NARC 2011. The empirical results indicated that the major sources of information that farmers obtained about NARC 2011 were research stations (83%), seed companies (7%) and fellow farmers (5%). Although production inputs were applied equally to both rust resistant NARC 2011 and rust susceptible wheat varieties the average yield of NARC 2011 (5,063 kg/ha) was superior to high yielding but rust susceptible varieties (4,446 kg/ha). Quality attributes of NARC 2011, including taste, color, dough kneading and chapatti making properties, were preferred by >70% of farmers). Seed availability and accessibility of NARC 2011 were major issues. Farmer awareness of rusts, especially the threat of exotic Pgt race Ug99, needs to be improved.
Stem rust is a potentially destructive fungal disease of wheat worldwide. In 1998 Pgt pathotype TTKSK virulent to Sr31 was detected in Uganda. The same pathotype was confirmed in Lorestan and Hamedan provinces of Iran in 2007. We used a derivative of race TTKSK to phenotype 62 Iranian wheat landraces (resistant to stripe rust in a previous study) at the seedling stage to this new pathotype (TTSSK). Twenty eight accessions were evaluated for the presence of resistance genes Sr2, Sr22, Sr24, Sr25, Sr26, Sr35, Sr36 and Srweb using SSR markers. None carried Sr2, Sr24 or Sr26, but the presence of Sr22, Sr25, Sr35 and Sr36 was indicated. Some susceptible landraces predicted to carry Sr2 by marker analysis require further investigation. To evaluate defense gene expression in compatible and incompatible stem rust interactions we sampled resistant and susceptible cultivars at 0, 12, 18, 24, 72 hours post-inoculation (hpi). ?-1,3 glucanase expression was studied using qGLU-S and qGLUU-AS primers and a real-time PCR step-one ABI machine, with ?-tubulin and EF1-? genes used as internal controls. In incompatible interactions defense gene expression was increased at 24 hpi, but in compatible interactions the highest level of expression occurred at 12 hpi and was significantly decreased at 18 hpi. The results revealed that expression of defense genes such as ?-1,3 glucanase was earlier in compatible than in incompatible interactions but the expression level was less in incompatible interactions. On the other hand, in susceptible genotypes the expression of defense genes increased immediately after inoculation and declined sharply after infection. In contrast defense gene expression in resistant genotypes began to increase after establishment of the pathogen.
Wheat landrace PI 177906 has seedling and field resistance to Pgt races TTKSK and TTKST. From a cross between PI 177906 and LMPG-6, 138 doubled haploid (DH) lines and 144 recombinant inbred lines (RILs) were developed and tested for seedling resistance to Pgt race TTKSK. Goodness-of-fit tests from both populations indicated that two dominant genes in PI 177906 conditioned resistance to race TTKSK. Parents and the 138 DH lines were evaluated in the field in two experiments in Kenya; one in the main season and one in the off-season. The 90K wheat iSelect SNP genotyping platform was used to genotype the parents and DH lines and data were used to construct a genetic linkage map. Two loci for seedling resistance were mapped to chromosomes 2BL and 4BL. Two major QTL for field resistance mapped to the same regions, a 14.4 cM interval on 2BL and an 8.5 cM interval on 4BL. The QTL on 2BL and 4BL explained, respectively, 31.9-32.3% and 18.2-19.1% of the variation in the off-season and 28.3-30.4% and 5.4-6.5% of the variation in the main-season. Based on the mapping results, race specificity, and the seedling infection types, the resistance gene in 2BL could be Sr28, whereas the gene on chromosome 4BL could be novel. The mapping results will be verified in the RIL population using the flanking SNP markers in KASP assays.
With ongoing threats of rust from both internal and international sources it has become a priority at CIMMYT and for Pakistan national programs to accelerate the rate of seed increase and to popularize new Pgt race Ug99 resistant varieties to avert future disasters. Seed of Ug99 resistant varieties NARC-11, Pak-2013, Dharabi-2011 and BARS-09 was produced under the Wheat Productivity Enhancement Program (WPEP). The country-wide participatory approach involves a partnership of farmers, seed companies and research institutes. In 2014 16,020 and 6,085 kg of seed of NARC-11 and Pak-13, respectively, were distributed all over the country, including Azad Jammu Kashmir and Gilgit-Baltistan. Comparative yields across Pakistan show that the rust resistant varieties are equal, or superior, to current stem rust susceptible varieties grown by farmers. Deployment and use of these varieties by farmers in Balochistan will have a significant impact not only on productivity, but may also avert the consequences of possible introduction of race Ug99.
Four Ug99 pathotypes occur in southern Africa. Although South African bread wheat cultivars and lines are regularly screened against representative isolates, the stem rust reactions of Zimbabwean germplasm to these variants were largely unknown. A collection of 49 wheat cultivars and lines, obtained from Seed-Co (Ltd.) and the Crop Breeding Institute in Zimbabwe, were tested as seedlings against pathotypes TTKSF, TTKSF+, TTKSP and PTKST. Twelve varieties and 21 experimental lines showed low infection types with all four pathotypes. Using molecular marker assays Sr31 was detected in 26 entries, Sr24 in five and Sr36 in one. The csSr2 marker suggested the presence of Sr2 in 20 entries. Screening of adult plants in the greenhouse using pathotype PTKST showed 34 entries with low infection types and 15 had high infection types. Stem rust field records in 2012 showed 5 susceptible entries with stem rust scores between 50S and 80S, whereas only 4 susceptible entries were identified in 2014 with scores ranging from 30S to 80S. Three lines were susceptible in both seasons. The study exposed the vulnerability of Zimbabwean wheat germplasm to Ug99 variants, but also identified suitable lines that can be used in breeding and possible commercialization.