Ethiopian institute of agricultural research
Dr. Netsanet B. Heyi, Dr. Getaneh W. Wolderufael, Tsegab T.
Stem rust caused by Puccinia graminis f. sp. tritici (Pgt) is a major production constraint in most wheat growing areas of Ethiopia. The stem rust pathogen is capable of rapidly developing new virulence to resistance genes. The highlands of Ethiopia are considered a hot spot for Pgt diversity. The present study was conducted to investigate the virulence diversity and spatial distribution of races of Pgt in the major wheat growing areas of Ethiopia. The physiologic races of Pgt were determined on seedlings of the standard wheat stem rust differentials following the international system of nomenclature. Stem rust race analyses were carried out both at Ambo Plant Protection Center and the Cereal Disease Laboratory in Minnesota. 426 stem rust samples were collected from major wheat growing of the country in the 2016 cropping season and 185 viable samples were analyzed. Stem rust races TKTTF, TTKSK, TTTTF, JRCQC and RRTTF were identified. Among the identified races, TKTTTF was dominant at a frequency of 78.7% followed by TTKSK (10.6%). Race TTTTF was found for the first time in Ethiopia in 2016. Only one resistance gene in the differential set, Sr24, was effective against all isolates. Stem rust resistance gene Sr31 was found to confer resistance to most of the races prevalent in Ethiopia with the exception of Ug99. Sr24 could be used in combination with other resistance genes in breeding for resistance to stem rust in Ethiopia.
University of Minnesota
Caixia Lan, Ravi Singh, Matthew Rouse, Muhammad Imtiaz, James Anderson
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.
University of Seville
Solis,Ignacio, , , , , , , , , , , , , , , , , , , , , , , , , , , ,
Leaf rust is an important worldwide disease on wheat caused by the fungus Puccinia triticina. Great infections on durum wheat occurred in Southern Spain in the 2000s but diminished in recent years due to deployment of resistant varieties and application of fungicides by farmers. A leaf rust survey was carried out from the 2009-15 period to monitor the virulence spectrum of the prevailing pathotypes. A total of 84 leaf rust isolates were collected on durum wheat fields. From those, single culture were obtained and used to inoculate a set of 27 differential isolines of the susceptible variety Thatcher. In addition 8 durum varieties with known Lr genes were also included.
The main highlight is that the resistance conferred by the popular Lr14a gene was broke up in 2013, but since then virulence to this gene is not widespread. In total, 23% of the isolates were virulent to the lines containing Lr14a. Lr1, Lr3, Lr3bg, Lr16, Lr24, Lr26, and Lr28 are very effective. Lines carrying Lr2c, Lr10, Lr14b, Lr20, Lr23, and LrB displayed susceptibility to most isolates. The durum varieties Jupare (Lr27+Lr31), Guayacan (Lr61), Storlom (Lr3+) and Camayo (LrCam) are also resistant against all isolates tested. Diversification of Lr genes is needed in the coming varieties to delay the appearance of new virulent races.
Jianping Zhang, Peng Zhang, Robert Park, Narayana Upadhyaya, Robert McIntosh, Sambasivam Periyannan, Brande Wulff, Burkhard Steuernagel, Evans Lagudah
Evolution of rust pathogens continues to pose challenges to global wheat production. Major resistance (R) genes, which encode proteins of the NBS-LRR (Nucleotide-binding site, leucine-rich repeat) family, have been a valuable resource for breeders to minimise yield losses from infection. Many wheat varieties harbor numerous R genes that could be identified and cloned in order to engineer more sustainable disease control. The advent of targeted gene enrichment and next-generation sequencing (NGS) has allowed rapid cloning of specific R genes, thus enhancing efforts to pyramid these genes and investigate their underlying resistance mechanisms. Several R genes present different phenotypes in certain genetic backgrounds, and cloning them would be an important step towards uncovering their interactions. Hybrid necrosis is one such phenotype observed in crosses of wheat genotypes involving the R gene Lr13 and complementary genes, Ne1 and Ne2, occurring in different allelic forms. It was recently concluded that Lr13 and an allele of Ne2 are actually the same gene based on genetic and mutational studies. The capability of Lr13 to confer both leaf rust resistance and hybrid necrosis cannot be answered without first cloning it. The lack of tightly linked markers coupled with the proximal 2BS chromosomal location of Lr13 does not make it easily amenable to map-based cloning. The NGS-based pipeline MutRenSeq (mutagenesis and R-gene enrichment sequencing) was used on EMS (Ethyl methanesulfonate) induced, susceptible Lr13 mutants along with support from comparative genomics to ascertain candidate gene sequences for Lr13, which are at advanced stages of screening and confirmation. Definite proof that a single gene is involved will only come with transformation studies when the cloned Lr13 candidate transformed into a susceptible line confers both a resistance phenotype in the transgenic line and a necrotic phenotype in the offspring of crosses between the transgenic line and a line possessing Ne1.
National Plant Protection Center
Namgay Om, Thinlay, Ugyen Yangchen
Wheat rusts are one of the important diseases that limit the production and downgrade wheat quality. Three rust diseases of wheat are stem rust caused by Puccinia graminis Pers. f. sp. tritici Eriks., stripe rust caused by Puccinia striiformis Westend. f. sp. tritici Eriks., and leaf rust caused by Puccinia triticina Eriks. This study was conducted to determine the reaction of wheat varieties to wheat rusts at different altitudes. Field experiments were conducted from December 2016 to March 2017 at Mendagang (27.5886°N, 89.8711°E, 1332 masl), Punakha Dzongkhag (district) for mid altitude and at Agriculture Research and Development Center (ARDC), Samtenling (26.9058°N, 90.4308°E, 378 masl), Sarpang Dzongkhag, Bhutan for low altitude. The experiment followed a RCBD with 15 treatments comprising of three Bhutanese released varieties, eight SAARC varieties, and four ICARDA varieties. Each treatment was replicated three times. Assessment of disease incidence and severity were performed three times starting from tillering to ripening stage, approximately at 60, 90 and 120 days after sowing (DAS). Disease severity was determined following the modified Cobb’s disease rating scale. Of the 15 varieties, only 11 germinated in both the sites. Among the three wheat rust diseases, only leaf rust was observed in both sites. Leaf rust incidences ranged from 2.5 to 10% and 2.5 to 16% at mid and low altitudes respectively. Disease severity of 5 to 20%, corresponding to field response of immune (5O) to moderately resistant (20MR), was observed at mid altitude, while 5 to 100%, with immune (5O) to susceptible (100S), was observed at low altitude. There was a significant difference in disease incidence by site (p=.038) but not in disease severity (p=.129). The variety, ICARDA 1, with 100% severity was highly susceptible (100S) to leaf rust at low altitude while Bajosokha Kaa remained immune (5O) in both the sites. The results indicate that leaf rust can occur in both low and mid altitudes; however selection of suitable varieties requires more extensive studies.
National Agronomic Institute of Tunisia
Sana Kamel, Elhem, Elfahem, Wissal Feriani, Hanen Sbei
In order to identify sources of resistance to tan spot caused by Pyrenophora tritici-repentis, 359 local wheat accessions were evaluated for reaction to the Oued-Mliz isolate in controlled conditions and in the field. Two and three assessments were carried out at the seedling and adult stages, respectively. There was a highly significant accession effect and 4.2% of accessions were highly resistant in both controlled conditions and the field. Assessments at the seedling stage were positively correlated with each other, and assessments in the adult stage were also positively correlated. However, assessments at the seedling stage were negatively correlated with those at the adult stage. One hundred and fifty five accessions with known origins (from 15 localities belonging to four districts) were projected on a graph defined by the two axes: reactions at the seedling stage and reactions at the adult stage. After placing the average reactions at the seedling and adult stages on the graph, four groups of accessions were obtained: accessions that were resistant to both stages, accessions that were resistant at the adult stage only, accessions that were resistant at the seedling stage only, and accessions that were susceptible at both stages. All four groups were found in each district. However, considering localities, reactions of accessions were highly variable. For example, accessions originating from Menzel Hbib were genetically variable and were represented in each of the four groups, whereas accessions from Sidi El Hani were all resistant at both stages. Further work is needed to study the genetic variability within and between localities and to better understand the resistant accessions.
Institute of Plant Protection, Chinese Academy of Agricultural Sciences
Jing Feng, Ruiming Lin, Fengtao Wang, Qiang Yao, Qingyun Guo, Shichang Xu
Wheat stripe rust is an important air borne disease caused by Puccinia striiformis f. sp. tritici, and seriously threatens the safety of wheat production. Breeding and utilization of resistant varieties is the most economical, safe and effective measure to control wheat stripe rust. Sifangmai is a landrace from the state of Guangxi, China, and maintains good resistance to the current epidemic species CYR34, CYR33, CYR32 and CYR29 in China. Sifangmai was crossed with Taichung 29 to obtain F1, F2 and F2:3 to analyze its character of inheritance. In the adult stage, the cross of Sifangmai /Taichung 29 was inoculated by CYR32. The genetic analysis showed that the resistance of Sifangmai to CYR32 was controlled by a dominant gene, named as YrSF. A mapping population of F2 was genotyped with simple sequence repeat (SSR) markers. SSR loci Xgpw8015, Xgpw4098, Xwmc73, Xgpw8092, Xgpw7309 and Xbarc89 on 5B chromosome showed polymorphic between Taichung 29, Sifangmai, and resistant and susceptible pools, indicating that the resistant gene in Sifangmai was located on the 5B chromosome. The linkage map of these SSR markers was constructed and the nearest SSR to the gene is Xgpw8015. A set of Chinese Spring nulli-tetrasomic lines was used to confirm YrSF on chromosome 5B. YrSF is different from known genes in chromosome 5B. Xgpw8015 can be used as a marker for detection of YrSF.
Ayele Badebo, Abebe Atilaw, Habtemariam Zegeye, Zerihun Tadesse, Wasihun Legesse, Terefe Fitta, Dawit Asnake
In Ethiopia, quality seed of improved varieties is the least expensive and most critical input for the sustainable production of wheat, a strategic food security crop grown by some 4.7 million households on 1.7 million hectares. Because wheat is self-pollinated, farmers can save and replant seed from their harvests for several years, without the variety losing its genetic identity. At the same time, recommended seed rates for wheat (150 to 200 kilograms per hectare) are significantly higher than those for tef (15 kg/ha) or maize (25 kg/ha), so some 255,000 tons of seed is required to sow Ethiopia's entire wheat area each year. Most of this still comes from informal seed systems; only four seed enterprises (ESE, ASE, OSE and SNNPSE) currently produce certified seed of various crops and they lack the capacity to supply enough high quality seed for the nation's approximately 20 million households. In collaboration with the Ethiopian Institute of Agricultural Research (EIAR) and through the USAID-funded project "Seed multiplication and delivery of high-yielding rust resistant bread and durum wheat varieties to Ethiopian farmers," the International Maize and Wheat Improvement Center (CIMMYT) is working to increase wheat farmers access to affordable, certified seed of improved varieties that are high-yielding and also feature durable resistance to the rust diseases. Approaches pursued include the fast-track evaluation and release of improved varieties, the pre-release or accelerated seed multiplication of released wheat varieties through formal and informal seed systems, and demonstrations and scaling up of improved wheat varieties. This paper describes best practices to address seed shortages faced by wheat farmers in 53 woredas.
The University of Agriculture, Peshawar, Pakistan
Muhammad Khan, Muhammad Imtiaz, Zahoor Swati, Annemarie Justesen, Sajid Ali
Yellow rust caused by Puccinia striiformis is an important disease in Pakistan. The population structure of P. striiformis in the North Eastern Himalayan region of Pakistan have been shown to be genotypically diverse with potential role of sexual recombination (Ali et al., 2014b), while lesser diversity in the Southern districts of Khyber Pakhtunkhwa (KP)(Khan et al., 2015). This study was designed for the first time to assess disease status and analyze population structure of P. striiformis across three distant parts of Northwestern Pakistan i.e., Bajaur in North Western Agency and Swat and Buner in Malakand Agency, and was compared with other Pakistani populations. Depending on the intensity of infections caused by the pathogen in the tested varieties and breeding lines, the severity of the disease ranged from 5% to 100% during 2015. Yellow rust severity was the maximum on Morocco (100%), Gomal (100%) and KPWYT-18 (80%) and moderate on Ghanimat-e-IBGE (10%) and PS-2008, PS-2013, Tatara and Millat with 20% severity. A total of 81 single lesion samples collected on infected varieties were genotyped with 18 microsatellite markers. From these, 63 distinct multilocus genotypes (MLGs) were detected; 15 single lesion samples collected from Buner produced 15 distinct MLGs signifying very high diversity. A high genotypic diversity with clear signature of recombination was detected across all the three locations. Buner (100%) had the maximum diversity followed by Swat (97%) and Bajaur (91%). The observed diversity was almost equal to other Northeastern Himalayan populations of Pakistan, while it was high when compared to some southern populations of KP (genotypic diversity of 0.895) and other worldwide clonal populations (Ali et al., 2014a). The high diversity and recombinant population structure suggested potential role of sexual reproduction in these areas, which needs to be further explored to establish the origin of diverse virulence pattern in Pakistan.
CREA-Research Centre for Cereal and Industrial Crops
Elisabetta Mazzucotelli, Oadi Matny, Antonietta Saccomanno, Raffaella Battaglia, Francesca Desiderio, Agata Gadaleta, Nicola Pecchioni, Pasquale De Vita, Giovanni Laido, Luigi Cattivelli, Brian Steffenson
The recent emergence of new widely virulent and aggressive strains of rusts (particularly stripe and stem rust) is threatening Italian durum wheat (Triticum turgidum L. var. durum) production, especially under the trend of higher temperature and humidity. A big effort has been undertaken to explore the genetic variability for resistance to these fungal pathogens and discovering novel resistance genes. In particular, a wide set of tetraploid wheat lines was genotyped with several thousands of SNP markers and used for association mapping. This large collection consisted of a group of durum wheat cultivars, produced from the beginning of the last century up to now, a collection of wild emmer wheats (T. dicoccoides), and lines belonging to other wild and domesticated tetraploid subspecies, as a large untapped source of genetic diversity. In a tight cooperation with the University of Minnesota, this collection was evaluated for reaction to several races of stem and stripe rust pathogens in both controlled greenhouse and field conditions. Among the genotypes belonging to the collection are parents of segregating populations which were used for the validation of mapping results. Novel resistance loci were identified, that can be incorporated into new durum varieties through breeding programs. The QTLs found in this study, together with those available in literature, were projected to the recently sequenced durum wheat genome in order to define more precisely the chromosome regions and candidate genes involved in resistance to rusts. Lines which were resistant to multiple races of rust pathogens were also found among both T. dicoccoides and durum wheat cultivars as a source of resistance genes, whose cloning will be undertaken based on the results here obtained.
This study was supported by the Italian Ministry of Foreign Affairs and International Cooperation, with the special grant RES-WHEAT.