Identification of markers closely linked with adult plant leaf rust resistance gene Lr48 in wheat
BGRI 2015 Poster Abstract Vallence Nsabiyera
The University of Sydney, Plant Breeding Institute, Australia
Leaf rust is endemic to all wheat-growing regions of the world. Resistance to leaf rust in wheat cultivars is controlled either by all stage resistance (ASR) or by adult plant resistance (APR) genes. Although deployment of single ASR genes can provide high levels of resistance, these are usually overcome by virulence in pathogen populations. In contrast, individual APR genes often provide low levels of resistance and combinations of three to four genes are necessary to achieve adequate resistance for crop protection. This kind of APR has proven to be durable. APR gene Lr48 in a single plant selection of Condor (CSP44) was mapped on chromosome 2BS and was flanked by markers gwm429b (6.1 cM, distal) and barc7 (7.3 cM, proximal) (Bansal et al. 2008, Theor. Appl. Genet. 117:307-312). The present study was planned to identify markers more closely linked to Lr48. Selective genotyping by 90K Infinium Assay identified 27 SNP markers linked with Lr48. The SNP sequences were used to design Kompetitive Allele-Specific Primers (KASP). Eleven KASP markers showing clear clustering were genotyped on a RIL population using the CFX96 Touch™ real-time PCR detection system (Biorad, USA). KASP marker IWB72894 co-segregated with Lr48.
Complementary resistance genes Yr73 and Yr74 (YrA) in wheat selection Avocet R confer resistance to the non-adapted barley grass stripe rust pathogen Puccinia striiformis f. sp. pseudohordei.
BGRI 2015 Poster Abstract Dracatos
The University of Sydney, Plant Breeding Institute, Australia
This is the first study on the inheritance and genetic mapping of resistance to the barley grass stripe rust pathogen (Puccinia striiformis f. sp. pseudohordei – Psph) in bread wheat. Psph, commonly infects barley grass (Hordeum leporinum, H. murinum), but about 10% of commercial barley varieties are also susceptible. We tested over 500 diverse wheat accessions and determined that less than 20% were susceptible at the seedling stage suggesting wheat is an ‘intermediate’ host to Psph. The Australian variety Teal is highly susceptible to Psph at the seedling stage, whereas selections Avocet S and Avocet R are highly resistant and resistant, respectively. We used the Teal/AvocetR doubled haploid (DH) population to characterize the resistance of Avocet R to Psph and determine whether the complementary genes Yr73 and Yr74 (YrA resistance) in Avocet R conferred resistance to Psph. Phenotypic comparison of the Teal/AvocetR DH lines in response to both Psph and Pst showed that all DH lines carrying YrA were also resistant to Psph; however, fewer DH lines were susceptible to Psph suggesting additional resistance genes. Marker-trait association analysis detected three DArT-Seq markers significantly associated with resistance to Psph, two mapping to chromosomes 3DL and 5BL in the same regions as Yr73 and Yr74 and the third mapping to chromosome 4A. Single gene stocks with the 4A gene and combinations of the 5BL and 3DL genes will be used for monitoring avirulence/virulence within Australian Psph population. Genetic analysis of seedling-susceptible T/AvR DH lines as adult plants in the greenhouse determined that Teal and Avocet R each carried at least one APR gene effective against Psph.
Molecular mapping of resistance to the Pgt race Ug99 group in spring wheat landrace PI 177906
BGRI 2015 Poster Abstract Babiker
USDA-ARS, Small Grains and Potato Germplasm Research Unit, USA
View babiker.pdf(941.79 KB)
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.
Rapid detection of micro-RNAs associated with APR to rust pathogens in wheat
BGRI 2015 Poster Abstract Singh
The University of Queensland, Queensland Alliance for Agriculture and Food Innovation, Australia
The identification of R-genes using traditional map-based approaches is a long, laborious process, not to mention the time required for subsequent development of cultivars incorporating the new resistances. Breeders seek to reduce the length of breeding cycles, and researchers require new tools to accelerate discovery and understanding of mechanisms associated with durable resistance, especially adult plant resistance (APR). A new method for rapid generation advancement, known as ‘speed breeding’, significantly reduces the length of breeding cycles, provide increased recombination during line development and enable selection in early generations. The speed breeding protocol uses controlled temperature regimes and 24h light to accelerate plant growth and development. Phenotyping methods adapted for use in the speed breeding system permit year-round evaluation of APR to rust pathogens within 5 weeks from time of sowing. RNA sequencing (RNA-Seq) technology has revolutionized gene expression profiling in plants. We previously used RNAseq to identify novel transcripts and miRNAs associated with seedling resistance (Lr28) leading to identification of transcription factors and miRNA families (e.g. miR36, miR37 and miR39) involved in signalling and defense response (Kumar et al. J. Nuc. Acids 2014:570176). In this study we report the application of speed breeding and RNAseq technologies for the purpose of rapidly identifying transcripts and miRNA associated with APR. Wheat landraces harbouring novel sources of resistance were grown under speed breeding conditions and sampled for RNA at key growth stages, before and after inoculation, which enabled discovery of differentially expressed miRNAs. Our next steps are aimed at validating these genetic factors associated with APR in order to better understand the signalling pathways and deliver tools to assist the assembly of robust wheat cultivars for the future.
Reactions of spring wheat genotypes in crossing block nursery to stem, leaf, and stripe rust
BGRI 2015 Poster Abstract Mert
The Central Research Institute for Field Crops, Turkey
Rusts (Puccinia spp.) are the most significant disease affecting wheat yield and quality in Turkey. Knowing the resistance status of wheat genotypes in crossing program is an important issue for breeding programs. The aim of the study was to determine of the resistance of the 106 wheat genotypes consisting of Crossing Block Spring Wheat (CBSW) nursery developed by the International Winter Wheat Improved Project (IWWIP). For this purpose, adult plant and seedling test were conducted for yellow rust while only seedling test were conducted for leaf and stem rust. Evaluations were carried out at the research facilities of CRIFC at İkizce and Yenimahalle in Ankara in the 2014 season. For adult plant reactions; the genotypes were inoculated with local Pst populations (virulent on Yr2,6,7,8,9,25,27,Sd,Su,Avs). Stripe rust development on each entry were scored using the modified Cobb scale when the susceptible check Little Club had reached 80S infection severity in June, 2014. Coefficients of infections were calculated and values below 20 were considered to be resistant. For seedling test; the seedling was inoculated with local Pgt (avirulent on Sr24, Sr26, Sr27, and Sr31), Pt (avirulent on Lr9, Lr19, Lr24, and Lr28) and Pst populations. Stripe, leaf and stem rust development on each entry were scored after 14 days with 0-4 and 0-9 scale for leaf-stem rust and yellow rust, respectively. In seedling stage, thirty nine (37%), 47 (44%), and 20 (19%) genotypes were resistant to local Pgt,Pt, and Pst populations, respectively. In adult plant test, 46 (43%) genotypes were resistant to Pst. The resistance genotypes to stem, leaf, and stripe rust were determined with this research.
The Stubbs Pst Culture Collection: Recovery, avirulence/virulence phenotyping and past population structure at a global scale
BGRI 2015 Poster Abstract Thach
Department of Agroecology, Aarhus University, Denmark
The "Stubbs Collection", began in 1956 by the late Dutch plant pathologist R.W. Stubbs, refers to a unique historic collection of urediniospore samples of Puccinia striiformis that had been stored in liquid nitrogen for decades. Since 2010 the collection has been maintained by the Global Rust Reference Center (GRRC) in Denmark. Part of the collection is now being in a study of past pathogen diversity. A subset of samples collected between 1958 and 1991 from 35 countries was investigated to assess recovery rate, race identity, and previously undetected virulences. A new method for recovery using an airbrush sprayer and NovecTM 7100 fluid as dispersal agent in inoculating host plants was highly successful, resulting in a 96% recovery from 231 isolates. Phenotyping on the World and European differential host sets and additional wheat genotypes revealed 181 apparently uniform isolates, of which race identities were confirmed for 102. Race identities were updated for additional isolates based on improved resolution due to updated and more informative differential lines. Additional virulences corresponding to Yr17, Yr25, and Yr27 were added, as these were not assayed earlier. The past population structure was investigated by genotyping 212 isolates using 19 multilocus microsatellites. Seven distinct populations were detected, including clonal populations and recombinant populations. These results were compared with recent studies and demonstrated an overall consistent population subdivision at the global scale with clear migration events between populations. The outcome of the study facilitates conclusions about long-term temporal dynamics and overall migration patterns within and among world-wide populations of Pst.
Global Pgt Initiative: An international genetic resource to combat stem rust
BGRI 2014 Plenary Abstract Les Szabo
USDA-ARS, Cereal Disease Laboratory
An important component of the management of wheat stem rust is an understanding of the population diversity of the pathogen, Puccinia graminis f. sp. tritici (Pgt). The discovery of “Ug99” resulted in renewed efforts on pathogen surveys, sample collections and pathotyping of Pgt, with a primary focus on Africa. In the last few years these efforts have been expanded to include other targeted regions, however a global effort is needed. The aims of the “Global Pgt Initiative” is: to capture and maintain living cultures that collectively reflect the entire global diversity of Pgt in the years 2014 - 2016; pathotype and genotype this collection; develop DNAbased diagnostic tools that will be able to rapidly detect shifts in Pgt populations, and provide an early warning system of the vulnerability of wheat to new virulent strains; and provide a genetic baseline for comparison of Pgt populations over time, both forward and backwards. This initiative will provide the wheat rust community with a geographically distributed, well characterized, living culture collection that represents the global diversity of Pgt; a global open access knowledge bank on Pgt pathotypes and genotypes; and advanced molecular diagnostic tools for rapid detection and tracking of Pgt populations. The Global Pgt Initiative represents the most comprehensive effort to capture and characterize the global diversity of Pgt and provide a unique resource to the global wheat rust community.
In 2010, Ethiopia experienced one of the largest stripe rust epidemics in recent history. Over 600,000 ha of wheat were affected, an estimated 60 million Ethiopian Birr ($US3.2 million) were spent on fungicides and large production losses were observed. Factors associated with the 2010 epidemic were conducive climatic conditions (prolonged rain and apparently optimal temperatures), large areas planted to susceptible cultivars, early infection and rapid spread of a virulent pathogen, a low level of awareness, and ineffective control measures. In 2013, highly favourable climatic conditions and early appearance of stripe rust showed remarkable similarity to the conditions observed in 2010, prompting fears of a similar major rust epidemic. However, no stripe rust epidemic developed in 2013. In contrast, only limited and localized outbreaks of stripe rust were observed in 2013; wheat crops remained in good condition and a good harvest was achieved. It seems that a series of positive and timely actions in Ethiopia contributed to the markedly different stripe rust situation in 2013 compared to 2010. The principle factors associated with the positive outcomes in 2013 are (i) effective promotion, plus rapid and widespread adoption of rust resistant wheat cultivars since 2010 - this dramatically reduced the vulnerability of the Ethiopian wheat crop; and (ii) timely and coordinated surveillance efforts, coupled to good information exchange amongst different stakeholders - this resulted in effective control and awareness campaigns that targeted emerging stripe rust outbreaks. A comparative analysis is presented which highlights the similarities and disparities between the 2010 and 2013 stripe rust situations in Ethiopia. The roles and contributions of different organisations are examined and an in-depth analysis of the biophysical conditions in the different years is presented.
There is emerging evidence that the geographical footprint of stripe rust is expanding, opening up prospects for an increase in economic losses attributable to this disease worldwide. Drawing on newly compiled data, along with insights obtained from a survey initiated at the BGRI meeting in New Delhi in August 2013, this talk will report on efforts to model the global occurrence and persistence of stripe rust in a geo-spatially sensitive fashion. Using the available data in conjunction with these newly developed climate suitability maps, I will present probabilistic crop production losses associated with the disease and place an economic value on the prospective losses. Given changes in the geographical spread of this disease, and the associated uncertainties about its likely wheat yield and economic effects, various scenarios will be assessed to inform and thereby help shape the research investment decisions regarding crop breeding and other options for ameliorating these prospective losses over the longer term.
Advances in breeding for resistance to stem rust caused by Ug99 and Ethiopian Pgt races in durum wheat
BGRI 2014 Plenary Abstract Karim Ammar
Stem rust (SR) resistance is required for CIMMYT durum germplasm to keep relevance in Ethiopia, where Ug99 and other Pgt races are a major yield-limiting constraint, and in countries along the possible dissemination paths of these races. Resistance to Ug99 is widespread in most durum germplasm groups when tested in Kenya, but resistance is lost when exposed to Ethiopian races; hence selection at the Debre Zeit site in Ethiopia is essential for durum wheat. Due to difficulties with shuttling segregating populations between Mexico and Ethiopia, we have adopted a strategy involving the identification of resistant/moderately resistant lines at Debre- Zeit, and inter-crossing in Mexico followed by selection for resistance to leaf rust and agronomic type and finally screening for SR reaction in the resulting F6 lines at Debre-Zeit at the same time as they are tested for yield and quality in preliminary yield trials in Mexico. This has generated a significant increase in the proportion of resistant and moderately resistant genotypes within outgoing CIMMYT germplasm, from less than 3% at the onset of the initiative in 2008 to 16% in 2011, and 38% in 2013. SR-resistant germplasm was characterized by similar frequency distributions to other traits in the overall germplasm such as yield potential, drought tolerance and industrial quality parameters. Advances have also been realized using marker-assisted selection (MAS) to introgress Sr22 from bread wheat and to combine it with Sr25, producing advanced lines with 2-gene stacks with confirmed outstanding resistance and superior quality attributes. Since the two genes are closely linked but from different sources bringing them together required a very rare recombination event finally detected via MAS among thousands of plants. They are now essentially inherited together with a very low likelihood of generating recombinant individuals with either gene. The yield potential and stability of these lines are under evaluation in Ethiopia and the best lines are being used in a second round of breeding.