P. triticina has a biotrophic relationship with wheat and needs certain elements from the wheat host for a successful life cycle. In recent years, several long lasting, minor resistance genes have been cloned, and their function suggests that the resistance is not due to a classic NB-LRR gene, but a gene that functions in a biotrophic pathway. The hypothesis was proposed that modification of a susceptibility gene can provide broad, long lasting resistance. To test this hypothesis, Thatcher was treated with EMS and screened for changes in susceptibility. M5 lines were evaluated in the greenhouse with BBBD Race 1 and 5 lines were identified. Also, M5 lines were planted in the field to verify the resistance and test the resistance effectiveness to natural infections of P. triticina. The same five lines were resistant in the field. Resistance ranged from few pustules, a race specific-like reaction, lesion mimics with few or no pustules, and near immunity. These lines were backcrossed to Thatcher, and resistant F2 plants were bulked and sequenced. Gene candidates will be identified and discussed.
College of Plant Protection, Northwest A&F University, China
Yuanyuan Zhao, Shuxia Zuo, Dan Zheng, Lili Huang, Zhengshen Kang
Wheat stripe rust, caused by basidiomycete fungus Puccinia striiformis f. sp. tritici (Pst), is a damaging disease worldwide. The recent discovery demonstrated the fungus depends on living wheat and aecial hosts, mainly barberry (Berberis) species, to complete its life cycle. In China, we determined that, under natural conditions, the sexual cycle of Pst occurs based on collections of Pst isolates from the diseased barberry in the past three years. However, no direct evidence to support whether barberry plays a role in spreading inoculums to wheat field to cause stripe rust was detected. In the present study, we recovered 103 Pst samples from natural-infected B. shensiana in the western Shaanxi in spring 2016, and also collected 107 Pst isolates from neighboring wheat fields. Phenotype and genotype of the two Pst populations were tested using a set of Chinese differential hosts for Pst and SSR markers, respectively. The phenotype tests showed that 57 race types produced from the barberry-derived Pst populations, consisting of 58 known races, such as CYR 34, CYR32, G22-14, and Su11-14-3, and 45 new races. Many of the two Pst populations shared the same race types. The genotype tests indicated the barberry-derived Pst population produced a rich genotype, obviously higher than the wheat-derived Pst populations. The seven same genotypes were found on 40 isolates of the former and 26 of the latter. Our results provide evidence to support that sexual cycle of Pst occurs regularly in nature in China and that barberry provides inoculums to neighboring wheat fields, triggering stripe rust infections in the spring. This could be a reason why the Chinese Pst populations represent extreme genetic diversity.
Laboratory of Molecular Plant Physiology, Biotechnology Center of Borj Cedria (CBBC)
Mahmoud Gargouri, Hesham A.Y Gibriel, Richard B. Todd, Michael F. Seidl, Gerrit H.J. Kema
Septoria tritici blotch disease, caused by the fungus Zymoseptoria tritici, is a major threat to global wheat production. With the recent advances in high-throughput DNA-based technologies, Z. tritici has become a powerful model system for the discovery of candidate determinants that underlie virulence and host specialization. Although a few important virulence/regulatory genes have been identified, a global understanding of the larger regulatory network has not been developed. Therefore, to uncover the transcriptional regulatory networks of the infection cycle and most particularly the regulatory hubs that control the switch between the biotrophic and necrotrophic phases, we applied an integrated approach combining transcriptomics, proteomics, and metabolomics analyses based on the identification of plant and fungal transcription factors and regulators, which we characterized from the newly annotated genome sequence of the reference isolate IPO323 (Grandaubert et al., 2015) and using datasets from Rudd et al. (2015). Bread wheat transcription factors and regulators were identified by querying the proteome and subsequent categorization from the Plant Transcription Factor database (PTFDB). Similarly, Z. tritici transcription factors and regulators were identified and categorized using the PFAM TF family databases, and following fungal transcription factor rules as outlined by Todd et al. (2014) and rules we developed for fungal transcription regulators. Insights into transcription factors and regulators will enable synthetic biology approaches to alter the Z. tritici-wheat interaction and lead to rewiring of the regulatory networks thereby turning off the fungal infection process. Beyond providing insights into the regulatory systems-levels involved in Z. tritici-wheat interaction, we believe that our dataset and approach sets the stage for an emerging series of studies that will decipher the dynamic regulatory networks in other plant-pathogen interactions.
QAAFI, The University of Queensland
Robert McIntosh, Peng Zhang, Sami Hoxha, Adnan Riaz, Burkhard Steuernagel, Brande Wulff, Evans Lagudah, Lee Hickey, Sambasivam Periyannan
Wheat is one of the most important staple food and agricultural crop cultivated worldwide. To meet the demands of the raising human population, global wheat production has to be increased which is however declined due to appearance of highly virulent strains of Puccinia striiformis f. sp. tritici (Pst) fungus causing stripe rust disease. Globally, the incidence of stripe rust is effectively managed through the deployment of host plant mediated genetic resistance. But as the resistance present in the current wheat cultivars are ineffective, new sources of resistance particularly from pathogen unexposed genetic resources are of urgent need to prevent stripe rust epidemics. Landrace collections with rich genetic diversity and being less exposed to prevalent pathogen are of valuable source for resistance to new pathogens. In this study, a total of 295 landrace accessions collected by the famous Russian botanist Vavilov was screened for stripe rust resistance using the two predominant lineage Pst strains of Australia. Six accessions with good resistance against the two aggressive Pst strains were selected for genetic characterization and for utilization in global wheat breeding. Characterisation of these novel resistance were undertaken using combination of conventional and advanced genetic tools. While the conventional approach involves the traditional map based gene cloning, the other tool is the recently identified rapid method based on mutagenesis, targeted gene capture and next generation sequencing called "MutRenSeq". Subsequently, the identified novel resistant traits were transferred into elite wheat cultivars through the combination of linked molecular markers and speed breeding techniques. Thus along with the identification of novel resistance, elite wheat cultivars with broad spectrum stripe rust resistance were also generated through the use state of art techniques to sustain global wheat production from the rapidly evolving stripe pathogens.
Sathguru Management Consultants
Kanan Vijayaraghavan, Venugopal Chintada, Rituparna Majumder, Richa Kapur, K. Aishwariya Varadan
South Asia has the highest "wheat dependent" low income community in the world. Stem rust and blast are recognized as the most damaging disease of wheat in the region producing 19% of the world's wheat. In order to combat the potential threat the national research centers were geared up to track the real time movement of wheat diseases, generate disease incidence data and create an enabling environment to boost wheat research in the region through streamlined efforts and enhanced SAARC tool box deployed six years ago.
Recent data (2016-17) from the tool box has shown a significant increase in the data records captured in this region compared to previous years. This has been possible because of heightened awareness amongst the scientists and with the continuous capacity building through pre-season and in-season surveillance trainings organized by Sathguru in collaboration with National Wheat Research Institutes at various levels.
The model is helping partner institutes to be self-sufficient for generating, maintaining wheat disease surveillance data in national and global databases and exchanging real time information with stakeholders. The application have been widely deployed and competently being used by 95% of rust surveillance teams in the wheat fields of SAARC region.
The study will focus on how national research center's judicious decision of carrying out diligent surveillance during the season contributed to safeguarding wheat crops in their respective nations through increased vigilance on emergence of new races and targeted introduction of regionally resistant varieties. Further using this data scientist's can aim to strategize their wheat research for identification of resistant varieties and eventually resulting in increased productivity addressing food security of the region.
Institute of Evolution and the Department of Evolutionary and Environmental Biology, University of Haifa, Israel
Dina Raats, Lin Huang, Valeria Bocharova, Jorge Dubcovsky, Abraham Korol, Tzion Fahima
Wild emmer wheat (Triticum dicoccoides, (DIC)) is an important source of resistance to stripe rust due to presence of Puccinia striiformis in its natural habitats with high humidity and relatively low temperatures that are favorable for stripe rust development. Previously, we showed that DIC accessions from northern Israel were highly resistant to stripe rust. According to the rust responses of three DIC accessions (G25, H52, G303) and mapping of the resistance to relatively close, but different, genetic positions on chromosome 1BS, three different resistance genes were assumed to be present. However, the development of additional critical recombinants and new higher resolution genetic maps for these three genes in subsequent work led us to place YrH52 and YrG303 in the same genetic interval as Yr15, suggesting that the three putative genes are allelic or identical. The recent cloning of Yr15 allowed us to test this hypothesis using an EMS mutagenesis approach. We sequenced the Yr15 locus in five yrH52 and three yrG303 susceptible mutants and identified missense point mutations associated with the susceptible phenotype in each one. Thus, YrH52 and YrG303 may not be new genes. Further work is under way to determine if these genes are allelic or identical.
Department of Field Crops, Ege University, Izmir, Turkey
Kumarse Nazari, Mehran Patpour, Davinder Singh, Aladdin Hamwieh
Rust diseases in wheat are the major threat to wheat production and yield gains. The breakdown in resistance of certain major genes and new emerging aggressive races of rusts are causing serious concerns in all main wheat growing areas of the world. Therefore, it is the need of the hour to search for new sources of resistance genes or QTL's for effective utilization in future breeding programs. In total 100 wheat genotypes were evaluated for seedling and adult-plant resistance to stem rust races TKTTF and TTKSK at Tel Hadya-Syria, and Njoro-Kenya, and Kelardasht-Iran. Evaluation to Yr27 virulent stripe rust race was carried out at Tel Hadya and Terbol-Lebanon research stations. In this study we used genome wide association studies (GWAS) to identify markers or QTLs linked to stem rust and stripe rust races using Diversity Arrays Technology (DArT?) in selected 35 Iranian wheat genotypes. The association of markers and phenotypes was carried out using a unified mixed-model approach (MLM) as implemented in the genome association and prediction integrated tool (GAPIT). Out of 3,072 markers, 986 were polymorphic and used for marker trait associations. A total of 44 DArT markers were identified to be significantly (p<=0.01) associated with studied traits in 16 genomic regions 1A, 1B, 2A, 4A, 6A, 7A, 1B.1R, 2B, 3B, 4B, 5B, 5B.7B, 6B, 7D and an unknown region. Among associated markers, 34 were linked to stem and nine to stripe rust. They were found on 16 genomic regions on chromosome arms 1A, 1B, 2A, 4A, 6A, 7A, 1B.1R, 2B, 3B, 4B, 5B, 5B.7B, 6B, 7D and an unknown region. Associated markers explained phenotypic variation ranging from 21 to 65%. In addition to validation of previously identified genes, this study revealed new QTL's linked to stem and stripe rust which will assist breeders to develop new resistant varieties.
Agriculture & Agri-Food Canada
Wheat cultivar McNair 701 carries resistance gene SrMcN and is used as a differential line to identify Pgt races using the international letter code nomenclature. The inheritance and location of the resistance gene has not been characterized. We developed a doubled haploid (DH) population from cross LMPG/McNair 701 to study the genetics and chromosomal location of SrMcN. A DH population inoculated with race QCCJB segregated 100 resistant : 94 susceptible, a 1:1 ratio (?2=0.186, P=0.666, NS) indicative of segregation at a single locus. This gene was mapped to chromosome 2DL using the Infinium 90k platform. The map position of SrMcN was similar to that of Sr54, one of two genes previously found in Norin 40. Comparison of stem rust seedling reactions using 12 diverse Pgt races indicated that McNair 701 and an Sr54 line derived from Norin 40 had an identical pattern of responses and similar low infection types (IT=12-) to races LCBNB and QCCJB. Based on the chromosomal location on 2DL and identical seedling responses to Sr54, it is likely that the resistance gene in McNair 701 formerly known as SrMcN is Sr54. This finding will be confirmed by a test of allelism.
Cereal Crops Research Institute Pirsabak Nowshera, Pakistan
Khilwat Afridi, Muhammad Ishaq, Irfan Shah, Ibne Khalil, Masood Jan
The Cereal Crops Research Institute (CCRI) is situated on the left bank of River Kabul, near village Pirsabak, 3 km east of Nowshera at an elevation of 288 m above sea level on the intersection of 74? E longitude and 32? N latitude. In July 2010, a devastating flood destroyed all the available germplasm, machineries, laboratories, and field equipment. After the flood research activities were restarted with full motivation, dedication and hard work in collaboration with PARC, ICARDA, CIMMYT, and with the help of wheat productivity enhancement program (WPEP). Developed new population of wheat via spring x spring, spring x facultative germplasm to elevate genetic diversity and lines selected from segregating populations for high yield and rust resistance are at advanced stage of testing.
Since the flood, the CCRI developed four new wheat cultivars: Pirsabak-2013 Pakhtunkhwa-2015 for irrigated areas and Shahkar-2013 and Pirsabak-2015 for rainfed areas of Khyber Pakhtunkhwa, Pakistan. Varietal maintenance and seed production of the released varieties has been undertaken by the wheat breeding team effectively. The seed of these newly developed wheat cultivars was multiplied on fast track basis through pre-released seed multiplication and now these four varieties are the most popular cultivars of Khyber Pakhtunkhwa, Pakistan. Three new candidate wheat lines (PR-106, PR-110 and PR-112) have been submitted to provincial seed council for approval as new wheat cultivars for Khyber Pakhtunkhwa, Pakistan. Two new candidate lines i.e. PR-115 and PR-118 got first position in National Uniform Wheat Yield Trials (NUWYT) on the basis of grain yield during 2016-17 under irrigated and rainfed conditions, respectively.
National Institute of Agricultural Research
Nsarellah Nasserlhaq, Wuletaw Tadesse, Ahmed Birouk
In the context of climate change, drought is one of the most important and complex abiotic stresses affecting crop production worldwide. The adoption of an appropriate technological package, principally drought tolerant varieties, may overcome these challenges to meet global food security needs for the rapidly growing human population, particularly in developing countries. Therefore, this research was carried out to identify efficient phenotypic and genetic selection criteria to identify drought tolerant wheat varieties. In this perspective, 200 diverse elite bread wheat lines from ICARDA and CIMMYT were evaluated under four Moroccan environments during the 2015 and 2016 seasons for yield and 15 agro-physiological traits. The same set of genotypes was genotyped using 15k SNPs. Significant environment and genotype environment interaction effects were observed for yield. Average yield reached 3.18t/ha and ranged from 2.45 to 4.27t/ha. The secondary traits were mostly dominated by the environment effect (p<0.001). Based on correlation and regression analysis between grain yield and phenotypic data, the biomass, grain number per m<sup>2</sup> and to a lesser extent fertile spikes number and thousand kernel weights (depending of drought scenarios) can be more reliable traits than yield for the identification of drought tolerant genotypes. Moreover, the ground cover and canopy temperature depression can be used as supplementary criteria for more accurate selection. Slow selection on the basis of phenotypic traits may be accelerated and improved by using molecular markers. The genetic analysis highlighted significant SNPs and identified new QTLs linked to yield and the most efficient phenotypic traits under drought conditions. These findings could be useful for breeding drought-resistant wheat cultivars using marker-assisted selection to accumulate these favorable alleles of SNPs associated with yield-related traits to increase grain yield.