Umesh Rosyara, Suchismita Mondal, Ravi Singh, Susanne Dreisigacker
Grain yield is the most important economic trait in wheat breeding. The detailed understanding of the genetic architecture of grain yield is crucial and the determining factor to optimize genomics-assisted selection strategies in wheat. First, we performed a marker and haplotype-based genome-wide association study (GWAS) for grain yield (GY) and yield stability coefficient (Pi) on 4,302 advanced breeding lines from five CIMMYT international bread wheat trails grown in multiple (optimally irrigated and stress) environments. All lines were genotyped using genotyping-by-sequencing. A haplotype map was built based on linkage disequilibrium between markers. Twenty-nine markers and 16 haplotypes were associated with GY and Pi across two and three germplasm trials with allelic effects ranging from 2 to 11% across environments. Secondly, we performed genomic prediction, testing eight different prediction models incorporating single markers (base model), haplotypes, epistatic interactions, and significant markers/haplotypes identified in GWAS. Initial results show that by including haplotypes and epistatic interactions among haplotypes (main effect and genome-wide), prediction accuracies range between 0.33-0.49 for GY, a 3 to 22.5% improvement over the base model. Despite the identification of significant marker/haplotype trait associations across traits and environments in GWAS, accounting for these markers in genomic prediction does not improve the prediction models. Our results suggest that the haplotype-based approach can increase prediction ability, but that the knowledge of the genetic architecture of grain yield might not have significant consequence on genomic-assisted selection.
Ebrahiem Babiker, Tyler Gordon, Sam Stoxen, Matthew Rouse, Yue Jin, Shiaoman Chao, John Bonman
Stem rust, caused by Puccinia graminis f. sp. tritici (Pgt), is a threat to wheat production worldwide. To manage this important disease, new sources of genetic resistance are needed and common wheat landraces are a potential source of such resistance. Landrace accessions from the USDA-ARS National Small Grains Collection were evaluated for seedling resistance to the Ug99 race group. To identify accessions most likely to carry novel resistance genes, a bulked segregant analysis (BSA) approach was used. Seven resistant accessions were crossed to a susceptible parent line and F3 families were tested against Pgt race TTKSK. The resistant plants were identified and grouped into two bulks per population. The bulks, along with the parents and F1 progeny, were genotyped with the 90K wheat iSelect SNP genotyping platform. Four of the populations appeared to segregate in a 1:1 phenotypic resistant/susceptible ratio, one in a 1:2 ratio, and two in 1:3 ratios. However, chi squared tests indicated the ratios were statistically the best fit for only two of the 1:1 segregating populations and one of the 1:3 segregating populations. Initial BSA results indicate the markers associated with reduced stem rust infection are located on wheat chromosomes 1DL and 2B. These mapping populations are being advanced for further evaluation to ascertain if novel resistance to the Ug99 stem rust race group is present.
Barani Agricultural Research Station, Kohat
Fida Mohammad, Muhammad Imtiaz
Stripe rust is one of the major limiting factors in wheat production. An objective-based breeding program was initiated at Barani Agricultural Research Station (BARS), Kohat in 2013/14 to transfer APR genes from CIMMYT and ICARDA spring wheat lines into wheat germplasm well adapted in Khyber Pakhtunkhwa (KPK). Nine high yielding but stripe rust susceptible KPK wheat varieties were crossed in various combination with 17 CIMMYT and ICARDA wheat lines carrying resistance genes. The resultant 79 F1s were backcrossed with respective susceptible parents followed by single plant selection in F2 generation. During 2015/16, 367 segregating populations/lines were screened in multi-environment stripe rust tests within Khyber Pakhtunkhwa. Sixty-nine out of 367 lines showing adequate resistance were again screened for strip rust resistance at hot spot and in yield trial at BARS, Kohat during 2016/17. Seventeen lines showed considerable resistance and were higher yielding than check cultivars. Lines exhibiting adequate resistance will be further tested in advanced yield trial at provincial and national level for possible release of new varieties in wheat.
Chris K. Sørensen, Rodrigo Labouriau, Annemarie Justesen, Mogens Hovmøller
Host vernalisation and temperature strongly affect the susceptibility of winter crops to pathogenic fungi. However, how the interaction of these environmental factors influence host susceptibility to Puccinia striiformis, the yellow (stripe) rust fungus, is poorly understood. An experimental system was developed to examine the effect of vernalisation, temperature regime (standard; 18 day/12 night °C and low; 12 day/6 night °C) and plant growth (seedling and adult plant stages) on changes in susceptibility of agronomically important winter wheat and triticale genotypes to P. striiformis races ('Warrior' and 'Kranich') highly predominant in several European countries. Host genotypes exposed to prolonged periods of low temperature, termed vernalisation, reduced disease susceptibility on specific winter host genotypes, although its effect differed considerably by the temperature regime and the P. striiformis race deployed. The influence of vernalisation on host susceptibility was more apparent at low temperature for the 'Warrior' race and at standard temperature for the 'Kranich' race. Triticale genotypes inoculated with the 'Kranich' race were particularly affected by the influence of vernalisation and temperature regime by displaying a shift towards reduced susceptibility at standard temperature. The effect of plant growth stage, i.e., vernalised seedlings versus adult plants, was most evident for the 'Warrior' race at standard temperature and at low temperature for the 'Kranich' race by revealing a lower infection type at the adult plant stage. The research findings presented here contributed to a better understanding of the role of environmental factors in host susceptibility. This, in fact, will aid in the development of more efficient early-warning systems and disease management strategies to the yellow rust fungus and in the decision making for the deployment of winter wheat and triticale genotypes.
Bogale Nigir, Cherinet Alem, Yosef G. Kidane, Mario Enrico Pè, Matteo Dell'Acqua
Septoria tritici blotch (STB) is a devastating fungal disease affecting durum and bread wheat cultivation worldwide. The search for resistance sources in untapped genetic resources may speed up breeding for STB resistance. Ethiopian durum wheat landraces represent a valuable source of allelic diversity for several traits, including disease resistance. In this study, we measure STB phenotypes under natural infection on two interconnected populations: i) a diversity panel comprising 318 Ethiopian durum wheat lines, mostly farmer varieties, and ii) a nested association mapping (NAM) population developed from a subset of the diversity panel. Phenology, yield and yield component traits were concurrently measured in the populations. We evaluated the distribution of STB resistance in Ethiopian genetic materials and the relationship existing between STB resistance and agronomic traits. STB resistance sources were found in landraces as well as in NAM lines. The genetic material was genotyped with more than 13 thousand genome-wide SNP markers to describe the linkage disequilibrium and genetic structure existing within the panels. The genotyping information was combined with phenotypes to identify marker-trait associations and loci involved in STB resistance. We identified several loci, each explaining up to 10% of the phenotypic variance for disease resistance. We developed KASP markers tagging the most interesting loci to allow the uptake of our results in a breeding perspective. Our results showed that the Ethiopian untapped allelic diversity bears a great value for studying the molecular basis of STB resistance and for breeding for resistance in local and international material.
Northwest A&F University
Dejun Han, Jia Guo, Manuel Spannagl, Jianhui Wu, Aizhong Cao, Peidu Chen, IWGSC, Lili Huang, Jun Guo, Klaus Mayer, Zhensheng, Kang
Wheat cultivation in many regions faces threats by devastating fungal infections. However, wheat cultivar 92R137 shows resistance to Puccinia striiformis infection. To isolate the stripe rust resistance gene Yr26, an integrated transcriptomic and comparative genomics approach was undertaken. Near-isogenic lines of wheat (carrying Yr26 or not) infected with two Puccinia striiformis f. sp. tritici (Pst) (Virulence or avirulence to Yr26) were analysed in a dual detailed time series RNA-seq study. The emerging IWGSC refseq v1.0 genome assembly sequence serves as a valuable template and was also used for comparative genomics studies of the gene candidate region with the genome sequences of close relatives and wheat progenitors. Using bulked segregant analysis (BSA) to identify polymorphic SNPs between parent and resistant DNA (R-bulk) and susceptible DNA (S-bulk), flanking markers for Yr26 were identified. These two markers were mapped to the Chinese spring reference genome sequence, spanning a region of about 250 kb. The synteny analysis of this candidate region in CS chr1B with chr1A, chr1D, Wild Emmer Wheat (Td_chr1A and Td_chr1B) and Barley (chr1H) identified three candidate Yr26 genes. To detect gene candidates a dual time series RNA-seq analysis was performed. Genes differently expressed between rust susceptible (NIL-S) host lines and rust resistant (NIL-R) lines, carrying the Yr26 candidate gene were analysed. Both lines were inoculated with Pst carrying different avirulence factors (Pst-CYR32 and Pst-V26), compatible or incompatible with the corresponding wheat lines. Differential gene expression analysis (DEG) between compatible and incompatible interaction revealed DEGs in the wheat genome and in the Pst genome. From a network analysis of both wheat and Pst genes, we inferred connected co-expressed modules. Resulting modules showed particular enrichments for disease resistance, defense response to fungus and cell wall components.
University of Sydney Plant Breeding Institute
Naeela Qureshi, Vallence Nsaiyera, Pakeer Kandiah, Mesfin Gesesse, Mandeep Randhawa, Mumta Chhetri, Bosco Chenayek, James Kolmer, Miroslav Valarik, Zaroslav Dolezel, Beat Keller, Matthew Hayden, Justin Faris, Harbans Bariana, Vanessa Wells
Dr. Norman Borlaug stated that rust never sleeps and this enables rust pathogens to produce new strains capable of putting rust resistance genes to rest. These pathogens continue to pose threats to global wheat production. Wheat breeders have made significant progress to control rust outbreaks using conventional selection technologies; however, some critical shifts in pathogen populations have let them down. Rapid evolution in molecular marker technologies in the last 15 years and refinement of phenomic facilities have expedited the process of discovery and characterisation of rust resistance genes to underpin the development and validation of markers closely linked with genetically diverse sources of resistance. A high proportion of the formally named rust resistance genes were characterized in the 21st century and markers closely linked with these genes have been developed and validated. The marker tagged sources of resistance to three rust diseases have equipped the wheat breeding community with tools to deploy combinations of all stage and adult plant resistance genes in future wheat cultivars. The question that whether we have enough resistance genes discovered to compete against the ever-awake rust pathogens. In our opinion, we cannot be complacent and discovery needs to continue to ensure food security. This presentation will discuss the role of advances in phenomic and genomic technologies to achieve durable rust control in wheat.
Ayub Agricultural Research Institute, Faisalabad
Different biotic and abiotic stresses are hampering wheat yield across different geographic regions. Among biotic stresses, wheat rusts are principal cause of yield reduction. Whereas among abiotic stresses, drought is the principle cause of reduction in growth and lowering yield potential. So developing rust resistance and drought tolerance in wheat germplasm is needed, which requires assessment of genetic potential of current cultivars against these stresses to identify variation among existing germplasm. Screening of genotypes under naturally prevailing races of rust species is the better and inexpensive approach. In the present study 65 genotypes including five checks (AARI-11, Chakwal- 50, Aas- 11, Morocco and Galaxy-13) were evaluated for adult plant response to wheat rusts and water deficit conditions. Experimental material was planted in four blocks each having new entries along with repetition of five checks in augmented design. Data was recorded on morphological traits including plant height, peduncle length, spike length, productive tillers per meter, flag leaf area, number of spikelet per spike, grains per spike, single head weight, 1000 grain weight, days to maturity and grain yield per acre. Significant variation was observed among genotypes for all the studied traits. On the basis of performance G39 and G36 were better than commercial drought check Chakwal-50 in almost all the traits. However rust screening under natural rust infestation revealed that although Morocco showed susceptible (S) response yet only six genotypes were susceptible to yellow rust whereas all others were resistant. In case of leaf rust 29 were completely resistance, 25 were moderately resistant, seven were moderately susceptible and only four were completely susceptible to currently active races of leaf rust. However, in the case of stem rust, 61 genotypes showed complete resistance to stem rust, two showed moderately resistance and two were moderately susceptible. Information obtained from this study would be favorable for breeding rust resistant and drought tolerant cultivars.
University of Eldoret
Julius Ochuodho, Ruth Wanyera, Sridhar Bhavani, Les Szabo
Stem rust Ug99 and related race group are one of the major constraints of wheat production in Kenya. The challenge has been largely due to rapid evolution of races within lineage defeating resistance genes resulting in boom and burst cycles. Understanding of the pathogen population structure in major wheat growing regions in Kenya gives comprehensive information of the predominant races as well as capturing new races which may have potential of causing epidemics. Such information can have significant impact on effective gene stewardship in breeding resistant varieties. Using 11 Pgt Simple Sequence Repeats (SSR) markers we analyzed 104 single uredinial-pustule samples. Allele frequency distribution ranged from 2 to 6 per locus with an average of 3.27 per locus. Observed heterozygosity ranging from 0.297-1.000 (mean HO=?0.809) was significantly different (P< 0.001) than the expected heterozygosity (0.264 to 0.507; mean HE=?0.407) indicating that the population is asexual. Analysis of molecular variance (AMOVA) showed that the majority of the variation occurred within the samples (98%) rather than between regions (2%). Analysis of 104 samples identified 21 multiple locus genotypes (MLGs). MLG.19 was observed across the three region analyzed that is Central Rift, North Rift and Mount Kenya while MLG.18 was predominant in Mount Kenya. Based on SSR genotypes of reference isolates, Pgt clade IV (race TKTTF) was associated with MLG.16 in Central Rift Kenya while clade I (race TTKSK) had a unique MLG.10. These results indicated two main groups corresponding to Clade I (Ug99 race group) and Clade IV (race TKTTF race group). This minimum spanning network analysis pattern points to the Pgt population being asexual due to mutation. These preliminary results suggest that Pgt population in Kenya is asexual in nature. Further analysis is being conducted to ascertain geographical structure as well as compare the results with the 2011 data.
CIMMYT International, South Asia, Regional Office, Kathmandu
Gurbinder Gill, Madan Bhatta, Andrew McDonald, Arun Joshi
The experience of seed system development works in Nepal shows that lack of business orientation is one of the key challenges for the growth of Nepalese seed enterprises (SEs). We implemented a business mentorship activity focusing on SEs in the project - Cereal System Initiative for South Asia. The business mentoring (BM) was implemented covering 10 SEs during 2014 to 2017 using discovery sensitization and facilitation mode. The key actions were, SWOT analysis, mentorship to develop strategic business plan, technical facilitation for seed quality improvement through maintenance breeding and on-farm farmer participatory trials as well as large plot demonstrations of recently released and pre-released wheat varieties. This resulted in 45% average increase in the volume of seed sold by individual SEs in three years. Five SEs started maintenance breeding in wheat and by third year, over 50% of the seed sold by the enterprises consisted of newly released varieties compared to 20% before mentoring. The improved performance of SEs prompted seed quality control authority of Nepal to issue a license to two of these private seed companies for maintenance breeding cum foundation seed production. As a consequence, foundation seed production of these entities increased by 60% in three years, and two of them have also started maintenance breeding in rice from 2017. The facilitation in developing business plan by SEs helped them to attract financial resources from different sources in upgrading seed processing, storage and laboratory facilities. Moreover, the National Wheat Research Program of Nepal has proposed the release of a new wheat variety BL 4341 through integration of on-farm trials and seed production data of these SEs. This new variety is resistant to major diseases, yields 7% higher than the popular cultivar NL 297 which was released 32 years ago and needed replacement due to susceptibility to wheat rusts