Sridhar Bhavani

The Emergence of Ug99 Races of the Stem Rust Fungus is a Threat to World Wheat Production

Race Ug99 of the fungus Puccinia graminis tritici that causes stem or black rust disease on wheat was first detected in Uganda in 1998. Seven races belonging to the Ug99 lineage are now known and have spread to various wheat-growing countries in the eastern African highlands, as well as Zimbabwe, South Africa, Sudan, Yemen, and Iran. Because of the susceptibility of 90% of the wheat varieties grown worldwide, the Ug99 group of races was recognized as a major threat to wheat production and food security. Its spread, either wind-mediated or human-aided, to other countries in Africa, Asia, and beyond is evident. Screening in Kenya and Ethiopia has identified a low frequency of resistant wheat varieties and breeding materials. Identification and transfer of new sources of race-specific resistance from various wheat relatives is underway to enhance the diversity of resistance. Although new Ug99-resistant varieties that yield more than current popular varieties are being released and promoted, major efforts are required to displace current Ug99 susceptible varieties with varieties that have diverse race-specific or durable resistance and mitigate the Ug99 threat.

Race non-specific resistance to rust diseases in CIMMYT spring wheats

Rust diseases continue to cause significant losses to wheat production worldwide. Although the life of effective race-specific resistance genes can be prolonged by using gene combinations, an alternative approach is to deploy varieties that posses adult plant resistance (APR) based on combinations of minor, slow rusting genes. When present alone, APR genes do not confer adequate resistance especially under high disease pressure; however, combinations of 4–5 such genes usually result in “near-immunity” or a high level of resistance. Although high diversity for APR occurs for all three rusts in improved germplasm, relatively few genes are characterized in detail. Breeding for APR to leaf rust and stripe rust in CIMMYT spring wheats was initiated in the early 1970s by crossing slow rusting parents that lacked effective race-specific resistance genes to prevalent pathogen populations and selecting plants in segregating populations under high disease pressure in field nurseries. Consequently most of the wheat germplasm distributed worldwide now possesses near-immunity or adequate levels of resistance. Some semidwarf wheats such as Kingbird, Pavon 76, Kiritati and Parula show high levels of APR to stem rust race Ug99 and its derivatives based on the Sr2-complex, or a combination of Sr2 with other uncharacterized slow rusting genes. These parents are being utilized in our crossing program and a Mexico-Kenya shuttle breeding scheme is used for selecting resistance to Ug99. High frequencies of lines with near-immunity to moderate levels of resistance are now emerging from these activities. After further yield trials and quality assessments these lines will be distributed internationally through the CIMMYT nursery system.

A consensus map for Ug99 stem rust resistance loci in wheat

Key messageThis consensus map of stem rust genes, QTLs, and molecular markers will facilitate the identification of new resistance genes and provide a resource of information for development of new markers for breeding wheat varieties resistant to Ug99.AbstractThe global effort to identify new sources of resistance to wheat stem rust, caused by Pucciniagraminis f. sp. tritici race group Ug99 has resulted in numerous studies reporting both qualitative genes and quantitative trait loci. The purpose of our study was to assemble all available information on loci associated with stem rust resistance from 21 recent studies on Triticum aestivum L. (bread wheat) and Triticum turgidum subsp. durum desf. (durum wheat). The software LPmerge was used to construct a stem rust resistance loci consensus wheat map with 1,433 markers incorporating Single Nucleotide Polymorphism, Diversity Arrays Technology, Genotyping-by-Sequencing as well as Simple Sequence Repeat marker information. Most of the markers associated with stem rust resistance have been identified in more than one population. Several loci identified in these populations map to the same regions with known Sr genes including Sr2, SrND643, Sr25 and Sr57 (Lr34/Yr18/Pm38), while other significant markers were located in chromosome regions where no Sr genes have been previously reported. This consensus map provides a comprehensive source of information on 141 stem rust resistance loci conferring resistance to stem rust Ug99 as well as linked markers for use in marker-assisted selection.

Genomic selection for quantitative adult plant stem rust resistance in wheat

Quantitative adult plant resistance (APR) to stem rust (Puccinia graminis f. sp. tritici) is an important breeding target in wheat (Triticum aestivum L.) and a potential target for genomic selection (GS). To evaluate the relative importance of known APR loci in applying GS, we characterized a set of CIMMYT germplasm at important APR loci and on a genome‐wide profile using genotyping‐by‐sequencing (GBS). Using this germplasm, we describe the genetic architecture and evaluate prediction models for APR using data from the international Ug99 stem rust screening nurseries. Prediction models incorporating markers linked to important APR loci and seedling phenotype scores as fixed effects were evaluated along with the classic prediction models: Multiple linear regression (MLR), Genomic best linear unbiased prediction (G‐BLUP), Bayesian Lasso (BL), and Bayes Cπ (BCπ). We found the Sr2 region to play an important role in APR in this germplasm. A model using Sr2 linked markers as fixed effects in G‐BLUP was more accurate than MLR with Sr2 linked markers (p‐value = 0.12), and ordinary G‐BLUP (p‐value = 0.15). Incorporating seedling phenotype information as fixed effects in G‐BLUP did not consistently increase accuracy. Overall, levels of prediction accuracy found in this study indicate that GS can be effectively applied to improve stem rust APR in this germplasm, and if genotypes at Sr2 linked markers are available, modeling these genotypes as fixed effects could lead to better predictions.

Haplotype diversity of stem rust resistance loci in uncharacterized wheat lines

Stem rust is one of the most destructive diseases of wheat worldwide. The recent emergence of wheat stem rust race Ug99 (TTKS based on the North American stem rust race nomenclature system) and related strains threaten global wheat production because they overcome widely used genes that had been effective for many years. Host resistance is likely to be more durable when several stem rust resistance genes are pyramided in a single wheat variety; however, little is known about the resistance genotypes of widely used wheat germplasm. In this study, a diverse collection of wheat germplasm was haplotyped for stem rust resistance genes Sr2, Sr22, Sr24, Sr25, Sr26, Sr36, Sr40, and 1A.1R using linked microsatellite or simple sequence repeat (SSR) and sequence tagged site (STS) markers. Haplotype analysis indicated that 83 out of 115 current wheat breeding lines from the International Maize and Wheat Improvement Center (CIMMYT) likely carry Sr2. Among those, five out of 94 CIMMYT spring lines tested had both Sr2 and Sr25 haplotypes. Five out of 22 Agriculture Research Service (ARS) lines likely have Sr2 and a few have Sr24, Sr36, and 1A.1R. Two out of 43 Chinese accessions have Sr2. No line was found to have the Sr26 and Sr40 haplotypes in this panel of accessions. DArT genotyping was used to identify new markers associated with the major stem resistance genes. Four DArT markers were significantly associated with Sr2 and one with Sr25. Principal component analysis grouped wheat lines from similar origins. Almost all CIMMYT spring wheats were clustered together as a large group and separated from the winter wheats. The results provide useful information for stem rust resistance breeding and pyramiding.

Genomic prediction of genetic values for resistance to wheat rusts

Durable resistance to the rust diseases of wheat (Triticum aestivum L.) can be achieved by developing lines that have race‐nonspecific adult plant resistance conferred by multiple minor slow‐rusting genes. Genomic selection (GS) is a promising tool for accumulating favorable alleles of slow‐rusting genes. In this study, five CIMMYT wheat populations evaluated for resistance were used to predict resistance to stem rust (Puccinia graminis) and yellow rust (Puccinia striiformis) using Bayesian least absolute shrinkage and selection operator (LASSO) (BL), ridge regression (RR), and support vector regression with linear or radial basis function kernel models. All parents and populations were genotyped using 1400 Diversity Arrays Technology markers and different prediction problems were assessed. Results show that prediction ability for yellow rust was lower than for stem rust, probably due to differences in the conditions of infection of both diseases. For within population and environment, the correlation between predicted and observed values (Pearsons correlation [ρ]) was greater than 0.50 in 90% of the evaluations whereas for yellow rust, ρ ranged from 0.0637 to 0.6253. The BL and RR models have similar prediction ability, with a slight superiority of the BL confirming reports about the additive nature of rust resistance. When making predictions between environments and/or between populations, including information from another environment or environments or another population or populations improved prediction.

Genetic gain from phenotypic and genomic selection for quantitative resistance to stem rust of wheat

Stem rust of wheat (Triticum aestivum L.) caused by Puccinia graminis f. sp. tritici Eriks. and E. Henn. is a globally important disease that can cause severe yield loss. Breeding for quantitative stem rust resistance (QSRR) is important for developing cultivars with durable resistance. Genomic selection (GS) could increase rates of genetic gain for quantitative traits, but few experiments comparing GS and phenotypic selection (PS) have been conducted. Our objectives were to (i) compare realized gain from GS based on markers only with that of PS for QSRR in spring wheat using equal selection intensities; (ii) determine if gains agree with theoretical expectations; and (iii) compare the impact of GS and PS on inbreeding, genetic variance, and correlated response for pseudo‐black chaff (PBC), a correlated trait. Over 2 yr, two cycles of GS were performed in parallel with one cycle of PS, with each method replicated twice. For GS, markers were generated using genotyping‐by‐sequencing, the prediction model was initially trained using historical data, and the model was updated before the second GS cycle. Overall, GS and PS led to a 31 ± 11 and 42 ± 12% increase in QSRR and a 138 ± 22 and 180 ± 70% increase in PBC, respectively. Genetic gains were not significant but were in agreement with expectations. Per year, gains from GS and PS were equal, but GS led to significantly lower genetic variance. This shows that while GS and PS can lead to equal rates of short‐term gains, GS can reduce genetic variance more rapidly. Further work to develop efficient GS implementation strategies in spring wheat is warranted.

Association mapping of North American spring wheat breeding germplasm reveals loci conferring resistance to Ug99 and other African stem rust races

The recently identified Puccinia graminis f. sp. tritici (Pgt) race TTKSK (Ug99) poses a severe threat to global wheat production because of its broad virulence on several widely deployed resistance genes. Additional virulences have been detected in the Ug99 group of races, and the spread of this race group has been documented across wheat growing regions in Africa, the Middle East (Yemen), and West Asia (Iran). Other broadly virulent Pgt races, such as TRTTF and TKTTF, present further difficulties in maintaining abundant genetic resistance for their effective use in wheat breeding against this destructive fungal disease of wheat. In an effort to identify loci conferring resistance to these races, a genome-wide association study was carried out on a panel of 250 spring wheat breeding lines from the International Maize and Wheat Improvement Center (CIMMYT), six wheat breeding programs in the United States and three wheat breeding programs in Canada. The lines included in this study were grouped into two major clusters, based on the results of principal component analysis using 23,976 SNP markers. Upon screening for adult plant resistance (APR) to Ug99 during 2013 and 2014 in artificial stem rust screening nurseries at Njoro, Kenya and at Debre Zeit, Ethiopia, several wheat lines were found to exhibit APR. The lines were also screened for resistance at the seedling stage against races TTKSK, TRTTF, and TKTTF at USDA-ARS Cereal Disease Laboratory in St. Paul, Minnesota; and only 9 of the 250 lines displayed seedling resistance to all the races. Using a mixed linear model, 27 SNP markers associated with APR against Ug99 were detected, including markers linked with the known APR gene Sr2. Using the same model, 23, 86, and 111 SNP markers associated with seedling resistance against races TTKSK, TRTTF, and TKTTF were identified, respectively. These included markers linked to the genes Sr8a and Sr11 providing seedling resistance to races TRTTF and TKTTF, respectively. We also identified putatively novel Sr resistance genes on chromosomes 3B, 4D, 5A, 5B, 6A, 7A, and 7B. Our results demonstrate that the North American wheat breeding lines have several resistance loci that provide APR and seedling resistance to highly virulent Pgt races. Using the resistant lines and the SNP markers identified in this study, marker-assisted resistance breeding can assist in development of varieties with elevated levels of resistance to virulent stem rust races including TTKSK.

Efficient Use of Historical Data for Genomic Selection: A Case Study of Stem Rust Resistance in Wheat

Genomic selection (GS) is a methodology that can improve crop breeding efficiency. To implement GS, a training population (TP) with phenotypic and genotypic data is required to train a statistical model used to predict genotyped selection candidates (SCs). A key factor impacting prediction accuracy is the relationship between the TP and the SCs. This study used empirical data for quantitative adult plant resistance to stem rust of wheat (Triticum aestivum L.) to investigate the utility of a historical TP (TPH) compared with a population‐specific TP (TPPS), the potential for TPH optimization, and the utility of TPH data when close relative data is available for training. We found that, depending on the population size, a TPPS was 1.5 to 4.4 times more accurate than a TPH, and TPH optimization based on the mean of the generalized coefficient of determination or prediction error variance enabled the selection of subsets that led to significantly higher accuracy than randomly selected subsets. Retaining historical data when data on close relatives were available lead to a 11.9% increase in accuracy, at best, and a 12% decrease in accuracy, at worst, depending on the heritability. We conclude that historical data could be used successfully to initiate a GS program, especially if the dataset is very large and of high heritability. Training population optimization would be useful for the identification of TPH subsets to phenotype additional traits. However, after model updating, discarding historical data may be warranted. More studies are needed to determine if these observations represent general trends.

Harnessing Diversity in Wheat to Enhance Grain Yield, Climate Resilience, Disease and Insect Pest Resistance and Nutrition Through Conventional and Modern Breeding Approaches

Current trends in population growth and consumption patterns continue to increase the demand for wheat, a key cereal for global food security. Further, multiple abiotic challenges due to climate change and evolving pathogen and pests pose a major concern for increasing wheat production globally. Triticeae species comprising of primary, secondary, and tertiary gene pools represent a rich source of genetic diversity in wheat. The conventional breeding strategies of direct hybridization, backcrossing and selection have successfully introgressed a number of desirable traits associated with grain yield, adaptation to abiotic stresses, disease resistance, and bio-fortification of wheat varieties. However, it is time consuming to incorporate genes conferring tolerance/resistance to multiple stresses in a single wheat variety by conventional approaches due to limitations in screening methods and the lower probabilities of combining desirable alleles. Efforts on developing innovative breeding strategies, novel tools and utilizing genetic diversity for new genes/alleles are essential to improve productivity, reduce vulnerability to diseases and pests and enhance nutritional quality. New technologies of high-throughput phenotyping, genome sequencing and genomic selection are promising approaches to maximize progeny screening and selection to accelerate the genetic gains in breeding more productive varieties. Use of cisgenic techniques to transfer beneficial alleles and their combinations within related species also offer great promise especially to achieve durable rust resistance.