Maria Newcomb

Emergence of Virulence to SrTmp in the Ug99 Race Group of Wheat Stem Rust, Puccinia graminis f. sp. tritici, in Africa

detected at two locations in Uganda (Rubaya and Muko in Kabale region) and at five locations in Rwanda (Kinigi, Rwerere, Rufungo, Gatebe, and Kamenyo). Three isolates derived from stem rust samples collected on cv. PBW343 (carrying Sr31) in Sakha in the Nile Delta region in Egypt were also typed as TTKTK. In addition, DNA from isolates of race TTKTK were analyzed using a diagnostic qPCR assay (Ug99 RG stage-1, Szabo, unpublished data), which confirmed that these samples belong to the Ug99 lineage. The identification of SrTmp virulence in the Ug99 race group in several countries in one year emphasizes the relevance of coordinated international surveillance efforts and utilization of diverse sources of resistance to control stem rust in wheat. Further studies are in progress to determine the detailed relationship of the newly emerged races and other Pgt isolates identified in the Ug99 group.

Kenyan Isolates of Puccinia graminis f. sp. tritici from 2008 to 2014: Virulence to SrTmp in the Ug99 Race Group and Implications for Breeding Programs

Frequent emergence of new variants in the Puccinia graminis f. sp. tritici Ug99 race group in Kenya has made pathogen survey a priority. We analyzed 140 isolates from 78 P. graminis f. sp. tritici samples collected in Kenya between 2008 and 2014 and identified six races, including three not detected prior to 2013. Genotypic analysis of 20 isolates from 2013 and 2014 collections showed that the new races TTHST, TTKTK, and TTKTT belong to the Ug99 race group. International advanced breeding lines were evaluated against an isolate of TTKTT (Sr31, Sr24, and SrTmp virulence) at the seedling stage. From 169 advanced lines from Kenya, 23% of lines with resistance to races TTKSK and TTKST were susceptible to TTKTT and, from two North American regional nurseries, 44 and 91% of resistant lines were susceptible. Three lines with combined resistance genes were developed to facilitate pathogen monitoring and race identification. These results indicate the increasing virulence and variability in the Kenyan P. graminis f. sp. tritici population and reveal vulnerabilities of elite germplasm to new races.

First Report of the Ug99 Race Group of Wheat Stem Rust, Puccinia graminis f. sp. tritici, in Egypt in 2014

American stem rust differential lines following standard race-typing procedure and infection type (IT) criteria determining virulence and avirulence (Jin et al. 2008). In addition, three supplemental tester lines of Siouxland (carrying Sr24+Sr31), Sisson (carrying Sr31+Sr36), and Triumph 64 (donor of SrTmp) were included to confirm virulence/avirulence to Sr24, Sr31, Sr36, and SrTmp. The experiments were repeated two to three times. Three races in the Ug99 race group were detected; TTKST (four isolates, IT 3+4 for Sr24, Sr31, and cv. Siouxland) from Al-Sharqia, TTKTK (13 isolates, IT 4 for Sr31, SrTmp, and cv. Triumph 64) from Sakha, and TTKSK (2 isolates, IT 4 for Sr31) from Nubaria. This is the first confirmation of races in the Ug99 race group in Egypt, thereby extending the geographical distribution of Ug99-related races. Since Egypt may play a role as green-bridge for P. graminis f. sp. tritici between East and North African countries and the wheat belts in the Middle East and Mediterranean regions, the rust surveillance efforts should be intensified in affected countries as well as in neighboring regions.

Characterization of Puccinia graminis f. sp. tritici isolates derived from an unusual wheat stem rust outbreak in Germany in 2013

An unusual stem rust infestation occurred in German wheat fields in summer 2013. This study analysed 48 isolates derived from 17 Puccinia graminis f. sp. tritici (Pgt) samples and six races were identified: TKTTF, TKKTF, TKPTF, TKKTP, PKPTF and MMMTF. Infection type and genotypic data confirmed that none of these races belonged to the TTKS (Ug99) race group. German isolates of race TKTTF are phenotypically different to the ones responsible for the stem rust epidemic in Ethiopia in 2013–2014. Forty isolates were genotyped using a custom SNP array. Phylogenetic analysis showed that these 40 isolates represented two distinct lineages (clade IV and clade V). Thirty-eight isolates clustered into clade IV, which previously was defined by Ethiopian isolates of race TKTTF. Race TKKTP is of special concern due to its combined virulence to stem rust resistance genes Sr24, SrTmp and Sr1RSAmigo. The vulnerability to race TKKTP in US and international winter wheat was confirmed as 55% of North American and international cultivars and breeding lines resistant to race TTKSK (Ug99) became susceptible to TKKTP. Races identified in Germany in 2013 confirmed the presence of virulence to important resistance genes that are effective against race TTKSK. This information should be useful for breeders to select diverse and effective resistance genes in order to provide more durable stem rust resistance and reduce the use of fungicides.

Factors contributing to seasonal fluctuations in rust severity on Ribes missouriense caused by Cronartium ribicola.

Cronartium ribicola, causal agent of white pine blister rust, is a macrocyclic heteroecious rust that cycles between white pines and members of the genus Ribes, which are typically wild plants in North America. To improve predictability of inoculum available for infection of ecologically and commercially important white pines, this research was conducted to identify the factors that influence the development and persistence of uredinia and telia on Ribes in their natural habitats. Numbers of infectious C. ribicola rust lesions (with potentially sporulating rust sori) on tagged Ribes missouriense plants in the woods fluctuated during the season. Changes in numbers of infectious rust lesions were related to rain that occurred 13 days earlier. In field experiments, supplemental leaf wetness provided for 2 days on Ribes shoots resulted in the development of rust lesions more frequently than on control shoots. Viable inoculum and susceptible hosts were present, and the environment was the limiting factor for disease development. Lesion necrosis and leaf abscission contributed to decreases in numbers of infectious rust lesions. Higher lesion density was significantly related to earlier leaf abscission. Telial fruiting bodies occurred in low numbers from early June throughout the remainder of the season.

Characterization and genome-wide association mapping of resistance to leaf rust, stem rust and stripe rust in a geographically diverse collection of spring wheat landraces

The challenge posed by rapidly changing wheat rust pathogens, both in virulence and in environmental adaptation, calls for the development and application of new techniques to accelerate the process of breeding for durable resistance. To expand the resistance gene pool available for germplasm improvement, a panel of 159 landraces plus old cultivars was evaluated for seedling and adult plant resistance (APR) to over 35 Australian pathotypes of Puccinia triticina, Puccinia graminis f. sp. tritici, and Puccinia striiformis f. sp. tritici. Known seedling resistance (SR) genes for leaf rust (Lr2a, Lr3a, Lr13, Lr23, Lr16, and Lr20), stem rust (Sr12, Sr13, Sr23, Sr30, and Sr36), and stripe rust (Yr3, Yr4, Yr5, Yr9, Yr10, Yr17, and Yr27) were postulated. The APR genes identified via field assessments and marker analyses included the pleiotropic genes (Lr34/Yr18/Sr57, Lr46/Yr29/Sr58, Lr67/Yr46/Sr55, and Sr2/Lr27/Yr30), Lr68, Lr74, and uncharacterized APR. A genome-wide association analysis using linear mixed models detected 79 single nucleotide polymorphism (SNP) markers significantly associated with rust resistance, which were mapped on chromosomes 1A, 1B, 1D, 2A, 2B, 3A, 3B, 3D, 4A, 5A, 5B, 6A, 6B, 6D, 7A, 7B and 7D. SNPs associated with multiple rust resistances probably indicate the presence of new pleiotropic or closely linked genes. SNPs were mapped on chromosome positions (1AL, 1DS, 2AL, 4AS, 5BS, 6DL, and 7AL) that have not been known to carry APR genes. This study revealed the presence of a range of possibly unidentified effective seedling and APRs among the landraces, which might represent new sources of rust resistance for the ongoing effort to develop improved wheat cultivars.

TERRA-REF Data Processing Infrastructure

The Transportation Energy Resources from Renewable Agriculture Phenotyping Reference Platform (TERRA-REF) provides a data and computation pipeline responsible for collecting, transferring, processing and distributing large volumes of crop sensing and genomic data from genetically informative germplasm sets. The primary source of these data is a field scanner system built over an experimental field at the University of Arizona Maricopa Agricultural Center. The scanner uses several different sensors to observe the field at a dense collection frequency with high resolution. These sensors include RGB stereo, thermal, pulse-amplitude modulated chlorophyll fluorescence, imaging spectrometer cameras, a 3D laser scanner, and environmental monitors. In addition, data from sensors mounted on tractors, UAVs, an indoor controlled-environment facility, and manually collected measurements are integrated into the pipeline. Up to two TB of data per day are collected and transferred to the National Center for Supercomputing Applications at the University of Illinois (NCSA) where they are processed. In this paper we describe the technical architecture for the TERRA-REF data and computing pipeline. This modular and scalable pipeline provides a suite of components to convert raw imagery to standard formats, geospatially subset data, and identify biophysical and physiological plant features related to crop productivity, resource use, and stress tolerance. Derived data products are uploaded to the Clowder content management system and the BETYdb traits and yields database for querying, supporting research at an experimental plot level. All software is open source2 under a BSD 3-clause or similar license and the data products are open access (currently for evaluation with a full release in fall 2019). In addition, we provide computing environments in which users can explore data and develop new tools. The goal of this system is to enable scientists to evaluate and use data, create new algorithms, and advance the science of digital agriculture and crop improvement.

Comparative Aerial and Ground Based High Throughput Phenotyping for the Genetic Dissection of NDVI as a Proxy for Drought Adaptive Traits in Durum Wheat

High-throughput phenotyping platforms (HTPPs) provide novel opportunities to more effectively dissect the genetic basis of drought-adaptive traits. This genome-wide association study (GWAS) compares the results obtained with two Unmanned Aerial Vehicles (UAVs) and a ground-based platform used to measure Normalized Difference Vegetation Index (NDVI) in a panel of 248 elite durum wheat (Triticum turgidum L. ssp. durum Desf.) accessions at different growth stages and water regimes. Our results suggest increased ability of aerial over ground-based platforms to detect quantitative trait loci (QTL) for NDVI, particularly under terminal drought stress, with 22 and 16 single QTLs detected, respectively, and accounting for 89.6 vs. 64.7% phenotypic variance based on multiple QTL models. Additionally, the durum panel was investigated for leaf chlorophyll content (SPAD), leaf rolling and dry biomass under terminal drought stress. In total, 46 significant QTLs affected NDVI across platforms, 22 of which showed concomitant effects on leaf greenness, 2 on leaf rolling and 10 on biomass. Among 9 QTL hotspots on chromosomes 1A, 1B, 2B, 4B, 5B, 6B, and 7B that influenced NDVI and other drought-adaptive traits, 8 showed per se effects unrelated to phenology.

Genetic Loci Conditioning Adult Plant Resistance to the Ug99 Race Group and Seedling Resistance to Races TRTTF and TTTTF of the Stem Rust Pathogen in Wheat Landrace CItr 15026

Wheat landrace CItr 15026 previously showed adult plant resistance (APR) to the Ug99 stem rust race group in Kenya and seedling resistance to Puccinia graminis f. sp. tritici races QFCSC, TTTTF, and TRTTF. CItr 15026 was crossed to susceptible accessions LMPG-6 and Red Bobs, and 180 double haploid (DH) lines and 140 recombinant inbred lines (RIL), respectively, were developed. The 90K wheat iSelect single-nucleotide polymorphism platform was used to genotype the parents and populations. Parents and 180 DH lines were evaluated in the field in Kenya for three seasons. A major quantitative trait locus (QTL) for APR was consistently detected on chromosome arm 6AS. This QTL was further detected in the RIL population screened in Kenya for one season. Parents, F1, and the two populations were tested as seedlings against races TRTTF and TTTTF. In addition, the DH population was tested against race QFCSC. Goodness-of-fit tests indicated that the TRTTF resistance in CItr 15026 was controlled by two complementary genes whereas the TTTTF and QFCSC resistance was conditioned by one dominant gene. The TRTTF resistance loci mapped to chromosome arms 6AS and 6DS, whereas the TTTTF and QFCSC resistance locus mapped to the same region on 6DS as the TRTTF resistance. The APR identified in CItr 15026 should be useful in developing cultivars with durable stem rust resistance.

Discovery of a Novel Stem Rust Resistance Allele in Durum Wheat that Exhibits Differential Reactions to Ug99 Isolates

Wheat stem rust, caused by Puccinia graminis f. sp. tritici Eriks. & E. Henn, can incur yield losses in susceptible cultivars of durum wheat, Triticum turgidum ssp. durum (Desf.) Husnot. Although several durum cultivars possess the stem rust resistance gene Sr13, additional genes in durum wheat effective against emerging virulent races have not been described. Durum line 8155-B1 confers resistance against the P. graminis f. sp. tritici race TTKST, the variant race of the Ug99 race group with additional virulence to wheat stem rust resistance gene Sr24. However, 8155-B1 does not confer resistance to the first-described race in the Ug99 race group: TTKSK. We mapped a single gene conferring resistance in 8155-B1 against race TTKST, Sr8155B1, to chromosome arm 6AS by utilizing Rusty/8155-B1 and Rusty*2/8155-B1 populations and the 90K Infinium iSelect Custom bead chip supplemented by KASP assays. One marker, KASP_6AS_IWB10558, cosegregated with Sr8155B1 in both populations and correctly predicted Sr8155B1 presence or absence in 11 durum cultivars tested. We confirmed the presence of Sr8155B1 in cultivar Mountrail by mapping in the population Choteau/Mountrail. The marker developed in this study could be used to predict the presence of resistance to race TTKST in uncharacterized durum breeding lines, and also to combine Sr8155B1 with resistance genes effective to Ug99 such as Sr13. The map location of Sr8155B1 cannot rule out the possibility that this gene is an allele at the Sr8 locus. However, race specificity indicates that Sr8155B1 is different from the known alleles Sr8a and Sr8b.