Watkins Collection Research Articles

Mapping of a Stripe Rust Resistance Gene Yr72 in the Common Wheat Landraces AUS27506 and AUS27894 from the Watkins Collection.

Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is among the major threats to global wheat production. The common wheat landraces AUS27506 and AUS27894 displayed stripe rust resistance against several commercially prevailing Pst pathotypes. These genotypes were crossed with a stripe-rust-susceptible landrace AUS27229 to understand the inheritance of resistance and to determine the genomic location(s) of underlying gene(s). F3 generations of crosses AUS27506/AUS27229 and AUS27894/AUS27229 showed monogenic segregation for stripe rust resistance under greenhouse conditions. The absence of segregation for stripe rust response among the AUS27506/AUS27894-derived F3 population suggested that both genotypes carry the same gene. The stripe rust resistance gene carried by AUS27506 and AUS27894 was tentatively named YrAW4. A bulked segregant analysis placed YrAW4 in the long arm of chromosome 2B. The AUS27506/AUS27229 F3 population was enhanced to develop an F6 recombinant inbred line (RIL) population for detailed mapping of chromosome 2BL. DArT-based SSR, STS and SNP markers were employed to enrich the 2BL map. DArT-based STS markers sun481 and SNP marker IWB12294 flanked YrAW4 proximally (1.8 cM) and distally (1.2 cM), respectively. Deletion mapping placed sun481 in the deletion bin 2BL-5. All stripe rust resistance genes, previously located on chromosome 2BL, neither express an infection type like YrAW4, nor are they mapped in the deletion bin 2BL-5. Hence, YrAW4 represented a new locus and was formally named Yr72. The usefulness of the markers IWB12294 and sun481 in marker-assisted selection was demonstrated by the amplification of alleles that are different to that linked with Yr72 in 19 common wheat and two durum wheat cultivars.

Mining the A.E. Watkins Wheat Landrace Collection for Variation in Starch Digestibility Using a New High-Throughput Assay.

Breeding for less digestible starch in wheat can improve the health impact of bread and other wheat foods. The application of forward genetic approaches has lately opened opportunities for the discovery of new genes that influence the digestibility of starch, without the burden of detrimental effects on yield or on pasta and bread-making quality. In this study we developed a high-throughput in vitro starch digestibility assay (HTA) for use in forward genetic approaches to screen wheat germplasm. The HTA was validated using standard maize and wheat starches. Using the HTA we measured starch digestibility in hydrothermally processed flour samples and found wide variation among 118 wheat landraces from the A. E. Watkins collection and among eight elite UK varieties (23.5 to 39.9% and 31.2 to 43.5% starch digested after 90 min, respectively). We further investigated starch digestibility in fractions of sieved wholemeal flour and purified starch in a subset of the Watkins lines and elite varieties and found that the matrix properties of flour rather than the intrinsic properties of starch granules conferred lower starch digestibility.

Discovery of the New Leaf Rust Resistance Gene Lr82 in Wheat: Molecular Mapping and Marker Development.

Breeding for leaf rust resistance has been successful worldwide and is underpinned by the discovery and characterisation of genetically diverse sources of resistance. An English scientist, Arthur Watkins, collected pre-Green Revolution wheat genotypes from 33 locations worldwide in the early part of the 20th Century and this collection is now referred to as the ‘Watkins Collection’. A common wheat genotype, Aus27352 from Yugoslavia, showed resistance to currently predominating Australian pathotypes of the wheat leaf rust pathogen. We crossed Aus27352 with a leaf rust susceptible wheat selection Avocet S and a recombinant inbred line (RIL) F6 population of 200 lines was generated. Initial screening at F3 generation showed monogenic segregation for seedling response to leaf rust in Aus27352. These results were confirmed by screening the Aus27352/Avocet S RIL population. The underlying locus was temporarily named LrAW2. Bulked segregant analysis using the 90K Infinium SNP array located LrAW2 in the long arm of chromosome 2B. Tests with molecular markers linked to two leaf rust resistance genes, Lr50 and Lr58, previously located in chromosome 2B, indicated the uniqueness of LrAW2 and it was formally designated Lr82. Kompetitive allele-specific polymerase chain reaction assays were developed for Lr82-linked SNPs. KASP_22131 mapped 0.8 cM proximal to Lr82 and KASP_11333 was placed 1.2 cM distal to this locus. KASP_22131 showed 91% polymorphism among a set of 89 Australian wheat cultivars. We recommend the use of KASP_22131 for marker assisted pyramiding of Lr82 in breeding programs following polymorphism check on parents.

Evaluation of the susceptibility of modern, wild, ancestral, and mutational wheat lines to Septoria tritici blotch disease

AbstractGlobally, bread wheat production is threatened by fungal diseases, including the devastating disease Septoria tritici blotch (STB). Given the global importance of STB, and the difficulty in identifying novel sources of resistance to this disease, we screened a variety of wheat genotypes, including wild, ancestral, and mutagenized lines, for their STB response. This delineated a panel of wild wheat relatives and Watkins collection lines with exceptional resistance to a range of Zymoseptoria tritici isolates, some of which are highly virulent on modern, elite wheat varieties. Additionally, we characterized the STB susceptibility of 500 lines of the wheat cultivar Cadenza TILLING population and developed backcross derivatives of two TILLING lines that show dominant partial resistance to STB. These backcross lines are partially resistant to multiple isolates of Z. tritici, and, with the wild and ancestral lines identified, provide a useful reservoir of STB‐resistant germplasm for use in wheat breeding programmes.

Mapping of Adult Plant Leaf Rust Resistance in Aus27506 and Validation of Underlying Loci by In-Planta Fungal Biomass Accumulation

Among the rust diseases, leaf rust of wheat caused by Puccinia triticina, is the most prevalent worldwide and causes significant yield losses. This study aimed to determine the genomic location of loci that control adult plant resistance (APR) to leaf rust in the pre-Green Revolution landrace accession, Aus27506, from the “Watkins Collection”. An Aus27506/Aus27229-derived F7 recombinant inbred line (RIL) population was screened under field conditions across three cropping seasons and genotyped with the iSelect 90K Infinium SNP bead chip array. One quantitative trait loci (QTL) on each of the chromosomes 1BL, 2B and 2DL explained most of the leaf rust response variation in the RIL population, and these were named QLr.sun-1BL, QLr.sun-2B and QLr.sun-2DL, respectively. QLr.sun-1BL and QLr.sun-2DL were contributed by Aus27506. QLr.sun-1BL is likely Lr46, while QLr.sun-2DL appeared to be a new APR locus. The alternate parent, Aus27229, carried the putatively new APR locus QLr.sun-2B. The comparison of average severities among RILs carrying these QTL in different combinations indicated that QLr.sun-2B does not interact with either of the other two QTL; however, the combination of QLr.sun-1BL and QLr.sun-2DL reduced disease severity significantly. In planta fungal quantification assays validated these results. The RILs carrying QLr.sun-1BL and QLr.sun-2DL did not differ significantly from the parent Aus27506 in terms of resistance. Aus27506 can be used as a source of adult plant leaf rust resistance in breeding programs.

Novel sources of variation in grain Zinc (Zn) concentration in bread wheat germplasm derived from Watkins landraces.

A diverse panel of 245 wheat genotypes, derived from crosses between landraces from the Watkins collection representing global diversity in the early 20th century and the modern wheat cultivar Paragon, was grown at two field sites in the UK in 2015–16 and the concentrations of zinc and iron determined in wholegrain using inductively coupled plasma-mass spectrometry (ICP-MS). Zinc concentrations in wholegrain varied from 24–49 mg kg-1 and were correlated with iron concentration (r = 0.64) and grain protein content (r = 0.14). However, the correlation with yield was low (r = -0.16) indicating little yield dilution. A sub-set of 24 wheat lines were selected from 245 wheat genotypes and characterised for Zn and Fe concentrations in wholegrain and white flour over two sites and years. White flours from 24 selected lines contained 8–15 mg kg-1 of zinc, which was positively correlated with the wholegrain Zn concentration (r = 0.79, averaged across sites and years). This demonstrates the potential to exploit the diversity in landraces to increase the concentration of Zn in wholegrain and flour of modern high yielding bread wheat cultivars.

Mining and genomic characterization of resistance to tan spot, Stagonospora nodorum blotch (SNB), and Fusarium head blight in Watkins core collection of wheat landraces

BackgroundIn the late 1920s, A. E. Watkins collected about 7000 landrace cultivars (LCs) of bread wheat (Triticum aestivum L.) from 32 different countries around the world. Among which 826 LCs remain viable and could be a valuable source of superior/favorable alleles to enhance disease resistance in wheat. In the present study, a core set of 121 LCs, which captures the majority of the genetic diversity of Watkins collection, was evaluated for identifying novel sources of resistance against tan spot, Stagonospora nodorum blotch (SNB), and Fusarium Head Blight (FHB).ResultsA diverse response was observed in 121 LCs for all three diseases. The majority of LCs were moderately susceptible to susceptible to tan spot Ptr race 1 (84%) and FHB (96%) whereas a large number of LCs were resistant or moderately resistant against tan spot Ptr race 5 (95%) and SNB (54%). Thirteen LCs were identified in this study could be a valuable source for multiple resistance to tan spot Ptr races 1 and 5, and SNB, and another five LCs could be a potential source for FHB resistance. GWAS analysis was carried out using disease phenotyping score and 8807 SNPs data of 118 LCs, which identified 30 significant marker-trait associations (MTAs) with -log10 (p-value) > 3.0. Ten, five, and five genomic regions were found to be associated with resistance to tan spot Ptr race 1, race 5, and SNB, respectively in this study. In addition to Tsn1, several novel genomic regions Q.Ts1.sdsu-4BS and Q.Ts1.sdsu-5BS (tan spot Ptr race 1) and Q.Ts5.sdsu-1BL, Q.Ts5.sdsu-2DL, Q.Ts5.sdsu-3AL, and Q.Ts5.sdsu-6BL (tan spot Ptr race 5) were also identified. Our results indicate that these putative genomic regions contain several genes that play an important role in plant defense mechanisms.ConclusionOur results suggest the existence of valuable resistant alleles against leaf spot diseases in Watkins LCs. The single-nucleotide polymorphism (SNP) markers linked to the quantitative trait loci (QTLs) for tan spot and SNB resistance along with LCs harboring multiple disease resistance could be useful for future wheat breeding.

A modified sequence capture approach allowing standard and methylation analyses of the same enriched genomic DNA sample

BackgroundBread wheat has a large complex genome that makes whole genome resequencing costly. Therefore, genome complexity reduction techniques such as sequence capture make re-sequencing cost effective. With a high-quality draft wheat genome now available it is possible to design capture probe sets and to use them to accurately genotype and anchor SNPs to the genome. Furthermore, in addition to genetic variation, epigenetic variation provides a source of natural variation contributing to changes in gene expression and phenotype that can be profiled at the base pair level using sequence capture coupled with bisulphite treatment. Here, we present a new 12 Mbp wheat capture probe set, that allows both the profiling of genotype and methylation from the same DNA sample. Furthermore, we present a method, based on Agilent SureSelect Methyl-Seq, that will use a single capture assay as a starting point to allow both DNA sequencing and methyl-seq.ResultsOur method uses a single capture assay that is sequentially split and used for both DNA sequencing and methyl-seq. The resultant genotype and epi-type data is highly comparable in terms of coverage and SNP/methylation site identification to that generated from separate captures for DNA sequencing and methyl-seq. Furthermore, by defining SNP frequencies in a diverse landrace from the Watkins collection we highlight the importance of having genotype data to prevent false positive methylation calls. Finally, we present the design of a new 12 Mbp wheat capture and demonstrate its successful application to re-sequence wheat.ConclusionsWe present a cost-effective method for performing both DNA sequencing and methyl-seq from a single capture reaction thus reducing reagent costs, sample preparation time and DNA requirements for these complementary analyses.

A new leaf rust resistance gene Lr79 mapped in chromosome 3BL from the durum wheat landrace Aus26582.

A new leaf rust resistance gene Lr79 has been mapped in the long arm of chromosome 3B and a linked marker was identified for marker-assisted selection. Aus26582, a durum wheat landrace from the A. E. Watkins Collection, showed seedling resistance against durum-specific and common wheat-specific Puccinia triticina (Pt) pathotypes. Genetic analysis using a recombinant inbred line (RIL) population developed from a cross between Aus26582 and the susceptible parent Bansi with Australian Pt pathotype showed digenic inheritance and the underlying loci were temporarily named LrAW2 and LrAW3. LrAW2 was located in chromosome 6BS and this study focused on characterisation of LrAW3 using RILs lacking LrAW2. LrAW3 was incorporated into the DArTseq map of Aus26582/Bansi and was located in chromosome 3BL. Markers linked with LrAW3 were developed from the chromosome survey sequence contig 3B_10474240 in which closely-linked DArTseq markers 1128708 and 3948563 were located. Although bulk segregant analysis (BSA) with the 90K Infinium array identified 51 SNPs associated with LrAW3, only one SNP-derived KASP marker mapped close to the locus. Deletion bin mapping of LrAW3-linked markers located LrAW3 between bins 3BL11-0.85-0.90 and 3BL7-0.63. Since no other all stage leaf rust resistance gene is located in chromosome 3BL, LrAW3 represented a new locus and was designated Lr79. Marker sun786 mapped 1.8cM distal to Lr79 and Aus26582 was null for this locus. However, the marker can be reliably scored as it also amplifies a monomorphic fragment that serves as an internal control to differentiate the null status of Aus26582 from reaction failure. This marker was validated among a set of durum and common wheat cultivars and was shown to be useful for marker-assisted selection of Lr79 at both ploidy levels.

High-density genotyping of the A.E. Watkins Collection of hexaploid landraces identifies a large molecular diversity compared to elite bread wheat.

SummaryThe importance of wheat as a food crop makes it a major target for agricultural improvements. As one of the most widely grown cereal grains, together with maize and rice, wheat is the leading provider of calories in the global diet, constituting 29% of global cereal production in 2015. In the last few decades, however, yields have plateaued, suggesting that the green revolution, at least for wheat, might have run its course and that new sources of genetic variation are urgently required. The overall aim of our work was to identify novel variation that may then be used to enable the breeding process. As landraces are a potential source of such diversity, here we have characterized the A.E. Watkins Collection alongside a collection of elite accessions using two complementary high‐density and high‐throughput genotyping platforms. While our results show the importance of using the appropriate SNP collection to compare diverse accessions, they also show that the Watkins Collection contains a substantial amount of novel genetic diversity which has either not been captured in current breeding programmes or which has been lost through previous selection pressures. As a consequence of our analysis, we have identified a number of accessions which carry an array of novel alleles along with a number of interesting chromosome rearrangements which confirm the variable nature of the wheat genome.

Searching for wheat resistance to aphids and wheat bulb fly in the historical Watkins and Gediflux wheat collections.

Insect pests can reduce wheat yield by direct feeding and transmission of plant viruses. Here we report results from laboratory and field phenotyping studies on a wide range of wheat, including landraces from the Watkins collection deriving from before the green revolution, more modern cultivars from the Gediflux collection (north‐western Europe) and modern UK Elite varieties, for resistance to the bird cherry‐oat aphid, Rhopalosiphum padi (Homoptera: Aphididae) and the English grain aphid, Sitobion avenae (Homoptera: Aphididae). A total of 338 lines were screened for R. padi and 340 lines for S. avenae. Field trials were also conducted on 122 Watkins lines to identify wheat bulb fly, Delia coarctata, preference on these landraces. Considerable variation was shown in insect performance among and within different wheat collections, with reduced susceptibility in a number of varieties, but phenotyping did not identify strong resistance to aphids or wheat bulb fly. Field trials showed within collection differences in aphid performance, with fewer aphids populating lines from the Watkins collection. This differs from development data in laboratory bioassays and suggests that there is a pre‐alighting cue deterring aphid settlement and demonstrates differences in aphid preference and performance on older plants in the field compared with seedlings in the laboratory, highlighting the need for phenotyping for aphid resistance at different plant growth stages. No association was identified between performance of the different insect species on individual varieties, potentially suggesting different nutritional requirements or resistance mechanisms.

Leaf photosynthesis and associations with grain yield, biomass and nitrogen-use efficiency in landraces, synthetic-derived lines and cultivars in wheat

Future genetic progress in wheat grain yield will depend on increasing above-ground biomass and this must be achieved without commensurate increases in N fertilizer inputs to minimise environmental impacts. Our objective was to quantify variation in grain yield, above-ground biomass and N-use efficiency (NUE) and associated traits in a panel of diverse hexaploid wheat germplasm comprising: (i) landraces from the AE Watkins collection, (ii) synthetic-derived hexaploid lines in a cv. Paragon spring wheat background and (iii) UK modern cultivars including cv. Paragon under low N and high N conditions. A field experiment was carried out in two seasons examining 15 genotypes (five landraces, five synthetic-derived (SD) hexaploid lines and five UK modern cultivars) under low N and high N conditions at Nottingham University farm, UK. Machine-harvested grain yield, above-ground biomass and NUE were measured. Physiological traits were assessed including flag-leaf light-saturated photosynthetic rate (Amax) and relative chlorophyll content (SPAD) under HN conditions; and flag-leaf senescence duration and rate and Normalized Difference Vegetative Index (NDVI) under LN and HN conditions. Under HN conditions, the modern cultivars overall produced higher grain yield than the SD lines (+9.7%) and the landraces (+60.4%); and the modern cultivars and SD lines also produced higher biomass than the landraces (30.3% and 28.4%, respectively). Under LN conditions, reduction in grain yield and biomass compared to HN conditions was least for the landraces (−1% and −8.6%, respectively), intermediate for the SD lines (−7.4 and −10.2%, respectively) and highest for the modern cultivars (−9.3 and −24.6%, respectively). As a result, the SD lines had higher biomass (+17%) than the modern cultivars under LN conditions. Under HN conditions the synthetic derivatives (23.8μmolm−2s−1) and modern cultivars (241.1μmolm−2s−1) had higher pre-anthesis Amax than the landraces (19.7μmolm−2s−1) (P<0.001). Pre-anthesis Amax was strongly positively linearly associated with above-ground biomass (R2=0.63, P<0.001) and grain yield (R2=0.75, P<0.001) amongst the 15 genotypes. Flag-leaf Amax was also positively linearly associated with flag-leaf relative chlorophyll content at anthesis (R2=0.74; P<0.001). Comparing the SD lines to the recurrent parent Paragon, under HN conditions one line (SD 22) had higher pre-anthesis flag-leaf Amax than Paragon (P<0.05). Under LN conditions one line (SD 24, +27%) had higher yield than Paragon (P<0.05) and two lines (SD 24 and SD 38, +32% and +31%, respectively) had more biomass than Paragon (P<0.05). Our results indicated that introgressing traits from synthetic-derived wheat and landraces into UK modern wheat germplasm offers scope to raise above-ground biomass and grain yield in moderate-to-low N availability environments.

Mapping of a new stripe rust resistance locus Yr57 on chromosome 3BS of wheat

A common wheat landrace, AUS27858, from the Watkins collection showed low seedling stripe rust response against Australian Puccinia striiformis f. sp. tritici pathotypes. Genetic analysis of stripe rust resistance indicated the involvement of two independent resistance loci YrAW1 and YrAW2. YrAW1 was mapped in chromosome 4AL and formally named Yr51. Ninety seeds of a heterozygous F3 line (HSB#5474; YrAW2yrAW2) were grown individually to produce a segregating population referred to as single gene segregating population #5474 (SGSP#5474) to map YrAW2. Monogenic segregation at the YrAW2 locus was confirmed among the SGSP#5474. YrAW2 was located in chromosome 3BS through DArT-based bulked segregant analysis. SGSP#5474 was advanced to F6 generation and was phenotyped for detailed mapping. Test of allelism with Yr4, previously located on chromosome 3BS, showed 5.2 ± 1.3 % recombination between YrAW2 and Yr4. Since there is no other stripe rust resistance gene located in the distal part of chromosome 3BS, YrAW2 was formally named Yr57. Markers gwm389 and BS00062676 flanked Yr57 at genetic distances of 2.0 and 2.3 cM, proximally and distally, respectively. These markers were genotyped on a set of Australian and Indian wheat cultivars and the absence of resistance-linked alleles of gwm389 and BS00062676 markers was shown in cultivars known to lack Yr57. These markers would be useful in marker-assisted pyramiding of Yr57 with other marker-tagged major and minor genes. The genetic stock carrying Yr57 singly has been deposited with the Australian Winter Cereal Collection, Tamworth, Australia, and it is accessioned as AUS91463.

Detection of puroindoline (Pina-D1 and Pinb-D1) allelic variation in wheat landraces

Allelic variation at the grain texture-controlling linked loci Pina-D1 and Pinb-D1 was studied among wheat landraces from the Watkins collection (pre-green revolution) using gene specific PCR based approaches. Of the 803 landraces, 593 carried the wild type alleles at these loci and were classified as soft textured. The remaining 210 genotypes were classified as hard-textured based on the presence of mutant Pin alleles known to be associated with grain hardness. Five allelic variants of Pinb-D1 were observed. Four allelic classes Pinb-D1a, Pinb-D1b, Pinb-D1c and Pinb-D1e resulted from separate single nucleotide substitutions leading to mis-sense changes, while a single base deletion at position 213 resulted in the Pinb-D1p allele exhibiting a frame shift and consequently stop codons. These five alleles occurred in six different allelic combinations with two Pina-D1 alleles (wild type and null), i.e. Pina-D1a/Pinb-D1a, Pina-D1b/Pinb-D1a, Pina-D1a/Pinb-D1b, Pina-D1a/Pina-D1c, Pina-D1a/Pinb-D1e and Pina-D1a/Pina-D1p. The alleles identified in this study provide additional resources for breeding programs for grain texture manipulations and may also have implications in microbial pathogen defence. Near-isogenic lines carrying different alleles are currently being produced to study the effect of this genetic variation on flour processing properties.

Establishing the A. E. Watkins landrace cultivar collection as a resource for systematic gene discovery in bread wheat.

Key message A high level of genetic diversity was found in the A. E. Watkins bread wheat landrace collection. Genotypic information was used to determine the population structure and to develop germplasm resources. In the 1930s A. E. Watkins acquired landrace cultivars of bread wheat (Triticum aestivum L.) from official channels of the board of Trade in London, many of which originated from local markets in 32 countries. The geographic distribution of the 826 landrace cultivars of the current collection, here called the Watkins collection, covers many Asian and European countries and some from Africa. The cultivars were genotyped with 41 microsatellite markers in order to investigate the genetic diversity and population structure of the collection. A high level of genetic diversity was found, higher than in a collection of modern European winter bread wheat varieties from 1945 to 2000. Furthermore, although weak, the population structure of the Watkins collection reveals nine ancestral geographical groupings. An exchange of genetic material between ancestral groups before commercial wheat-breeding started would be a possible explanation for this. The increased knowledge regarding the diversity of the Watkins collection was used to develop resources for wheat research and breeding, one of them a core set, which captures the majority of the genetic diversity detected. The understanding of genetic diversity and population structure together with the availability of breeding resources should help to accelerate the detection of new alleles in the Watkins collection.Electronic supplementary materialThe online version of this article (doi:10.1007/s00122-014-2344-5) contains supplementary material, which is available to authorized users.

Genomic prediction for rust resistance in diverse wheat landraces.

We have demonstrated that genomic selection in diverse wheat landraces for resistance to leaf, stem and strip rust is possible, as genomic breeding values were moderately accurate. Markers with large effects in the Bayesian analysis confirmed many known genes, while also discovering many previously uncharacterised genome regions associated with rust scores. Genomic selection, where selection decisions are based on genomic estimated breeding values (GEBVs) derived from genome-wide DNA markers, could accelerate genetic progress in plant breeding. In this study, we assessed the accuracy of GEBVs for rust resistance in 206 hexaploid wheat (Triticum aestivum) landraces from the Watkins collection of phenotypically diverse wheat genotypes from 32 countries. The landraces were genotyped for 5,568 SNPs using an Illumina iSelect 9 K bead chip assay and phenotyped for field-based leaf rust (Lr), stem rust (Sr) and stripe rust (Yr) responses across multiple years. Genomic Best Linear Unbiased Prediction (GBLUP) and a Bayesian Regression method (BayesR) were used to predict GEBVs. Based on fivefold cross-validation, the accuracy of genomic prediction averaged across years was 0.35, 0.27 and 0.44 for Lr, Sr and Yr using GBLUP and 0.33, 0.38 and 0.30 for Lr, Sr and Yr using BayesR, respectively. Inclusion of PCR-predicted genotypes for known rust resistance genes increased accuracy more substantially when the marker was diagnostic (Lr34/Sr57/Yr18) for the presence-absence of the gene rather than just linked (Sr2). Investigation of the impact of genetic relatedness between validation and reference lines on accuracy of genomic prediction showed that accuracy will be higher when each validation line had at least one close relationship to the reference lines. Overall, the prediction accuracies achieved in this study are encouraging, and confirm the feasibility of genomic selection in wheat. In several instances, estimated marker effects were confirmed by published literature and results of mapping experiments using Watkins accessions.

Mining the Watkins collection of wheat landraces for novel sources of eyespot resistance

Eyespot is an economically important stem base disease of wheat caused by the soilborne fungal pathogens Oculimacula yallundae and Oculimacula acuformis. The most effective method of controlling the disease is host resistance. However, there are only three genetically characterized resistances in wheat varieties and further sources of resistance are required. Previous studies have identified resistances in wild relatives, but use of these resistances has been limited by linkage drag with deleterious traits exacerbated by low rates of recombination. Therefore, the identification of novel resistances in hexaploid wheat germplasm is desirable. The Watkins collection currently consists of 1056 hexaploid wheat landraces that represent global wheat diversity at the time of its collection in the 1920s and 1930s. As such, it may contain beneficial agronomic traits such as eyespot resistance. The Watkins collection was screened for resistance to O. yallundae based on a glasshouse test of all 1056 accessions and a polytunnel test of 44 accessions selected from a previous field trial. Resistant lines identified in these tests were retested against both O. yallundae and O. acuformis. This identified 17 accessions with resistance to one or both of the pathogen species. From these, two accessions (1190094.1 and 1190736.3) provided a high level of resistance to both pathogen species. An F4 population derived from accession 1190736.3 indicated that the resistance to O. acuformis in this accession is conferred by a single gene and therefore would be suitable for introgression into elite wheat varieties to provide an alternative source of eyespot resistance.

Molecular mapping of stripe rust resistance gene Yr51 in chromosome 4AL of wheat

This manuscript describes the chromosomal location of a new source of stripe rust resistance in wheat. DNA markers closely linked with the resistance locus were identified and validated. A wheat landrace, AUS27858, from the Watkins collection showed high levels of resistance against Australian pathotypes of Puccinia striiformis f. sp. tritici. It was reported to carry two genes for stripe rust resistance, tentatively named YrAW1 and YrAW2. One hundred seeds of an F3 line (HSB#5515; YrAW1yrAW1) that showed monogenic segregation for stripe rust response were sown and harvested individually to generate monogenically segregating population (MSP) #5515. Stripe rust response variation in MSP#5515 conformed to segregation at a single locus. Bulked segregant analysis using high-throughput DArT markers placed YrAW1 in chromosome 4AL. MSP#5515 was advanced to F6 and phenotyped for detailed mapping. Novel wheat genomic resources including chromosome-specific sequence and genome zipper were employed to develop markers specific for the long arm of chromosome 4A. These markers were used for further saturation of the YrAW1 carrying region. YrAW1 was delimited by 3.7cM between markers owm45F3R3 and sun104. Since there was no other stripe rust resistance gene located in chromosome 4AL, YrAW1 was formally named Yr51. Reference stock for Yr51 was lodged at the Australian Winter Cereal Collection, Tamworth, Australia and it was accessioned as AUS91456. Marker sun104 was genotyped on a set of Australian and Indian wheat cultivars and was shown to lack the resistance-linked sun104-225bp allele. Marker sun104 is currently being used for marker-assisted backcrossing of Yr51 in Australian and Indian wheat backgrounds.

Characterization of stem rust resistance in old tetraploid wheat landraces from the Watkins collection

Stem rust is one of the important diseases of tetraploid wheats worldwide. One hundred and five landraces from the Watkins collection were assessed for seedling and adult plant stem rust response variation. Seedling resistance genes Sr8a, Sr8b, Sr9e, Sr9g, Sr12, Sr13, Sr17 and Sr23 were postulated in 28 genotypes using Australian and Indian Puccinia graminis f. sp. tritici pathotypes. Four genotypes possessed either uncharacterized seedling stem rust resistance gene(s) or combinations of known stem rust resistance genes with compensating avirulences among pathotypes used. Adult plant stem rust response assessments were made on 73 seedling susceptible genotypes in Australia and Ethiopia. Adult plant stem rust responses varied from 1 (very resistant) to 7 (moderately susceptible) on a 1–9 scale. The Ethiopian nursery was exposed to Ug99 (TTKSK) and JRCQC (virulent on Sr9e and Sr13). Some genotypes that exhibited high responses in Australia were scored low in Ethiopia. These results demonstrated the effectiveness of some resistance gene(s) that were ineffective in Australia. The opposite trend was also noted. Over 50 % genotypes exhibited commercially acceptable rust responses varying between 2 and 5 across sites and years. Genotyping with the Sr2-linked molecular marker csSr2 did not detect Sr2 in any of the genotypes. The marker gwm533 however detected the presence of Sr2 in eight genotypes. Stem rust resistant genotypes that carried varying levels of APR could carry new genes. Genetic analysis and deployment of these uncharacterized sources of seedling and APR in new cultivars will ensure durable stem rust control through increased diversity.

Characterisation of a new stripe rust resistance gene Yr47 and its genetic association with the leaf rust resistance gene Lr52

Two Iranian common wheat landraces AUS28183 and AUS28187 from the Watkins collection showed high levels of seedling resistance against Australian pathotypes of leaf rust and stripe rust pathogens. Chi-squared analyses of rust response segregation among F(3) populations derived from crosses of AUS28183 and AUS28187 with a susceptible genotype AUS27229 revealed monogenic inheritance of leaf rust and stripe rust resistance. As both genotypes produced similar leaf rust and stripe rust infection types, they were assumed to carry the same genes. The genes were temporarily named as LrW1 and YrW1. Molecular mapping placed LrW1 and YrW1 in the short arm of chromosome 5B, about 10 and 15cM proximal to the SSR marker gwm234, respectively, and the marker cfb309 mapped 8-12cM proximal to YrW1. LrW1 mapped 3-6cM distal to YrW1 in two F(3) populations. AUS28183 corresponded to the accession V336 of the Watkins collection which was the original source of Lr52. Based on the genomic location and accession records, LrW1 was concluded to be Lr52. Because no other seedling stripe rust resistance gene has previously been mapped in chromosome 5BS, YrW1 was permanently named as Yr47. A combination of flanking markers gwm234 and cfb309 with phenotypic assays could be used to ascertain the presence of Lr52 and Yr47 in segregating populations. This investigation characterised a valuable source of dual leaf rust and stripe rust resistance for deployment in new wheat cultivars. Transfer of Lr52 and Yr47 into current Australian wheat backgrounds is in progress.