A. Lehmensiek

Curation of wheat maps to improve map accuracy and QTL detection

Three Australian doubled haploid populations were used to illustrate the importance of map curation in order to improve the quality of linkage maps and quantative trait locus (QTL) detection. The maps were refined and improved by re-examining the order of markers, inspection of the genetic maps in relation to a consensus map, editing the marker data for double crossovers, and determining estimated recombination fractions for all pairs of markers. The re-ordering of markers and replacing genotypes at double crossovers with missing values resulted in an overall decrease in the length of the maps. Fewer apparent genotyping errors, associated with the presence of double recombinants, were identified with restriction fragment length polymorphisms (RFLPs) than with other types of markers used in this study. The complications that translocations may cause in the ordering of markers and subsequent QTL analysis were investigated. QTL analysis using both the original and revised maps indicated that QTL peaks were more sharply located or had improved log-likelihood (LOD) scores in the revised maps. An accurate indication of the QTL peak and a significant LOD score are both essential for the identification of markers suitable for marker-assisted selection. Recommendations are provided for the improvement of the quality of linkage maps.

Molecular Mapping of Durable Rust Resistance in Wheat and its Implication in Breeding

Genetic characterization of sources of durable resistance enables their strategic deployment in breeding programs. Genomic locations of uncharacterized adult plant resistance (APR) sources to leaf rust and stripe rust diseases of wheat were determined. Two genomic regions, 3DS (Halberd) and 5DS (Cranbrook) controlled APR to both leaf rust and stripe rust. Chromosomes 6B (Cranbrook) and 7B (Halberd) reduced leaf rust severity. Chromosomes 2DS, 3BS and 7A also reduced stripe rust severities in at least one crop season. Stem rust resistance genes Sr2 (3BS) and Sr30 (5DL) from Cranbrook explained stem rust response variation. Regression analysis also indicated strong positive interaction of these two loci in controlling stem rust. Expression of Sr2-linked psuedo black chaff (Pbc) was controlled by a major gene on chromosome 3BS and three modifiers located on chromosomes 6A, 3D and 7A. The chromosome 7A located region was not consistent across all seasons and sites. QTLs detected consistently in different experiments were temporarily designated as QYr/Lr3D, QYr/Lr5D, QLr6B and QLr7B

An investigation of genetic variation among Australian isolates of Bipolaris sorokiniana from different cereal tissues and comparison of their abilities to cause spot blotch on barley

Bipolaris sorokiniana (teleomorph: Cochliobolus sativus), the causal agent of common rootrot (CRR) and foliar spot blotch (SB) diseases in barley and wheat, is an economically important fungal pathogen worldwide. However, the relationship between these two diseases is poorly understood. Differences within Australian B. sorokiniana populations were revealed by cluster analysis of amplified fragment length polymorphisms in genomic DNA of 48 B. sorokiniana isolates collected from the northern grain-growing region of Australia. Isolates collected from SB infections clustered apart from isolates collected from CRR infections. A subset of 31 B. sorokiniana isolates was assessed for their abilities to cause SB infections on barley leaves using a differential set of 15 barley genotypes and three other cereal species. The pathogen samples included 14 isolates from CRR infections of either wheat or barley and 14 isolates from SB infections of barley. Phenotypic experiments revealed that isolates of B. sorokiniana collected from barley SB infections showed a high level of pathogenic variability across the differential set. In contrast, isolates from CRR infections produced significantly less SB disease on inoculated barley leaves. Cluster analysis of the phenotypic infection response scores grouped isolates into three pathogenicity clusters demonstrating low, intermediate or high pathogenicity. The results of this study suggest divergence within Australian populations of B. sorokiniana in relation to host tissue specificity.

Mapping spot blotch resistance genes in four barley populations

Bipolaris sorokiniana (teleomorph: Cochliobolus sativus) is the fungal pathogen responsible for spot blotch in barley (Hordeum vulgare L.) and occurs worldwide in warmer, humid growing conditions. Current Australian barley varieties are largely susceptible to this disease and attempts are being made to introduce sources of resistance from North America. In this study we have compared chromosomal locations of spot blotch resistance reactions in four North American two-rowed barley lines; the North Dakota lines ND11231-12 and ND11231-11 and the Canadian lines TR251 and WPG8412-9-2-1. Diversity arrays technology-based PCR, expressed sequence tag and SSR markers have been mapped across four populations derived from crosses between susceptible parental lines and these four resistant parents to determine the location of resistance loci. Quantitative trait loci (QTL) conferring resistance to spot blotch in adult plants (APR) were detected on chromosomes 3HS and 7HS. In contrast, seedling resistance (SLR) was controlled solely by a locus on chromosome 7HS. The phenotypic variance explained by the APR QTL on 3HS was between 16 and 25% and the phenotypic variance explained by the 7HS APR QTL was between 8 and 42% across the four populations. The SLR QTL on 7HS explained between 52 and 64% of the phenotypic variance. An examination of the pedigrees of these resistance sources supports the common identity of resistance in these lines and indicates that only a limited number of major resistance loci are available in current two-rowed germplasm.

Mapping of adult plant resistance to net form of net blotch in three Australian barley populations

Net form of net blotch (NFNB), caused by Pyrenophora teres Drechs. f. teres Smedeg., is a serious disease problem for the barley industry in Australia and other parts of the world. Three doubled haploid barley populations, Alexis/Sloop, WI2875-1/Alexis, and Arapiles/Franklin, were used to identify genes conferring adult plant resistance to NFNB in field trials. Quantitative trait loci (QTLs) identified were specific for adult plant resistance because seedlings of the parental lines were susceptible to the NFNB isolates used in this study. QTLs were identified on chromosomes 2H, 3H, 4H, and 7H in both the Alexis/Sloop and WI2875-1/Alexis populations and on chromosomes 1H, 2H, and 7H in the Arapiles/Franklin population. Using QTLNetwork, epistatic interactions were identified between loci on chromosomes 3H and 6H in the Alexis/Sloop population, between 2H and 4H in the WI2875-1/Alexis population, and between 5H and 7H in the Arapiles/Franklin population. Comparisons with earlier studies of NFNB resistance indicate the pathotype-dependent nature of many resistance QTLs and the importance of establishing an international system of pathotype nomenclature and differential testing.

Flour yield QTLs in three Australian doubled haploid wheat populations

Flour yield quantitative trait loci (QTLs) were identified in 3 Australian doubled haploid populations, Sunco × Tasman, CD87 × Katepwa, and Cranbrook × Halberd. Trial data from 3 to 4 sites or years were available for each population. QTLs were identified on chromosomes 2BS, 4B, 5AL, and 6BL in the Sunco × Tasman population, on chromosomes 4B, 5AS, and 6DL in the CD87 × Katepwa population, and on chromosomes 4DS, 5DS, and 7AS in the Cranbrook × Halberd population. In the Sunco × Tasman cross the highest genetic variance was detected with the QTL on chromosome 2B (31.3%), in the CD87 × Katepwa cross with the QTL on chromosome 4B (23.8%), and in the Cranbrook × Halberd cross with the QTL on chromosome 5D (18%). Only one QTL occurred in a similar location in more than one population, indicating the complexity of the flour yield character across different backgrounds.

Chromosome composition of an F2 Triticum aestivum × T. turgidum spp. durum cross analysed by DArT markers and MCFISH

This study has employed multicolour fluorescence in situ hybridisation (MCFISH) and Diversity Arrays Technology (DArT) markers to determine the segregation of parental A, B and D genome material into the progeny of a cross between a hexaploid bread wheat (Triticum aestivum L. var. 2-49) and a tetraploid durum wheat [T. turgidum L. spp. durum (Desf.) var. Bellaroi]. In the F2 progeny from a 2-49/Bellaroi cross, 82 out of 83 F2 plants investigated with DArT analysis carried some D genome material, principally as entire chromosomes, while 40 plants included at least one complete copy of all seven D genome chromosomes. Twelve plants containing partial D chromosomes were identified. MCFISH analysis of 26 additional F2 plants of the same cross showed that all 26 plants contained varying amounts of D genome material of which three carried single A-D translocations. In addition two telocentric D genome chromosomes were detected. The D genome content of each line and the breakpoint positions of the three A-D translocations were confirmed with DArT marker analysis. Overall results indicate a random recombination of A and B genome loci from the hexaploid female parent and the tetraploid male parent in this F2 population and a significant retention of the maternal D genome material. This study illustrates that the combined application of the MCFISH and DArT techniques provides a powerful approach for the analysis of crosses between cereal genotypes of different ploidy.

Simple sequence repeat markers associated with three quantitative trait loci for black point resistance can be used to enrich selection populations in bread wheat

Black point in wheat has the potential to cost the Australian industry

Genetic Mapping in the Triticeae

A30.4 million a year. It is difficult and expensive to screen for resistance, so the aim of this study was to validate 3 previously identified quantitative trait loci (QTLs) for black point resistance on chromosomes 2B, 4A, and 3D of the wheat variety Sunco. Black point resistance data and simple sequence repeat (SSR) markers, linked to the resistance QTLs and suited to high-throughput assay, were analysed in the doubled haploid population, Batavia (susceptible) × Pelsart (resistant). Sunco and Pelsart both have Cook in their pedigree and both have the Triticum timopheevii translocation on 2B. SSR markers identified for the 3 genetic regions were gwm319 (2B, T. timopheevii translocation), wmc048 (4AS), and gwm341 (3DS). Gwm319 and wmc048 were associated with black point resistance in the validation population. Gwm341 may have an epistatic influence on the trait because when resistance alleles were present at both gwm319 and wmc048, the Batavia-derived allele at gwm341 was associated with a higher proportion of resistant lines. Data are presented showing the level of enrichment achieved for black point resistance, using 1, 2, or 3 of these molecular markers, and the number of associated discarded resistant lines. The level of population enrichment was found to be 1.83-fold with 6 of 17 resistant lines discarded when gwm319 and wmc048 were both used for selection. Interactions among the 3 QTLs appear complex and other genetic and epigenetic factors influence susceptibility to black point. Polymorphism was assessed for these markers within potential breeding material. This indicated that alternative markers to wmc048 may be required for some parental combinations. Based on these results, marker-assisted selection for the major black point resistance QTLs can increase the rate of genetic gain by improving the selection efficiency and may facilitate stacking of black point resistances from different sources.

Genomic regions associated with common root rot resistance in the barley variety Delta

Genetic maps are the fundamental tools to identify features of phenotypes that are linked to specific genetic loci and eventually DNA sequences or genes. The major use of genetic linkage maps has, therefore, been to identify quantitative trait loci (QTL). Genetic maps are also essential for marker-assisted selection, comparative mapping, high-resolution mapping and map-based cloning. To date, over 40 maps with at least 300 markers have been published for different Triticeae populations. The quality of genetic maps can be affected by a number of factors and map curation ensures that map quality issues are identified and, where possible, resolved. We report on the issues involved in the production of quality genetic linkage maps by inspection of marker genotype data after map construction.