Paul Eckermann

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.

QTL analysis of malting quality traits in two barley populations

Selection for malting quality traits is a major breeding objective for barley breeding programs. With molecular markers linked to loci affecting these traits, this selection can be undertaken at an earlier stage of the breeding program than is possible using conventional tests. Quantitative trait loci (QTLs) associated with malting quality traits were mapped in 2 populations derived from parents with elite malting quality. Progeny from an Arapiles/Franklin population grown in 4 environments and an Alexis/Sloop population grown in 5 environments were tested for grain protein percentage, α-amylase activity, diastatic power, hot water extract, wort viscosity, wort β-glucan, β-glucanase, and free α-amino acids. QTL analysis was performed using a one-stage approach, which allowed for modelling of spatial variation in the field, and in each phase of the malting quality analysis in the laboratory. QTLs for malting quality traits were detected on all chromosomes and for both populations. Few of these QTLs were significant in all of the environments, indicating that QTL × environment interactions were important. There were many coincident QTLs for traits that are expected to be related such as diastatic power and α-amylase activity, wort β-glucan and wort viscosity and for some traits that are not expected to be related such as hot water extract and malt viscosity.

The analysis of quantitative trait loci in multi-environment trials using a multiplicative mixed model

A new approach for multi-environment quantitative trait locus (QTL) analysis based on an appropriate genetic model is presented. To accommodate a multi-environment analysis, the size of a QTL effect is assumed to be a random effect. The approach results in a multiplicative mixed model for QTL × environment interaction of the factor analytic type. The full genetic model may also include a factor analytic model for the residual genotype × environment interaction, whereas the environmental model for the non-genetic variation involves local, global, and extraneous variation. The approach is used to determine QTLs for yield in the Arapiles × Franklin doubled haploid population of the National Barley Molecular Marker Program. Analysis leads to the determination of 8 QTLs. Many of these QTLs are associated with other traits.

The analysis of quantitative traits in wheat mapping populations

This paper discusses the analysis of quantitative trait loci (QTLs) using molecular markers from a doubled haploid wheat mapping population arising from the Cranbrook Halberd cross. Two field trials are used to provide phenotypic information on the trait of interest, which is grain percentage protein. Methods for QTL analysis are reviewed together with methods for the analysis of field trials. The aim of the paper is to examine different approaches for QTL analysis, namely the conventional approach available in standard software, which ignores field variation, a 2-stage approach that provides adjusted phenotypic effects for a subsequent QTL analysis, and a joint marker and spatial analysis. The major effect, however, is the maturity class of the doubled haploid lines. Maturity and percent protein appear highly correlated genetically so QTL analysis shows marked changes if maturity is included as a covariate. More subtle changes occur due to field variation but this may not be the standard situation.

A DNA-based method for studying root responses to drought in field-grown wheat genotypes

Root systems are critical for water and nutrient acquisition by crops. Current methods measuring root biomass and length are slow and labour-intensive for studying root responses to environmental stresses in the field. Here, we report the development of a method that measures changes in the root DNA concentration in soil and detects root responses to drought in controlled environment and field trials. To allow comparison of soil DNA concentrations from different wheat genotypes, we also developed a procedure for correcting genotypic differences in the copy number of the target DNA sequence. The new method eliminates the need for separation of roots from soil and permits large-scale phenotyping of root responses to drought or other environmental and disease stresses in the field.

New eSSR and gSSR markers added to Australian barley maps

To enhance genetic maps of barley previously developed in Australia for identifying markers useable in molecular breeding, a new set of simple sequence repeat (SSR) and indel markers was added to the maps. These markers were developed through (i) database mining of barley expressed sequence tag (EST) sequences, (ii) comparative barley-rice genome analysis, and (iii) screening of a genomic library with SSR probes. The primer set selected for this study comprised 216 EST-SSR (eSSR) and 25 genomic SSR (gSSR) markers, which were screened for polymorphism on 4 doubled haploid (DH) or recombinant inbred line (RIL) populations. In total, 81 new markers were added to the maps, with good coverage on all 7 chromosomes, except 6H, which only had 2 new markers added. The marker order of previously published maps was re-evaluated by comparing recombination fractions calculated by 2 methods to discover the best position for each marker. The new SSR markers were then added to the updated maps. Several of these new markers are linked to important barley disease resistance genes such as those for cereal cyst nematode, spot form of net blotch, and leaf scald resistance, and are readily useable for marker-assisted barley breeding. The new maps are available on-line at www.genica.net.au.

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.

Optical and physical mapping with local finishing enables megabase-scale resolution of agronomically important regions in the wheat genome

BackgroundNumerous scaffold-level sequences for wheat are now being released and, in this context, we report on a strategy for improving the overall assembly to a level comparable to that of the human genome.ResultsUsing chromosome 7A of wheat as a model, sequence-finished megabase-scale sections of this chromosome were established by combining a new independent assembly using a bacterial artificial chromosome (BAC)-based physical map, BAC pool paired-end sequencing, chromosome-arm-specific mate-pair sequencing and Bionano optical mapping with the International Wheat Genome Sequencing Consortium RefSeq v1.0 sequence and its underlying raw data. The combined assembly results in 18 super-scaffolds across the chromosome. The value of finished genome regions is demonstrated for two approximately 2.5 Mb regions associated with yield and the grain quality phenotype of fructan carbohydrate grain levels. In addition, the 50 Mb centromere region analysis incorporates cytological data highlighting the importance of non-sequence data in the assembly of this complex genome region.ConclusionsSufficient genome sequence information is shown to now be available for the wheat community to produce sequence-finished releases of each chromosome of the reference genome. The high-level completion identified that an array of seven fructosyl transferase genes underpins grain quality and that yield attributes are affected by five F-box-only-protein-ubiquitin ligase domain and four root-specific lipid transfer domain genes. The completed sequence also includes the centromere.