Wheat Doubled Haploid Population Research Articles

Mapping QTLs for seedling root traits in a doubled haploid wheat population under different water regimes

A deep and thick root system has a positive effect on wheat yield, particularly in drought environments. A doubled haploid (DH) population of 150 lines derived from the cross Hanxuan 10 × Lumai 14 was used to map QTLs for seedling root characteristics. The DH lines were cultivated in an agarose gel-chamber under well-watered (WW) and water-stressed (WS) regimes. Water stress was simulated by adding mannitol to the agarose gel. The seminal root traits, including maximum root length (MRL), seminal root number, total root length, project root area, root surface area, and seminal root angle were measured after 6 days of seedling development. Grain yields (GY) were measured in a field experiment. A total of 29 QTLs were identified for seedlings cultured under WW regimes, and 23 QTLs under WS regimes. Individual QTL accounted for phenotypic variations ranging from 4.98 to 24.31 %. The QTLs were distributed on 17 chromosomes, except 1D, 4D, 6B and 6D. Seven consistently expressed QTLs were detected for all the traits tested except MRL under both water regimes. The QTLs for root traits were unevenly distributed among chromosomes, and clustered in eight loci on seven chromosomes, showing pleiotropic effects on target traits. One region in the interval Xgwm644.2–P6901.2 on chromosome 3B contained 9 QTLs affecting most root traits. The present data provide an insight into the genetic basis of seedling root development under different water regimes and may benefit breeding programs using marker-assisted selection (MAS) for root traits.

Genetic Analysis of Carbon Isotope Discrimination and its Relation to Yield in a Wheat Doubled Haploid Population

Carbon isotope discrimination (Δ(13)C) is considered a useful indicator for indirect selection of grain yield (GY) in cereals. Therefore, it is important to evaluate the genetic variation in Δ(13)C and its relationship with GY. A doubled haploid (DH) population derived from a cross of two common wheat varieties, Hanxuan 10 (H10) and Lumai 14 (L14), was phenotyped for Δ(13)C in the flag leaf, GY and yield associated traits in two trials contrasted by water availability, specifically, rain-fed and irrigated. Quantitative trait loci (QTLs) were identified by single locus and two locus QTL analyses. QTLs for Δ(13)C were located on chromosomes 1A, 2B, 3B, 5A, 7A and 7B, and QTLs for other traits on all chromosomes except 1A, 4D, 5A, 5B and 6D. The population selected for high Δ(13)C had an increased frequency of QTL for high Δ(13)C, GY and number of spikes per plant (NSP) when grown under rain-fed conditions and only for high Δ(13)C and NSP when grown under irrigated conditions, which was consistent with agronomic performance of the corresponding trait values in the high Δ(13)C progeny; that is, significantly greater than that in the low Δ(13)C. Therefore, selection for Δ(13)C was beneficial in increasing grain yield in rain-fed environments.

Genetic analysis of disease susceptibility contributed by the compatible Tsn1–SnToxA and Snn1–SnTox1 interactions in the wheat-Stagonospora nodorum pathosystem

Stagonospora nodorum is a foliar pathogen of wheat that produces several host-selective toxins (HSTs) and causes the disease Stagonospora nodorum blotch (SNB). The wheat genes Snn1 and Tsn1 confer sensitivity to the HSTs SnTox1 and SnToxA, respectively. The objectives of this study were to dissect, quantify, and compare the effects of compatible Snn1-SnTox1 and Tsn1-SnToxA interactions on susceptibility in the wheat-S. nodorum pathosystem. Inoculation of a wheat doubled haploid population that segregates for both Snn1 and Tsn1 with an S. nodorum isolate that produces both SnTox1 and SnToxA indicated that both interactions were strongly associated with SNB susceptibility. The Snn1-SnTox1 and Tsn1-SnToxA interactions explained 22 and 28% of the variation in disease, respectively, and together they explained 48% indicating that their effects are largely additive. The Snn1-SnTox1 interaction accounted for 50% of the variation when the population was inoculated with an S. nodorum strain where the SnToxA gene had been mutated, eliminating the Tsn1-SnToxA interaction. These results support the theory that the wheat-S. nodorum pathosystem is largely based on multiple host-toxin interactions that follow an inverse gene-for-gene scenario at the host-toxin interface, but disease exhibits quantitative variation due to the mainly additive nature of compatible interactions. The elimination of either Snn1 or Tsn1 toxin sensitivity alleles resulted in decreased susceptibility, but the elimination of both interactions was required to obtain high levels of resistance. We propose the use of molecular markers to select against Snn1, Tsn1, and other toxin sensitivity alleles to develop wheat varieties with high levels of SNB resistance.

Genetics of late maturity α-amylase in a doubled haploid wheat population

Late maturity α-amylase (LMA) in wheat is a defect where high-isoelectric point (pI) α-amylase accumulates in the ripening grain. Wheat genotypes vary in expression from zero to high levels of α-amylase, the latter with detrimental consequences on their use for value-added end products. Expression in each genotype is characterised by varying numbers of grains affected and different levels in each grain. Analysis of a doubled haploid (DH) population (188 lines) from WW1842 × Whistler has identified significant QTL on chromosomes 2DL, 3A, 3B, 3D, 4B, 4D, 5DS and 5BL. The 4B LMA allele (P < 0.0001) from Whistler is closely linked to the QTL for the ‘tall’ allele (P < 0.0001) of the Rht-B1 gene. The 4D LMA QTL (P < 0.0001) in WW1842 co-locates with the QTL for the ‘tall’ allele (P < 0.0001) of the Rht-D1 gene. This study has shown for the first time that a DH cross between two semi-dwarf cultivars with low or no LMA produces ~25% of progeny lines of the ‘tall’ genotypes with a high frequency of LMA. This is attributed to the large additive positive effects from the combination of one recessive ‘tall’ Rht-B1 gene and one recessive ‘tall’ Rht-D1 gene. High-yielding semi-dwarf genotypes with different combinations of Rht-B1 and Rht-D1 alleles which have very low or non-existent LMA expression (e.g. WW1842 and Whistler) may meet industry criteria for registration as commercial wheat varieties. However, when they are used as breeding lines, the cross produces some progeny genotypes with severe levels of LMA. These LMA genotypes comprise the gibberellic acid-sensitive ‘tall’ progenies and a very small proportion of semi-dwarfs. Thus, it is of paramount importance to screen the defect in wheat breeding programs. The suite of QTL identified for LMA will enable the use of marker assisted selection in the pyramiding of the beneficial QTL to maximise yield and minimise (or eliminate) LMA in semi-dwarf genotypes.

Identification of novel tan spot resistance QTLs using an SSR-based linkage map of tetraploid wheat

Durum wheat (Triticum turgidum L. subsp. durum ,2 n= 4x = 28, AABB) is an important cereal used for making pasta products. Compared with bread wheat, durum wheat receives less attention in genetic and genomic studies. In this research, a tetraploid wheat doubled haploid (DH) population derived from the cross between the durum wheat cultivar 'Lebsock' and the T. turgidum subsp. carthlicum (2n = 4x = 28, AABB) accession PI 94749 was developed. The population consisted of 146 lines and was used to construct linkage maps of all 14 chromosomes. The maps consisted of 280 SSR markers and spanned 2,034.1 cM with an average density of one marker per 7.2 cM. The DH population and the whole genome linkage maps were then used to identify QTLs associ- ated with tan spot resistance. The DH population was inoculated separately with two Ptr ToxA-producing isolates (Pti2 and 86-124) representing races 1 and 2, respectively, of Pyrenophora tritici-repentis, and five resistance QTLs were detected on chromosome arms 3AS, 3BL, 5AL and 7BL. Together, the QTLs explained a total of 46 and 41% of the phenotypic variation for reaction to Pti2 and 86-124, respectively. The Tsn1-Ptr ToxA interaction was not a significant factor in tan spot development in this population, and none of the QTLs corresponded to previously identified loci known to confer insensitivity to host-selective toxins (HSTs) produced by P. tritici-repentis. This result, together with those of other similar studies, indicates that the wheat-P. tritici-repentis pathosystem involves more factors than currently published host-toxin interactions. The DH population and genetic maps reported here will be useful for genetic dissection of important agronomic traits as well as the identification and development of markers for marker-assisted selection (MAS).

Identification of novel QTLs for seedling and adult plant leaf rust resistance in a wheat doubled haploid population

Pyramiding of genes that confer partial resistance is a method for developing wheat (Triticum aestivum L.) cultivars with durable resistance to leaf rust caused by Puccinia triticina. In this research, a doubled haploid population derived from the cross between the synthetic hexaploid wheat (SHW) (xAegilotriticum spp.) line TA4152-60 and the North Dakota breeding line ND495 was used for identifying genes conferring partial resistance to leaf rust in both the adult plant and seedling stages. Five QTLs located on chromosome arms 3AL, 3BL, 4DL, 5BL and 6BL were associated with adult plant resistance with the latter four representing novel leaf rust resistance QTLs. Resistance effects of the 4DL QTL were contributed by ND495 and the effects of the other QTLs were contributed by the SHW line. The QTL on chromosome arm 3AL had large effects and also conferred seedling resistance to leaf rust races MJBJ, TDBG and MFPS. The other major QTL, which was on chromosome arm 3BL, conferred seedling resistance to race MFPS and was involved in a significant interaction with a locus on chromosome arm 5DS. The QTLs and the associated molecular markers identified in this research can be used to develop wheat cultivars with potentially durable leaf rust resistance.

A new PCR-based marker on chromosome 4AL for resistance to pre-harvest sprouting in wheat (Triticum aestivum L.)

Pre-harvest sprouting (PHS) is a complex trait controlled by multiple genes with strong interaction between environment and genotype that makes it difficult to select breeding materials by phenotypic assessment. One of the most important genes for pre-harvest sprouting resistance is consistently identified on the long arm of chromosome 4A. The 4AL PHS tolerance gene has therefore been targeted by Australian white-grained wheat breeders. A new robust PCR marker for the PHS QTL on wheat chromosome 4AL based on candidate genes search was developed in this study. The new marker was mapped on 4AL deletion bin 13-0.59-0.66 using 4AL deletion lines derived from Chinese Spring. This marker is located on 4AL between molecular markers Xbarc170 and Xwg622 in the doubled-haploid wheat population Cranbrook × Halberd. It was mapped between molecular markers Xbarc170 and Xgwm269 that have been previously shown to be closely linked to grain dormancy in the doubled haploid wheat population SW95-50213 × Cunningham and was co-located with Xgwm269 in population Janz × AUS1408. This marker offers an additional efficient tool for marker-assisted selection of dormancy for white-grained wheat breeding. Comparative analysis indicated that the wheat chromosome 4AL QTL for seed dormancy and PHS resistance is homologous with the barley QTL on chromosome 5HL controlling seed dormancy and PHS resistance. This marker will facilitate identification of the gene associated with the 4A QTL that controls a major component of grain dormancy and PHS resistance.

Whole genome mapping in a wheat doubled haploid population using SSRs and TRAPs and the identification of QTL for agronomic traits

Genetic maps are useful for detecting quantitative trait loci (QTL) associated with quantitative traits and for marker-assisted selection (MAS) in breeding. In this research, we used the wheat × maize method to develop a doubled haploid (DH) population derived from the synthetic hexaploid wheat (SHW) line TA4152-60 and the North Dakota hard red spring wheat line ND495. The population consisted of 213 lines, of which a subset of 120 lines was randomly selected and used to construct linkage maps of all 21 chromosomes and for QTL detection. The whole genome maps consisted of 632 markers including 410 SSRs, 218 TRAPs, 1 RFLP, and 3 phenotypic markers, and spanned 3,811.5 cM with an average density of one marker per 6.03 cM. Telomere sequence-based TRAPs allowed us to define the ends of seven linkage groups. Analysis revealed major QTLs associated with the traits of days to heading on chromosomes 5A and 5B, plant height on chromosomes 4D and 5A, and spike characteristics on chromosomes 3D, 4A, 4D, 5A and 5B. The DH population and genetic map will be a useful tool for the identification of disease resistance QTL and agronomically important loci, and will aid in the identification and development of markers for MAS.

Matrix‐assisted laser desorption/ionization time‐of‐flight based wheat gliadin protein peaks are useful molecular markers for wheat genetic study

Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) instrumentation has been used to analyze wheat seed gliadins as an alternative to other established methods, including sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), capillary electrophoresis (CE), high-performance liquid chromatography (HPLC), etc. The MALDI-TOF approach has shown to have many advantages such as high resolution, cost effectiveness and high throughput. MALDI-TOF-based gliadin profiles have been used for fast wheat cultivar identification. However, the genetic information represented by individual gliadin peaks has not been utilized. In this study a wheat doubled haploid population with a genetic linkage map of good coverage was used to assay individual gliadin peaks from MALDI-TOF profiles as molecular markers. Eight segregating peaks in the population were scored as polymorphic across the population. The 1 to 1 segregating ratios validated the scoring of the peaks and all peaks were mapped to the expected chromosomes or linkage groups on the available linkage map: 1 peak on chromosome 1A, 1 on 6A, 4 on 6B and 2 on 6D.

Identification of novel QTL for resistance to crown rot in the doubled haploid wheat population ‘W21MMT70’ × ‘Mendos’

AbstractCrown rot (causal agent Fusarium pseudograminearum) is a fungal disease of major significance to wheat cultivation in Australia. A doubled haploid wheat population was produced from a cross between line ‘W21MMT70’, which displays partial seedling and adult plant (field) resistance to crown rot, and ‘Mendos’, which is moderately susceptible in seedling tests but partially resistant in field trials. Bulked segregant analysis (BSA) based on seedling trial data did not reveal markers for crown rot resistance. A framework map was produced consisting of 128 microsatellite markers, four phenotypic markers, and one sequence tagged site marker. To this map 331 previously screened AFLP markers were then added. Three quantitative trait loci (QTL) were identified with composite interval mapping across all of the three seedling trials conducted. These QTL are located on chromosomes 2B, 2D and 5D. The 2D and 5D QTL are inherited from the line ‘W21MMT70’, whereas the 2B QTL is inherited from ‘Mendos’. These loci are different from those associated with crown rot resistance in other wheat populations that have been examined, and may represent an opportunity for pyramiding QTL to provide more durable resistance to crown rot.

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.

Genetic characterisation of dough rheological properties in a wheat doubled haploid population: additive genetic effects and epistatic interactions

Doubled haploid lines (n = 160) from a cross between wheat cultivars 'Cranbrook' (high dough extensibility) and 'Halberd' (low dough extensibility) were grown at three Australian locations. The parents differ at all high- and low-molecular-weight glutenin loci. Dough rheological parameters were measured using small-scale testing procedures, and quantitative trait locus (QTL) mapping procedures were carried out using an existing well-saturated genetic linkage map for this cross. Genetic parameters were estimated using three software packages: QTLCartographer, Epistat and Genstat. Results indicated that environmental factors are a major determinant of dough extensibility across the three trial sites, whereas genotypic factors are the major determinants of dough strength. Composite interval mapping analysis across the 21 linkage groups revealed that as expected, the main additive QTLs for dough rheological properties are located at the high- and low-molecular-weight glutenin loci. A new QTL on chromosome 5A for M-extensibility (a mixograph-estimated measure of extensibility) was detected. Analysis of epistatic interactions revealed that there were significant conditional epistatic interactions related with the additive effects of glutenin loci on dough rheological properties. Therefore, the additive genetic effects of glutenins on dough rheological properties are conditional upon the genetic background of the wheat line. The molecular basis of the interactions with the glutenin loci may be via proteins that modify or alter the gluten protein matrix or variations in the expression level of the glutenin genes. Reverse-phase high performance liquid chromatography analysis of the molar number of individual glutenin subunits across the population showed that certain conditional epistases resulted in increased expression of the affected glutenin. The epistatic interactions detected in this study provide a possible explanation of the variable genetic effects of some glutenins on quality attributes in different genetic backgrounds and provide essential information for the accurate prediction of glutenin related variance in marker-assisted wheat breeding.

Mapping of genes expressed inFusarium graminearum-infected heads of wheat cultivar 'Frontana'

The isolation, physical, and genetic mapping of a group of wheat genes expressed in infected heads of Triticum aestivum 'Frontana' resistant to Fusarium head blight is reported. A cDNA library was built from heads of 'Frontana' through suppressive subtractive hybridization, to enrich for sequences induced by the pathogen Fusarium graminearum during infection. A group of 1794 clones was screened by dot blot hybridization for differential gene expression following infection. Twenty of these clones showed a strong difference in intensity of hybridization between infected and mock-inoculated wheat head samples, suggesting that they corresponded to genes induced during infection. The 20 clones were sequenced and used for mapping analysis. We determined a precise chromosomal location for 14 selected clones by using series of chromosome deletion stocks. It was shown that the 14 clones detected 90 fragments with the use of the restriction enzyme EcoRI; 52 bands were assigned to chromosome bins, whereas 38 fragments could not be assigned. The selected clones were also screened for polymorphisms on a 'Wuhan' x 'Maringa' wheat doubled haploid mapping population. One clone, Ta01_02b03, was related to a quantitative trait locus for type II resistance located on chromosome 2AL, as determined with simple sequence repeat markers on another mapping population, but did not map in the same location on our population. Another clone, Ta01_06f04, was identified by BLAST (basic local alignment search tool) search in public databases to code for a novel beta-1,3-glucanase, homologous to a major pathogenesis-related protein. This clone mapped to chromosomal regions on chromosome 3, including 3BL and 3DL, where B glucanase gene clusters are known to exist. Seven other clones, including 1 coding for an ethylene-response element binding protein and 3 for ribosomal proteins, and 4 clones corresponding to proteins with unknown function, were also mapped.

Development of a doubled haploid mapping population and linkage map for the bread wheat cross Kariega × Avocet S

A doubled haploid wheat population of 150 lines was constructed from the F1 of a cross between cultivars Kariega and Avocet S using the wheat-maize technique. Segregation and linkage analysis of 203 DNA markers, two storage protein markers and the stem rust resistance gene Sr26 yielded 31 linkage groups of which 28 were chromosome-anchored. These linkage groups covered the entire genome of 21 chromosomes. SSR markers were advantageous for chromosome anchoring and AFLP markers for consolidation of the various linkage groups. A significant feature of the linkage map is a relatively low level of polymorphism for markers on the D genome, viz, 18.5% of all markers mapped. The Kariega × Avocet S doubled haploid population and linkage maps have emerged as valuable resources for further genetic studies of qualitative and quantitative traits of economic importance in bread wheat.

Annual Meeting, Ottawa, Ontario, 2004

Annual Meeting, Ottawa, Ontario, 2004

Flour proteins linked to quality traits in an Australian doubled haploid wheat population

The Cranbrook/Halberd doubled haploid population has provided a unique opportunity to examine in detail the contributions made by a number of different high molecular weight (HMW) and low molecular weight (LMW) glutenin alleles to the dough properties in a set of homogeneous lines of wheat. A range of different instruments was employed, including Farinograph, Extensograph, Do-Corder, Resistograph, and GRL/EasyMix, to study the dough rheology of the lines from 3 sites over 2 years. Correlation studies showed that 2 basic parameters (dough strength and extensibility) were measured by these different instruments. The results presented are mainly from the Extensograph, which is a major Australian standard for determining release and marketing classification of Australian wheats. Approaches to investigate the data include bulk segregant analysis, distribution of protein alleles in the population, and multiple linear regression. As expected, the HMW glutenin alleles made a major contribution to dough strength, with a minor, but not insignificant, contribution from the LMW glutenin alleles. From a knowledge of their glutenin alleles, a glutenin strength score (GSS) was devised to allow breeders to rank the dough strength of various lines. The GSS scoring system is based on both HMW and LMW glutenin alleles, adding to a total out of 10. Extensibility, on the other hand, was predominantly influenced by protein levels in the flour and environmental conditions such as site and season. However, the LMW glutenin alleles make a significant genetic contribution to the extensibility, which can be assessed by using a glutenin extensibility score. These two glutenin quality scores currently include only the alleles present in the parents, Cranbrook and Halberd, but this could be expanded to include a wider range of alleles by analysis of the quality data from other doubled haploid populations. These quality scores would then be an extremely useful tool for assessing the potential quality of parental and early generation germplasm in wheat breeding programs, by a knowledge of the allelic composition of their HMW and LMW glutenins.

Segregation of glutenins in wheat × maize-derived doubled haploid wheat populations

The segregation of both high and low molecular weight glutenin subunits across 7 F1 wheat (Triticum aestivum L.) × maize (Zea mays L.) derived doubled haploid populations was examined. The F1 wheats used in each population were produced from parents of interest to Australian wheat breeding programs. The parents varied by up to 5 glutenin subunit loci. Examination of subunits individually within each population using a chi-square analysis revealed that all but 2 of the 26 pairs of alleles analysed fitted the expected 1 : 1 segregation ratio. Glutenin profiles were examined for each cross individually and all but one (Sonalika/Hartog) fitted the expected Mendelian segregation pattern. The analysis of allele distribution of the 6 glutenin loci across all 7 crosses showed all falling well within expected segregation ratios. Closer examination of parental lines and populations revealed irregularities which conflict with original assumptions and provide a valid explanation for the few segregation distortions observed. It is concluded that wheat × maize-derived doubled haploid populations represent a unbiased assortment of parental gametes on both arms of Group 1 chromosomes.