Chromosome 4AL Research Articles

Adult-plant resistance to Septoria tritici blotch in hexaploid spring wheat.

New QTL for Septoria tritici blotch detected in hexapoid spring wheat under field conditions across diverse environments. Septoria tritici blotch caused by the ascomycete fungus Zymoseptoria tritici presents a serious and consistent challenge to global wheat production. In particular the augmented use of soil management practices that leave large amounts of wheat stubble on the soil surface and global warming increases the chance of Septoria tritici blotch epidemics to emerge more frequently including in developing countries. Two recombinant inbred line populations developed from a cross between the susceptible Moroccan spring bread wheat variety 'NASMA' and the CIMMYT resistant lines, 'IAS20*5/H567.71' and 'RPB709.71/COC' were used to study the genetics and map adult-plant resistance to Septoria tritici blotch under field conditions in different environments. Resistance to Septoria tritici blotch in both populations was quantitative and overall, five across environment consistent resistance loci on chromosomes 1BS, 3AL, 5AL and 7AS were detected in the two populations. The QTL on chromosome 1BS and 7AS are likely to be allelic with the known Septoria tritici blotch genes Stb3 and Stb11. All identified QTL were additive and explained between 4 and 27% of the phenotypic variation. Epistatic interaction was not observed. Low cost KASP assays were developed as flanking markers for all five QTL that will facilitate molecular breeding. Our study represents the first mapping effort under field conditions utilizing two spring bread wheat resistant sources evaluated over multiple environments.

Molecular Characterization of TaFAR1 Involved in Primary Alcohol Biosynthesis of Cuticular Wax in Hexaploid Wheat.

Cuticular waxes are complex mixtures of very long chain (VLC) fatty acids and their derivatives in which primary alcohols are the most abundant components in the leaf surface of common wheat (Triticum aestivum L.). However, the genes involved in primary alcohol biosynthesis in wheat are still largely unknown. Here we identified, via a homology-based approach, the TaFAR1 gene belonging to the fatty acyl-CoA reductases (FARs) from wheat. Heterologous expression of TaFAR1 in yeast (Saccharomyces cerevisiae) and in the Arabidopsis (Arabidopsis thaliana) cer4-3 mutant afforded production of C22 primary alcohol and C22-C24 primary alcohols, respectively, and transgenic expression of TaFAR1 in tomato (Solanum lycopersicum) cv MicroTom leaves and fruits resulted in the accumulation of C26-C30 primary alcohols and C30-C34 primary alcohols, respectively. The TaFAR1 protein was localized to the endoplasmic reticulum (ER) in rice (Oryza sativa L.) leaf protoplasts. Moreover, the TaFAR1 expression pattern across various organs correlated with the levels of primary alcohols accumulating in corresponding waxes, and with the presence of platelet-shaped epicuticular wax crystals formed by primary alcohols. A nullisomic-tetrasomic wheat line lacking TaFAR1 had significantly reduced levels of primary alcohols in its leaf blade and anther wax. TaFAR1 was located on chromosome 4AL and appeared to be highly conserved, with only one haplotype among 32 wheat cultivars. Finally, TaFAR1 expression was induced by drought and cold stress in an ABA-dependent manner. Taken together, our results show that TaFAR1 is an active enzyme forming primary alcohols destined for the wheat cuticle.

Fine mapping of powdery mildew resistance genes PmTb7A.1 and PmTb7A.2 in Triticum boeoticum (Boiss.) using the shotgun sequence assembly of chromosome 7AL.

A novel powdery mildew resistance gene and a new allele of Pm1 were identified and fine mapped. DNA markers suitable for marker-assisted selection have been identified. Powdery mildew caused by Blumeria graminis is one of the most important foliar diseases of wheat and causes significant yield losses worldwide. Diploid A genome species are an important genetic resource for disease resistance genes. Two powdery mildew resistance genes, identified in Triticum boeoticum (A(b)A(b)) accession pau5088, PmTb7A.1 and PmTb7A.2 were mapped on chromosome 7AL. In the present study, shotgun sequence assembly data for chromosome 7AL were utilised for fine mapping of these Pm resistance genes. Forty SSR, 73 resistance gene analogue-based sequence-tagged sites (RGA-STS) and 36 single nucleotide polymorphism markers were designed for fine mapping of PmTb7A.1 and PmTb7A.2. Twenty-one RGA-STS, 8 SSR and 13 SNP markers were mapped to 7AL. RGA-STS markers Ta7AL-4556232 and 7AL-4426363 were linked to the PmTb7A.1 and PmTb7A.2, at a genetic distance of 0.6 and 6.0cM, respectively. The present investigation established that PmTb7A.1 is a new powdery mildew resistance gene that confers resistance to a broad range of Bgt isolates, whereas PmTb7A.2 most probably is a new allele of Pm1 based on chromosomal location and screening with Bgt isolates showing differential reaction on lines with different Pm1 alleles. The markers identified to be linked to the two Pm resistance genes are robust and can be used for marker-assisted introgression of these genes to hexaploid wheat.

Mapping quantitative trait loci for peroxidase activity and developing gene-specific markers for TaPod-A1 on wheat chromosome 3AL.

Three novel QTL for peroxidase activity were mapped, and gene-specific markers for TaPod-A1 were developed and validated using RILs derived from the Doumai/Shi 4185 cross and 281 wheat cultivars. TaPod-A1 is within one of the three QTL. Peroxidase (POD) activity in grain is an important factor determining the color of flour and end-use products of wheat, such as noodles and steamed bread. Mapping QTL for POD activity, characterization of POD genes and development of gene-specific markers are important for molecular marker-assisted selection in wheat breeding. Quantitative trait loci (QTL) for POD activity in common wheat were mapped using a recombinant inbred line (RIL) population derived from a Doumai/Shi 4185 cross grown in four environments and genotyped using the wheat 90 K iSelect assay. Three novel QTL for POD activity, QPod.caas-3AL, QPod.caas-4BS and QPod.caas-5AS, were identified on chromosomes 3AL, 4BS and 5AS, explaining 5.3-21.2% of phenotypic variance across environments. The full-length genomic DNA (gDNA) sequence of a POD gene, designated TaPod-A1, on chromosome 3A was characterized by homolog cloning and PCR verification. Two complementary dominant sequence-tagged site (STS) markers, POD-3A1 and POD-3A2, were developed based on single nucleotide polymorphisms (SNPs) between two alleles at the TaPod-A1 locus, amplifying 291- and 766-bp fragments in cultivars with lower and higher POD activities, respectively. The two gene-specific markers were mapped on chromosome 3AL using a set of Chinese Spring (CS) nulli-tetrasomic lines, and ditelosomic lines 3AL and 3AS. QTL analysis indicated that QPod.caas-3AL co-segregated with the gene-specific markers POD-3A1 and POD-3A2. POD-3A1 and POD-3A2 were verified on 281 wheat cultivars and advanced lines, and showed significant (P < 0.05) associations with POD activities. POD-3A1 and POD-3A2 may be useful as markers for improving color attributes in wheat breeding programs.

Identification of a member of the catalase multigene family on wheat chromosome 7A associated with flour b* colour and biological significance of allelic variation.

Carotenoids (especially lutein) are known to be the pigment source for flour b* colour in bread wheat. Flour b* colour variation is controlled by a quantitative trait locus (QTL) on wheat chromosome 7AL and one gene from the carotenoid pathway, phytoene synthase, was functionally associated with the QTL on 7AL in some, but not all, wheat genotypes. A SNP marker within a sequence similar to catalase (Cat3-A1snp) derived from full-length (FL) cDNA (AK332460), however, was consistently associated with the QTL on 7AL and implicated in regulating hydrogen peroxide (H2O2) to control carotenoid accumulation affecting flour b* colour. The number of catalase genes on chromosome 7AL was investigated in this study to identify which gene may be implicated in flour b* variation and two were identified through interrogation of the draft wheat genome survey sequence consisting of five exons and a further two members having eight exons identified through comparative analysis with the single catalase gene on rice chromosome 6, PCR amplification and sequencing. It was evident that the catalase genes on chromosome 7A had duplicated and diverged during evolution relative to its counterpart on rice chromosome 6. The detection of transcripts in seeds, the co-location with Cat3-A1snp marker and maximised alignment of FL-cDNA (AK332460) with cognate genomic sequence indicated that TaCat3-A1 was the member of the catalase gene family associated with flour b* colour variation. Re-sequencing identified three alleles from three wheat varieties, TaCat3-A1a, TaCat3-A1b and TaCat3-A1c, and their predicted protein identified differences in peroxisomal targeting signal tri-peptide domain in the carboxyl terminal end providing new insights into their potential role in regulating cellular H2O2 that contribute to flour b* colour variation.

Forcing the shift of the crossover site to proximal regions in wheat chromosomes.

Terminal deletions obligate the first crossover to be formed in more proximal positions. This increases the recombination rate in intercalary intervals but not in the proximity of the centromere. Crossovers are not uniformly distributed along chromosomes in wheat. They take place preferentially in distal positions. The effect of the chromosomal architecture on crossover positioning has been analyzed from the chiasmate bonds at metaphase I formed by the truncated arms of 51 terminal deletion lines of eight wheat chromosomes. Chromosome 4A and the B genome chromosomes, in their standard or truncated conformation, and their arms, were identified by C-banding. Chromosomes studied show a similar chiasma distribution. Reduction of the size of the truncated arms is accompanied by a gradual decrease of the chiasma frequency in chromosome arms 1BL, 3BS, 3BL, 4BL, 5BS, 5BL, 6BL, 7BS, 7BL and 4AL. In chromosome arm 1BS, most chiasmata are concentrated in the distal half of the satellite and, in 4AS, in the distal 24 %. The arms 2BS, 2BL and 6BS do not show a simple decreasing gradient of the recombination rate, the chiasma frequency increases in subdistal intervals compared to more distal regions. Although terminal deletions usually induce an increase of chiasma frequency in intercalary regions, the level of intact chromosome arms is maintained in only a few deletion lines. Truncated arms containing only the 20 % proximal of the intact arm do not form chiasmata. The relationships of chiasma positioning with chromatin structure and genome organization is discussed.

Marker Assisted Transfer of Two Powdery Mildew Resistance Genes PmTb7A.1 and PmTb7A.2 from Triticum boeoticum (Boiss.) to Triticum aestivum (L.).

Powdery mildew (PM), caused by Blumeria graminis f. sp. tritici, is one of the important wheat diseases, worldwide. Two PM resistance genes, designated as PmTb7A.1 and PmTb7A.2, were identified in T. boeoticum acc. pau5088 and mapped on chromosome 7AL approximately 48cM apart. Two resistance gene analogue (RGA)-STS markers Ta7AL-4556232 and 7AL-4426363 were identified to be linked to the PmTb7A.1 and PmTb7A.2, at a distance of 0.6cM and 6.0cM, respectively. In the present study, following marker assisted selection (MAS), the two genes were transferred to T. aestivum using T. durum as bridging species. As many as 12,317 florets of F1 of the cross T. durum /T. boeoticum were pollinated with T. aestivum lines PBW343-IL and PBW621 to produce 61 and 65 seeds, respectively, of three-way F1. The resulting F1s of the cross T. durum/T. boeoticum//T. aestivum were screened with marker flanking both the PM resistance genes PmTb7A.1 and PmTb7A.2 (foreground selection) and the selected plants were backcrossed to generate BC1F1. Marker assisted selection was carried both in BC1F1 and the BC2F1 generations. Introgression of alien chromatin in BC2F1 plants varied from 15.4 - 62.9 percent. Out of more than 110 BC2F1 plants showing introgression for markers linked to the two PM resistance genes, 40 agronomically desirable plants were selected for background selection for the carrier chromosome to identify the plants with minimum of the alien introgression. Cytological analysis showed that most plants have chromosome number ranging from 40-42. The BC2F2 plants homozygous for the two genes have been identified. These will be crossed to generate lines combining both the PM resistance genes but with minimal of the alien introgression. The PM resistance gene PmTb7A.1 maps in a region very close to Sr22, a stem rust resistance gene effective against the race Ug99. Analysis of selected plants with markers linked to Sr22 showed introgression of Sr22 from T. boeoticum in several BC2F1 plants. Thus, in addition to PM resistance, these progeny might also carry resistance to stem rust race Ug99.

Genetic evidence for differential selection of grain and embryo weight during wheat evolution under domestication.

Wheat is one of the Neolithic founder crops domesticated ~10 500 years ago. Following the domestication episode, its evolution under domestication has resulted in various genetic modifications. Grain weight, embryo weight, and the interaction between those factors were examined among domesticated durum wheat and its direct progenitor, wild emmer wheat. Experimental data show that grain weight has increased over the course of wheat evolution without any parallel change in embryo weight, resulting in a significantly reduced (30%) embryo weight/grain weight ratio in domesticated wheat. The genetic factors associated with these modifications were further investigated using a population of recombinant inbred substitution lines that segregated for chromosome 2A. A cluster of loci affecting grain weight and shape was identified on the long arm of chromosome 2AL. Interestingly, a novel locus controlling embryo weight was mapped on chromosome 2AS, on which the wild emmer allele promotes heavier embryos and greater seedling vigour. To the best of our knowledge, this is the first report of a QTL for embryo weight in wheat. The results suggest a differential selection of grain and embryo weight during the evolution of domesticated wheat. It is argued that conscious selection by early farmers favouring larger grains and smaller embryos appears to have resulted in a significant change in endosperm weight/embryo weight ratio in the domesticated wheat. Exposing the genetic factors associated with endosperm and embryo size improves our understanding of the evolutionary dynamics of wheat under domestication and is likely to be useful for future wheat-breeding efforts.

Transcriptomic analysis of wheat near-isogenic lines identifies PM19-A1 and A2 as candidates for a major dormancy QTL.

BackgroundNext-generation sequencing technologies provide new opportunities to identify the genetic components responsible for trait variation. However, in species with large polyploid genomes, such as bread wheat, the ability to rapidly identify genes underlying quantitative trait loci (QTL) remains non-trivial. To overcome this, we introduce a novel pipeline that analyses, by RNA-sequencing, multiple near-isogenic lines segregating for a targeted QTL.ResultsWe use this approach to characterize a major and widely utilized seed dormancy QTL located on chromosome 4AL. It exploits the power and mapping resolution afforded by large multi-parent mapping populations, whilst reducing complexity by using multi-allelic contrasts at the targeted QTL region. Our approach identifies two adjacent candidate genes within the QTL region belonging to the ABA-induced Wheat Plasma Membrane 19 family. One of them, PM19-A1, is highly expressed during grain maturation in dormant genotypes. The second, PM19-A2, shows changes in sequence causing several amino acid alterations between dormant and non-dormant genotypes. We confirm that PM19 genes are positive regulators of seed dormancy.ConclusionsThe efficient identification of these strong candidates demonstrates the utility of our transcriptomic pipeline for rapid QTL to gene mapping. By using this approach we are able to provide a comprehensive genetic analysis of the major source of grain dormancy in wheat. Further analysis across a diverse panel of bread and durum wheats indicates that this important dormancy QTL predates hexaploid wheat. The use of these genes by wheat breeders could assist in the elimination of pre-harvest sprouting in wheat.Electronic supplementary materialThe online version of this article (doi:10.1186/s13059-015-0665-6) contains supplementary material, which is available to authorized users.

Genome-wide association mapping of preharvest sprouting resistance in a diversity panel of European winter wheats.

Global wheat production will benefit from cultivars showing genetic resistance to preharvest sprouting (PHS). Working on PHS resistance is still challenging due to the lack of simple protocols for the provocation of symptoms for appropriate trait differentiation under highly variable environmental conditions. Therefore, the availability of molecular markers for enhancing PHS resistance in breeding lines is of utmost importance. Genome-wide association mapping was performed to unravel the genetics of PHS resistance in a diversity panel of 124 winter wheat genotypes using both random and targeted marker locus approaches. Data for grain germination tests, spike wetting treatments, and field sprouting damage measurements of grains were collected in 11, 12, and four environments, respectively. Twenty-two quantitative trait loci (QTL) linked with 40 markers were detected for the three traits commonly used for assessing the PHS resistance of cultivars. All but five QTL on chromosomes 1B, 1D (two QTL), 3D, and 5D showed locations similar to previous studies, including prominent QTL on chromosomes 2BS, 3AS, and 4AL. The highest retrieval rate across environments was found for QTL on chromosomes 1D, 2BS, 3D, 4AL, and 7B. The study identified genomic signatures useful for marker-assisted improvement of PHS resistance not only in European breeding programs, but of global significance.

Identification of a stem rust resistance locus effective against Ug99 on wheat chromosome 7AL using a RAD-Seq approach.

A locus of major effect for stem rust resistance, effective against Ug99 and possibly a target of a suppressor on chromosome arm 7DL in wheat cultivar Canthatch, was mapped to 7AL. Wheat stem rust, caused by Puccinia graminis f. sp. tritici (Pgt), is responsible for major production losses around the world. The development of resistant cultivars is an effective and environmentally friendly way to manage the disease, but outbreaks can occur when new pathogen races overcome the existing host resistance genes. Ug99 (race TTKSK) and related Pgt races are virulent to the majority of existing cultivars, which presents a potential threat to global wheat production. The hexaploid wheat cultivar Canthatch has long been known to carry a suppressor of stem rust resistance on chromosome arm 7DL. Multiple "non-suppressor" mutants of Canthatch are reported to have gained resistance to Pgt races, including Ug99 (TTKSK) and related races TTKST and TTTSK. To genetically map the suppressor locus, a mapping population was developed from a cross between the susceptible cultivar Columbus, thought to possess the suppressor, and Columbus-NS766, a resistant, near-isogenic line believed to contain a mutant non-suppressor allele introgressed from Canthatch. Genetic mapping using a 9K SNP genotyping assay and restriction site-associated DNA sequencing (RAD-Seq) on bulked segregants led to the identification of markers linked to a locus of stem rust resistance. Surprisingly, genomic sequence information revealed the markers to be located on 7AL instead of 7DL, indicating that the resistance phenotype was due to a new resistance locus, rather than the inactivated suppressor. We suggest that the 7AL locus of resistance is most likely suppressed by the 7DL suppressor.

Genotyping-by-sequencing (GBS) identified SNP tightly linked to QTL for pre-harvest sprouting resistance.

Using a GBS-SNP map, a QTL for pre-harvest sprouting resistance on 4AL of Totoumai A was delimited to 2.9-cM interval, and SNP closely linked to several other QTL were identified. Pre-harvest sprouting (PHS) of wheat is a major constraint to wheat production in many wheat-growing areas worldwide, because it reduces both wheat grain yield and the end-use quality. To identify markers tightly linked to the quantitative trait loci (QTL) for PHS resistance and seed dormancy (SD), we evaluated 155 recombinant inbred lines (RIL) derived from a cross between a PHS-resistant parent 'Tutoumai A' and a PHS-susceptible parent 'Siyang 936' for single-nucleotide polymorphisms (SNP) using genotyping-by-sequencing (GBS), and for PHS resistance and SD using both field and greenhouse grown plants. Two SNP, GBS109947 and GBS212432, were mapped to a major QTL region for PHS resistance and SD on chromosome 4AL, and delimited the QTL to a 2.9-cM interval. Two and nine additional SNP were mapped to minor QTL regions for SD on chromosome 5B and 5A, respectively. Critical SNP in these QTL regions were converted into KBioscience Competitive Allele-Specific PCR (KASP) assays that can be easily used for marker-assisted selection to improve PHS resistance.

Genome-wide DArT and SNP scan for QTL associated with resistance to stripe rust (Puccinia striiformis f. sp. tritici) in elite ICARDA wheat (Triticum aestivum L.) germplasm.

Identified DArT and SNP markers including a first reported QTL on 3AS, validated large effect APR on 3BS. The different genes can be used to incorporate stripe resistance in cultivated varieties. Stripe rust [yellow rust, caused by Puccinia striiformis f. sp. tritici (Pst)] is a serious disease in wheat (Triticum aestivum). This study employed genome-wide association mapping (GWAM) to identify markers linked to stripe rust resistance genes using Diversity Arrays Technology (DArT(®)) and single-nucleotide polymorphism (SNP) Infinium 9K assays in 200 ICARDA wheat genotypes, phenotyped for seedling and adult plant resistance in two sites over two growing seasons in Syria. Only 25.8% of the genotypes showed resistance at seedling stage while about 33 and 44% showed moderate resistance and resistance response, respectively. Mixed-linear model adjusted for false discovery rate at p<0.05 identified 12 DArT and 29 SNP markers on chromosome arms 3AS, 3AL, 1AL, 2AL, 2BS, 2BL, 3BS, 3BL, 5BL, 6AL, and 7DS significantly linked to Pst resistance genes. Of these, the locus on 3AS has not been previously reported to confer resistance to stripe rust in wheat. The QTL on 3AS, 3AL, 1AL, 2AL, and 2BS were effective at seedling and adult plant growth stages while those on 3BS, 3BL, 5BL, 6AL and 7DS were effective at adult plant stage. The 3BS QTL was validated in Cham-6×Cham-8 recombinant inbred line population; composite interval analysis identified a stripe resistance QTL flanked by the DArT marker, wPt-798970, contributed by Cham-6 parent which accounted for 31.2% of the phenotypic variation. The DArT marker "wPt-798970" lies 1.6cM away from the 3BS QTL detected within GWAM. Epistatic interactions were also investigated; only the QTL on 1AL, 3AS and 6AL exhibited interactions with other loci. These results suggest that GWAM can be an effective approach for identifying and improving resistance to stripe rust in wheat.

Multi-location wheat stripe rust QTL analysis: genetic background and epistatic interactions.

Epistasis and genetic background were important influences on expression of stripe rust resistance in two wheat RIL populations, one with resistance conditioned by two major genes and the other conditioned by several minor QTL. Stripe rust is a foliar disease of wheat (Triticum aestivum L.) caused by the air-borne fungus Puccinia striiformis f. sp. tritici and is present in most regions around the world where commercial wheat is grown. Breeding for durable resistance to stripe rust continues to be a priority, but also is a challenge due to the complexity of interactions among resistance genes and to the wide diversity and continuous evolution of the pathogen races. The goal of this study was to detect chromosomal regions for resistance to stripe rust in two winter wheat populations, 'Tubbs'/'NSA-98-0995' (T/N) and 'Einstein'/'Tubbs' (E/T), evaluated across seven environments and mapped with diversity array technology and simple sequence repeat markers covering polymorphic regions of ≈1480 and 1117cM, respectively. Analysis of variance for phenotypic data revealed significant (P<0.01) genotypic differentiation for stripe rust among the recombinant inbred lines. Results for quantitative trait loci/locus (QTL) analysis in the E/T population indicated that two major QTL located in chromosomes 2AS and 6AL, with epistatic interaction between them, were responsible for the main phenotypic response. For the T/N population, eight QTL were identified, with those in chromosomes 2AL and 2BL accounting for the largest percentage of the phenotypic variance.

Molecular Mapping and Validation of SrND643: A New Wheat Gene for Resistance to the Stem Rust Pathogen Ug99 Race Group.

This study reports the identification of a new gene conferring resistance to the Ug99 lineage of races of Puccinia graminis f. sp. tritici in wheat (Triticum aestivum L.). Because the virulent races of stem rust pathogen continue to pose a serious threat in global wheat production, identification and molecular characterization of new resistance genes remains of utmost important to enhance resistance diversity and durability in wheat germplasm. Advanced wheat breeding line 'ND643/2*Weebill1' carries a stem rust resistance gene, temporarily designated as SrND643, effective against the Ug99 group of P. graminis f. sp. tritici races at both seedling and adult growth stages. This study was conducted to map the chromosomal location of SrND643 and identify closely linked molecular markers to allow its selection in breeding populations. In total, 123 recombinant inbred lines, developed by crossing ND643/2*Weebill1 with susceptible line 'Cacuke', were evaluated for stem rust response in field nurseries at Njoro, Kenya, during two growing seasons in 2010, and were genotyped with DNA markers, including Diversity Arrays Technology, simple sequence repeats (SSR), and single-nucleotide polymorphisms. Linkage mapping tagged SrND643 at the distal end of chromosome 4AL, showing close association with SSR markers Xgwm350 (0.5 centimorgans [cM]), Xwmc219 (4.1 cM), and Xwmc776 (2.9 cM). The race specificity of SrND643 is different from that of Sr7a and Sr7b, indicating that the resistance is conferred by a gene at a new locus or by a new allele of Sr7. The flanking markers Xgwm350 and Xwmc219 were predictive of the presence of SrND643 in advanced germplasm, thus validating the map location and their use in marker-assisted selection.

Genetic mapping and development of near-isogenic lines for genes governing a liguleless phenotype in tetraploid wheat

The liguleless phenotype in wheat lacks ligules and auricles on all plant leaves. Two duplicate dominant genes (Lg 1 and Lg 3 ) principally control the presence of ligules in tetraploid wheat. Although the Lg 3 gene was tentatively located in chromosome 2A, a genetic stock of Triticum durum Desf. (2n = 4x = 28, BBAA genome) having genotype lg 1 lg 1 Lg 3 Lg 3 has not been available. We speculated that a wild emmer (T. dicoccoides Korn., 2n = 4x = 28, BBAA genome) could be a possible source of the Lg 3 gene. In the present study near-isogenic lines (NIL’s) ANW 12E for genotype lg 1 lg 1 Lg 3 Lg 3 and ANW 12F for genotype Lg 1 Lg 1 lg 3 lg 3 were developed from a cross between ANW 12A (lg 1 lg 1 lg 3 lg 3 ), a liguleless NIL of T. durum LD222, and LDN(DIC 2A) (Lg 1 Lg 1 Lg 3 Lg 3 ) through backcrossing to ANW 12A. LDN(DIC 2A) is a chromosome substitution line in which a pair of T. durum cv. Langdon (LDN) 2A chromosomes is replaced with a pair of chromosomes 2A from T. dicoccoides (DIC) line Israel A. Molecular mapping using the F2 of liguleless durum PI 370751/ANW 12F revealed that the Lg 1 gene was 17.5 cM distal to Xgwm47 in chromosome 2BL. In the F2 of PI 370751/ANW 12E the Lg 3 gene was located 18.6 cM distal to Xgwm312 in chromosome 2AL. These results confirmed that the Lg 3 locus was present in chromosome 2A, and dominant alleles at the homoeologous Lg loci determine the presence of ligules in polyploid wheat.

Chromosome mapping of four novel mutants in bread wheat (Triticum aestivum L.)

Induced mutants constitute an important resource for mapping and cloning of genes for specific traits. Five loci underlying four induced mutants in wheat were mapped using phenotypic data and simple sequence repeat (SSR) genotyping data collected on F2 and F3 populations. The four mutant traits included the following: (1) axillary branching, (2) early leaf senescence, (3) reduced plant height and (4) reduced number of nodes. The ‘axillary branching’ locus (Axb) was mapped near Xcfd152 at a distance of 7.5 cM on chromosome 3DS. The ‘early leaf senescence’ gene (els) was mapped on 2DL at a distance of 28.6 cM from the nearest marker Xgwm539. A wheat expressed sequence tag (EST) TaSAG1 representing a candidate for els gene for early leaf senescence was also identified. The ‘reduced plant height’ locus was mapped on chromosome 2BL close to Xwmc361, which is tightly linked with Xwmc317 associated with a ‘GA-responsive dwarfing gene’ Rht4; hence, the ‘dwarf mutant-3’ used in the present study may be a phenotypic manifestation of an another allele of the gene Rht4. The ‘reduced number of node’ mutant was mapped on two loci (rnd1 and rnd2) on chromosomes 2DS and 5AL. The marker locus Xgwm261-2D linked with rnd1 was earlier reported to be closely associated with Rht8 gene, suggesting that rnd1 may perhaps represent a mutation in Rht8 gene; the second locus rnd2 on 5AL represents a novel gene. These results may prove useful in explaining the genetics of four mutant traits and may later also facilitate cloning the genes for these traits.

Exploring the synthetic hexaploid wheat for novel sources of tolerance to excess boron

Excessive levels of boron (B) in the subsoil can severely limit crop yields, particularly under dryland conditions where crops often have to depend on rainfall and available water stored in the soil during fallow. In bread wheat (Triticumaestivum L.), boron tolerance is controlled by at least two loci located on chromosomes 4AL (Bo4) and 7BL (Bo1). In this study, we sought to determine whether novel genomic regions can be identified in wheat primary synthetic hexaploids (SHWs). The study used data from 333 SHW lines imported into Australia in different shipments between 2003 and 2007. On average, the SHWs were uniformly more tolerant to boron toxicity than the sensitive check, Meering, and the top 5 % showed tolerance levels that were superior (P ≤ 0.05) to that of Halberd, the most tolerant wheat check cultivar. At a genome-wide significance threshold of −log(P) ≥ 2.8, association analyses using different algorithms consistently identified three DArT markers, two on chromosome 1AL and one on 4AL. The 4AL region was localised close to the deletion bin location of Bo4, the root-specific boron transporter gene. On the other hand, the two loci on chromosome 1A represent novel regions, which when validated will increase the options of achieving tolerance beyond that conferred by Bo1 and Bo4 alone in breeding programmes.

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.

Identification, Validation of a SSR Marker and Marker Assisted Selection for the Goat Grass Derived Seedling Resistance Gene Lr28 in Wheat

The goat grass (Aegilops speltoides) derived seedling leaf rust resistance gene Lr28 is effective in providing resistance against infection to leaf rust including its most virulent strain, 77-5 (121R63-1) of the pathogen. A polymorphic SSR marker specific to Lr28 was identified by employing bulk segregant analysis on an F2 population derived from the cross between PBW343-Lr28, a leaf rust resistant near isogenic line of the most cultivated variety PBW343 and CSP44-Lr48, the Australian cultivar Condor derived CSP44 line carrying the APR gene Lr48. The marker amplified a polymorphic fragment which was particular to the presence of the seedling resistance gene and it was mapped at a distance of 2.9 cM from the Lr28 resistance locus on chromosome 4AL. It was also validated on a set of 42 NILs which carried other potent leaf rust resistance genes of diverse origin. Such a polymorphic codominant SSR marker will be useful in wheat breeding programmes to differentiate plants homozygous at the Lr28 locus from those that are heterozygous.