Marker-assisted pyramiding of two cereal cyst nematode resistance genes from <Emphasis Type="Italic">Aegilops variabilis</Emphasis> in wheat

The cereal cyst nematode (CCN) Heterodera avenae, is a significant pathogen of wheat. The wild grass Aegilops variabilis Accession No.1 has been found to be resistant to pathotypes of CCN; at least two genes transferred to wheat, designated as CreX and CreY, are involved in the resistance response. The CreY gene may be the same as Rkn-mn1, which confers resistance to root knot nematode (RKN) Meloidogyne naasi. The objective of this work was to pyramid the two CCN resistance genes in a wheat background through marker-assisted selection. As a first step, molecular markers flanking CreX were identified. The completely linked RAPD marker of Rkn-mn1 (CreY), OpY16-_1065, previously obtained, was converted into a SCAR. All these dominant markers were used to incorporate in the same genotype the two Ae. variabilis chromosome segments carrying the two genes for resistance. CCN bioassays with the Ha12 pathotype showed that the level of resistance of the pyramided line was significantly higher than that of CreX and CreY single introgression lines, but lower than that of Ae. variabilis. This study thus illustrates the utilization of molecular markers in breeding for host resistance.     

The molecular bases for resistance to acetyl co-enzyme A carboxylase (ACCase) inhibiting herbicides in two target-based resistant biotypes of annual ryegrass (Lolium rigidum)

Acetyl-CoA carboxylase (ACCase) (EC.6.4.1.2) is an essential enzyme in fatty acid biosynthesis and, in world agriculture, commercial herbicides target this enzyme in plant species. In nearly all grass species the plastidic ACCase is strongly inhibited by commercial ACCase inhibiting herbicides [aryloxyphenoxypropionate (APP) and cyclohexanedione (CHD) herbicide chemicals]. Many ACCase herbicide resistant biotypes (populations) of L. rigidum have evolved, especially in Australia. In many cases, resistance to ACCase inhibiting herbicides is due to a resistant ACCase enzyme. Two ACCase herbicide resistant L. rigidum biotypes were studied to identify the molecular basis of ACCase inhibiting herbicide resistance. The carboxyl-transferase (CT) domain of the plastidic ACCase gene was amplified by PCR and sequenced. Amino acid substitutions in the CT domain were identified by comparison of sequences from resistant and susceptible plants. The amino acid residues Gln-102 (CAG codon) and Ile-127 (ATA codon) were substituted with a Glu residue (GAG codon) and Leu residue (TTA codon), respectively, in both resistant biotypes. Amino acid positions 102 and 127 within the fragment sequenced from L. rigidum corresponded to amino acid residues 1756 and 1781, respectively, in the A. myosuroides full ACCase sequence. Allele-specific PCR results further confirmed the mutations linked with resistance in these populations. The Ile-to-Leu substitution at position 1781 has been identified in other resistant grass species as endowing resistance to APP and CHD herbicides. The Gln-to-Glu substitution at position 1756 has not previously been reported and its role in herbicide resistance remains to be established.     

Hypothesis: Transgene establishment in wild relatives of wheat can be prevented by utilizing the Ph1 gene as a senso stricto chaperon to prevent homoeologous recombination

Durum and bread wheat need transgenic traits such as herbicide and disease resistance due to recent evolution of herbicide resistant grass weeds and an intractable new strain of stem rust. Transgenic wheat varieties have not been commercialized partly due to potential transgene movement to wild/weedy relatives, which occurs naturally to closely related Aegilops and other spp. Recombination does not occur in the F₁ hybrid between wheat and its relatives due to the presence of the Ph1 gene on wheat chromosome arm 5BL, which acts as a chaperone, preventing promiscuous homoeologous pairing to similar, but not homologous chromosomes of the wild/weedy species. Thus recombination must occur during backcrossing after the wheat Ph1 gene has been eliminated. Based on these findings, we speculate that Ph1 could be used to prevent gene introgression into weedy relatives. We propose two methods to prevent such transgene establishment: (1) link the transgene in proximity to the wheat Ph1 gene and (2) insert the transgene in tandem with the lethal barnase on any chromosome arm other than 5BL, and insert barstar, which suppresses barnase on chromosome arm 5BL in proximity to Ph1. The presence of Ph1 in backcross plants containing 5BL will prevent the homoeologous establishment of barnase coupled to the desired transgene in the wild population. 5BL itself will be eliminated during repeated backcrossing to the wild parent, and progeny bearing the desired transgene in tandem with barnase but without the Ph1-barstar complex will die.     

Molecular markers for wheat leaf rust resistance gene Lr41

Leaf rust, caused by Puccinia triticina Eriks., is an important foliar disease of common wheat (Triticum aestivum L.) worldwide. Pyramiding several major rust-resistance genes into one adapted cultivar is one strategy for obtaining more durable resistance. Molecular markers linked to these genes are essential tools for gene pyramiding. The rust-resistance gene Lr41 from T. tauschii has been introgressed into chromosome 2D of several wheat cultivars that are currently under commercial production. To discover molecular markers closely linked to Lr41, a set of near-isogenic lines (NILs) of the hard winter wheat cultivar Century were developed through backcrossing. A population of 95 BC₃F_2:6 NILs were evaluated for leaf rust resistance at both seedling and adult plant stages and analyzed with simple sequence repeat (SSR) markers using bulked segregant analysis. Four markers closely linked to Lr41 were identified on chromosome 2DS; the closest marker, Xbarc124, was about 1 cM from Lr41. Physical mapping using Chinese Spring nullitetrasomic and ditelosomic genetic stocks confirmed that markers linked to Lr41 were on chromosome arm 2DS. Marker analysis in a diverse set of wheat germplasm indicated that primers BARC124, GWM210, and GDM35 amplified polymorphic bands between most resistant and susceptible accessions and can be used for marker-assisted selection in breeding programs.     

Quantitative Trait Loci for Aluminum Resistance in Wheat Cultivar Chinese Spring

Aluminum (Al) toxicity is one of the major constrains for wheat production in many wheat growing areas worldwide. Further understanding of inheritance of Al resistance may facilitate improvement of Al resistance of wheat cultivars (Triticum aestivum L.). A set of ditelosomic lines derived from the moderately Al-resistant wheat cultivar Chinese Spring was assessed for Al resistance. The root growth of ditelosomic lines DT5AL, DT7AL, DT2DS and DT4DS was significantly lower than that of euploid Chinese Spring under Al stress, suggesting that Al-resistance genes might exist on the missing chromosome arms of 5AS, 7AS, 2DL and 4DL of Chinese Spring. A population of recombinant inbred lines (RILs) from the cross Annong 8455 × Chinese Spring-Sumai 3 7A substitution line was used to determine the effects of these chromosome arms on Al resistance. A genetic linkage map consisting of 381 amplified fragment length polymorphism (AFLP) markers and 168 simple sequence repeat (SSR) markers was constructed to determine the genetic effect of the quantitative trait loci (QTLs) for Al resistance in Chinese Spring. Three QTLs, Qalt.pser-4D, Qalt.pser-5A and Qalt.pser-2D, were identified that enhanced root growth under Al stress, suggesting that inheritance of Al resistance in Chinese Spring is polygenic. The QTL with the largest effect was flanked by the markers of Xcfd23 and Xwmc331 on chromosome 4DL and most probably is multi-allelic to the major QTL identified in Atlas 66. Two additional QTLs, Qalt.pser-5A and Qalt.pser-2D on chromosome 5AS and 2DL, respectively, were also detected with marginal significance in the population. Some SSR markers identified in this study would be useful for marker-assisted pyramiding of different QTLs for Al resistance in wheat cultivars.     

Phenotypic assessment and mapped markers for<Emphasis Type="Italic"> H31,</Emphasis> a new wheat gene conferring resistance to Hessian fly (Diptera: Cecidomyiidae)

A new source of resistance to the highly virulent and widespread biotype L of the Hessian fly, Mayetiola destructor (Say), was identified in an accession of tetraploid durum wheat, Triticum turgidum Desf., and was introgressed into hexaploid common wheat, Triticum aestivum L. Genetic analysis and deletion mapping revealed that the common wheat line contained a single locus for resistance, H31, residing at the terminus of chromosome 5BS. H31 is the first Hessian fly-resistance gene to be placed on 5BS, making it unique from all previously reported sources of resistance. AFLP analysis identified two markers linked to the resistance locus. These markers were converted to highly specific sequence-tagged site markers. The markers are being applied to the development of cultivars carrying multiple genes for resistance to Hessian fly biotype L in order to test gene pyramiding as a strategy for extending the durability of deployed resistance.Communicated by J. Dvorak     

Mapping the Sw-5 locus for tomato spotted wilt virus resistance in tomatoes using RAPD and RFLP analyses

The Sw-5 locus confers dominant resistance to tomato spotted wilt virus (TSWV). To map the location and facilitate the identification of markers linked to Sw-5 we developed a pair of near-isogenic lines (NILs) and an F₂ Lycopersicon esculentum x L. pennellii population segregating for resistance to TSWV. DNA from the NILs was analyzed using 748 random 10-mer oligonucleotides to discern linked molecular markers using a random amplified polymorphic DNA (RAPD) approach. One random primer (GAGCACGGGA) was found to produce a RAPD band of about 2200 bp that demonstrates linkage to Sw-5. Data from co-segregation of resistance and restriction fragment length polymorphisms (RFLPs) in a F₂ interspecific population position Sw-5 between the markers CT71 and CT220 near the telomere of the long arm of chromosome 9.     

Development of an STS marker tightly linked to <Emphasis Type="Italic">Yr26</Emphasis> against wheat stripe rust using the resistance gene-analog polymorphism (RGAP) technique

The gene Yr26 confers resistance to all races of Puccinia striiformis f. sp. tritici (PST), the casual pathogen of wheat stripe rust in China. Here, we report development of a molecular marker closely linked to Yr26 using a resistance gene-analog polymorphism (RGAP) technique. A total of 787 F₂ plants and 165 F₃ lines derived from the cross Chuanmai 42/Taichung 29 were used for linkage analysis. Eighteen near-isogenic lines (NILs) and 18 Chinese wheat cultivars and advanced lines with different genes for stripe rust resistance were employed for the validation of STS markers. A total of 1,711 RGAP primer combinations were used to test the parents and resistant and susceptible bulks. Five polymorphic RGAP markers were used for genotyping all F₂ plants. Linkage analysis showed that the five RGAP markers were closely linked to Yr26 with genetic distances ranging from 0.5 to 2.9 cM. These markers were then converted into STS markers, one, CYS-5, of which was located 0.5 cM to Yr26 and was closely associated with the resistance gene when validated over 18 NILs and 18 Chinese wheat cultivars and lines. The results indicated that CYS-5 can be used in marker-assisted selection targeted at pyramiding Yr26 and other genes for stripe rust resistance.     

Chromosomal localization and molecular-marker tagging of the powdery mildew resistance gene (Lv) in tomato

We report the tagging of a powdery mildew [Leveillula taurica (Lév.) Arnaud.] resistance gene (Lv) in tomato using RAPD and RFLP markers. DNA from a resistant (cv Laurica) and a susceptible cultivar were screened with 300 random primers that were used to amplify DNA of resistant and susceptible plants. Four primers yielded fragments that were unique to the resistant line and linked to the resistance gene in an F₂ population. One of these amplified fragments, OP248, with a molecular weight of 0.7 kb, was subsequently mapped to chromosome 12, 1 cM away from CT134. Using RFLP markers located on chromosome 12, it was shown that approximately one half of chromosome 12 (about 42 cM), in the resistant variety is comprised of foreign DNA, presumably introgressed with the resistance gene from the wild species L. chilense. Further analysis of a backcross population revealed that the Lv gene lies in the 5.5-cM interval between RFLP markers, CT211 and CT219. As a prelude to map-based cloning of the Lv gene, we are currently enriching the density of markers in this region by a combination of RAPD primers and other techniques.     

Hessian fly resistance gene H13 is mapped to a distal cluster of resistance genes in chromosome 6DS of wheat

H13 is inherited as a major dominant resistance gene in wheat. It was previously mapped to chromosome 6DL and expresses a high level of antibiosis against Hessian fly (Hf) [Mayetiola destructor (Say)] larvae. The objective of this study was to identify tightly linked molecular markers for marker-assisted selection in wheat breeding and as a starting point toward the map-based cloning of H13. Fifty-two chromosome 6D-specific microsatellite (simple sequence repeat) markers were tested for linkage to H13 using near-isogenic lines Molly (PI 562619) and Newton-207, and a segregating population consisting of 192 F_2:3 families derived from the cross PI 372129 (Dn4) × Molly (H13). Marker Xcfd132 co-segregated with H13, and several other markers were tightly linked to H13 in the distal region of wheat chromosome 6DS. Deletion analysis assigned H13 to a small region closely proximal to the breakpoint of del6DS-6 (FL 0.99). Further evaluation and comparison of the H13-linked markers revealed that the same chromosome region may also contain H23 in KS89WGRC03, an unnamed H gene (H_WGRC4) in KS89WGRC04, the wheat curl mite resistance gene Cmc4, and a defense response gene Ppo for polyphenol oxidase. Thus, these genes comprise a cluster of arthropod resistance genes. Marker analysis also revealed that a very small intercalary chromosomal segment carrying H13 was transferred from the H13 donor parent to the wheat line Molly.Mention of commercial or proprietary product does not constitute an endorsement by the USDA.     

Hessian fly resistance genes <Emphasis Type="Italic">H16</Emphasis> and <Emphasis Type="Italic">H17</Emphasis> are mapped to a resistance gene cluster in the distal region of chromosome 1AS in wheat

Hessian fly [Mayetiola destructor (Say)] is one of the major insect pests of wheat (Triticum aestivum L.) worldwide. Hessian fly (Hf)-resistance genes H16 and H17 were reported to condition resistance to Hf biotype L that is prevalent in many wheat-growing areas of eastern USA, and both of them were previously assigned to wheat chromosome 5A by their linkage to H9. The objectives in this study were to (1) map H16 and H17 independent of their linkage with H9 and (2) identify DNA markers that co-segregate with H16 or H17, and that are useful for selection of these genes in segregating populations and to combine these genes with other Hf-resistance genes in wheat cultivars. Contrary to previously reported locations, H16 and H17 did not show linkage with the molecular markers on chromosome 5A. Instead, both of them are linked with the molecular markers on the short arm of chromosome 1A (1AS). The simple sequence repeat (SSR) marker Xpsp2999 and EST-derived SSR (eSSR) marker Xwem6b are two flanking markers that are linked to H16 at genetic distances of 3.7 and 5.5 cM, respectively. Similarly, H17 is located between markers Xpsp2999 and Xwem6b at genetic distances of 6.2 and 5.1 cM, respectively. Five other SSR and eSSR markers including Xcfa2153, Xbarc263, Xwem3a, Xwmc329, and Xwmc24 were also linked to H16 and H17 at close genetic distances. These closely linked molecular markers should be useful for pyramiding H16 and H17 with other Hessian fly resistance genes in a single wheat genotype. In addition, using Chinese Spring deletion line bin mapping we positioned all of the linked markers and the Hf-resistance genes (H16 and H17) to the distal 14% of chromosome 1AS, where Hf-resistance genes H9, H10, and H11 are located. Our results together with previous studies suggest that Hf-resistance genes H9, H10, H11, H16, and H17 along with the pathogen resistance genes Pm3 and Lr10 appear to occupy a resistance gene cluster in the distal region of chromosome 1AS in wheat. Contribution from Purdue Univ. Agric. Res. Programs Journal Article No. 2007-18105.     

Microsatellite tagging of the leaf rust resistance gene <Emphasis Type="Italic">Lr16</Emphasis> on wheat chromosome 2BSc

Leaf rust, caused by Puccinia triticina, is one of the most damaging diseases of wheat worldwide. Lr16 is a widely deployed leaf rust resistance gene effective at the seedling stage. Although virulence to Lr16 exists in the Canadian P. triticina population, Lr16 provides a level of partial resistance in the field. The primary objective of this study was to identify markers linked to Lr16 that are suitable for marker-assisted selection (MAS). Lr16 was tagged with microsatellite markers on the distal end of chromosome 2BS in three mapping populations. Seven microsatellite loci mapped within 10 cM of Lr16, with the map distances varying among populations. Xwmc764 was the closest microsatellite locus to Lr16, and mapped 1, 9, and 3 cM away in the RL4452/AC Domain, BW278/AC Foremost, and HY644/McKenzie mapping populations, respectively. Lr16 was the terminal locus mapped in all three populations. Xwmc764, Xgwm210, and Xwmc661 were the most suitable markers for selection of Lr16 because they had simple PCR profiles, numerous alleles, high polymorphism information content (PIC), and were tightly linked to Lr16. Twenty-eight spring wheat lines were evaluated for leaf rust reaction with the P. triticina virulence phenotypes MBDS, MBRJ, and MGBJ, and analyzed with five microsatellite markers tightly linked to Lr16. There was good agreement between leaf rust infection type (IT) data and the microsatellite allele data. Microsatellite markers were useful for postulating Lr16 in wheat lines with multiple leaf rust resistance genes.     

The expression of a maize glutathione S-transferase gene in transgenic wheat confers herbicide tolerance,both in planta and in vitro

Maize (Zea mays), in common with a number of other important crop species, has several glutathione S-transferase (GST) isoforms that have been implicated in the detoxification of xenobiotics via glutathione conjugation. A cDNA encoding the maize GST subunit GST-27, under the control of a strong constitutive promoter, was introduced into explants of the wheat (Triticum aestivum L.) lines cv. Florida and L88-31 via particle bombardment, using the phosphinothricin acetyltransferase (pat) gene as a selectable marker. All six independent transgenic wheat lines recovered expressed the GST-27 gene. T₁ progeny of these wheat lines were germinated on solid medium containing the chloroacetanilide herbicide alachlor, and tolerance to this herbicide was correlated with GST-27 expression levels. In glasshouse sprays, homozygous T₂ plants were resistant not only to alachlor but also to the chloroacetanilide herbicide dimethenamid and the thiocarbamate herbicide EPTC. These additional GST-27 activities, demonstrated via over-expression in a heterologous host, have not been described previously. T₂ plants showed no enhanced tolerance to the herbicides atrazine (an s-triazine) or oxyfluorfen (a diphenyl ether). In further experiments, T₂ wheat plants were recovered from immature transgenic scutella cultured on medium containing 100 mg/l alachlor, a concentration which killed null segregant and wild-type scutella. These data indicate the potential of the maize GST-27 gene as a selectable marker in wheat transformation.     

Saturation and comparative mapping of a major Fusarium head blight resistance QTL in tetraploid wheat

Fusarium head blight (FHB) is a devastating disease of cultivated wheat worldwide. Partial resistance to FHB has been identified in common wheat (Triticum aestivum L.). However, sources of effective FHB resistance have not been found in durum wheat (T. turgidum L. var. durum). A major FHB resistance quantitative trait loci (QTL), Qfhs.ndsu-3AS, was identified on chromosome 3A of T. dicoccoides, a wild relative of durum wheat. Here, we saturated the genomic region containing the QTL using EST-derived target region amplified polymorphism (TRAP), sequence tagged site (STS), and simple sequence repeat (SSR) markers. A total of 45 new molecular marker loci were detected on chromosome 3A and the resulting linkage map consisted of 55 markers spanning a genetic distance of 277.2 cM. Qfhs.ndsu-3AS was positioned within a chromosomal interval of 11.5 cM and is flanked by the TRAP marker loci, Xfcp401 and Xfcp397.2. The average map distance between the marker loci within this QTL region was reduced from 4.9 cM in the previous study to 3.5 cM in the present study. Comparative mapping indicated that Qfhs.ndsu-3AS is not homoeologous to Qfhs.ndsu-3BS, a major FHB QTL derived from the common wheat cultivar Sumai 3. These results facilitate our efforts toward map-based cloning of Qfhs.ndsu-3AS and utilization of this QTL in durum wheat breeding via marker-assisted selection.     

A “one-marker-for-two-genes” approach for efficient molecular discrimination of Pm12 and Pm21 conferring resistance to powdery mildew in wheat

Powdery mildew, caused by Blumeria graminis f. sp. tritici, is one of the most important wheat diseases worldwide. Pyramiding different resistance genes into single cultivar has been proposed as one remedy to provide durable resistance. Powdery mildew resistance genes Pm12 (T6BS-6SS.6SL), transferred from Aegilops speltoides to wheat cv. Wembley, and Pm21 (T6VS.6AL), introduced from Dasypyrum villosum to wheat cv. Yangmai5, conferred broad-spectrum resistance to B. graminis f. sp. tritici. Both Pm12 and Pm21 genes are located on the short arms of homologous group six involved translocated chromosomes 6SS.6BL and 6VS.6AL, respectively. Simple sequence repeat motifs of wheat simple sequence repeat (SSR) and expressed sequence tag (EST) sequences on the short arm of homologous group six chromosomes were analyzed to develop molecular markers for discriminating chromosome arms 6AS, 6BS, 6DS, 6VS, and 6SS. One EST–SSR marker, Xcau127, was polymorphic, and therefore can be used to distinguish the two resistance genes and the respective susceptible alleles. This marker allowed us to develop an efficient “one-marker-for-two-genes” procedure for identifying powdery mildew resistance genes Pm12 and Pm21 for marker-assisted selection and gene pyramiding in wheat breeding programs. Wei Song and Chaojie Xie contributed equally to this work     

A major QTL controlling seed dormancy and pre-harvest sprouting resistance on chromosome 4A in a Chinese wheat landrace

Wheat pre-harvest sprouting (PHS) can cause significant reduction in yield and end-use quality of wheat grains in many wheat-growing areas worldwide. To identify a quantitative trait locus (QTL) for PHS resistance in wheat, seed dormancy and sprouting of matured spikes were investigated in a population of 162 recombinant inbred lines (RILs) derived from a cross between the white PHS-resistant Chinese landrace Totoumai A and the white PHS-susceptible cultivar Siyang 936. Following screening of 1,125 SSR primers, 236 were found to be polymorphic between parents, and were used to screen the mapping population. Both seed dormancy and PHS of matured spikes were evaluated by the percentage of germinated kernels under controlled moist conditions. Twelve SSR markers associated with both PHS and seed dormancy were located on the long arm of chromosome 4A. One QTL for both seed dormancy and PHS resistance was detected on chromosome 4AL. Two SSR markers, Xbarc 170 and Xgwm 397, are 9.14 cM apart, and flanked the QTL that explained 28.3% of the phenotypic variation for seed dormancy and 30.6% for PHS resistance. This QTL most likely contributed to both long seed dormancy period and enhanced PHS resistance. Therefore, this QTL is most likely responsible for both seed dormancy and PHS resistance. The SSR markers linked to the QTL can be used for marker-assisted selection of PHS-resistant white wheat cultivars. Shi-Bin Cai and Cui-Xia Chen contributed equally to this work.     

Phytotoxicity of isoxaflutole to Phalaris minor Retz.

Littleseed canarygrass (Phalaris minor Retz.) is a major weed in wheat fields, and has developed resistance to the commonly used herbicide isoproturon. This study explores the potential use of isoxaflutole, a pre-emergence herbicide, to control littleseed canarygrass. Greenhouse studies were carried out to determine the phytotoxicity of isoxaflutole in relation to shoot height, fresh shoot biomass and leaf chlorophyll concentration of wheat and littleseed canarygrass. Electron microscopy was used to examine any damage to leaf chloroplast at ultrastructural level. Results indicate that isoxaflutole (0.5 mg/L) significantly reduced the shoot height of littleseed canarygrass (39.6%), but no significant reduction in the shoot height of wheat was observed (9.6%) when compared to control. None of the concentrations (0.05, 0.1, 0.5 and 1 mg/L) of isoxaflutole altered soil chemistry in relation to pH, organic matter, macro or micro inorganic ions. While untreated littleseed canarygrass leaves had elongated chloroplast, starch grains and small number of plastoglobuli; treated littleseed canarygrass leaves had swollen chloroplast, large number of plastoglobuli, and a lack of starch grains. We conclude that isoxaflutole can be an effective herbicide for controlling littleseed canarygrass.     

Chromosomal location of a race-specific resistance gene to <Emphasis Type="Italic">Mycosphaerella graminicola</Emphasis> in the spring wheat ST6

Septoria tritici blotch, caused by Mycosphaerella graminicola, is a serious foliar disease of wheat worldwide. Qualitative, race-specific resistance sources have been identified and utilized for resistant cultivar development. However, septoria tritici blotch resistant varieties have succumbed to changes in virulence of M. graminicola on at least three continents. The use of resistance gene pyramids may slow or prevent the breakdown of resistance. A clear understanding of the genetics of resistance and the identification of linked PCR-based markers will facilitate the recovery of wheat lines carrying multiple septoria tritici blotch resistance genes. The resistance gene in ST6 to isolate MG2 of M. graminicola was mapped with microsatellite markers in two populations, ST6/Erik and ST6/Katepwa. Bulk segregant analysis identified a marker on chromosome 4AL putatively linked to the resistance gene. A large linkage group was identified in each population using additional microsatellite markers mapping to chromosome 4AL. The resistance gene in ST6 mapped to the distal end of chromosome 4AL in each mapping population and was designated Stb7. Three of the microsatellite loci, Xwmc313, Xwmc219 and Xgwm160, mapped within 3.5 cM of Stb7; however, none flanked Stb7. Xwmc313 was the closest and mapped 0.3 and 0.5 cM from Stb7 in the crosses ST6/Katepwa and ST6/Erik, respectively. WMC313 will be very useful for marker-assisted selection of Stb7 in Canadian breeding programs because the ST6 allele of Xwmc313 was not identified in any of the Canadian common wheat cultivars tested.Communicated by P. Langridge     

Molecular mapping and physical location of major gene conferring seedling resistance to Septoria tritici blotch in wheat

Septoria tritici blotch (STB) caused by Mycosphaerella graminicola (anamorph: Septoria tritici), is one of the most important foliar diseases of wheat. We assessed three doubled-haploid (DH) populations derived from Chara (STB-susceptible)/WW2449 (STB-resistant), Whistler (STB-susceptible)/WW1842 (STB-resistant) and Krichauff (STB susceptible)/WW2451 (STB-resistant) for resistance to a single-pycnidium isolate 79.2.1A of M. graminicola at the seedling stage. STB resistance in each of the three DH populations was conditioned by a single major gene designated as StbWW2449, StbWW1842 and StbWW2451. Linkage analyses and physical mapping indicated that the StbWW loci were located on the short arm of chromosome 1B (IBS). Four simple sequence repeat (SSR) markers linked with STB resistance: Xwmc230, Xbarc119b, Xksum045 and Xbarc008 were located to the distal bin of 1BS.sat1BS-4 (FL: 0.52–1.00) in the 1BS physical map. Xwmc230, Xbarc119b and Xksum045 markers, mapped within 7 cM from StbWW were validated for their linkage and predicted the STB resistance with over 94% accuracy in the 79 advanced breeding lines having WW2449 as one of the parents. The marker interval Xwmc230/Xksum045-Xbarc119b also explained up to 38% of the phenotypic variance at the adult plant stage in all three DH mapping populations. These results have proven that SSR markers are useful in monitoring STB resistance both at seedling and adult plant stages and hence are suitable for routine marker-assisted selection in the wheat breeding programs. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.