Fusarium head blight resistance in hexaploid wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis>)-<Emphasis Type="Italic">Lophopyrum</Emphasis> genetic lines and tagging of the alien chromatin by PCR markers

The objective of this research was to identify Fusarium head blight (FHB) resistance in wheat (Triticum aestivum)-Lophopyrum genetic lines that might complement FHB resistance in common wheat; and to identify DNA markers that can be used to tag the resistance gene in the alien chromatin (E or el₂ genome) for the development of improved wheat cultivars. FHB resistance was evaluated in 19 Chinese Spring-Lophopyrum elongatum (EE) substitution lines, two Thatcher-L. ponticum (el₁ and el₂) substitution lines, and four Thatcher-L. ponticum translocation lines. Significant resistance was identified in the substitution lines 7E(7A), 7E(7B), and 7E(7D). The homoeologous chromosome, 7el₂,also showed resistance in the Thatcher genetic background. Both the Thatcher-7el₁ substitution and translocation lines were susceptible, like Thatcher, indicating that there is no resistance gene on the 7el₁ chromosome. Simple sequence repeat (SSR) and cleaved amplified polymorphic sequences (CAPS) in homoeologous group 7 chromosomes were used to identify DNA markers located on 7E and 7el₂. As expected, the transferability of wheat SSR markers to Lophopyrum is low. Of the 52 SSR markers that we tested, only five were found to be co-dominant on 7E of L. elongatum versus 7A, 7B, and 7D, one of which is also positive on 7el₂. A CAPS marker, derived from the RFLP probe PSR129, can serve as a dominant marker for 7el₂ chromatin.Communicated by J. Dvorak     

Molecular mapping of <Emphasis Type="Italic">Thinopyrum</Emphasis>-derived Fusarium head blight resistance in common wheat

Resistance to Fusarium head blight (FHB) caused by Fusarium graminearum Schwabe in wheat (Triticum aestivum L.) was identified in disomic chromosome substitution and translocation lines, into which chromosome 7el₂ had been introgressed from wheatgrass, Thinopyrum ponticum. In this study, two chromosome substitution lines with different origins (designated as el₁ and el₂) and with different reactions to infection by F. graminearum were crossed to develop a segregating mapping population. The objectives of this study were to determine the effectiveness of this type II resistance and map it on chromosome 7el₂. Type II resistance to FHB was characterized in the F₂, F_2:3 families, F_4:5 plants and F_5:6 recombinant inbred lines developed by single-seed descent; and the population was characterized in the F₂ and F₅ with DNA markers along the long arm of 7el. Composite interval mapping revealed a FHB resistance QTL, designated Qfhs.pur-7EL, located in the distal region of the long arm of 7el₂ and delimited with flanking markers XBE445653 and Xcfa2240. Additive effects of Qfhs.pur-7EL reduced the number of diseased spikelets per spike following inoculation of one floret in four experiments by 1.5–2.6 and explained 15.1–32.5% of the phenotypic variation in the populations. Several STS-derived and EST-derived PCR or CAPS markers were developed in this chromosomal region, and showed the specificity of 7el₂ compared to an array of wheat lines possessing other sources of FHB resistance. These markers are useful in an effort to shorten the chromosome segment of 7el₂ and to use for marker-assisted introgression of this resistance into wheat.     

Genotyping-by-sequencing to remap QTL for type II Fusarium head blight and leaf rust resistance in a wheat–tall wheatgrass introgression recombinant inbred population

Fusarium graminearum Schwabe (Fusarium head blight, FHB) and Puccinia triticina Eriks (leaf rust) are two major fungal pathogens posing a continuous threat to the wheat crop; consequently, identifying resistance genes from various sources is always of importance to wheat breeders. We identified tightly linked single nucleotide polymorphism (SNP) markers for the FHB resistance quantitative trait locus (QTL) Qfhs.pur-7EL and the leaf rust resistance locus Lr19 using genotyping-by-sequencing (GBS) in a wheat–tall wheatgrass introgression-derived recombinant inbred line (RIL) population. One thousand and seven hundred high-confidence SNPs were used to conduct the linkage and QTL analysis. Qfhs.pur-7EL was mapped to a 2.9 cM region containing four markers within a 43.6 cM segment of wheatgrass chromosome 7el₂ that was translocated onto wheat chromosome 7DL. Lr19 from 7el₁ was mapped to a 1.21 cM region containing two markers in the same area, in repulsion. Five lines were identified with the resistance-associated SNP alleles for Qfhs.pur-7EL and Lr19 in coupling. Two SNP markers in the Qfhs.pur-7EL region were converted into PCR-based KASP markers. Investigation of the genetic characteristics of the parental lines of this RIL population indicated that they are translocation lines in two different wheat cultivar genetic backgrounds instead of 7E–7D substitution lines in Thatcher wheat background, as previously reported in the literature.     

Identification of microsatellite markers linked to leaf rust resistance gene <Emphasis Type="Italic">Lr25</Emphasis> in wheat

The leaf rust resistance gene Lr25, transferred from Secale cereale L. into wheat and located on chromosome 4B, imparts resistance to all pathotypes of leaf rust in South-East Asia. In an F₂-derived F₃ population, created by crossing TcLr25 that carries the gene Lr25 for leaf rust resistance with leaf rust-susceptible parent Agra Local, three microsatellite markers located on the long arm of chromosome 4B were found to be linked to the Lr25 locus. The donor parent TcLr25 is a near-isogenic line derived from the variety Thatcher. The most virulent pathotype of leaf rust in the South-East Asian region, designated 77–5 (121R63-1), was used for challenging the population under artificially controlled conditions. The marker Xgwm251 behaved as a co-dominant marker placed 3.8 cM away from the Lr25 locus on 4BL. Two null allele markers, Xgwm538 and Xgwm6, in the same linkage group were located at a distance of 3.8 cM and 16.2 cM from the Lr25 locus, respectively. The genetic sequence of Xgwm251, Lr25, Xgwm538, and Xgwm6 covered a total length of 20 cM on 4BL. The markers were validated for their specificity to Lr25 resistance in a set of 43 wheat genetic stocks representing 43 other Lr genes.     

AFLP and STS tagging of Lr19, a gene conferring resistance to leaf rust in wheat

Amplified fragment length polymorphism (AFLP) markers were used to enrich the map of the wheat chromosomal region containing the Thinopyrum-derived Lr19 leaf rust resistance gene. The region closest to Lr19 was targeted through the use of deletion and recombinant lines of the translocated segment. One of the AFLP bands thus identified was converted into a sequence-tagged-site (STS) marker. This assay generated a 130-bp PCR fragment in all Lr19-carrying lines tested, except for one deletion mutant, while non-carrier template failed to amplify any product. This sequence represents the first marker to map on the distal side of Lr19 on chromosome 7el₁. The conversion process of AFLP fragments to STS markers was technically difficult, mainly because of the presence of contaminating fragments. Various approaches were taken to reduce the frequency of false positives and to identify the correct clone. We were able to formulate a general verification strategy prior to clone sequencing. Various other factors causing problems with converting AFLP bands to an STS assays are also discussed. Received: 15 September 2000 / Accepted: 5 January 2001     

Molecular characterization of durum and common wheat recombinant lines carrying leaf rust resistance (Lr19) and yellow pigment (Y) genes from Lophopyrum ponticum

Chromosome 7E from Lophopyrum ponticum carries a valuable leaf rust resistant gene designated Lr19. This gene has not been widely used in common wheat breeding because of linkage with the yellow pigment gene Y. This gene tints flour yellow, reducing its appeal in bread making. However, a high level of yellow pigment is desirable in durum wheat breeding. We produced 97 recombinant chromosomes between L. ponticum transfer 7D.7E#1 and its wheat homoeologues, using the ph1b mutation that promotes homoeologous pairing. We characterized a subset of 37 of these lines with 11 molecular markers and evaluated their resistance to leaf rust and the abundance of yellow pigment. The Lr19 gene was mapped between loci Xwg420 and Xmwg2062, whereas Y was mapped distal to Xpsr687, the most distal marker on the long arm of chromosome 7. A short terminal 7EL segment translocated to 7A, including Lr19 and Y (line 1-23), has been transferred to durum wheat by backcrossing. The presence of this alien segment significantly increased the abundance of yellow pigment. The Lr19 also conferred resistance to a new durum leaf rust race from California and Mexico that is virulent on most durum wheat cultivars. The new durum lines with the recombinant 7E segment will be useful parents to increase yellow pigment and leaf rust resistance in durum wheat breeding programs. For the common wheat breeding programs, we selected the recombinant line 1-96, which has an interstitial 7E segment carrying Lr19 but not Y. This recombinant line can be used to improve leaf rust resistance without affecting flour color. The 7EL/7DL 1-96 recombinant chromosome did not show the meiotic self-elimination previously reported for a 7EL/7BL translocation.     

Microsatellite mapping of adult-plant leaf rust resistance gene <Emphasis Type="Italic">Lr22a</Emphasis> in wheat

This study was conducted to identify microsatellite markers (SSR) linked to the adult-plant leaf rust resistance gene Lr22a and examine their cross-applicability for marker-assisted selection in different genetic backgrounds. Lr22a was previously introgressed from Aegilops tauschii Coss. to wheat (Triticum aestivum L.) and located to chromosome 2DS. Comparing SSR alleles from the donor of Lr22a to two backcross lines and their recurrent parents showed that between two and five SSR markers were co-introgressed with Lr22a and the size range of the Ae. tauschii introgression was 9–20 cM. An F₂ population from the cross of 98B34-T4B × 98B26-N1C01 confirmed linkage between the introgressed markers and Lr22a on chromosome 2DS. The closest marker, GWM296, was 2.9 cM from Lr22a. One hundred and eighteen cultivars and breeding lines of different geographical origins were tested with GWM296. In total 14 alleles were amplified, however, only those lines predicted or known to carry Lr22a had the unique Ae. tauschii allele at GWM296 with fragments of 121 and 131 bp. Thus, GWM296 is useful for selecting Lr22a in diverse genetic backgrounds. Genotypes carrying Lr22a showed strong resistance to leaf rust in the field from 2002 to 2006. Lr22a is an ideal candidate to be included in a stack of leaf rust resistance genes because of its strong adult-plant resistance, low frequency of commercial deployment, and the availability of a unique marker. An erratum to this article can be found at     

Genetic dissection of a major Fusarium head blight QTL in tetraploid wheat

The devastating effect of Fusarium head blight (FHB) caused by Fusarium graminearum has led to significant financial losses across the Upper Midwest of the USA. These losses have spurred the need for research in biological, chemical, and genetic control methods for this disease. To date, most of the research on FHB resistance has concentrated on hexaploid wheat (Triticum aestivum L.) lines originating from China. Other sources of resistance to FHB would be desirable. One other source of resistance for both hexaploid wheat and tetraploid durum wheat (T. turgidum L. var. durum) is the wild tetraploid, T. turgidum L. var. dicoccoides (T. dicoccoides). Previous analysis of the `Langdon'-T. dicoccoides chromosome substitution lines, LDN(Dic), indicated that the chromosome 3A substitution line expresses moderate levels of resistance to FHB. LDN(Dic-3A) recombinant inbred chromosome lines (RICL) were used to generate a linkage map of chromosome 3A with 19 molecular markers spanning a distance of 155.2 cM. The individual RICL and controls were screened for their FHB phenotype in two greenhouse seasons. Analysis of 83 RICL identified a single major quantitative trait locus, Qfhs.ndsu-3AS, that explains 37% of the phenotypic or 55% of the genetic variation for FHB resistance. A microsatellite locus, Xgwm2, is tightly linked to the highest point of the QTL peak. A region of the LDN (Dic-3A) chromosome associated with the QTL for FHB resistance encompasses a 29.3 cM region from Xmwg14 to Xbcd828.     

Development and characterization of wheat lines carrying stem rust resistance gene Sr43 derived from Thinopyrum ponticum

Key message

Wheat lines carrying Ug99-effective stem rust resistance gene Sr43 on shortened alien chromosome segments were produced using chromosome engineering, and molecular markers linked to Sr43 were identified for marker-assisted selection.

Abstract

Stem rust resistance gene Sr43, transferred into common wheat (Triticum aestivum) from Thinopyrum ponticum, is an effective gene against stem rust Ug99 races. However, this gene has not been used in wheat breeding because it is located on a large Th. ponticum 7el₂ chromosome segment, which also harbors genes for undesirable traits. The objective of this study was to eliminate excessive Th. ponticum chromatin surrounding Sr43 to make it usable in wheat breeding. The two original translocation lines KS10-2 and KS24-1 carrying Sr43 were first analyzed using simple sequence repeat (SSR) markers and florescent genomic in situ hybridization. Six SSR markers located on wheat chromosome arm 7DL were identified to be associated with the Th. ponticum chromatin in KS10-2 and KS24-1. The results confirmed that KS24-1 is a 7DS·7el₂L Robertsonian translocation as previously reported. However, KS10-2, which was previously designated as a 7el₂S·7el₂L-7DL translocation, was identified as a 7DS-7el₂S·7el₂L translocation. To reduce the Th. ponticum chromatin carrying Sr43, a BC₂F₁ population (Chinese Spring//Chinese Spring ph1bph1b*2/KS10-2) containing ph1b-induced homoeologous recombinants was developed, tested with stem rust, and genotyped with the six SSR markers identified above. Two new wheat lines (RWG33 and RWG34) carrying Sr43 on shortened alien chromosome segments (about 17.5 and 13.7 % of the translocation chromosomes, respectively) were obtained, and two molecular markers linked to Sr43 in these lines were identified. The new wheat lines with Sr43 and the closely linked markers provide new resources for improving resistance to Ug99 and other races of stem rust in wheat.     

Development and characterization of wheat- Leymus racemosus translocation lines with resistance to Fusarium Head Blight

Wheat scab (Fusarium Head Blight, FHB) is a destructive disease in the warm and humid wheat-growing areas of the world. Finding diverse sources of FHB resistance is critical for genetic diversity of resistance for wheat breeding programs. Leymus racemosus is a wild perennial relative of wheat and is highly resistant to FHB. Three wheat- L. racemosus disomic addition (DA) lines DA5Lr#1, DA7Lr#1 and DALr.7 resistant to FHB were used to develop wheat- L.racemosus translocation lines through irradiation and gametocidal gene-induced chromosome breakage. A total of nine wheat-alien translocation lines with wheat scab resistance were identified by chromosome C-banding, GISH, telosomic pairing and RFLP analyses. In line NAU614, the long arm of 5Lr#1 was translocated to wheat chromosome 6B. Four lines, NAU601, NAU615, NAU617, and NAU635, had a part of the short arm of 7Lr#1 transferred to different wheat chromosomes. Four other lines, NAU611, NAU634, NAU633, and NAU618, contained translocations involving Leymus chromosome Lr.7 and different wheat chromosomes. The resistance level of the translocation lines with a single alien chromosome segment was higher than the susceptible wheat parent Chinese Spring but lower than the alien resistant parent L. racemosus. At least three resistance genes in L. racemosus were identified. One was located on chromosome Lr.7, and two could be assigned to the long arm of 5Lr#1 and the short arm of 7Lr#1.     

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.     

<Emphasis Type="Italic">Lr41, Lr39,</Emphasis> and a leaf rust resistance gene from<Emphasis Type="Italic"> Aegilops cylindrica</Emphasis> may be allelic and are located on wheat chromosome 2DS

The leaf rust resistance gene Lr41 in wheat germplasm KS90WGRC10 and a resistance gene in wheat breeding line WX93D246-R-1 were transferred to Triticum aestivum from Aegilops tauschii and Ae. cylindrica, respectively. The leaf rust resistance gene in WX93D246-R-1 was located on wheat chromosome 2D by monosomic analysis. Molecular marker analysis of F₂ plants from non-critical crosses determined that this gene is 11.2 cM distal to marker Xgwm210 on the short arm of 2D. No susceptible plants were detected in a population of 300 F₂ plants from a cross between WX93D246-R-1 and TA 4186 (Lr39), suggesting that the gene in WX93D246-R-1 is the same as, or closely linked to, Lr39. In addition, no susceptible plants were detected in a population of 180 F₂ plants from the cross between KS90WGRC10 and WX93D246-R-1. The resistance gene in KS90WGRC10, Lr41, was previously reported to be located on wheat chromosome 1D. In this study, no genetic association was found between Lr41 and 51 markers located on chromosome 1D. A population of 110 F₃ lines from a cross between KS90WGRC10 and TAM 107 was evaluated with polymorphic SSR markers from chromosome 2D and marker Xgdm35 was found to be 1.9 cM proximal to Lr41. When evaluated with diverse isolates of Puccinia triticina, similar reactions were observed on WX93D246-R-1, KS90WGRC10, and TA 4186. The results of mapping, allelism, and race specificity test indicate that these germplasms likely have the same gene for resistance to leaf rust.Contribution number 03-348-J from the Kansas Agricultural Experimental Station, Manhattan, KansasCommunicated by J. Dvorak     

Mapping genes Lr53 and Yr35 on the short arm of chromosome 6B of common wheat with microsatellite markers and studies of their association with Lr36

The rust resistance genes Lr53 and Yr35, transferred to common wheat from Triticum dicoccoides, were reported previously to be completely linked on chromosome 6B. Four F ₃ families were produced from a cross between a line carrying Lr53 and Yr35 (98M71) and the leaf rust and stripe rust susceptible genotype Avocet “S” and were rust tested using Puccinina triticina pathotype 53-1,(6),(7),10,11 and Puccinia striiformis f. sp. tritici pathotype 110 E143 A+. The homozygous resistant lines produced infection types of “;1−” and “;N” to these pathotypes, respectively. The Chi-squared tests indicated goodness-of-fit of the data for one leaf rust gene and one stripe rust gene segregation. Linkage analysis using this population demonstrated recombination of 3% between the genes. Microsatellite markers located on the short arm of chromosome 6B were used to map the genes, with the markers cfd1 and gwm508 being mapped approximately 1.1 and 4.5 cM, respectively, proximal to Lr53. Additional studies of the relationship between Lr36, also located on the short arm of chromosome 6B, and Lr53 indicated that the two genes were independent.     

Identification and validation of molecular markers linked to the leaf rust resistance gene Lr19 in wheat

A leaf rust resistance gene Lr19 on the chromosome 7DL of wheat derived from Agropyron elongatum was tagged with random amplified polymorphic DNA (RAPD) and microsatellite markers. The F₂ population of 340 plants derived from a cross between the leaf rust resistant near-isogenic line (NIL) of Thatcher (Tc + Lr19) and leaf rust susceptible line Agra Local that segregated for dominant monogenic leaf rust resistance was utilized for generating the mapping population. The molecular markers were mapped in the F₂ derived F₃ homozygous population of 140 seedlings. Sixteen RAPD markers were identified as linked to the alien gene Lr19 among which eight were in a coupling phase linkage. Twelve RAPD markers co-segregated with Lr19 locus. Nine microsatellite markers located on the long arm of chromosome 7D were also mapped as linked to the gene Lr19, including 7 markers which co-segregated with Lr19 locus, thus generating a saturated region carrying 25 molecular markers linked to the gene Lr19 within 10.2 ± 0.062 cM on either side of the locus. Two RAPD markers S265₅₁₂ and S253₇₃₇ which flanked the locus Lr19 were converted to sequence characterized amplified region markers SCS265₅₁₂ and SCS253₇₃₆, respectively. The marker SCS265₅₁₂ was linked with Lr19 in a coupling phase and the marker SCS253₇₃₆ was linked in a repulsion phase, which when used together mimicked one co-dominant marker capable of distinguishing the heterozygous resistant seedlings from the homozygous resistant. The molecular markers were validated on NILs mostly in Thatcher background isogenic for 44 different Lr genes belonging to both native and alien origin. The validation for polymorphism in common leaf rust susceptible cultivars also confirmed the utility of these tightly linked markers to the gene Lr19 in marker-assisted selection.     

A study of modified forms of the Lr19 translocation of common wheat

 Following the induction of allosyndetic pairing between the Thinopyrum-derived Lr19 translocation in ‘Indis’ wheat and homoeologous wheat chromatin, eight suspected recombinants for the Lr19 region were recovered. These selections were characterised for marker loci that were previously used to construct a physical map of the Lr19 segment. At the same time near-isogenic lines were developed for some of the selected segments and tested for seedling leaf-rust resistance in order to confirm the presence of Lr19. It appeared that three of the four white-endosperm selections do not possess Lr19 and only one, 88M22-149, is a true Lr19 recombinant. The resistance gene in the three non-Lr19 selections resides on chromosome 6B, appears to derive from ‘Indis’, and was selected unintentionally during backcrossing. The pedigree of ‘Indis’ is suspect and it is believed that the Lr19 translocation in ‘Indis’ is in reality the Th. ponticum-derived (T4) segment rather than being of Th. distichum origin as was believed earlier. The white-endosperm recombinant, 88M22-149, retained the complete Lr19 resistance and was apparently re-located to chromosome arm 7BL in a double-crossover event. 88M22-149 has lost the Sd1 gene and often shows strong self-elimination in translocation heterozygotes. This effect may result from additional gametocidal loci or from an altered chromosome structure following re-location of the segment. 88M22-149 in fact contains a duplicated region involving the Wsp-B1 locus. Three selections had partially white endosperms and expressed Lr19 and other Thinopyrum marker alleles. Polymorphisms for the available markers confirmed that the translocated segment in at least one of them had been shortened through recombination with chromosome arm 7DL. Further markers need to be studied in order to determine whether the translocation in the remaining two partially white recombinants had also undergone recombination with wheat. The eighth selection has yellow endosperm and appears to self-eliminate in certain translocation heterozygotes. No evidence of recombination could be found with the markers used. If the latter selections are in fact recombinants they may prove useful in attempts to unravel the complex segregation distortion mechanism. Received: 8 August 1996 / Accepted: 10 January 1997     

Molecular mapping of leaf rust resistance gene <Emphasis Type="Italic">LrZH84</Emphasis> in Chinese wheat line Zhou 8425B

Leaf rust, caused by Puccinia triticina, is one of the most widespread diseases in common wheat (Triticum aestivum L.) worldwide. With the objective of identifying and mapping new genes for resistance to leaf rust, F₁, F₂ plants and F₃ lines from a cross between resistant line Zhou 8425B and susceptible line Chinese Spring were inoculated with Chinese P. triticina races THTT and MBHP in the greenhouse. A total of 793 pairs of SSR primers were used to test the parents and resistant and susceptible bulks. Seven polymorphic chromosome 1B markers were used for genotyping the F₂ and F₃ populations. Zhou 8425B carried a single dominant resistance gene, temporarily designated LrZH84, linked to SSR markers gwm582 and barc8 with genetic distances of 3.9 and 5.2 cM, respectively. The Xbarc8 allele co-segregated with Lr26 in the F₃ population. The Xgwm582 allele associated with LrZH84 was identified as a leaf rust resistance gene and shown to be present in the Predgornaia 2 parent of Zhou 8425B. The seedling reaction pattern of LrZH84 was different from those of lines with Lr26, Lr33, Lr44 and Lr46, all of which are located in chromosome 1B. It was concluded that LrZH84 is likely to be a new leaf rust resistance gene.     

Molecular mapping of adult-plant race-specific leaf rust resistance gene <Emphasis Type="Italic">Lr12</Emphasis> in bread wheat

Wheat (Triticum aestivum) gene Lr12 provides adult-plant race-specific resistance to leaf rust caused by Puccinia triticina. It is completely linked or identical to Lr31, which confers seedling resistance only when the complementary gene Lr27 is also present. F₂ and F₂-derived F₃ families were developed from a cross between the susceptible variety Thatcher and TcLr12, an isoline carrying Lr12. Of 230 F₃ families, 55 were homozygous resistant, 115 were segregating for resistance, and 60 were susceptible to P. triticina, fitting a monogenic 1:2:1 segregation ratio. Lr12 was mapped on chromosome arm 4BL and was flanked by markers Xgwm251 and Xgwm149 at distances of 0.9 and 1.9 cM, respectively. Using linked markers and wheat deletion stocks, Lr12 was located in deletion bin 4BL-5, FL = 0.86–1.0, comprising the terminal 14% of 4BL. The markers will be useful for following Lr12/Lr31 in crosses and for further mapping studies.     

A candidate for Lr19, an exotic gene conditioning leaf rust resistance in wheat

Lr19, one of the few widely effective genes conferring resistance to leaf rust in wheat, was transferred from the wild relative Thinopyrum ponticum to durum wheat. Since Lr19 confers a hypersensitive response to the pathogen, it was considered likely that the gene would be a member of the major nucleotide-binding site (NBS)-leucine-rich repeat (LRR) plant R gene family. NBS profiling, based on PCR amplification of conserved NBS motifs, was applied to durum wheat–Th. ponticum recombinant lines involving different segments of the alien 7AgL chromosome arm, carrying or lacking Lr19. Differential PCR products were isolated and sequenced. From one such sequence (AG15), tightly linked to Lr19, a 4,121-bp full-length cDNA was obtained. Its deduced 1,258 amino acid sequence has the characteristic NBS-LRR domains of plant R gene products and includes a coiled-coil (CC) region typical of monocots. The genomic DNA sequence showed the presence of two exons and a short intron upstream of the predicted stop codon. Homology searches revealed considerable identity of AG15 with the cloned wheat resistance gene Pm3a and a lower similarity with wheat Lr1, Lr21, and Lr10. Quantitative PCR on leaf-rust-infected and non-infected Lr19 carriers proved AG15 to be constitutively expressed, as is common for R genes. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Trigenomic chromosomes by recombination of <Emphasis Type="Italic">Thinopyrum intermedium </Emphasis>and <Emphasis Type="Italic">Th. ponticum </Emphasis>translocations in wheat

Rusts and barley yellow dwarf virus (BYDV) are among the main diseases affecting wheat production world wide for which wild relatives have been the source of a number of translocations carrying resistance genes. Nevertheless, along with desirable traits, alien translocations often carry deleterious genes. We have generated recombinants in a bread wheat background between two alien translocations: TC5, ex-Thinopyrum (Th) intermedium, carrying BYDV resistance gene Bdv2; and T4m, ex-Th. ponticum, carrying rust resistance genes Lr19 and Sr25. Because both these translocations are on the wheat chromosome arm 7DL, homoeologous recombination was attempted in the double hemizygote (TC5/T4m) in a background homozygous for the ph1b mutation. The identification of recombinants was facilitated by the use of newly developed molecular markers for each of the alien genomes represented in the two translocations and by studying derived F₂, F₃ and doubled haploid populations. The occurrence of recombination was confirmed with molecular markers and bioassays on families of testcrosses between putative recombinants and bread wheat, and in F₂ populations derived from the testcrosses. As a consequence it has been possible to derive a genetic map of markers and resistance genes on these previously fixed alien linkage blocks. We have obtained fertile progeny carrying new tri-genomic recombinant chromosomes. Furthermore we have demonstrated that some of the recombinants carried resistance genes Lr19 and Bdv2 yet lacked the self-elimination trait associated with shortened T4 segments. We have also shown that the recombinant translocations are fixed and stable once removed from the influence of the ph1b. The molecular markers developed in this study will facilitate selection of individuals carrying recombinant Th. intermedium–Th. ponticum translocations (Pontin series) in breeding programs. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Molecular mapping of leaf rust resistance gene <Emphasis Type="Italic">LrBi16</Emphasis> in Chinese wheat cultivar Bimai 16

Leaf rust, caused by Puccinia triticina, is one of the most widespread diseases in common wheat (Triticum aestivum L.) globally. With the objective of identifying and mapping new genes for resistance to leaf rust, F₁, F₂ plants and F₃ lines from a cross between resistant cultivar Bimai 16 and susceptible cultivar Thatcher were inoculated with Chinese Puccinia triticina pathotypes FHTT and PHTS in the greenhouse. In the first seedling test, Bimai 16, Thatcher, 20 F₁ plants, 359 F₂ plants and 298 F₃ lines were inoculated with pathotype FHTT. A set of 1,255 simple sequence repeat (SSR) primer pairs were used to test the parents, and resistant and susceptible bulks. Seven polymorphic markers on chromosome 7BL were used for genotyping the F₂ and F₃ populations. The results indicated that Bimai 16 carried a single dominant resistance gene, temporarily designated LrBi16, closely linked to SSR markers Xcfa2257 and Xgwm344, with genetic distances of 2.8 and 2.9 cM, respectively. In the second seedling test, two dominant resistance genes were identified in Bimai 16 based on seedling reactions of 254 F₂ plants inoculated with pathotype PHTS. One of the genes was LrBi16, and the other was likely to be LrZH84, which is located in chromosome 1BL. The seedling reaction pattern of plants with LrBi16 was different from that of the Thatcher lines, with Lr14a and Lr14b located on chromosome 7BL. It was concluded that LrBi16 is likely to be a new leaf rust resistance gene.