Inheritance and mapping of powdery mildew resistance gene <Emphasis Type="Italic">Pm43</Emphasis> introgressed from <Emphasis Type="Italic">Thinopyrum</Emphasis><Emphasis Type="Italic">intermedium</Emphasis> into wheat

Powdery mildew resistance from Thinopyrum intermedium was introgressed into common wheat (Triticum aestivum L.). Genetic analysis of the F₁, F₂, F₃ and BC₁ populations from powdery mildew resistant line CH5025 revealed that resistance was controlled by a single dominant allele. The gene responsible for powdery mildew resistance was mapped by the linkage analysis of a segregating F₂ population. The resistance gene was linked to five co-dominant genomic SSR markers (Xcfd233, Xwmc41, Xbarc11, Xgwm539 and Xwmc175) and their most likely order was Xcfd233–Xwmc41–Pm43–Xbarc11–Xgwm539–Xwmc175 at 2.6, 2.3, 4.2, 3.5 and 7.0 cM, respectively. Using the Chinese Spring nullisomic-tetrasomic and ditelosomic lines, the polymorphic markers and the resistance gene were assigned to chromosome 2DL. As no powdery mildew resistance gene was previously assigned to chromosome 2DL, this new resistance gene was designated Pm43. Pm43, together with the identified closely linked markers, could be useful in marker-assisted selection for pyramiding powdery mildew resistance genes. Runli He and Zhijian Chang contributed equally to this work.     

High-density genetic and physical bin mapping of wheat chromosome 1D reveals that the powdery mildew resistance gene <Emphasis Type="Italic">Pm24</Emphasis> is located in a highly recombinogenic region

Genetic maps of wheat chromosome 1D consisting of 57 microsatellite marker loci were constructed using Chinese Spring (CS) × Chiyacao F₂ and the International Triticeae Mapping Initiative (ITMI) recombinant inbred lines (RILs) mapping populations. Marker order was consistent, but genetic distances of neighboring markers were different in two populations. Physical bin map of 57 microsatellite marker loci was generated by means of 10 CS 1D deletion lines. The physical bin mapping indicated that microsatellite marker loci were not randomly distributed on chromosome 1D. Nineteen of the 24 (79.2%) microsatellite markers were mapped in the distal 30% genomic region of 1DS, whereas 25 of the 33 (75.8%) markers were assigned to the distal 59% region of 1DL. The powdery mildew resistance gene Pm24, originating from the Chinese wheat landrace Chiyacao, was previously mapped in the vicinity of the centromere on the short arm of chromosome 1D. A high density genetic map of chromosome 1D was constructed, consisting of 36 markers and Pm24, with a total map length of 292.7 cM. Twelve marker loci were found to be closely linked to Pm24. Pm24 was flanked by Xgwm789 (Xgwm603) and Xbarc229 with genetic distances of 2.4 and 3.6 cM, respectively, whereas a microsatellite marker Xgwm1291 co-segregated with Pm24. The microsatellite marker Xgwm1291 was assigned to the bin 1DS5-0.70-1.00 of the chromosome arm 1DS. It could be concluded that Pm24 is located in the ‘1S0.8 gene-rich region’, a highly recombinogenic region of wheat. The results presented here would provide a start point for the map-based cloning of Pm24.     

Molecular mapping of powdery mildew resistance gene <Emphasis Type="Italic">PmHNK</Emphasis> in winter wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) cultivar Zhoumai 22

The Chinese winter wheat cultivar Zhoumai 22 is highly resistant to powdery mildew. The objectives of this study were to map a powdery mildew resistance gene in Zhoumai 22 using molecular markers and investigate its allelism with Pm13. A total of 278 F₂ and 30 BC₁ plants, and 143 F₃ lines derived from the cross between resistant cultivar Zhoumai 22 and susceptible cultivar Chinese Spring were used for resistance gene tagging. The 137 F₂ plants from the cross Zhoumai 22/2761-5 (Pm13) were employed for the allelic test of the resistance genes. Two hundred and ten simple sequence repeat (SSR) markers were used to test the two parents, and resistant and susceptible bulks. Subsequently, seven polymorphic markers were used for genotyping the F₂ and F₃ populations. The results indicated that the powdery mildew resistance in Zhoumai 22 was conferred by a single dominant gene, designated PmHNK tentatively, flanked by seven SSR markers Xgwm299, Xgwm108, Xbarc77, Xbarc84, Xwmc326, Xwmc291 and Xwmc687 on chromosome 3BL. The resistance gene was closely linked to Xwmc291 and Xgwm108, with genetic distances of 3.8 and 10.3 cM, respectively, and located on the chromosome bin 3BL-7-0.63-1.0 in the test with a set of deletion lines. Seedling tests with seven isolates of Blumeria graminis f. sp. tritici (Bgt) and allellic test indicated that PmHNK is different from Pm13, and Pm41 seems also to be different from PmHNK due to its origin from T. dicoccoides and molecular evidence. These results indicate that PmHNK is likely to be a novel powdery mildew resistance gene in wheat.     

Development and evaluation of broadly applicable markers for <Emphasis Type="Italic">Restorer-of-fertility</Emphasis> in pepper

The Restorer-of-fertility (Rf) gene is used for efficient hybrid seed production in chili pepper. Although molecular markers linked to Rf in pepper are available, their applications have been limited by lack of agreement between marker genotype and phenotype. To overcome this limitation, we developed new molecular markers using an Rf-segregating population for which most of previously developed markers are not suitable, because of lack of polymorphism. The petunia Rf gene was used as a candidate for marker development. First of all, a pepper bacterial artificial chromosome (BAC) library was screened using a pepper homolog of the petunia Rf gene. The 52 selected BAC clones were classified into three contig groups and each contig group was mapped to chromosome 6. Three markers were developed using the three groups; their genetic distances from the Rf locus were 1.4, 3.2 and 14 cM, respectively. In the second place, an Rf-linked marker was developed from the sequence of a tomoto BAC clone containing three genes which are homologous to petunia Rf gene. Genetic distance between this marker and Rf gene was 1.4 cM. When newly-, and previously-developed molecular markers linked to Rf were applied to 55 pepper breeding lines, one marker named CRF-SCAR was found to be the most broadly applicable, based on correct determination of phenotypes. In the present study, we demonstrate that previously cloned Rf genes can be used as candidate genes for development of new markers for the reliable detection of restorer lines. We expect that the newly-developed markers and information obtained from application of markers will be useful for reliable detection of restorer lines.     

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.     

Genetic mapping of <Emphasis Type="Italic">Eef1</Emphasis>, a major effect QTL for early flowering in <Emphasis Type="Italic">Eucalyptus grandis</Emphasis>

An early flowering mutant plant of Eucalyptus grandis with normal vegetative growth was found in a nursery in northern Brazil. This mutant plant flowers at approximately 90 days from germination. A cross between a wild-type (normal flowering) tree and the mutant was carried out, generating a progeny of 88 individuals where early flowering segregated in an approximate 1:1 ratio. A genome scan with 100 microsatellite markers distributed across the genome was carried out using bulk segregant analysis (BSA) on two contrasting bulks of 15 plants each. Linkages (LOD>3.0) with a major effect early flowering quantitative trait locus (QTL) were detected and confirmed by a full scale cosegregation analysis for markers EMBRA27, EMBRA60, EMBRA164, EMBRA158, EMBRA91, and EMBRA65. A localized linkage map involving the six loci and the early flowering QTL named Eucalyptus early flowering 1 (Eef1) was constructed belonging to linkage group #2 in the existing microsatellite reference map. The Eef1 locus was mapped between markers EMBRA27 and EMBRA164, with distances of 21.8 and 6.4 cM, respectively. In introgression experiments, these two markers could be successfully used with an expected precision of 98% to select plants carrying the Eef1 mutant allele, assuming no recombination interference in the genomic segment. Early flowering could be a very useful trait both in breeding as well as experimental genetics of Eucalyptus.     

<Emphasis Type="Italic">Pm37</Emphasis>, a new broadly effective powdery mildew resistance gene from <Emphasis Type="Italic">Triticum </Emphasis><Emphasis Type="Italic">timopheevii</Emphasis>

Powdery mildew is an important foliar disease in wheat, especially in areas with a cool or maritime climate. A dominant powdery mildew resistance gene transferred to the hexaploid germplasm line NC99BGTAG11 from T. timopheevii subsp. armeniacum was mapped distally on the long arm of chromosome 7A. Differential reactions were observed between the resistance gene in NC99BGTAG11 and the alleles of the Pm1 locus that is also located on chromosome arm 7AL. Observed segregation in F_2:3 lines from the cross NC99BGTAG11 × Axminster (Pm1a) demonstrate that germplasm line NC99BGTAG11 carries a novel powdery mildew resistance gene, which is now designated as Pm37. This new gene is highly effective against all powdery mildew isolates tested so far. Analyses of the population with molecular markers indicate that Pm37 is located 16 cM proximal to the Pm1 complex. Simple sequence repeat (SSR) markers Xgwm332 and Xwmc790 were located 0.5 cM proximal and distal, respectively, to Pm37. In order to identify new markers in the region, wheat expressed sequence tags (ESTs) located in the distal 10% of 7AL that were orthologous to sequences from chromosome 6 of rice were targeted. The two new EST-derived STS markers were located distal to Pm37 and one marker was closely linked to the Pm1a region. These new markers can be used in marker-assisted selection schemes to develop wheat cultivars with pyramids of powdery mildew resistance genes, including combinations of Pm37 in coupling linkage with alleles of the Pm1 locus.     

Molecular mapping of resistance gene to English grain aphid (<Emphasis Type="Italic">Sitobion avenae</Emphasis> F.) in <Emphasis Type="Italic">Triticum durum</Emphasis> wheat line C273

The English grain aphid, Sitobion avenae (Fabricius), is one of the most important insect pests causing substantial yield losses in wheat production in China and other grain-growing areas in the world. The efficient utilization of wheat genes for resistance to English grain aphid (EGA) provides an efficient, economic and environmentally sound approach to reduce the yield losses. In the present study, the wheat line C273 (Triticum durum AABB, 2n = 4x = 28), is resistant to EGA in greenhouse and field tests. To identify the resistance gene, designated RA-1 temporarily, C273 was crossed with susceptible genotype Poland 305 (T. polonicum, AABB, 2n = 4x = 28). The F₁, F₂ and F_2:3 lines were tested with EGA in the field and greenhouse. The results indicated that RA-1 is a single dominant gene, closely linked to the microsatellite markers (SSR) Xwmc179, Xwmc553 and Xwmc201 on chromosome 6AL at genetic distances of 3.47, 4.73 and 7.57 cM, respectively. The three SSR markers will be valuable in marker-assisted selection for resistance to EGA as well as for cloning this gene in the future.     

Molecular mapping of <Emphasis Type="Italic">Stb1</Emphasis>, a potentially durable gene for resistance to septoria tritici blotch in wheat

Septoria tritici blotch (STB), caused by the ascomycete Mycosphaerella graminicola (anamorph Septoria tritici), was the most destructive disease of wheat in Indiana and adjacent states before deployment of the resistance gene Stb1 during the early 1970s. Since then, Stb1 has provided durable protection against STB in widely grown wheat cultivars. However, its chromosomal location and allelic relationships to most other STB genes are not known, so the molecular mapping of Stb1 is of great interest. Genetic analyses and molecular mapping were performed for two mapping populations. A total of 148 F₁ plants (mapping population I) were derived from a three-way cross between the resistant line P881072-75-1 and the susceptible lines P881072-75-2 and Monon, and 106 F₆ recombinant-inbred lines (mapping population II) were developed from a cross between the resistant line 72626E2-12-9-1 and the susceptible cultivar Arthur. Bulked-segregant analysis with random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), and microsatellite or simple-sequence repeat (SSR) markers was conducted to identify those that were putatively linked to the Stb1 gene. Segregation analyses confirmed that a single dominant gene controls the resistance to M. graminicola in each mapping population. Two RAPD markers, G7₁₂₀₀ and H19₅₂₀, were tightly linked to Stb1 in wheat line P881072-75-1 at distances of less than 0.68 cM and 1.4 cM, respectively. In mapping population II, the most closely linked marker was SSR Xbarc74, which was 2.8 cM proximal to Stb1 on chromosome 5BL. Microsatellite loci Xgwm335 and Xgwm213 also were proximal to Stb1 at distances of 7.4 cM and 8.3 cM, respectively. The flanking AFLP marker, EcoRI-AGC/MseI-CTA-1, was 8.4 cM distal to Stb1. The two RAPD markers, G7₁₂₀₀ and H19₅₂₀, and AFLP EcoRI-AGC/MseI-CTA-1, were cloned and sequenced for conversion into sequence-characterized amplified region (SCAR) markers. Only RAPD allele H19₅₂₀ could be converted successfully, and none of the SCAR markers was diagnostic for the Stb1 locus. Analysis of SSR and the original RAPD primers on several 5BL deletion stocks positioned the Stb1 locus in the region delineated by chromosome breakpoints at fraction lengths 0.59 and 0.75. The molecular markers tightly linked to Stb1 could be useful for marker-assisted selection and for pyramiding of Stb1 with other genes for resistance to M. graminicola in wheat.     

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<Emphasis Type="Italic"> Fusarium oxysporum</Emphasis> f. sp.<Emphasis Type="Italic"> ciceris</Emphasis> race 3 resistance gene in chickpea

Sequence-tagged microsatellite site (STMS) and sequence-tagged site (STS) markers linked closely to Fusarium oxysporum f. sp. ciceris race 3 resistance gene in chickpea were identified, and linkage between three wilt resistance genes was elucidated. The resistance to race 3 in chickpea germplasm accession WR-315 was inherited as a single gene, designated foc-3, in 100 F₇ recombinant inbred lines derived from the cross of WR-315 (resistant) × C-104 (susceptible). The foc-3 gene was mapped 0.6 cM from STMS markers TA96 and TA27 and STS marker CS27A. Another STMS marker, TA194, at 14.3 cM, flanked the gene on the other side. Linkage between foc-3 and two other chickpea wilt resistance genes, foc-1 (syn. h ₁ ) and foc-4, was established. foc-3 was mapped 9.8 cM from foc-1 and 8.7 cM from foc-4, whereas foc-1 and foc-4 are closely linked at 1.1 cM. The identification of closely linked markers to resistance genes will facilitate marker-assisted selection for introgression of the race 3 resistance gene to susceptible chickpea lines.Communicated by H.C. Becker     

Genetic characterization and mapping of major gene resistance to potato leafroll virus in <Emphasis Type="Italic">Solanum</Emphasis><Emphasis Type="Italic">tuberosum</Emphasis> ssp. <Emphasis Type="Italic">andigena</Emphasis>

Major gene inheritance of resistance to Potato leafroll virus (PLRV) was demonstrated in a parthenogenic population derived from the highly resistant tetraploid andigena landrace, LOP-868. This major gene or chromosome region seems to control a single mechanism for resistance to infection and virus accumulation in this source. About 149 dihaploid lines segregated in a ratio of 107 resistant to 32 susceptible, fitting the expected ratio for inheritance of a duplex gene under random chromatid segregation. A tetraploid AFLP map was constructed using as reference the ultra high density (UHD) map. All AFLP markers associated with PLRV resistance mapped to the same linkage group. Map position was confirmed by analysis of previously-mapped SSR markers. Rl _adg is located on the upper arm of chromosome V, at 1 cM from its most closely linked AFLP marker, E35M48.192. This marker will be used to develop allele-specific primers or a pair of flanking PCR-based markers for their use in marker assisted selection.     

Genetic mapping of the novel <Emphasis Type="Italic">Turnip mosaic virus</Emphasis> resistance gene <Emphasis Type="Italic">TuRB03</Emphasis> in <Emphasis Type="Italic">Brassica napus</Emphasis>

A new source of resistance to the pathotype 4 isolate of Turnip mosaic virus (TuMV) CDN 1 has been identified in Brassica napus (oilseed rape). Analysis of segregation of resistance to TuMV isolate CDN 1 in a backcross generation following a cross between a resistant and a susceptible B. napus line showed that the resistance was dominant and monogenic. Molecular markers linked to this dominant resistance were identified using amplified fragment length polymorphism (AFLP) and microsatellite bulk segregant analysis. Bulks consisted of individuals from a BC₁ population with the resistant or the susceptible phenotype following challenge with CDN 1. One AFLP and six microsatellite markers were associated with the resistance locus, named TuRB03, and these mapped to the same region on chromosome N6 as a previously mapped TuMV resistance gene TuRB01. Further testing of TuRB03 with other TuMV isolates showed that it was not effective against all pathotype 4 isolates. It was effective against some, but not all pathotype 3 isolates tested. It provided further resolution of TuMV pathotypes by sub-dividing pathotypes 3 and 4. TuRB03 also provides a new source of resistance for combining with other resistances in our attempts to generate durable resistance to this virus.     

Genetic linkage maps of barfin flounder (<Emphasis Type="Italic">Verasper moseri</Emphasis>) and spotted halibut (<Emphasis Type="Italic">Verasper variegatus</Emphasis>) based on AFLP and microsatellite markers

Barfin flounder (Verasper moseri) and spotted halibut (Verasper variegatus) are two economically important marine fish species for aquaculture in China, Korea and Japan. Construction of genetic linkage maps is an interesting issue for molecular marker-assisted selection (MAS) and for better understanding the genome structure. In the present study, we constructed genetic linkage maps for both fish species using AFLP and microsatellite markers based on an interspecific F₁ hybrid family (female V. moseri and male V. variegatus). The female genetic map comprised 98 markers (58 AFLP markers and 40 microsatellite markers), distributing in 27 linkage groups, and spanning 637 cM with an average resolution of 8.9 cM. Whereas the male genetic map consisted of 86 markers (48 AFLP and 38 microsatellite markers) in 24 linkage groups, covering a length of 625 cM with an average marker spacing of 10 cM. The expected genome length was 1,128 cM in female and 1,115 cM in male, and the estimated coverage of genome was 56% for both genetic maps. Moreover, five microsatellite markers were observed to be common to both genetic maps. This is the first time to report the genetic linkage maps of V. moseri and V. variegatus that could serve as the basis for genetic improvement and selective breeding, candidate genes cloning, and genome structure research.     

High-density AFLP map of nonbrittle rachis 1 (<Emphasis Type="Italic">btr1</Emphasis>) and 2 (<Emphasis Type="Italic">btr2</Emphasis>) genes in barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.)

Wild relatives of barley disperse their seeds at maturity by means of their brittle rachis. In cultivated barley, brittleness of the rachis was lost during domestication. Nonbrittle rachis of occidental barley lines is controlled by a single gene (btr1) on chromosome 3H. However, nonbrittle rachis of oriental barley lines is controlled by a major gene (btr2) on chromosome 3H and two quantitative trait loci on chromosomes 5HL and 7H. This result suggests multiple mutations of the genes involved in the formation of brittle rachis in oriental lines. The btr1 and btr2 loci did not recombine in the mapping population analyzed. This result agrees with the theory of tight linkage between the two loci. A high-density amplified fragment-length polymorphism (AFLP) map of the btr1/btr2 region was constructed, providing an average density of 0.08 cM/locus. A phylogenetic tree based on the AFLPs showed clear separation of occidental and oriental barley lines. Thus, barley consists of at least two lineages as far as revealed by molecular markers linked to nonbrittle rachis genes.Electronic Supplementary Material Supplementary material is available for this article at An erratum to this article can be found at     

Chromosomal location of <Emphasis Type="Italic">Pm35</Emphasis>, a novel <Emphasis Type="Italic">Aegilops tauschii</Emphasis> derived powdery mildew resistance gene introgressed into common wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

A single gene controlling powdery mildew resistance was identified in the North Carolina germplasm line NC96BGTD3 (NCD3) using genetic analysis of F₂ derived lines from a NCD3 X Saluda cross. Microsatellite markers linked to this Pm gene were identified and their most likely order was Xcfd7, 10.3 cM, Xgdm43, 8.6 cM, Xcfd26, 11.9 cM, Pm gene. These markers and the Pm gene were assigned to chromosome 5DL by means of Chinese Spring Nullitetrasomic (Nulli5D-tetra5A) and ditelosomic (Dt5DL) lines. A detached leaf test showed a distinctive disease reaction to six pathogen isolates among the NCD3 Pm gene, Pm2 (5DS) and Pm34 (5DL). An allelism test showed independence between Pm34 and the NCD3 Pm gene. Together, the tests provided strong evidence for the presence of a novel Pm gene in NCD3, and this gene was designated Pm35.     

<Emphasis Type="Italic">Anthocyaninless1</Emphasis> gene of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> encodes a UDP-glucose:flavonoid-3-<Emphasis Type="Italic">O</Emphasis>-glucosyltransferase

We isolated several mutants of Arabidopsis thaliana (L.) Heynh. that accumulated less anthocyanin in the plant tissues, but had seeds with a brown color similar to the wild-type. These mutants were allelic with the anthocyaninless1 (anl1) mutant that has been mapped at 15.0 cM of chromosome 5. We performed fine mapping of the anl1 locus and determined that ANL1 is located between the nga106 marker and a marker corresponding to the MKP11 clone. About 70 genes are located between these two markers, including three UDP-glucose:flavonoid-3-O-glucosyltransferase-like genes and a glutathione transferase gene (TT19). A mutant of one of the glucosyltransferase genes (At5g17050) was unable to complement the anl1 phenotype, showing that the ANL1 gene encodes UDP-glucose:flavonoid-3-O-glucosyltransferase. ANL1 was expressed in all tissues examined, including rosette leaves, stems, flower buds and roots. ANL1 was not regulated by TTG1.     

High-resolution mapping of the <Emphasis Type="Italic">Alp</Emphasis> locus and identification of a candidate gene <Emphasis Type="Italic">HvMATE</Emphasis> controlling aluminium tolerance in barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.)

Aluminium (Al) tolerance in barley is conditioned by the Alp locus on the long arm of chromosome 4H, which is associated with Al-activated release of citrate from roots. We developed a high-resolution map of the Alp locus using 132 doubled haploid (DH) lines from a cross between Dayton (Al-tolerant) and Zhepi 2 (Al-sensitive) and 2,070 F₂ individuals from a cross between Dayton and Gairdner (Al-sensitive). The Al-activated efflux of citrate from the root apices of Al-tolerant Dayton was 10-fold greater than from the Al-sensitive parents Zhepi 2 and Gairdner. A suite of markers (ABG715, Bmag353, GBM1071, GWM165, HvMATE and HvGABP) exhibited complete linkage with the Alp locus in the DH population accounting 72% of the variation for Al tolerance evaluated as relative root elongation. These markers were used to map this genomic region in the Dayton/Gairdner population in more detail. Flanking markers HvGABP and ABG715 delineated the Alp locus to a 0.2 cM interval. Since the HvMATE marker was not polymorphic in the Dayton/Gairdner population we instead investigated the expression of the HvMATE gene. Relative expression of the HvMATE gene was 30-fold greater in Dayton than Gardiner. Furthermore, HvMATE expression in the F_2:3 families tested, including all the informative recombinant lines identified between HvGABP and ABG715 was significantly correlated with Al tolerance and Al-activated citrate efflux. These results identify HvMATE, a gene encoding a multidrug and toxic compound extrusion protein, as a candidate controlling Al tolerance in barley.     

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.     

Genetic linkage map of the loach <Emphasis Type="Italic">Misgurnus anguillicaudatus</Emphasis> (Teleostei: Cobitidae)

In the present study, the first genetic linkage map of the loach Misgurnus anguillicaudatus was constructed with 164 microsatellite markers and a color locus, and it included 155 newly developed markers. A total of 159 microsatellite markers and a color locus were mapped in 27 linkage groups (LGs). The female map covered 784.5 cM with 153 microsatellite markers and a color locus, whereas the male map covered 662.2 cM with 119 microsatellite markers. The centromeric position in each LG was estimated by marker-centromere mapping based on half-tetrad analysis. In 4 LGs (LG2, LG3, LG4, and LG5), the centromere was estimated at the intermediate region. In LG1, LG11, and LG12, the centromere was estimated to shift from the sub-intermediate region to the end (telomeric). The number of these LGs (7) was identical to the collective number of bi-arm metacentric (5) and sub-metacentric chromosome (2) of the haploid chromosome set (n = 5) of the loach. In the other LGs, the position of the centromere was estimated at the end or outside. These results indicate satisfactory compliance between the linkage map and the chromosome set. Our map would cover approximately almost the entire loach genome because most markers were successfully mapped.