An STS marker distinguishing the rye-derived powdery mildew resistance alleles at the Pm8/Pm17 locus of common wheat

A sequence‐tagged site marker has been developed from restriction fragment length polymorphism marker probe IAG95 for the rye‐derived powdery mildew resistance Pm8/Pm17 locus of common wheat. This polymerase chain reaction marker enables the amplification of DNA fragments with different sizes from T1AL.1RS and T1BL.1RS wheat‐rye translocation cultivars with chromatin from ‘Insave’ and ‘Petkus’ rye, respectively, and therefore will be very useful in distinguishing Pm8‐carrying cultivars from Pm17‐carrying cultivars. Results obtained with that marker were compared with resistance tests performed on detached primary leaves of 29 wheat lines from two populations derived from doubled haploid production. The molecular assay corresponded well with the resistance tests in all the lines, and therefore will be helpful for the identification of Pm17 in lines in which other Pm genes or quantitative trait loci are present.     

Identification of powdery-mildew-resistance genes in common wheat (Triticum aestivum L.). VI. Wheat cultivars grown in China

A total of 26 common wheat cultivars and advanced breeding lines grown in China were tested with a set of 11 differential powdery-mildew isolates. Seven cultivars were susceptible. Another seven cultivars showed the response pattern of resistance gene Pm2, either individually or in combination with genes Pm3d or Pm4a. Five cultivars expressed the resistance of gene Pm4b singly or in combination with Pm6. Another four cultivars exhibited the response patterns of genes Pm5, Pm6 and Pm8, respectively. Three cultivars, which included one breeding line with a pair of substituted chromosomes from Haynaldia villosa, presumably carrying the resistance gene Pm21, showed resistance-response patterns to all the isolates tested.     

Microsatellite mapping of powdery mildew resistance allele <Emphasis Type="Italic">Pm5d</Emphasis> from common wheat line IGV1-455

The powdery mildew resistance allele Pm5d in the backcross-derived wheat lines IGV1-455 (CI10904/7*Prins) and IGV1-556 (CI10904/7*Starke) shows a wide spectrum of resistance and virulent pathotypes have not yet been detected in Germany. Although this allele may be distinguished from the other documented Pm5 alleles by employing a differential set of Blumeria graminis tritici isolates, the use of linked molecular markers could enhance selection, especially for gene pyramiding. Pm5d was genetically mapped relative to six microsatellite markers in the distal part of chromosome 7BL using 82 F₃ families of the cross Chinese Spring × IGV1-455. Microsatellite-based deletion line mapping placed Pm5d in the terminal 14% of chromosome 7BL. The closely linked microsatellite markers Xgwm577 and Xwmc581 showed useful variation for distinguishing the different Pm5 alleles except the ones originating from Chinese wheat germplasm. Their use, however, would be limited to particular crosses because they are not functional markers. The occurrence of resistance genes closely linked to the Pm5 locus is discussed. Ghazaleh Nematollahi and Volker Mohler equally contributed to this work.     

Race Specific Powdery Mildew Resistance in 31 Northwest European Wheat Cultivars

Race specific powdery mildew resistance in 23 winter wheat cultivars, eight spring wheat cultivars, and 14 lines/cultivars possessing known powdery mildew resistance genes, has been studied by analyzing host/pathogen interactions. The cultivars were tested as intact seedlings, and as detached primary leaf segments on water agar; both methods revealed reproducible and concordant results. The 45 cultivars/lines were divided into 24 resistance spectra according to the patterns of reaction to the powdery mildew isolates used. Of the 31 cultivars investigated, eight did not possess any of the resistance genes detected, and the remaining 23 were divided in 16 resistance spectra. The race specific resistance of nine cultivars was conferred by the single resistance genes Pm2, Pm4b, Pm5/Ml-i: or Pm6, while the race specific resistance of 14 cultivars was conferred by 2, 3, 4 or 5 genes in combination.     

Chromosome location of genes for resistance to powdery mildew in common wheat (Triticum aestivum L.) 1. Mlk and other alleles at the Pm3 locus

Summary Several wheat cultivars/lines were inoculated with isolates of Erysiphe graminis tritici to identify new genes/alleles for resistance. The wheats were tested with 13 isolates that had been characterized from responses on differential lines with known resistance genes. Gene Mlk which occurs in cultivars Kolibri, Syros, Ralle and several other European common wheats was found to be an allele at the Pm3 locus and is now designated Pm3d. The mildew resistance in an old Australian wheat, W150, is conferred by a single gene also allelic to Pm3 and now designated Pm3e. The near-isogenic line Michigan Amber/8^*Cc possesses another allele now designated Pm3f. A Syrian land variety of common wheat shows mildew resistance that is conditioned by the combination of genes Pm1 and Pm3a. Finally, two accessions of Triticum aestivum ssp. sphaerococcum appeared to possess the Pm3c allele.     

12个小麦品种(系)白粉病抗性的遗传分析

利用17个不同来源和毒力的白粉菌菌株对12个小麦品种(系)进行苗期抗性鉴定和抗病性遗传分析,同时利用Pm2和Pm8基因的特异分子标记检测了相应基因。供试的12个品种至少能够抗11个白粉菌菌株。用E09、E20和Bg2菌株接种F₂群体,抗感植株分离比例和适合性测验证明这12个品种对不同白粉菌菌株的抗性均受1对显性基因控制。抗谱分析和基因紧密连锁分子标记(Xcfd81)分析表明良星66很可能含有Pm2或其等位基因。ω-黑麦碱基因(1RS染色体)和Glu-B1基因(1BS染色体)特异分子标记分析结果证明,山农20和郑麦9962含有T1BL·1RS易位染色体,即可能携带Pm8基因。由于Pm8基因对大多数菌株表现感病,所以这2个品种除Pm8外,还具有其他抗病基因。偃展4110与天民668对参试菌株的反应型表现一致,其他材料对不同菌株的反应型表现不同。     

Genes for Powdery Mildew Resistance in Cultivars of Spring Wheat

Twenty-three cultivars of spring wheat were inoculated with nineteen different powdery mildew isolates; their ruction patterns hive been compared with those of twenty-two cultivars/lines carrying identified powdery mildew resistance genes. Applying the gene-for-gene hypothesis, it is evident that three cultivars have none of the resistance genes used, seven others (including ‘Solo’) may carry Pm4b, only. The resistance pattern of ‘Selpek’ is identical to A/-1 resistant cultivars of winter wheat and may be explained by the presence of Pm5. The resistance pattern of Pm5 (Mt-i) cultivars is very different from a number of ‘Kolibri’-related cultivars of spring wheat. Since either all or nothing of that specific pattern has been transferred to all cross progenies of ‘Kolibri’, a single gene is assumed to oe responsible for it, preliminarily designated as Ml-k. The cultivar ‘Mephisto’ carries the ‘Normandie’ resistance (Pwl 2, 9). In five cultivars to spring wheat the combined effects of at least two of the above-mentioned sources have been found. Despite the fact that ‘Normandie’ and ‘Sappo’ are not closely related. ‘Sappo’ shows the complete ‘Normandie’ resistance pattern plus that of Pm4b. The same is true for ‘Planet’ and ‘Walter’.     

Identification of Powdery Mildew Resistance Genes in Common Wheat (Triticum aestivum L.)

Major genes for resistance to powdery mildew were analysed in 24 Czechoslovakian wheat cultivars and, in part, in their parents. For this purpose individual isolates of the pathogen, able to differentiate host lines with known resistance genes, were selected. Eight of nineteen winter wheat cultivars do not possess any major resistance gene. Three cultivars have one and seven have two genes. One cultivar carries a combination of three genes (Pm2, Pm4b, Pm8). The most common resistance genes are Pm4b, Pm5 and Pm8. Pm2 is once combined with Pm6. Only one of five spring cultivars lacked a major resistance gene. Mlk is once present alone and twice combined with Pm5. There is one spring cultivar with a novel combination of three genes: Pm1, Pm5 and another gene needing further characterization. The observations are discussed with additional results of parent lines and further information on pedigrees.     

Location of Pm3g, a powdery mildew resistance allele in wheat, by using a monosomic analysis and by identifying associated molecular markers

A segregating population of doubled-haploid lines issued from the cross between the wheat (Triticum aestivum L. em. Thell) cultivars Courtot, resistant to several isolates of powdery mildew (Blumeria graminis DC. f. sp. tritici Em. Marchal), and Chinese Spring (susceptible) was used to map Mlar, a gene carried by Courtot and conferring resistance to this pathogen. The assignation of Mlar using monosomic lines of Courtot was confirmed by the mapping analysis. Mlar was located on the short arm of the chromosome 1A, in the vicinity of the locus XGli-A5 coding for storage proteins. This result was in accordance with those demonstrating that Mlar was an allele of the Pm3 locus (Pm3g), a gene also involved in the resistance to powdery mildew. This revised version was published online in August 2006 with corrections to the Cover Date.     

Identification of powdery mildew resistance genes in common wheat (Triticum aestivum L. em Thell.). IX. Cultivars, land races and breeding lines grown in China

The objective of the study was to provide information about the occurrence and distribution of resistance genes in wheat cultivars, including old cultivars, land races and advanced breeding lines grown in China. Ninety-four accessions were analysed with a set of 11 differential powdery mildew isolates. Forty-four cultivars did not possess any major mildew resistance genes. Thirty cultivars revealed the response pattern of individual resistance genes. The most frequently encountered gene was Pm8, which occurred singly in 11 cultivars, combined either with Pm4a in three cultivars or with Pm4b in another three cultivars. However, 12 cultivars possessing the wheat-rye translocated chromosome pair T1BL-1RS did not express Pm8. Gene Pm2 was found in four cultivars and in combination with Pm6 in one cultivar. Genes Pm4a and Pm4b were observed in four and five cultivars, respectively. Another six cultivars carried Pm5. A gene combination of Pm2+Pm4b+Pm6 was found in one cultivar. Twelve cultivars and breeding lines exhibited a response pattern that could not be assigned to resistance genes or gene combinations present in the differential cultivars. Five out of these 12 cultivars/lines showed resistance to all the isolates tested. There is an urgent need to search for novel sources of mildew resistance in order to sustain resistance to existing and emerging powdery mildew pathogens.     

Identification of resistance gene analogue markers closely linked to wheat powdery mildew resistance gene Pm31

Specific oligonucleotide primers, designed for the sequences of known plant disease resistance genes, were used to amplify resistance gene analogues (RGAs) from wheat genomic DNA. This method was applied in a bulked segregant analysis to screen for the RGA markers linked to the powdery mildew resistance gene Pm31, introgressed into common wheat from wild emmer. Two RGA markers (RGA200 and RGA390) were found to be closely linked to Pm31 and completely co‐segregating with the marker allele of Xpsp3029 linked to Pm31, with a genetic distance of 0.6 cM. These two RGA markers were then integrated into the formerly established microsatellite map of Pm31 region. The result showed the effectiveness of the RGA approach for developing molecular markers linked to disease resistance genes and demonstrated the efficiency of denaturing polyacrylamide‐gel electrophoresis for detecting polymerase chain reaction polymorphism.     

A major QTL effect controlling resistance to powdery mildew in winter wheat at the adult plant stage

Powdery mildew is one of the major diseases of wheat in regions with a maritime or semi‐continental climate which can strongly affect grain yield. The objective of the study was to identify and compare quantitative resistance to powdery mildew of line RE9001 at the adult plant and vernalized seedling stages. RE9001 has no known Pm gene and shows a high level of adult plant resistance in the field. Using 104 recombinant inbred lines (RILs) of an RE9001 × ‘Courtot’ F8 population, a genetic map was developed with 363 markers distributed over 26 linkage groups and covering 3825 cM. The global map density was 1 locus/10.3 cM. RILs were assessed under field and tunnel greenhouse conditions for 2 years in two locations. Eleven quantitative trait loci (QTL) were detected at the adult stage and they explained 63% of the variation, depending on the environment. Three QTLs were found, at least, in the two environments. One QTL from RE9001, mapped on chromosome 2B, was stable in each environment. This QTL, QPm.inra.2B, explained 10.3–36.6% of the variation and could be mapped in the vicinity of the Pm6 gene. At the vernalized seedling stage, one QTL detected by the isolate 93‐27 could be an allele of the Pm3g gene present in ‘Courtot’. No residual effect of the Pm3g gene was detected at either stage. Markers flanking the QTL 2B could be useful tools to combine resistance to powdery mildew in wheat cultivars.     

Molecular mapping determines that Hessian fly resistance gene H9 is located on chromosome 1A of wheat

Hessian fly [Mayetiola destructor (Say)] is one of the major insect pests of wheat (Triticum aestivum L.) worldwide. Hessian fly resistance gene H9 was previously reported to condition resistance to Hessian fly biotype L that is prevalent in many wheat‐growing areas of eastern USA and an RAPD marker, OPO05₁₀₀₀, linked to H9 in wheat was developed using wheat near‐isogenic lines (NILs). However, marker‐assisted selection (MAS) with RAPD markers is not always feasible. One of the objectives in this study was to convert an RAPD marker linked to the gene H9 into a sequence characterized amplified region (SCAR) marker to facilitate MAS and to map H9 in the wheat genome. The RAPD fragment from OPO05₁₀₀₀ was cloned, sequenced, and converted into a SCAR marker SOPO05₉₀₉, whose linkage relationship with H9 was subsequently confirmed in two F₂ populations segregating for H9. Linkage analysis identified one sequence tagged site (STS) marker, STS‐Pm3, and the eight microsatellite markers Xbarc263, Xcfa2153, Xpsp2999, Xgwm136, Xgdm33, Xcnl76, Xcnl117 and Xwmc24 near the H9 locus on the distal region of the short arm of chromosome 1A, contrary to the previously reported location of H9 on chromosome 5A. Locus Xbarc263 was 1.2 cM distal to H9, which itself was 1.7 cM proximal to loci Xcfa2153, Xpsp2999 and Xgwm136. The loci Xgwm136, Xcfa2153 and SOPO05₉₀₉ were shown to be specific to H9 and not diagnostic to several other Hessian fly resistance genes, and therefore should be useful for pyramiding H9 with other Hessian fly resistance genes in a single genotype.     

Identification of Wheat Powdery Mildew Resistance Genes by Analysing Host-Pathogen Interactions

Fifty-nine winter wheat cultivars and thirteen lines possessing known powdery mildew resistance genes were inoculated with eleven different isolates. By comparing their resistance patterns the responsible major resistance genes of the above-mentioned cultivars have been determined. The so-called “Blaukorn” resistance is conditioned by gent Pm4b. The resistance patterns of Ml-i and Pm5 being similar, the relationship between them has to be analysed by segregating populations.     

Allelic variation for the rust resistance gene <Emphasis Type="Italic">Lr34</Emphasis>/<Emphasis Type="Italic">Yr18</Emphasis> in Canadian wheat cultivars

The wheat (Triticum aestivum L.) gene Lr34/Yr18 conditions resistance to leaf rust, stripe rust, and stem rust, along with other diseases such as powdery mildew. This makes it one of the most important genes in wheat. In Canada, Lr34 has provided effective leaf rust resistance since it was first incorporated into the cultivar Glenlea, registered in 1972. Recently, molecular markers were discovered that are either closely linked to this locus, or contained within the gene. Canadian wheat cultivars released from 1900 to 2007, breeding lines and related parental lines, were tested for sequence based markers caSNP12, caIND11, caIND10, caSNP4, microsatellite markers wms1220, cam11, csLVMS1, swm10, csLV34, and insertion site based polymorphism marker caISBP1. Thirty different molecular marker haplotypes were found among the 375 lines tested; 5 haplotypes had the resistance allele for Lr34, and 25 haplotypes had a susceptibility allele at this locus. The numbers of lines in each haplotype group varied from 1 to 140. The largest group was represented by the leaf rust susceptible cultivar “Thatcher” and many lines derived from “Thatcher”. The 5 haplotypes that had the resistance allele for Lr34 were identical for the markers tested within the coding region of the gene but differed in the linked markers wms1220, caISBP1, cam11, and csLV34. The presence of the resistance or susceptibility allele at the Lr34 locus was tracked through the ancestries of the Canadian wheat classes, revealing that the resistance allele was present in many cultivars released since the 1970s, but not generally in the older cultivars.     

A PCR-based DNA marker for detection of mutant and normal alleles of the Wx-D1 gene of wheat

To assist waxy wheat breeding a DNA marker was developed to discriminate mutant and normal alleles at the Wx‐D1 locus. This polymerase chain reaction‐based marker distinguishes the mutant from the normal allele by targeting the previously reported deletion basis of the mutant. The marker codominantly identifies the normal allele of the Wx‐D1 gene from the mutant allele originated from the Chinese landrace ‘Baihoumai’. However, attempts with a number of primer combinations targeting this deletion failed to amplify the corresponding fragment from an unrelated wheat line (NP150) that has a mutant null allele at the same locus. This indicates that NP150 has a different mutant allele from that of ‘Baihoumai’. This marker is a useful tool to identify wheat cultivars with mutant and normal alleles of the Wx‐D1 gene, and is used in marker‐assisted selection of the Wx‐D1 gene in our waxy wheat breeding programme.     

Development of SCAR markers linked to the Pm21 gene conferring resistance to powdery mildew in common wheat

Powdery mildew is an important disease in most of the wheat production areas of the world. The resistance gene Pm21 (6AL/6VS trans-location) derived from Haynaldia villosa confers resistance to all available isolates of Erysiphe (Blumeria) graminis f. sp. tritici in China and Europe. The objective of this study was to develop fast and reliable sequence characterized amplified region (SCAR) markers linked to the Pm21 gene. A random amplified polymorphic DNA (RAPD) marker for Pm21, OPH17₁₄₀₀, was converted to SCAR markers after sequencing the two ends of the polymorphic DNA fragment. Two SCAR markers, SCAR₁₂₆₅ and SCAR₁₄₀₀, were developed to detect the Pm21 gene in different genetic backgrounds. The specific SCAR₁₂₆₅ marker enable large-scale accurate screening for the presence/absence of Pm21 allele.     

Development of two multiplex PCR assays targeting improvement of bread-making and noodle qualities in common wheat

Wheat quality properties are genetically determined by the compositions of high and low molecular weight glutenin subunits, grain hardness, polyphenol oxidase (PPO) activity and starch viscosity. Two multiplex PCR assays were developed and validated using 70 cultivars and advanced lines from Chinese autumn‐sown wheat regions. Multiplex PCR I includes molecular markers for genes/loci ω‐secalin, Glu‐B1‐2a (By8), Glu‐D1‐1d (Dx5), Glu‐A3d, Glu‐B3 (for non‐1B·1R type) and Pinb‐D1b targeting improved gluten parameters and pan bread quality. Multiplex PCR II comprises markers for genes/loci Ppo‐A1, Ppo‐D1 and Wx‐B1b targeting improved noodle quality. The results were consistent with those achieved by SDS‐PAGE and RP‐HPLC, indicating that the two multiplex assays were highly effective, with good repeatability and low costs enabling their use in wheat breeding programmes. In total, nine alleles (subunits) at locus Glu‐B1, four at Glu‐D1 and five at Glu‐A3 locus were identified, and the alleles (subunits) Glu‐B1b (7 + 8), Glu‐B1c (7 + 9), Glu‐D1a (2 + 12), Glu‐D1d (5 + 10), Glu‐A3a, Glu‐A3c and Glu‐A3d were most frequently present in the cultivars and lines tested. The 1B·1R translocation was present in 28 (40.0%) lines, whereas the Wx‐B1 null allele for better noodle quality was present in only seven (10.0%) cultivars and advanced lines, and 37 (52.9%) lines had Pinb‐D1b associated with hard grains. The allele Ppo‐A1b on chromosome 2AL associated with lower PPO activity was present in 38 (54.3%) genotypes, whereas the less effective allele Ppo‐D1a on chromosome 2DL, also associated with low PPO activity was present in 45 (64.3%) of genotypes. These two multiplex PCR assays should be effective in marker assisted selection targeting improved pan bread‐making and noodle qualities.     

Identification,distribution and effects on agronomic traits of the semi-dwarfing <Emphasis Type="Italic">Rht</Emphasis> alleles in Bulgarian common wheat cultivars

Bulgarian common wheat cultivars released in the period 1925–2003 were studied using the gibberellic acid (GA) test and microsatellite analysis of the Xgwm261 locus on chromosome 2DS to identify the semi-dwarfing (Rht) genes. The old cultivars, isolated through selection from landraces, carried rare alleles (211- and 215-bp) at Xgwm261 locus, and those developed by hybridisation to foreign cultivars, carried the 165- and 174-bp alleles. Forty-two (55.3%) of 76 modern cultivars were GA-responsive. The 192-bp allele, diagnostic for Rht8, was observed in 64 (84.2%) modern cultivars, of which 37 carry Rht8 alone, and 27 possess a combination of Rht8 and a GA-insensitive allele viz. Rht-B1d (17); Rht-D1b (6) and Rht-B1b (4). The 174-bp allele is present in seven cultivars, only one of which is photoperiod-sensitive, and the rest are day-length insensitive. The 203-bp allele was found in six modern cultivars. Cultivars carrying the Rht8 allele are the most widespread and some of them have been cultivated for a long period. Cultivars with the `Saitama 27' allele (Rht-B1d) are the most productive and are second in distribution in the country. The recently observed trend for increasing the proportion of cultivars with GA-insensitive Rht genes is probably due to their combination with the 192-bp allele of Xgwm261 locus tightly linked to the Ppd-D1, to the break of the link between the 174-bp allele and ppd-D1, and to the introduction of other genes influencing flowering time.     

Identification of powdery-mildew-resistance genes in common wheat (Triticum aestivum L. em. Thell.). V. Old German cultivars and cultivars released in the former GDR

A total of 59 old wheat cultivars grown in Germany prior to 1960 were tested for mildew response using a collection of 12 differential isolates of Erysiphe graminis DC f. sp. tritici Marchal (Blumeria graminis (DC) Speer f. sp. tritici). Nineteen cultivars did not possess any major resistance gene and 25 were characterized by susceptible or intermediate responses. Fifteen cultivars revealed isolate-specific response patterns that could not be attributed to known major resistance genes or gene combinations. Many of the old German cultivars inherited a mildew-resistance gene from the Canadian cultivar ‘Garnet’ which is tentatively designated M1-Ga. Cultivars ‘Bretonischer Bartweizen’ (designated M1-Br) and ‘Adlungs Alemannen’ (designated M1-Ad) appeared to carry unknown resistance genes. Among 18 winter wheat cultivars released in the former GDR. eight showed susceptibility to all isolates used. Cv. “Borenos” carries resistance gene Pm3c. Five cultivars possess gene Pm4b. two cultivars gene pm5 and one cultivar a combination of genes Pm2 and Pm4b. Cultivar ‘Zentos’ was resistant to almost all isolates used. Its resistance might be conditioned by different unknown major resistance genes.