Comparison of a modified assay method for the endopeptidase marker Ep-D1b with the Sequence Tag Site marker XustSSR2001–7DL for strawbreaker foot rot resistance in wheat

The endopeptidase marker Ep‐D1b and Sequence Tag Site (STS) marker XustSSR2001–7DL were reported to be closely associated with the most effective resistance gene (Pch1) in wheat (Triticum aestivum L.) for strawbreaker foot rot [Pseudocercosporella herpotrichoides (Fron) Deighton]. Our objectives were to: (i) develop an efficient assay method for Ep‐D1b in wheat; (ii) correlate endopeptidase zymograms to strawbreaker foot rot reactions of various wheat genotypes; and (iii) compare the utility of Ep‐D1b and XustSSR2001–7DL for predicting disease response. An improved method of assaying for the Ep‐D1b marker using roots from a single seedling was developed, which is a 2.5‐fold improvement over the previous method. Thirty‐eight wheat genotypes with known reactions to strawbreaker foot rot were analysed for Ep‐D1b and the STS marker. Six distinct endopeptidase zymograms were identified among these 38 genotypes tested, and three of these patterns were novel. The endopeptidase marker was 100% accurate for predicting strawbreaker foot rot disease response, whereas the STS marker predicted the correct phenotype with approximately 90% accuracy. The endopeptidase marker Ep‐D1b was more effective and was more economical for use in marker‐assisted selection strategies for Pch1 in our laboratory compared with the STS marker.     

Inheritance of resistance of wheat to eyespot at the adult stage

Moderate resistance to eyespot was first incorporated in the variety Cappelle‐Desprez (CD). Later the gene Pch1, which could confer a higher level of resistance, was introduced from Aegilops ventricosa. However Pch1‐carrying varieties can sustain significant eyespot‐induced yield losses in severe attack situations. A strategy to further enhance the resistance of wheat is by pyramiding Pch1 and the genes for resistance in CD. The first requirement to achieve this is a better understanding of the genetics of resistance in CD. The resistance of the 21 Cappelle‐Desprez (Bezostaya) disomic substitution lines was evaluated. Chromosome 7A was confirmed as carrying a major gene for resistance to eyespot at the seedling stage. However, this study demonstrates that this chromosome has no effects at the adult stage. Chromosome 5A was shown to carry a major gene for resistance to eyespot at the adult stage, which was stably expressed each year of testing. Chromosomes 1A and 2B had significant effects for only two years among four.     

Amplified fragment length polymorphism-derived microsatellite sequence linked to the Pch1 and Ep-D1 loci in common wheat

Amplified fragment length polymorphism (AFLP) markers linked to the Aegilops ventricosa‐derived chromosome segment in ‘VPM1’ on which the eyespot resistance gene, Pch1, and the endopeptidase gene, Ep‐D1b, occur were identified. One marker was isolated from the gel, cloned and sequenced. Sequence analysis revealed a microsatellite repeat motif. Sequence‐specific primers were designed to amplify a product containing the repeat motif, and the microsatellite marker was tested for cosegregation with the Ep‐D1b allele. Distinct alleles were produced by the Pch1 sources, normal wheat and wheat containing the Lr19 translocation. A recombination frequency of 0.02 was calculated between the microsatellite marker and Ep‐D1.     

A New Source of Resistance to Pseudocercosporella herpotrichoides, Cause of Eyespot Disease of Wheat, Located on Chromosome 4V of Dasypyrum villosum

Resistance to Pseudocercosporella herpotrichoides in five wheat cultivars, accession W6 7283 of Dasypyrum villosum, and ‘Chinese Spring’ disomic addition lines of the D. villosum chromosomes IV, 2V, 4V, 5V, 6V and 7V, was evaluated in seedlings by measuring disease progress 6 weeks after inoculation with a β—glucuronidase—transformed strain of the pathogen and by visual estimates of disease severity. D. villosum and the disomic addition line of chromosome 4V were as resistant as wheat cultivars ‘VPM—1’ and ‘Cappelle Desprez’, but less resistant than ‘Rendezvous’. Resistance of the chromosome 4V disomic addition line was equivalent to that of D. villosum.‘Chinese Spring’ and disomic addition lines of IV, 2V, 5V, 6V and 7V were all susceptible. These results confirm Sparaguee's (1936) report of resistance in D. villosum to P. herpotrichoides and establish the chromosomal location for the genes controlling resistance. The presence of chromosome 4V in the addition line and its homocology to chromosome 4 in wheat were confirmed by Southern analysis of genomic DNA using chromosome group 4-specific clones. This genetic locus is not homoeologous with other known genes for resistance to P. herpotrichoides located on chromosome group 7, and thus represents a new source of resistance to this pathogen.     

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.     

Variation of resistance to Pseudocercosporella herpotrichoides (Fron) Deighton in wheat genotypes carrying the gene Pch-1

The effect of the gene Pch-1 on the resistance of wheat to Pseudocercosporella herpotrichoides was studied at four growth stages. The germplasm used consisted of adapted cultivars, genotypes provided by European plant breeders, near-homozygous lines and double haploid lines developed from our own breeding projects. The resistance was measured by ELISA. At all growth stages, genotypes carrying Pch-1 differed significantly in resistance. At early growth stages, there was a strong effect of the gene in most genotypes, but later the effect decreased and significant genotypes-environment interactions appeared. In addition, minor genes became more important and determined the level of adult plant resistance that proved to be inherited quantitatively. Pch-1 was of minor importance for this type of resistance. It is concluded that a high and long-lasting resistance level could be attained if the two genetically different sources of resistance were combined (resistance at juvenile stages, induced by Pch-1, and quantitative resistance at adult stages).     

Development of molecular markers linked to the wheat powdery mildew resistance gene Pm4b and marker validation for molecular breeding

Powdery mildew, caused by Blumeria graminis (DC.) E.O. Speer f. sp. tritici, is an important disease in wheat (Triticum aestivum L.). Bulk segregant analysis (BSA) was employed to identify SRAP (sequence‐related amplified polymorphism), sequence tagged site (STS) and simple sequence repeat (SSR) markers linked to the Pm4b gene, which confers good resistance to powdery mildew in wheat. Out of 240 SRAP primer combinations tested, primer combinations Me8/Em7 and Me12/Em7 yielded 220‐bp and 205‐bp band, respectively, each of them associated with Pm4b. STS‐₂₄₁ also linked to Pm4b with a genetic distance of 4.9 cM. Among the eight SSR markers located on wheat chromosome 2AL, Xgwm382 was found to be polymorphic and linked to Pm4b with a genetic distance of 11.8 cM. Further analysis was carried out using the four markers to investigate marker validation for marker‐assisted selection (MAS). The results showed that a combination of the linked markers STS_?241, Me8/Em7_?220 and Xgwm382 could be used for marker‐assisted selection of the resistance gene Pm4b in wheat breeding programmes.     

Analysis of diallel crosses between wheat genotypes with genetically different resistance to Pseudocercosporella herpotrichoides (Fron) Deighton

Two diallels were analysed for general combining ability (GCA) and specific combining ability (SCA) to study the resistance of crosses‐between wheat genotypes, advanced to the F₅ generation, to Pseudocer‐cosporella herpotrichoides. The parents either carried the resistance‐gene Pch‐1 or had different levels of quantitative resistance, one genotype was susceptible. At medium milk‐ripening, significant effects were‐found for GCA and SCA. GCA effects were the more important. Diallel crosses between genotypes, all carrying Pch‐1, revealed interactions‐of the gene with the genotypic background. Some combinations had a‐higher level of resistance than the best parent. In these populations'CH‐75417’ was involved as a parent. Both ‘CH‐75417’ and ‘F–210.13.4.42’ had significant GCA effects. Crosses between quantitatively resistant parents yielded populations that transgressed both parents. The increased resistance level was associated with ‘Cappelle‐Desprez’, distinguished by its high GCA. In some crosses SCA contributed significantly to an increase in resistance level. Selection for resistance within the best advanced populations is recommended since it‐takes advantage of additive gene action and the high heritability estimates based on ELISA values in plant progenies.     

黄淮麦区Fhb1基因的育种应用

小麦赤霉病是由禾谷镰孢菌引起的一种世界性重要病害,严重威胁小麦生产安全。黄淮麦区作为我国小麦主产区,赤霉病危害日趋严重,因缺乏半冬性抗源,抗赤霉病育种进展缓慢。Fhb1基因是迄今发现的效应最大、抗性最稳定,也是被广泛应用于全球小麦赤霉病抗性育种的主效基因,但Fhb1基因在黄淮麦区尚未被广泛应用。本研究以感病品种矮抗58为轮回亲本, H35为Fhb1基因供体亲本,通过有限回交和分子标记辅助选择,同时利用双单倍体育种和传统系谱选育两种方法,培育出了一批综合性状较好、具有Fhb1基因的优良新品系,其中徐麦DH9和徐麦17252经多年鉴定均达到中抗水平。在以徐麦36和徐麦2023为杂交父本的后代品系中,含Fhb1基因的家系赤霉病平均抗性明显优于感病对照。Fhb1基因的导入显著提高了赤霉病抗性,但部分家系对赤霉病仍旧表现出高感水平,说明赤霉病抗性还受到Fhb1基因以外其他遗传因素的显著影响。本研究为Fhb1基因在黄淮麦区抗赤霉病小麦育种中的应用提供了成功的经验。     

Amplified fragment length polymorphism- and simple sequence repeat-based molecular tagging and mapping of greenbug resistance gene Gb3 in wheat

The greenbug, Schizaphis graminum (Rondani), is the most economically damaging aphid pest of wheat in the southern Great Plains of the USA. In this study, the single, dominant greenbug resistance gene, Gb3, was molecularly tagged and genetically mapped using amplified fragment length polymorphism (AFLP) and simple sequence repeat(SSR) markers. Three AFLP loci were associated with the Gb3 locus in linkage analysis with 75 F_2:3 families from the cross between two near‐isogenic lines (NILs) for Gb3,‘TXGBE273’ and ‘TXGBE281′. Two of these loci, XMgcc Pagg and Xmagg Patg cosegregate with Gb3 in the population analysed. Further analysis indicated that XMgcc Pagg and Xmagg Patg are specific for the Gb3 locus in diverse genetic backgrounds. Two SSR markers, Xgwm111 and Xgwm428 previously mapped in wheat chromosome 7D, were shown to be linked with Gb3, 22.5 cM and 33.1 cM from Gb3, respectively, in an F₂ population of ‘Largo’בTAM 107’, suggesting that Gb3 is located in the long arm of chromosome 7D. The two AFLP markers cosegregating with Gb3 are valuable tools in developing molecular markers for marker‐assisted selection of greenbug resistance in wheat breeding.     

QTL mapping and marker-assisted selection for Fusarium head blight resistance in wheat: a review

During the past decade, numerous studies have been published on molecular mapping of Fusarium head blight (FHB) resistance in wheat. We summarize the relevant findings from 52 quantitative trait loci (QTL) mapping studies, nine research articles on marker-assisted selection and seven on marker-assisted germplasm evaluation. QTL for FHB resistance were found on all wheat chromosomes except chromosome 7D. Some QTL were found in several independent mapping studies indicating that such QTL are stable and therefore useful in breeding programmes. We summarize and update current knowledge on the genetics of FHB resistance in wheat resulting from QTL mapping investigations and review and suggest FHB breeding strategies based on the available information and DNA markers.     

Linkage between a major gene for powdery mildew resistance and an RFLP marker on chromosome 1R of rye

DNA samples from an F₂ progeny which segregated for resistance to powdery mildew were bulked for resistant and susceptible individuals. In a segregant analysis, genomic rye probes which had been localized previously in a linkage map of rye were systematically screened for polymorphisms between these bulks. An RFLP marker located on linkage group 1RS was found to be tightly linked to a dominant mildew resistance gene. This is the first publication mapping a major gene for mildew resistance in rye.     

Development of SCAR markers for identification of stem rust resistance gene Sr31 in the homozygous or heterozygous condition in bread wheat

The stem rust resistance gene Sr31, transferred from rye (Secale cereale) into wheat (Triticum aestivum L.) imparts resistance to all the virulent pathotypes of stem rust (Puccinia graminis f. sp. tritici) found in India. Wheat genotypes including carriers and non‐carriers of the Sr31 gene were analysed using arbitrary primed polymerase chain reaction (AP‐PCR). AP‐PCR markers viz. SS30.2_580(H) associated with the Sr31 gene and SS26.1₁₁₀₀ associated with the allele for susceptibility were identified. Linkage between the markers and phenotypes was confirmed by analysing an F₂ population obtained from a cross between a resistant and a susceptible genotype. The markers were tightly linked to the respective alleles. Both the AP‐PCR markers were converted into sequence characterized amplified region (SCAR) markers, viz. SCSS30.2₅₇₆ and SCSS26.1₁₁₀₀ respectively. The markers were validated in two more segregating populations and 49 wheat genotypes. Using both markers it was possible to distinguish the homozygous from the heterozygous carriers of the Sr31 gene in the F₂ generation. The markers developed in this study can be used for pyramiding of the Sr31 gene with other rust resistance genes and in marker‐assisted selection.     

Location of a Gene for Resistance to Eyespot (Pseudocercosparella herpotrichoides) on Chromosome 7D of Bread Wheat

Chromosome 7D of the wheat line VPM1 derived from a cross of Aegilops ventricosa with wheat confers resistance to the facultative fungal parasite Pseudocercosporella herpotrichoides. To determine the number of genes responsible fur this resistance, homozygous recombinant lines were developed from an F₁ between the wheat variety ‘Hobbit sib’ and a substitution line carrying chromosome 7D of VPM1 in a ‘Hobbit sib’ background. Resistance to Pseudocercosporella herpotrichoides is shown to be determined by a single gene located distally on the long arm of chromosome 7D. EpD1b, a unique allele of a gene encoding the readily detectable isoenzyme — endopeptidase, maps without recombination to Pch1 suggesting for two separate genes a maximum recombination value of 0.03 (P 0.05). Resistance to Pherpotrichoides could alter-natively be a product of Ep-D1b. Pch1 is also mapped against a gene for adult plant resistance to brown rust (Puccinia recondita), to Rc3 which confers coleoptile colour, and to α-Amy-D2, an isozyme that encodes α-amylase production.     

Fusarium head blight resistance derived from Lophopyrum elongatum chromosome 7E and its augmentation with Fhb1 in wheat

The objective of this study was to assess the effectiveness of Fusarium head blight (FHB) resistance derived from wheatgrass Lophopyrum elongatum chromosome 7E and to determine whether this resistance can augment resistance in combination with other FHB resistance quantitative trait loci (QTL) or genes in wheat. The ‘Chinese Spring’–Lophopyrum elongatum disomic substitution line 7E(7B) was crossed to three wheat lines: ‘Ning 7840’, L3, and L4. F₂ populations were evaluated for type II resistance with the single‐floret inoculation method in the greenhouse. Simple sequence repeat markers associated with Fhb1 in ‘Ning 7840’ and L4 and markers located on chromosome 7E were genotyped in each population. Marker–trait association was analysed with one‐way or two‐way analysis of variance. The research showed that, in the three populations, the average number of diseased spikelets (NDS) in plants with chromosome 7E is 1.2, 3.1 and 3.2, vs. NDS of 3.3, 7.2 and 9.1 in plants without 7E, a reduction in NDS of 2.1, 4.1 and 5.9 in the respective populations. The QTL on 7E and the Fhb1 gene augment disease resistance when combined. The effect of the QTL on 7E was greater than that on 3BS in this experiment. Data also suggest that the FHB resistance gene derived from L. elongatum is located on the long arm of 7E.     

Molecular markers linked to the Aegilops variabilis-derived root-knot nematode resistance gene Rkn-mn1 in wheat

Aegilops variabilis no. 1 is the only known source of resistance to the root‐knot nematode Meloidogyne naasi in wheat. Previous studies showed that a dominant gene, Rkn‐mn1, was transferred to a wheat translocation line from the donor Ae. variabilis. Random amplified polymorphic DNA (RAPD) analysis was performed on the wheat cultivar ‘Lutin’, on Ae. variabilis, on a resistant disomic addition line and on a resistant translocation line. For genetic and molecular studies, 114‐117 BC₃F₂ plants and F₃‐derived families were tested. Five DNA and one isozyme marker were linked to Rkn‐mn1. Three RAPD markers flanking the Rkn‐mn1 locus were mapped at 0 cM (OpY16_‐1065), 0.8 cM (OpB12_‐1320) and 1.7 cM (OpN20_‐1235), respectively. Since the Rkn‐mn1 gene remained effective, its introduction into different wheat cultivars by marker‐assisted selection is suggested.     

A specific PCR assay for resistance to biotypes 1 and 2 of the rosy leaf curling aphid in apple based on an RFLP marker closely linked to the Sd1 gene

This report describes the conversion of a restriction fragment length polymorphism (RFLP) marker (the 2B12a locus). linked to the Sd₁ aphid resistance gene, to a polymerase chain reaction (PCR) based marker. A section of the 2BI2 probe was sequenced and two primers were designed lo amplify this sequence in the cultivars‘Prima’and‘Fiesta’: all the amplification products were the same size. After sequencing. two specific 24-mer oligonueleotides were synthesized (DdARM-5¹ and DdAR.M-3²) to exploit a single base-pair difference. These primers were used to screen 44 plants from the‘Prima’x‘Fiesta’family and generated a single amplification product (196bp). in approximately half of the seedlings, which was linked to the resistance gene Sd₁,. The DdARM primer combination was used to evaluate a range of apple cultivars and selections, including some varieties derived from‘Cox’and alternative sources of resistance reported in the literature. In parallel with this work, the phenotypic response of the same genotypes was either confirmed or determined in replicated glasshouse tests. The sequence characterized amplified regions (.SCAR) marker was amplified in all the resistant plants, with the exception of‘Northern Spy’and 3760 (the sources of Sd₂ and Sd₃ resistance, respectively), but never in the susceptible plants. The possible role of this marker in a marker-assisted breeding strategy, and its compatibility with a SCAR marker linked to the I, gene for resistance to apple scab. is discussed.     

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.     

The control of adult-plant resistance to yellow rust by the translocated chromosome 5BS-7BS of bread wheat

The reciprocal translocation 5BL-7BL and 5BS-7BS was widespread in West European wheats 30 years ago, and is probably present in many of their descendants today. In varieties with a history of durable adult-plant resistance to yellow rust and carrying this translocation, removal of the 5BS-7BS chromosome gave adult plants which were much more susceptible. It was suggested that this chromosome might therefore carry the gene(s) responsible for a major part of their resistance and possibly their durability. To test this, a series of lines was developed in which 5BS-7BS chromosomes from both resistant and susceptible varieties were substituted into a number of the durably resistant varieties. In every case, the substituted 5BS-7BS chromosome, irrespective of origin, was found to produce the resistant phenotype, indicating that background chromosomes were responsible for the differences between the varieties. The resistance and durability of the resistant varieties cannot therefore be due solely to the translocated chromosome. In similar experiments, the 5BS and 7BS arms from varieties not carrying the translocation were substituted into a variety carrying the translocation. In each instance, the lines with the substituted arms were much more susceptible than their recipient, confirming the major effect of the 5BS-7BS chromosome on resistance. The complete correlation between the translocation and resistance and between increased susceptibility and its absence suggests that the gene(s) for adult-plant resistance, located on the 5BS-7BS chromosome, may be closely linked to the break point. Alternatively, it may be a consequence of the close relatedness of some of the varieties. The presence of this gene(s) might be a factor explaining the prevalence of this translocation in some West European wheats.