Partial Resistance Genes Research Articles

Identification of Quantitative Trait Loci Conditioning Partial Resistance to Phytophthora sojae in Soybean PI 407861A

ABSTRACTImproving resistance for Phytophthora root and stem rot is an important goal in soybean [Glycine max (L.) Merr.] breeding. The objective of this study was to identify quantitative trait loci (QTLs) conferring partial resistance to Phytophthora sojae in an OX20‐8 × PI 407861A cross, in which PI 407861A was used as the source of partial resistance. One hundred fifty‐seven F7–derived recombinant inbred lines (RILs) were evaluated for partial resistance to P. sojae isolate OH25 using a tray test. Composite interval mapping identified three QTLs on chromosomes 8, 13, and 15 by a genomewide logarithm of odds (LOD) threshold and six QTLs on chromosomes 3, 4, 10, and 18 by chromosomewide LOD threshold. The phenotypic variance explained by each QTL ranged from 2.4 to 8.6%, with a total of 41.6%. Quantitative trait loci on chromosomes 3 and 8 are novel QTLs first reported for partial resistance to P. sojae in this study. All nine QTLs were localized near known resistance gene rich regions or those previously reported for resistance to other soil‐borne pathogens. Genomewide distribution of the QTLs in this population was distinct from the QTLs identified from previous studies with ‘Conrad’ but many were in common with those identified with another South Korean soybean accession. This indicates that the underlying genes for partial resistance in these exotic accessions may be distinct from genes controlling variation for partial resistance observed in Conrad. Plant Introduction 407861A is a promising genetic source of alleles that can be used to increase the genetic diversity and enhance levels of partial resistance to P. sojae in North American soybean cultivars.

Prospects for advancing defense to cereal rusts through genetical genomics.

Rusts are one of the most severe threats to cereal crops because new pathogen races emerge regularly, resulting in infestations that lead to large yield losses. In 1999, a new race of stem rust, Puccinia graminis f. sp. tritici (Pgt TTKSK or Ug99), was discovered in Uganda. Most of the wheat and barley cultivars grown currently worldwide are susceptible to this new race. Pgt TTKSK has already spread northward into Iran and will likely spread eastward throughout the Indian subcontinent in the near future. This scenario is not unique to stem rust; new races of leaf rust (Puccinia triticina) and stripe rust (Puccinia striiformis) have also emerged recently. One strategy for countering the persistent adaptability of these pathogens is to stack complete- and partial-resistance genes, which requires significant breeding efforts in order to reduce deleterious effects of linkage drag. These varied resistance combinations are typically more difficult for the pathogen to defeat, since they would be predicted to apply lower selection pressure. Genetical genomics or expression Quantitative Trait Locus (eQTL) analysis enables the identification of regulatory loci that control the expression of many to hundreds of genes. Integrated deployment of these technologies coupled with efficient phenotyping offers significant potential to elucidate the regulatory nodes in genetic networks that orchestrate host defense responses. The focus of this review will be to present advances in genetical genomic experimental designs and analysis, particularly as they apply to the prospects for discovering partial disease resistance alleles in cereals.

Evidence for a Minor Gene–for–Minor Gene Interaction Explaining Nonhypersensitive Polygenic Partial Disease Resistance

ABSTRACT Partial resistance is a quantitative type of resistance that, by definition of Parlevliet, is not based on hypersensitivity. It is largely pathotype nonspecific, although some minor isolate-specific responses have been reported. In order to elucidate the isolate specificity of individual genes for partial resistance, three barley recombinant inbred line mapping populations were analyzed for resistance to the leaf rust fungus Puccinia hordei. The mapping populations were inoculated with one isolate avirulent and two isolates virulent to resistance gene Rph7g. Six significant quantitative trait loci (QTLs) were detected. Of these, two (Rphq3 and Rphq11) were detected with only the avirulent isolate (1.2.1.) and one (Rphq18) only with both virulent isolates (CO-04 and 28.1). The effectiveness of these QTLs was tested with 14 isolates, using a tester set of genotypes containing alleles for resistance or susceptibility for these QTLs. QTL Rphq18 was effective to only two isolates, CO-04 and 28.1, whereas Rphq3 and Rphq11 were ineffective to CO-04 and 28.1 but effective to all other isolates, except one. This resulted in a significant Person's differential interaction, which is a hallmark of a gene-for-gene interaction. The minor gene-for-minor gene interaction is not based on hypersensitivity and there is no evidence that the resistance is based on genes belonging to the nucleotide-binding leucine-rich repeat class.

Mapping and validation of QTL which confer partial resistance to broadly virulent post-2000 North American races of stripe rust in hexaploid wheat.

A mapping population of 186 recombinant inbred lines developed from a cross between UC1110, an adapted California spring wheat, and PI610750, a synthetic derivative from CIMMYT's Wide Cross Program, was evaluated for its response to current California races of stripe rust (Puccinia striiformis f. sp. tritici) in replicated field trials over four seasons (2007-2010) in the northern Sacramento Valley. A genetic map was constructed consisting of 1,494 polymorphic probes (SSRs, DArTs, and ESTs) mapped to 558 unique loci, and QTL analysis revealed the presence of four stripe rust resistance QTL segregating in this population, two from UC1110 (on chromosomes 3BS and 2BS) and two from PI610750 (5AL and 2AS). The two QTL of largest effects (on 3BS and 5AL) were validated in independent populations and their intervals narrowed to 2.5 and 5.3 cM, respectively. The 3BS QTL was shown, by allelism test and genotype, to carry a gene different from the Yr30/Sr2 complex. Mapped position also suggests that the 3BS QTL is associated with a gene different from either Yrns-B1 or YrRub, two stripe rust resistance genes mapped to this region in other studies. The 5AL QTL carries a previously unreported partial stripe rust resistance gene, designated here as Yr48. This paper discusses the individual contributions to resistance of these four QTL, their epistatic interactions, and their potential in durable resistance breeding strategies based on combinations of partial resistance genes.

Durable resistance to the wheat rusts: Integrating systems biology and traditional phenotype-based research methods to guide the deployment of resistance genes.

Genes which confer partial resistance to the rusts in wheat figure prominently in discussions of potential durable resistance strategies. The positional cloning of the first of these genes, Lr34/Yr18 and Yr36, has revealed different protein structures, suggesting that the category of partial resistance genes, as defined by phenotype, likely groups together suites of functionally heterogenous genes. With the number of mapped partial rust resistance genes increasing rapidly as a result of ongoing advances in marker and sequencing technologies, breeding programs needing to select and prioritize genes for deployment confront a fundamental question: which genes or gene combinations are more likely to provide durable protection against these evolving pathogens? We argue that a refined classification of partial rust resistance genes is required to start answering this question, one based not merely on disease phenotype but also on gene cloning, molecular functional characterization, and interactions with other host and pathogen proteins. Combined with accurate and detailed disease phenotyping and standard genetic studies, an integrated wheat-rust interactome promises to provide the basis for a functional classification of partial resistance genes and thus a conceptual framework for their rational deployment.

The phenotypic expression of QTLs for partial resistance to barley leaf rust during plant development

Partial resistance is generally considered to be a durable form of resistance. In barley, Rphq2, Rphq3 and Rphq4 have been identified as consistent quantitative trait loci (QTLs) for partial resistance to the barley leaf rust pathogen Puccinia hordei. These QTLs have been incorporated separately into the susceptible L94 and the partially resistant Vada barley genetic backgrounds to obtain two sets of near isogenic lines (NILs). Previous studies have shown that these QTLs are not effective at conferring disease resistance in all stages of plant development. In the present study, the two sets of QTL-NILs and the two recurrent parents, L94 and Vada, were evaluated for resistance to P. hordei isolate 1.2.1 simultaneously under greenhouse conditions from the first leaf to the flag leaf stage. Effect of the QTLs on resistance was measured by development rate of the pathogen, expressed as latency period (LP). The data show that Rphq2 prolongs LP at the seedling stage (the first and second leaf stages) but has almost no effect on disease resistance in adult plants. Rphq4 showed no effect on LP until the third leaf stage, whereas Rphq3 is consistently effective at prolonging LP from the first leaf to the flag leaf. The changes in the effectiveness of Rphq2 and Rphq4 happen at the barley tillering stage (the third to fourth leaf stages). These results indicate that multiple disease evaluations of a single plant by repeated inoculations of the fourth leaf to the flag leaf should be conducted to precisely estimate the effect of Rphq4. The present study confirms and describes in detail the plant development-dependent effectiveness of partial resistance genes and, consequently, will enable a more precise evaluation of partial resistance regulation during barley development.

Effect of combining two genes for partial resistance to Barley yellow dwarf virus‐PAV (BYDV‐PAV) derived from Thinopyrum intermedium in wheat

In this study two sources of resistance to Barley yellow dwarf virus (BYDV), both originating from Thinopyrum intermedium, were combined in a single genotype, line ZT, using GISH (genomic in situ hybridization) and a new molecular marker of the partial resistance gene Bdv2. Susceptible wheat cv. Sunstar, the translocation line TC14 carrying Bdv2, the addition line ZH with the group‐2 chromosome arm carrying resistance, and line ZT with both resistances were inoculated with five strains of Barley yellow dwarf virus‐PAV. The tests confirmed the resistance of TC14 and ZH and revealed an additive effect of the two sources of resistance in ZT. The resistance of line ZT was characterized by a proportion of infected plants significantly lower than the parental lines TC14 and ZH (42% vs. a mean of 76% for the parents) and a very low virus titre (area under the virus concentration progress curve of 1·2 vs. a mean on 6·3 for the parents).

Contributions of disease resistance and escape to the control of septoria tritici blotch of wheat

The contributions of disease escape and disease resistance to the responses of wheat to septoria tritici leaf blotch (STB) were analysed in a set of 226 lines, including modern cultivars, breeding lines and their progenitors dating back to the origin of scientific wheat breeding. Field trials were located in the important wheat‐growing region of eastern England and were subject to natural infection by Mycosphaerella graminicola. STB scores were related to disease‐escape traits, notably height, leaf spacing, leaf morphology and heading date, and to the presence of known Stb resistance genes and isolate‐specific resistances. The Stb6 resistance gene was associated with a reduction of 19% in the level of STB in the complete set of 226 lines and with a 33% reduction in a subset of 139 lines of semidwarf stature. Greater plant height was strongly associated with reduced STB in the full set of lines, but only weakly in the semidwarf lines. Shorter leaf length was also associated with reduced STB, but, in contrast to earlier reports, lines with more prostrate leaves had more STB on average, probably because they tended to have longer leaves. Several lines, notably cvs Pastiche and Exsept, had low mean levels of STB which could not be explained by either escape traits or specific resistance genes, implying that they have unknown genes for partial resistance to STB.

Oculimacula yallundae lifestyle revisited: relationships between the timing of eyespot symptom appearance, the development of the pathogen and the responses of infected partially resistant wheat plants

Possible relationships between the timing of eyespot symptom appearance, the development of Oculimacula yallundae and the responses of infected wheat plants were investigated. By comparing, for 45 days after inoculation (d.a.i.), eyespot‐ and mock‐inoculated coleoptiles and leaf sheaths of a wheat line carrying the Pch1 partial resistance gene, this study showed that cell death, used as a criterion for symptom appearance, occurred on the first leaf sheath around 30 d.a.i. Monitoring of O. yallundae development by optical and scanning electron microscopy revealed that the fungus colonized the coleoptile within 30 d.a.i. It then attacked the first leaf sheath and underwent morphological and growth modifications. For 30 d.a.i., infected plants responded by local accumulations of H2O2 and callose in coleoptile epidermal cells and by up‐regulation of pathogenesis‐related (PR) genes. From 30 d.a.i., natural browning occurred in the first leaf sheath epidermis and the expression level of PR genes, determined by quantitative PCR, significantly increased in infected plants. These results show that the wheat/O. yallundae interaction is characterized by two distinct phases: the asymptomatic phase and the symptomatic phase. The possibility that O. yallundae should be considered hemibiotrophic rather than necrotrophic is discussed.

Quantitative disease resistance assessment by real-time PCR using the Stagonospora nodorum-wheat pathosystem as a model

Measuring disease resistance accurately and reproducibly is a key requirement for the introgression of partial resistance genes into breeding lines. Here, a qPCR protocol is used to measure fungal biomass, using the wheat-Stagonospora nodorum pathosystem as a model. Seven cultivars of differing reported resistance levels were used. Fungal biomass taken at 220°C thermal days after inoculation accurately predicts the final grain weight loss. It is concluded that a test based on qPCR methods is specific, quantitative, rapid and objective. Such tests could provide useful and economic tools in the development of robustly resistant crop cultivars.

Resistance and virulence interactions between two white pine species and blister rust in a 30-year field trial

We monitored infection and mortality of full-sib families of sugar pine (SP) and western white pine (WWP) selected for different mechanisms of resistance to white pine blister rust for more than 30 years in a field test in northern California. Natural infection was enhanced by interplanting alternate host Ribes spp. among test seedlings. Parents of the families were from three geographic provinces, representing the northern Rocky Mountains of Idaho (WWP), the Cascade Range of Oregon and Washington (WWP), and the Siskiyou Mountains of California and Oregon (SP). Several families of each pine species had major gene resistance (MGR), or genes for partial resistance (PR), or both types. Under the chronic epidemic conditions that prevailed, susceptible controls of both species became infected and died within a few years. Families with MGR segregated in expected Mendelian ratios (susceptibility/resistance), but each R gene—Cr1 in sugar pine and Cr2 in western white pine—was eventually defeated by biotypes of the rust with specific virulence to it (vcr1 and vcr2, respectively). WWP families from Idaho, which lacked Cr2, performed the best overall, ranging from 44 to 64% rust-free; a few WWP families from the Cascades, both with and without Cr2, performed comparably. Families with PR were not specifically vulnerable to vcr1 or vcr2. PR was less frequent in SP than WWP, and SP families had steeper infection rate curves than WWP. Combining PR in pedigrees with MGR may prolong protection of MGR by inhibiting selection and epidemic increase of vcr genes in the rust.

Genetic and Physical Mapping of the Partial Resistance Gene, Pi34, to Blast in Rice

ABSTRACT Partial resistance to rice blast in the Oryza sativa japonica group cv. Chubu 32 is controlled by Pi34, a major quantitative trait locus (QTL) on chromosome 11, and several uncharacterized QTLs. The objectives of the study were (i) high-resolution genetic and physical mapping of Pi34 and (ii) identification of new QTL imparting resistance to rice blast. Chubu 32 was crossed with a susceptible chromosomal segment substitution line (CSSL) of cv. Koshihikari. From 4,012 of segregating individuals, 213 recombinants in the Pi34 region were screened by using polymerase chain reaction-based markers and tested resistance in the field and greenhouse. The Pi34 locus is located in the 54.1-kb region on the genomic sequence of cv. Nipponbare. We constructed a bacterial artificial chromosome (BAC) library of Chubu 32, selected the clone containing Pi34, and sequenced it. The Pi34 locus consequently was located on an interval of 65.3 kb containing 10 predicted open reading frames (ORFs). Two of these ORFs were predicted only in Chubu 32 and encoded transposable elements. The other eight ORFs were found in both Chubu 32 and Nipponbare and one of them, which encoded an unknown protein, showed significantly different amino acid sequences between two cultivars. The new QTL, Piq6(t), was detected on the short arm of chromosome 6 and the genetic distance of flanking markers was 16.9 centimorgans in Nipponbare. Pi34 and Piq6(t) acted additively on resistance to rice blast but the effect of Piq6(t) was relatively small compared with Pi34.

Field trial of serially passaged isolates of BYDV‐PAV overcoming resistance derived from Thinopyrum intermedium in wheat

AbstractBarley yellow dwarf disease (BYDD) is one of the main viral diseases of small grain cereals. This disease, reported on numerous plant species of the Poaceae family, is caused by a complex of viral species including the species Barley yellow dwarf virus‐PAV (BYDV‐PAV, family Luteoviridae, genus Luteovirus), frequently found in western Europe. Resistance sources towards BYDD are scarce. Indeed, breeding‐resistant genotypes is a long and expensive process. Thus, estimating the durability of the resistance genes before the achievement of selection would be an asset for breeders. One isolate of BYDV‐PAV has been serially passaged on two hosts, ‘Zhong ZH’ and ‘TC14’, carrying a gene for partial resistance. The resulting viral population showed an increase of the speed of development of the infection in controlled conditions. In this study, these viral populations were evaluated in a 3‐year field trial, including a susceptible host, ‘Rendezvous’, and a host carrying the resistance gene of ‘TC14’ in a ‘Rendezvous’ background, to assess the effect of serial passages in field conditions. Results indicate that isolates issued from serial passages on hosts carrying a gene for partial resistance induced increased damage in field conditions when compared with the initial isolate. Yield losses are mainly due to a decrease of the number of kernels per square metre. The interest on using partial resistance gene to control BYDD is discussed.

Pi34-AVRPi34: a new gene-for-gene interaction for partial resistance in rice to blast caused by Magnaporthe grisea

The japonica rice (Oryza sativa) cultivar Chubu 32 has a high level of partial resistance to blast, which is mainly controlled by a dominant resistance gene located on chromosome 11. The partial resistance to the rice blast fungus (Magnaporthe grisea) in Chubu 32 has isolate specificity; isolate IBOS8-1-1 is more aggressive on Chubu 32 than are other isolates. We hypothesized that the gene-for-gene relationship fits this case of a partial resistance gene in Chubu 32 against the avirulence gene in the pathogen. The partial resistance gene in Chubu 32 was mapped between DNA markers C1172 (and three other co-segregated markers) and E2021 and was designated Pi34. In the 32 F3 lines from the cross between a chromosome segment substitution line (Pi34−) from Koshihikari/Kasalath and Chubu 32, the lines with high levels of partial resistance to the M. grisea isolate Y93-245c-2 corresponded to the presence of Pi34 estimated by graphic genotyping. This indicated that Pi34 has partial resistance to isolate Y93-245c-2 in compatible interactions. The 69 blast isolates from the F1 progeny produced by the cross between Y93-245c-2 and IBOS8-1-1 were tested for aggressiveness on Chubu 32 and rice cultivar Koshihikari (Pi34−). The progeny segregated at a 1 : 1 ratio for strong to weak aggressiveness on Chubu 32. The results suggested that Y93-245c-2 has one gene encoding avirulence to Pi34 (AVRPi34), and IBOS8-1-1 is extremely aggressive on Chubu 32 because of the absence of AVRPi34. This is the first report of a gene-for-gene relationship between a fungal disease resistance gene associated with severity of disease and pathogen aggressiveness.

Loci controlling partial resistance to rice blast do not show marked QTL × environment interaction when plant nitrogen status alters disease severity

Plant disease susceptibility is often increased by nitrogen (N) application. Therefore, it is important to know if resistance loci are effective in different plant N environments. One-hundred lines of the Bala x Azucena rice (Oryza sativa) mapping population were grown in two N treatments and tested for partial resistance to blast (Magnaporthe grisea) isolate CD100. Disease severity (DS), the number and size of lesions and plant N and C concentrations were measured and the results subject to quantitative trait loci (QTL) and QTL x environment analysis. There was a 66% higher plant N concentration in the high N treatment and DS increased significantly, mostly as a result of increased numbers of lesions. Nine regions contained QTL for disease traits but only one showed evidence of statistically significant QTL x treatment interaction. This was a large effect quantitative trait locus at marker R1933 on chromosome 12 which was less effective at high N. Apparently, blast disease is increased by higher plant N, but the efficacy of partial resistance genes is not greatly affected by N application.

Leaf Rust and Stem Rust Resistance in Triticum dicoccoides Populations in Israel.

A total of 742 single plant accessions of Triticum dicoccoides were collected from 26 locations in Israel. All accessions were evaluated for leaf rust (Puccinia triticina) resistance in field plots at Tel Aviv, and subsets of 284 and 468 accessions were tested in the greenhouse in Tel Aviv and St. Paul, MN, respectively, for seedling resistance to leaf rust; 460 accessions were also tested for seedling resistance to stem rust (Puccinia graminis f. sp. tritici) in St. Paul. One accession was highly resistant to leaf rust in seedling tests in Tel Aviv, and 21 others had moderately susceptible to moderately resistant seedling resistance. Four accessions were highly resistant to leaf rust in seedling tests in St. Paul, and 11 were resistant to at least one stem rust race. Adult resistance to leaf rust was more common than seedling resistance among the accessions; 21 accessions had less than 25% leaf rust severity in field plots compared with 80 to 90% severity for highly susceptible accessions. Most of the accessions with effective adult plant resistance came from two nearby locations in Upper Galilee, a region where populations of T. dicoccoides are most extensive and genetically diverse. These accessions may provide valuable new partial resistance genes for durable protection against leaf rust in cultivated wheat.

Quantitative trait loci analysis suggests that partial resistance to rice blast is mostly determined by race–specific interactions

• The race specificity of quantitative trait loci (QTL) for partial resistance to blast (Magnaporthe grisea) disease was tested by using isolates for which no major resistance gene segregated in a mapping population based on rice (Oryza sativa) varieties Bala × Azucena. • Recombinant inbred lines were repeatedly inoculated with isolates CD100, CM28 and PH19 and scored for lesion type, lesion size and number of lesions. Composite interval mapping was employed to identify QTL. • Eighteen main effect QTL were detected. Of these, eight were detected in only one isolate, seven detected in two isolates and only three in all three isolates. Diallelic epistasis was identified as an important genetic component but had little discernible pattern. Fourteen main-effect QTL mapped to previously identified QTL for blast resistance and 10 to previously identified major resistance genes. • It is concluded that the majority of QTL detected are race-specific. The results also support the hypothesis that partial resistance genes might be defeated major genes, with residual effectiveness and race specificity.

Field Resistance to Bemisia tabaci in Cucumis melo

The whitefly Bemisia tabaci (Gennadius) B-Biotype is a major pest on cucurbits in the Caribbean islands. Five field trials were conducted to identify resistance among 80 genotypes of Cucumis melo L. from diverse geographic origins. We focused on resistant rather than tolerant genotypes by counting adults on the abaxial side of two leaves of each plant at least three times in each trial, and larval density of under leaf disk samples at least twice in each trial. On the basis of insect density, three Indian accessions, PI 414723, PI 164723, and 90625, and one Korean accession, PI 161375, had field resistance. On those accessions, we observed 3.6 to 6 times fewer adults than on the most susceptible genotypes (`AR Top Mark' or `Délice de Table') and 11 to 29 times fewer larvae than on `AR Top Mark' or B66-5. Those levels of resistance may be sufficient to significantly reduce pesticide use in Guadeloupe (Lesser Antilles) where B. tabaci populations are lower than in the Southern United States or in the Mediterranean Basin. Higher levels of resistance are needed for genetic control, and may be achieved by a combination of different partial resistance genes.

Beneficial Effects of Fungicide Seed Treatments for Soybean Cultivars with Partial Resistance to Phytophthora sojae.

Phytophthora sojae is a yield-limiting soybean pathogen in areas where soils remain saturated for long periods of time. P. sojae has been successfully managed with single dominant resistance genes (Rps genes). The proportion of fields with populations of P. sojae capable of causing susceptible interactions with many of the Rps genes has increased in number. The fungicides metalaxyl and mefenoxam have been used both as in-furrow and seed treatments to provide protection against damping-off caused by P. sojae. To determine the plant age when partial resistance and Rps genes are effective against P. sojae, we evaluated a greenhouse assay in which soybean seeds were planted and inoculated with a zoospore suspension to compare the disease reaction of soybean seeds and seedlings. Efficacy of different fungicide rates also was evaluated using the cultivar with partial resistance with this inoculation technique. Seeds and seedlings of a cultivar with high levels of partial resistance were susceptible to infection by P. sojae while those of a cultivar with an Rps gene were resistant. For the cultivar with partial resistance, reductions in percent emergence and the number of damped-off seedlings were significantly higher for plants inoculated at the day of planting compared to inoculations of plants with unifoliates present (5 days after planting). Results also indicate that fungicide seed treatment on cultivars with partial resistance may be beneficial when the environmental conditions that favor P. sojae infections occur prior to soybean emergence. This greenhouse assay appears to be useful in examining overall fungicide efficacy; however, it did not detect consistent and quantifiable differences in rates of seed treatment fungicides.

Components of partial resistance to powdery mildew in wheat mutants

Components of resistance to powdery mildew (latent period, pustule density and conidium production) were analysed on glasshouse-grown artificially inoculated plants of wheat mutants (n = 11) derived from either induced mutagenesis or adventitious regeneration of cv. Guardian. These mutants had previously been shown to be partially resistant to the disease in the field over a two-year period. Analyses were carried out at three stages of development: seedling, tillering and heading. None of the mutants exhibited a latent period significantly different from that of the susceptible parent ‘Guardian’ at any stage of development tested. With respect to pustule density, one mutant (M61) was significantly more resistant than ‘Guardian’ at all stages of development, two mutants (M19 and SC100) were more resistant at tillering and heading, and two (M156 and SC267) were more resistant only at heading. Significantly reduced conidium production per pustule (as a measure of pustule size) over a 24 h period was observed on three mutants (M61, SC100 and SC231) at tillering and heading, while mutant SC68 exhibited this trait at heading only. On the basis of resistance components and relative grain yield in the presence of mildew, the eleven mutants were categorised into five categories. Tests for associations between field resistance score and components of resistance measured in the glasshouse, both measured at heading, revealed a significant positive correlation only between pustule density and field score. To assess the effects of combining different components (i.e. pyramiding different partial resistance genes), mutants were crossed. Transgressive segregation for at least one component was observed in the F2 generation in crosses between mutants with complementary modes of action (i.e. involving different components of partial resistance), but not in cases where the parents exhibited the same component of resistance.