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Global food security depends on cereal crops with durable disease resistance. Most cereals are colonized by rust fungi, which are pathogens of major significance for global agriculture1. Cereal rusts display a high degree of host specificity and one rust species or forma specialis generally colonizes only one cereal host2. Exploiting the non-host status and transferring non-host resistance genes between cereal crop species has been proposed as a strategy for durable rust resistance breeding. The molecular determinants that define the host status to rusts, however, are largely unknown. Here, we show that orthologous genes at the Rphq2 locus for quantitative leaf rust resistance from cultivated barley3 and Rph22 from wild bulbous barley4 affect the host status to leaf rusts. Both genes encode lectin receptor-like kinases. We transformed Rphq2 and Rph22 into an experimental barley line that has been bred for susceptibility to non-adapted leaf rusts, which allowed us to quantify resistance responses against various leaf rust species. Rphq2 conferred a much stronger resistance to the leaf rust of wild bulbous barley than to the leaf rust adapted to cultivated barley, while for Rph22 the reverse was observed. We hypothesize that adapted leaf rust species mitigate perception by cognate host receptors by lowering ligand recognition. Our results provide an example of orthologous genes that connect the quantitative host with non-host resistance to cereal rusts. Such genes provide a basis to exploit non-host resistance in molecular breeding.

Three barley mapping populations sharing the parental line SusPtrit suffered from an unknown condition affecting grain quality and germination. Grains showed dark blotches or even whole-grain discoloration and shrivelling. Symptoms were similar to kernel discoloration as reported in wheat and barley. Some covered-seed lines affected by this quality condition produced grains that were either not covered at all or in which partial hull loss was observed, a phenotype that resembles that of grain skinning. No fungal or bacterial agent was identified as causal organism, and we demonstrate that the poor quality phenotype is not transmitted to the next generation – therefore unlikely due to a seed borne pathogen. A major effect QTL on chromosome 6H was mapped independently in all three populations, with the poor quality allele contributed by SusPtrit. A second, minor effect QTL, was mapped on chromosome 2H in the SusPtrit x Golden Promise population, with Golden Promise as the parent donor of the allele conferring poor quality. The grain quality disorder is not linked to the nud gene on chromosome 7H for the naked/covered seed trait.

Key message The nonhost resistance of wild lettuce to lettuce downy mildew seems explained by four components of a putative set of epistatic genes. Abstract The commonplace observation that plants are immune to most potential pathogens is known as nonhost resistance (NHR). The genetic basis of NHR is poorly understood. Inheritance studies of NHR require crosses of nonhost species with a host, but these crosses are usually unsuccessful. The plant-pathosystem of lettuce and downy mildew, Bremia lactucae, provides a rare opportunity to study the inheritance of NHR, because the nonhost wild lettuce species Lactuca saligna is sufficiently cross-compatible with the cultivated host Lactuca sativa. Our previous studies on NHR in one L. saligna accession led to the hypothesis that multi-locus epistatic interactions might explain NHR. Here, we studied NHR at the species level in nine accessions. Besides the commonly used approach of studying a target trait from a wild donor species in a cultivar genetic background, we also explored the opposite, complementary approach of cultivar introgression in a wild species background. This bidirectional approach encompassed (1) nonhost into host introgression: identification of L. saligna derived chromosome regions that were overrepresented in highly resistant BC1 plants (F1 × L. sativa), (2) host into nonhost introgression: identification of L. sativa derived chromosome regions that were overrepresented in BC1 inbred lines (F1 × L. saligna) with relatively high infection levels. We demonstrated that NHR is based on resistance factors from L. saligna and the genetic dose for NHR differs between accessions. NHR seemed explained by combinations of epistatic genes on three or four chromosome segments, of which one chromosome segment was validated by the host into nonhost approach. Electronic supplementary material The online version of this article (10.1007/s00122-018-3112-8) contains supplementary material, which is available to authorized users.

Key message Resistance factors against non-adapted powdery mildews were mapped in barley. Some QTLs seem effective only to non-adapted mildews, while others also play a role in defense against the adapted form. The durability and effectiveness of nonhost resistance suggests promising practical applications for crop breeding, relying upon elucidation of key aspects of this type of resistance. We investigated which genetic factors determine the nonhost status of barley (Hordeum vulgare L.) to powdery mildews (Blumeria graminis). We set out to verify whether genes involved in nonhost resistance have a wide effectiveness spectrum, and whether nonhost resistance genes confer resistance to the barley adapted powdery mildew. Two barley lines, SusBgtSC and SusBgtDC, with some susceptibility to the wheat powdery mildew B. graminis f.sp. tritici (Bgt) were crossed with cv Vada to generate two mapping populations. Each population was assessed for level of infection against four B. graminis ff.spp, and QTL mapping analyses were performed. Our results demonstrate polygenic inheritance for nonhost resistance, with some QTLs effective only to non-adapted mildews, while others play a role against adapted and non-adapted forms. Histology analyses of nonhost interaction show that most penetration attempts are stopped in association with papillae, and also suggest independent layers of defence at haustorium establishment and conidiophore formation. Nonhost resistance of barley to powdery mildew relies mostly on non-hypersensitive mechanisms. A large-effect nonhost resistance QTL mapped to a 1.4 cM interval is suitable for map-based cloning. Electronic supplementary material The online version of this article (10.1007/s00122-018-3055-0) contains supplementary material, which is available to authorized users.

Blast disease, caused by the Magnaporthe oryzae/grisea species complex, occurs in a wide range of wild and cultivated gramineous plant species including rice, wheat and barley. We inoculated a collection of cultivated (Hordeum vulgare ssp. vulgare L.) and wild (ssp. spontaneum) barley accessions with M. oryzae Oryza pathotype (MoO), Triticum pathotype (MoT) and Pennisetum pathotype (MsP) to quantify the host status of barley, and to identify sources of blast resistance. Unlike wheat, the barley gene pool is rich with sources of complete and partial resistance against Magnaporthe. Cultivated barley appeared a nonhost to MsP, whereas wild barley showed some degree of susceptibility. All 153 tested rice accessions were resistant to the MoT isolate, suggesting that rice is nonhost to this pathotype. Inoculation of L94/Vada and Vada/SusPtrit RIL mapping populations with MoO and/or MoT isolates of M. oryzae indicated one large effect QTL, designated as Rmoq1, on the short arm of chromosome 7H against the MoT isolate PY 67.1 in both populations. Resistance in L94 to the MoO isolate was due to a different QTL, located at 5H. An association mapping panel of West European barley cultivars also suggested that most QTLs were pathotype specific. Six blast resistance genes found in the biparental and association mapping studies coincided with map positions of powdery mildew resistance genes viz. Mlt, Mla6, Mlg, mlo, Mlj, and Mlhb genes. Our QTL and association mapping analyses do not support the association of the mlo resistance gene with enhanced susceptibility to M. oryzae as reported in literature.

Global food security depends on cereal crops with durable disease resistance. Most cereals are colonized by rust fungi, which are pathogens of major significance for global agriculture

The main objective of this research was to analyze whether selection under conventional conditions (indirect selection) is as effective as selection under organic conditions (direct selection) to develop varieties suitable for organic farming systems. Two F3 barley (Hordeum vulgare L.) populations ‘Primus’/‘Idumeja’ (P/I) and ‘Anni’/‘Dziugiai’ (A/Dz) targeted for organic variety development were selected in two organically and two conventionally managed environments during F3 to F3:6. From there, the performance of the five best F3:6 lines selected in each of the four environments from each cross (in total, 40 lines) were compared at all four sites during two years. For obtaining varieties adapted to organic conditions for the P/I cross, it did not matter at which condition the selection was performed. For A/Dz, the best selection results in terms of yield, combined with other traits included for an organic ideotype, were achieved under well-managed organic conditions. We conclude that direct and indirect selection in early breeding stages are equally suitable for the development of cultivars for organic conditions if (i) care is taken that selection considers not only yield, but also other traits important for organic growing conditions, (ii) selection is not performed under too stressful conditions, and (iii) testing in later stages of the breeding program is conducted under various organic farming conditions for the best recommended varieties for organic agriculture.

A disease test at different leaf layers (plant stages) of homologous rust and heterologous rust species were studied. The result from homologous rust species showed that those quantitative trait loci (QTLs) (Rphq2 and Rphq11) which were effective at seedling stage were also effective across all plant stages with gradually decreasing effect as plants grew older. Rphq3 which had consistent effect in all leaf layers confirmed the same result, that it is a plant stage independent QTL. For heterologous rusts, the effect of Rnhq-V was studied on three rust species; P. hordei-murini (Phm), P. hordei-secalini (Phs) and P. triticina isolate ‘Flamingo’ at three stages (leaf layers). Infection frequencies are higher at seedling stage and dramatically decrease as plants grow older in all three rust species tested on both SusPtrit and Su-Rnhq-V. The difference between lines tends to be reduced with higher leaf layer in all three tested inappropriate rust species. However, this would be not because of less effectiveness of the Rnhq. Key words: Plant stage, homologous rust, heterologous rust.

Scabiosa columbaria (Dipsacaceae) is a popular perennial ornamental in the United States. It is native to Europe and was introduced to North America by nursery trade only recently. In the spring of 2006, symptoms of powdery mildew infection were observed on overwintered plants of S. columbaria cv. Butterfly Blue in a nursery in Cutchogue, NY. White powdery mildew mycelia with abundant sporulation were observed on upper and lower leaf surfaces. The portions of leaves with powdery mildew colonies often showed purplish discoloration. Conidia were cylindric to doliiform, measured 20 to 33 × 10 to 15 μm, and were produced singly on 60 to 130 μm long conidiophores consisting of a foot-cell measuring 20 to 50 × 6 to 10 μm, followed by one to three, 12 to 40 μm long cells. Hyphal appressoria were lobed or multilobed. The teleomorph stage was not found. On the basis of these characteristics, the pathogen was identified as an Oidium sp. belonging to the subgenus Pseudoidium. Recently, an anamorphic powdery mildew fungus with similar morphological characteristics, identified as Erysiphe knautiae, was reported on S. columbaria cv. Butterfly Blue in Washington (2). E. knautiae is a common powdery mildew species of dipsacaceous plants such as Scabiosa spp. and Knautia spp. in Europe and Asia (1). To determine whether the fungus reported here was E. knautiae, DNA was extracted from its mycelium, and the internal transcribed spacer (ITS) region of the ribosomal DNA was amplified and sequenced as described earlier (4). No ITS sequences are available in public DNA databases for E. knautiae, thus, we determined this sequence in a specimen of E. knautiae collected from Knautia arvensis in The Netherlands. Herbarium specimens of the Oidium sp. infecting S. columbaria in New York and E. knautiae from the Netherlands were deposited at the U.S. National Fungus Collections under accession numbers BPI 878259 and BPI 878258, respectively. The ITS sequence from Oidium sp. infecting S. columbaria in New York (GenBank Accession No. EU377474) differed in two nucleotides from that of E. knautiae infecting K. arvensis in the Netherlands (GenBank Accession No. EU377475). These two ITS sequences were also more than 99% similar to those of some newly emerged anamorphic powdery mildew fungi: Oidium neolycopersici and other Oidium spp. infecting Chelidonium majus, Passiflora caerulea, and some crassulaceous plants (3,4). Thus, it is unclear whether the fungus reported here was E. knautiae known from Eurasia or an Oidium sp. that has acquired pathogenicity to S. columbaria. To our knowledge, this is the first report of powdery mildew on S. columbaria in New York. References: (1) U. Braun. Beih. Nova Hedwigia 89:1, 1987. (2) D. A. Glawe and G. G. Grove. Online publication. doi:10.1094/PHP-2005-1024-01-BR. Plant Health Progress, 2005. (3) B. Henricot. Plant Pathol. 57:779, 2008. (4) T. Jankovics et al. Phytopathology 98:529, 2008.

Interspecific crosses can result in progeny with reduced vitality or fertility due to genetic incompatibilities between species, a phenomenon known as hybrid incompatibility (HI). HI is often caused by a bias against deleterious allele combinations, which results in transmission ratio distortion (TRD). Here, we determined the genome-wide distribution of HI between wild lettuce, Lactuca saligna, and cultivated lettuce, L. sativa, in a set of backcross inbred lines (BILs) with single introgression segments from L. saligna introgressed into a L. sativa genetic background. Almost all BILs contained an introgression segment in a homozygous state except a few BILs, for which we were able to obtain only a single heterozygous introgression. Their inbred progenies displayed severe TRD with a bias toward the L. sativa allele and complete nontransmission of the homozygous L. saligna introgression, i.e., absolute HI. These HI might be caused by deleterious heterospecific allele combinations at two loci. We used an multilocus segregating interspecific F2 population to identify candidate conspecific loci that can nullify the HI in BILs. Segregation analysis of developed double-introgression progenies showed nullification of three HI and proved that these HI are explained by nuclear pairwise incompatibilities. One of these digenic HI showed 29% reduced seed set and its pattern of TRD pointed to a sex-independent gametophytic barrier. Namely, this HI was caused by complete nontransmission of one heterospecific allele combination at the haploid stage, surprisingly in both male and female gametophytes. Our study shows that two-locus incompatibility systems contribute to reproductive barriers among Lactuca species.

Powdery mildew caused by Blumeria graminis f. sp. hordei is a foliar disease with highly negative impact on yield and grain quality in barley. Thus, breeding for powdery mildew resistance is an important goal and requires constantly the discovery of new sources of natural resistance. Here, we report the high resolution genetic and physical mapping of a dominant race-specific powdery mildew resistance locus, originating from an Ethiopian spring barley accession ‘HOR2573,’ conferring resistance to several modern mildew isolates. High-resolution genetic mapping narrowed down the interval containing the resistance locus to a physical span of 850 kb. Four candidate genes with homology to known disease resistance gene families were identified. The mapped resistance locus coincides with a previously reported resistance locus from Hordeum laevigatum, suggesting allelism at the same locus in two different barley lines. Therefore, we named the newly mapped resistance locus from HOR2573 as MlLa-H. The reported co-segregating and flanking markers may provide new tools for marker-assisted selection of this resistance locus in barley breeding.

Quantitative resistance (QR) refers to a resistance that is phenotypically incomplete and is based on the joined effect of several genes, each contributing quantitatively to the level of plant defense. Often, QR remains durably effective, which is the primary driver behind the interest in it. The various terms that are used to refer to QR, such as field resistance, adult plant resistance, and basal resistance, reflect the many properties attributed to it. In this article, we discuss aspects connected to those attributions, in particular the hypothesis that much of the QR to biotrophic filamentous pathogens is basal resistance, i.e., poor suppression of PAMP-triggered defense by effectors. We discuss what role effectors play in suppressing defense or improving access to nutrients. Based on the functions of the few plant proteins identified as involved in QR, vesicle trafficking and protein/metabolite transportation are likely to be common physiological processes relevant to QR.

Wheat stem rust, a devastating disease of wheat and barley caused by the fungal pathogen Puccinia graminis f. sp. tritici, was largely eradicated in Western Europe during the mid-to-late twentieth century. However, isolated outbreaks have occurred in recent years. Here we investigate whether a lack of resistance in modern European varieties, increased presence of its alternate host barberry and changes in climatic conditions could be facilitating its resurgence. We report the first wheat stem rust occurrence in the United Kingdom in nearly 60 years, with only 20% of UK wheat varieties resistant to this strain. Climate changes over the past 25 years also suggest increasingly conducive conditions for infection. Furthermore, we document the first occurrence in decades of P. graminis on barberry in the UK . Our data illustrate that wheat stem rust does occur in the UK and, when climatic conditions are conducive, could severely harm wheat and barley production. Clare Lewis et al. report the first identification in nearly 60 years of a cultivated wheat plant infected with the fungal pathogen Puccinia graminis f.sp. tritici (wheat stem rust) in the United Kingdom. They find that only 20% of UK wheat varieties are resistant to this strain and urge growers to resume resistance breeding programs.

Barley (Hordeum vulgare L.) mapping populations have been developed that are useful to study the inheritance of quantitative resistance to adapted and unadapted rust fungi. In a recent host range study, we found that the parents of those mapping populations also differed in their resistance to the crown rust Puccinia coronata (PcE) of couch grass (Elymus repens), as well as three isolates of P. striiformis, representing formae speciales hordei (Psh), tritici (Pst) and bromi (Psb). Available mapping populations were phenotyped at the seedling stage to map the genes conferring resistance to these rust isolates. Resistances to PcE, Psb and Pst inherit quantitatively. This contrasted with reports that barley nonhost resistance to unadapted formae speciales of P. striiformis is based on major genes. We mapped QTLs effective against PcE using relative latency period and relative infection frequency. Some QTLs for resistance were contributed by ‘Vada’ and ‘Cebada Capa’, others by SusPtrit. One PcE-resistance QTL on 3H, contributed by ‘Cebada Capa’, co-localised with a QTL effective against four unadapted grass rust species, indicating either a single gene with broad-spectrum effectiveness or a cluster of rather specific genes. Chromosome arm 7HL from Vada seems particularly rich in genes for resistance to rust fungi. Resistance to Pst and Psb, measured as the number of uredinia, tended to co-localise with each other and mapped to 1HS, 2HL and 7HL. The nearly complete resistance of Ethiopian line L94 to Psh was due to a major gene mapped on chromosome 4H. That gene is likely the recessive gene rpsGZ, which was previously reported in the Ethiopian line Grannenlose Zweizeilige.

Barley yellow dwarf disease (BYD) is transmitted by aphids and is caused by different strains of Barley yellow dwarf virus (BYDV) and Cereal yellow dwarf virus (CYDV). Economically it is one of the most important diseases of cereals worldwide. Besides chemical control of the vector, growing of tolerant/resistant cultivars is an effective way of protecting crops against BYD. The Ryd3 gene in barley (Hordeum vulgare L.) confers tolerance to BYDV-PAV and BYDV-MAV and the locus was previously mapped on the short arm of barley chromosome 6H near the centromere. We applied a strategy for high-resolution mapping and marker saturation at the Ryd3 locus by exploiting recent genomic tools available in barley. In a population of 3,210 F2 plants, 14 tightly linked markers were identified, including 10 that co-segregated with Ryd3. The centromeric region where Ryd3 is located suffers suppressed recombination or reduced recombination rate, suggesting potential problems in achieving (1) map-based cloning of Ryd3 and (2) marker selection of the resistance in breeding programmes without the introduction of undesirable traits via linkage drag.

Barley is a near-nonhost to numerous heterologous (nonadapted) rust pathogens because a small proportion of genotypes are somewhat susceptible. We assessed 66 barley accessions and three mapping populations (Vada × SusPtrit, Cebada Capa × SusPtrit, and SusPtrit × Golden Promise) for response to three Swedish oat stem rust (Puccinia graminis f. sp. avenae) fungal isolates and determined that barley is a near-nonhost to P. graminis f. sp. avenae and that resistance was polygenically inherited. The parental genotypes Vada and Golden Promise were immune to all three isolates, whereas Cebada Capa was immune to two isolates and moderately resistant to the third. Phenotypic data from the Vada × SusPtrit mapping population and the barley accessions tested also demonstrated isolate-specific resistance. In particular, the SusPtrit parent and several other accessions allowed sporulation by isolate Ingeberga but were resistant to isolate Evertsholm. Nine chromosomal regions carried quantitative trait loci (QTL) (Rpgaq1 to Rpgaq9) of varying effect, most of which colocated to previously identified QTL for resistance to other heterologous rust pathogens. Rpgaq1 on chromosome 1H (Vada and Golden Promise) was effective toward all isolates tested. Microscopic examination indicated that resistance was prehaustorial in Vada whereas, in SusPtrit, both pre- and posthaustorial mechanisms play a role.

Rusts and powdery mildews employ different strategies to suppress defense during penetration. We observed that a compatible interaction of barley-Puccinia hordei induced increased penetration resistance to a challenge infection by powdery mildew. This induced resistance is local and its level is not determined by the virulence spectrum of the challenger isolate. Our data suggest that the inducer effect is due to rust-stoma communication during penetration, to the presence of the rust hyphae in the apoplast, or to penetration resistance mounted by the rust attacked mesophyll cells. We hypothesized that the rust “primes” the basal defense prior to the mildew infection.

Both Mexico and South America are rich in Solanum species that might be valuable sources of resistance (R) genes to late blight (Phytophthora infestans). Here, we focus on an R gene present in the diploid Bolivian species S. avilesii. The genotype carrying the R gene was resistant to eight out of 10 Phytophthora isolates of various provenances. The identification of a resistant phenotype and the generation of a segregating population allowed the mapping of a single dominant R gene, Rpi-avl1, which is located in an R gene cluster on chromosome 11. This R gene cluster is considered as an R gene “hot spot”, containing R genes to at least five different pathogens. High resolution mapping of the Rpi-avl1 gene revealed a marker co-segregating in 3890 F1 individuals, which may be used for marker assisted selection in breeding programs and for further cloning of Rpi-avl1.

Barley yellow dwarf virus (BYDV) is an economically important pathogen of barley, which may become even more important due to global warming. In barley, several loci conferring tolerance to BYDV-PAV-ASL-1 are known, e.g. Ryd2, Ryd3 and a quantitative trait locus (QTL) on chromosome 2H. The aim of the present study was to get information whether the level of tolerance against this isolate of BYDV in barley can be improved by combining these loci. Therefore, a winter and a spring barley population of doubled haploid (DH) lines were genotyped by molecular markers for the presence of the susceptibility or the resistance encoding allele at respective loci (Ryd2, Ryd3, QTL on chromosome 2H) and were tested for their level of BYDV-tolerance after inoculation with viruliferous (BYDV-PAV-ASL-1) aphids in field trials. In DH-lines carrying the combination Ryd2 and Ryd3, a significant reduction of the virus titre was detected as compared to lines carrying only one of these genes. Furthermore, spring barley DH-lines with this allele combination also showed a significantly higher relative grain yield as compared to lines carrying only Ryd2 or Ryd3. The QTL on chromosome 2H had only a small effect on the level of tolerance in those lines carrying only Ryd2, or Ryd3 or a combination of both, but the effect in comparison to lines carrying no tolerance allele was significant. Overall, these results show that the combination of Ryd2 and Ryd3 leads to quantitative resistance against BYDV-PAV instead of tolerance.

Basal resistance of barley to powdery mildew is a quantitatively inherited trait that limits the growth and sporulation of barley powdery mildew pathogen by a non-hypersensitive mechanism of defense. Two experimental barley lines were developed with a very high (ErBgh) and low (EsBgh) level of basal resistance to powdery mildew by cycles of convergent crossing and phenotypic selection between the most resistant and between the most susceptible lines, respectively, from four mapping populations of barley. Phenotypic selection in convergent crossing was highly effective in producing contrasting phenotypes for basal resistance and susceptibility. In ErBgh, almost 90% of infection units failed to form a primary haustorium in the epidermal cells in association with papilla formation, but in EsBgh only 33% of infection units failed to form a primary haustorium. The contrast between ErBgh and EsBgh for successful formation of secondary and subsequent haustoria was much less obvious (69% versus 79% successful secondary haustorium formation). In an earlier investigation, we determined seven QTLs for basal resistance in the four mapping populations. Checking the peak markers of these QTLs indicated that only four out of seven QTLs were confirmed to be present in the selected resistant lines and only four QTLs for susceptibility were confirmed to be present in the selected susceptible lines. Surprisingly, none of the expected QTLs could be detected in the resistant line ErBgh. We discuss some reasons why marker aided selection might be less efficient in raising levels of basal resistance than phenotypic selection. The very resistant and susceptible lines developed here are valuable material to be used in further experiments to characterize the molecular basis of basal resistance to powdery mildew.

Plants face continuous attacks from a broad range of pathogens and have evolved effective defence mechanisms that are initiated upon pathogen attack. Invading oomycete pathogens secrete effectors, molecules that manipulate host cell defence and thereby enable colonization. However, plant species evolved resistance (R) genes to most specialized pathogen species. The R proteins can detect effectors, termed avirulence (AVR) proteins, and thus confer immunity to pathogens. Effectors and their interacting genes in the plant play a central role in the co-evolution of pathogens with their hosts. In this review, we discuss the role that effectors play in the pathogenesis and lifestyles of oomycetes. Particularly intriguing features emerge for (hemi-)biotrophic oomycetes, which establish an intimate contact with the host by forming haustoria. At this interface, effectors with an RXLR motif are translocated into the cytoplasm, where they reprogramme the host towards susceptibility. Such interactions between effectors and host targets are highly specific and are considered a result of tight co-evolution. In addition, we elaborate on the Phytophthora infestans-Solanum pathosystem, from which various R and Avr genes were cloned recently. We discuss a rationale for exploiting molecular insights into R-Avr interactions for developing more durable resistance strategies to control late blight in agriculture.

This study used the pathosystem of lettuce (Lactuca spp.) and downy mildew (Bremia lactucae) as a model to investigate the inheritance of nonhost resistance, and focused on the contribution of quantitative trait loci (QTLs) to nonhost resistance at various developmental stages in the lettuce life cycle. A set of 28 backcross inbred lines (BILs) of L. saligna CGN05271 (nonhost) introgressions in a L. sativa cv. Olof (host) background identified 16 introgressions that contributed to resistance at various plant developmental stages: seedlings, young plants, adult plants in the greenhouse and adult plants in the field. This paper provisionally considered these introgressions to be 16 QTLs. Of these 16 QTLs, seven were identified previously and nine were new. For 15 QTLs (Rbq1, Rbq2, rbq3–7 and Rbq8–15), the resistance alleles were derived from the nonhost L. saligna; the resistance allele of the other QTL (Rbq16) was from the susceptible L. sativa cv. Olof. Of the 15 QTLs in L. saligna, only two, rbq5 and rbq7, were found to be effective at every plant developmental stage; the other 13 QTLs were only effective at certain developmental stages. Experiments with seven B. lactucae races did not provide evidence that any QTL was race-specific. The data suggest that nonhost resistance in L. saligna is the result of cumulative effects of many resistance QTLs operating at various developmental stages.

Rphq2 , a minor gene for partial resistance to Puccinia hordei , was physically mapped in a 188 kbp introgression with suppressed recombination between haplotypes of rphq2 and Rphq2 barley cultivars. Partial and non-host resistances to rust fungi in barley (Hordeum vulgare) may be based on pathogen-associated molecular pattern (PAMP)-triggered immunity. Understanding partial resistance may help to understand non-host resistance, and vice versa. We constructed two non-gridded BAC libraries from cultivar Vada and line SusPtrit. Vada is immune to non-adapted Puccinia rust fungi, and partially resistant to P. hordei. SusPtrit is susceptible to several non-adapted rust fungi, and has been used for mapping QTLs for non-host and partial resistance. The BAC libraries help to identify genes determining the natural variation for partial and non-host resistances of barley to rust fungi. A major-effect QTL, Rphq2, for partial resistance to P. hordei was mapped in a complete Vada and an incomplete SusPtrit contig. The physical distance between the markers flanking Rphq2 was 195 Kbp in Vada and at least 226 Kbp in SusPtrit. This marker interval was predicted to contain 12 genes in either accession, of which only five genes were in common. The haplotypes represented by Vada and SusPtrit were found in 57 and 43 %, respectively, of a 194 barley accessions panel. The lack of homology between the two haplotypes probably explains the suppression of recombination in the Rphq2 area and limit further genetic resolution in fine mapping. The possible candidate genes for Rphq2 encode peroxidases, kinases and a member of seven-in-absentia protein family. This result suggests that Rphq2 does not belong to the NB-LRR gene family and does not resemble any of the partial resistance genes cloned previously.

Cell walls and cellular turgor pressure shape and suspend the bodies of all vascular plants. In response to attack by fungal and oomycete pathogens, which usually breach their host's cell walls by mechanical force or by secreting lytic enzymes, plants often form local cell wall appositions (papillae) as an important first line of defence. The involvement of cell wall biosynthetic enzymes in the formation of these papillae is still poorly understood, especially in cereal crops. To investigate the role in plant defence of a candidate gene from barley (Hordeum vulgare) encoding cellulose synthase-like D2 (HvCslD2), we generated transgenic barley plants in which HvCslD2 was silenced through RNA interference (RNAi). The transgenic plants showed no growth defects but their papillae were more successfully penetrated by host-adapted, virulent as well as avirulent nonhost isolates of the powdery mildew fungus Blumeria graminis. Papilla penetration was associated with lower contents of cellulose in epidermal cell walls and increased digestion by fungal cell wall degrading enzymes. The results suggest that HvCslD2-mediated cell wall changes in the epidermal layer represent an important defence reaction both for nonhost and for quantitative host resistance against nonadapted wheat and host-adapted barley powdery mildew pathogens, respectively.

A consensus map of barley was constructed based on three reference doubled haploid (DH) populations and three recombinant inbred line (RIL) populations. Several sets of microsatellites were used as bridge markers in the integration of those populations previously genotyped with RFLP or with AFLP markers. Another set of 61 genic microsatellites was mapped for the first time using a newly developed fluorescent labelling strategy, referred to as A/T labelling. The final map contains 3,258 markers spanning 1,081 centiMorgans (cM) with an average distance between two adjacent loci of 0.33 cM. This is the highest density of markers reported for a barley genetic map to date. The consensus map was divided into 210 BINs of about 5 cM each in which were placed 19 quantitative trait loci (QTL) contributing to the partial resistance to barley leaf rust (Puccinia hordei Otth) in five of the integrated populations. Each parental barley combination segregated for different sets of QTLs, with only few QTLs shared by any pair of cultivars. Defence gene homologues (DGH) were identified by tBlastx homology to known genes involved in the defence of plants against microbial pathogens. Sixty-three DGHs were located into the 210 BINs in order to identify candidate genes responsible for the QTL effects. Eight BINs were co-occupied by a QTL and DGH(s). The positional candidates identified are receptor-like kinase, WIR1 homologues and several defence response genes like peroxidases, superoxide dismutase and thaumatin.

Mitogen-activated protein kinases (MAPKs) have been shown to act as key regulators of stress responses in model plant and crop species. So far, however, the MAPK family has not been systematically studied in barley. Herein, we identified 16 HvMAPKs (Hv—Hordeum vulgare) based on computational analysis of barley transcriptomics and genomics databases. HvMAPKs contain all canonical MAPK domains, except for HvMPK2, which lacks a MAPK domain signature. In addition, five HvMAPKs harbor TEY and ten HvMAPKs harbor TDY dual phosphorylation motif in the activation loop. Interestingly, HvMPK2 contains a MEY instead of TEY phosphorylation motif. We classified HvMAPKs into four major plant MAPK clades based on phylogeny reconstruction and anchored all HvMAPK genes to five out of seven barley chromosomes. Furthermore, we inoculated seedlings of susceptible barley line L94 and its isolines L94-Rph3 and L94-Rph7 with rust fungus Pucciniahordei and analyzed the expression of 16 HvMAPK genes using qRT-PCR at 1–4.5 days post inoculation. In total, six HvMAPK genes exhibited significantly altered expression by P. hordei infection. The expression of HvMPK5, HvMPK6, HvMPK7 and HvMPK12 (set one genes) was strongly induced especially during effector-triggered immunity (ETI), whereas the expression of HvMPK2 and HvMPK17 (set two genes) was specifically downregulated during ETI. Yet the expression of HvMPK8 was also specifically but weakly downregulated during ETI. Overall, the expression patterns suggest that set one genes positively regulate ETI in barley–P. hordei pathosystem, whereas set two genes negatively regulate ETI and/or programmed cell death in this pathosystem.

A set of 148 modern spring barley cultivars was explored for the extent of linkage disequilibrium (LD) between genes governing traits and nearby marker alleles. Associations of agronomically relevant traits (days to heading, plant height), resistance traits (leaf rust, barley yellow dwarf virus (BYD)), and morphological traits (rachilla hair length, lodicule size) with AFLP markers and SSR markers were found. Known major genes and QTLs were confirmed, but also new putative QTLs were found. The LD mapping clearly indicated the common occurrence of Rph3, a gene for hypersensitivity resistance against Puccinia hordei, and also confirmed the QTL Rphq2 for prolonging latency period of P. hordei in seedlings. We also found strong indication for a hitherto not reported gene for resistance or tolerance to BYD on chromosome 2, linked to SSR marker HVM054. Our conclusion is that LD mapping is a valuable additional tool in the search for applicable marker associations with major genes and QTLs.

Tomato powdery mildew caused by Oidium neolycopersici has become a globally important disease of tomato (Lycopersicon esculentum). To study the defense responses of tomato triggered by tomato powdery mildew, we first mapped a set of resistance genes to O. neolycopersici from related Lycopersicon species. An integrated genetic map was generated showing that all the dominant resistance genes (Ol-1, Ol-3, Ol-4, Ol-5, and Ol-6) are located on tomato chromosome 6 and are organized in three genetic loci. Then, near-isogenic lines (NIL) were produced that contain the different dominant Ol genes in a L. esculentum genetic background. These NIL were used in disease tests with local isolates of O. neolycopersici in different geographic locations, demonstrating that the resistance conferred by different Ol genes was isolate-dependent and, hence, may be race-specific. In addition, the resistance mechanism was analyzed histologically. The mechanism of resistance conferred by the dominant Ol genes was associated with hypersensitive respo-nse, which varies in details depending on the Ol-gene in the NIL, while the mechanism of resistance governed by the recessive gene ol-2 on tomato chromosome 4 was associated with papillae formation.

In this second paper on the use of secondary genepools in barley improvement, we describe the char- acterisation of leaf rust resistant recombinant lines (RLs) derived from Hordeum vulgare × H. bulbosum crosses. Twelve RLs were inoculated with leaf rust and the early stages of disease development were observed. Several RLs showed complete resistance to the pathogen, but others had a high level of partial resistance, which may be durable. Some of these RLs and others were tested in yield trials to determine the effects of introgressed chro- matin from H. bulbosum on yield and quality. We conclude that there are no major adverse effects that cannot be overcome through normal breeding techniques.

Summary Specialised plant pathogens are in many ways adapted to exploit their host plants. Infective propagules should reach the appropriate plant tissue, gain access to the tissue and negate or suppress various kinds of constitutive and inducible resistance mechanisms. The resistance type most frequently deployed in plant breeding is the racespecific resistance, where a hypersensitive response of plant tissue is elicited by an avirulence factor produced by the pathogen. The great disadvantage of this type of resistance is, that it is often ephemeral. Detailed screening of germplasm may result in the discovery of alternative defence mechanisms not associated with hypersensitivity, that may be durable. Avoidance mechanisms may reduce the chance of infection. Upright plant habit has been reported to decrease spore deposition in cereals. Crop architecture may also affect aspects such as humidity and aeration in the crop, and hence the chances for successful infection by pathogen propagules. Other examples of avoidance are leaf surface properties that interfere with leaf wettability, germ tube orientation and finding of stomata to enter the leaf. Stomata in some accessions of Hordeum chilense are excessively covered by cuticular wax that prevent rust fungal germ tubes from perceiving the stomata, resulting in failure of penetration of the pathogen into the leaf. There is evidence that in compatible host species, biotrophic pathogenic fungi induce basic compatibility by suppressing defence mechanisms. Failure of this induction results in abortion of the infection attempt. Several cases of apparently durable resistance are discussed that are based on failure of haustorium formation, and are not associated with hypersensitivity. They may represent cases where the pathogen has problems in establishing basic compatibility.

The three diploid wheat species Triticum monococcum, Triticum boeoticum and Triticum urartu differ in their reaction to wheat leaf rust, Puccinia triticina. In general, T. monococcum is resistant while T. boeoticum and T. urartu are susceptible. However, upon screening a large collection of diploid wheat accessions, 1% resistant T. boeoticum accessions and 16% susceptible T. monococcum accessions were found. In the present study these atypical accessions were compared with 49 typical T. monococcum, T. boeoticum and T. urartu accessions to gain insight into the host-status of the diploid wheat species for wheat leaf rust. Cluster analysis of morphological data and AFLP fingerprints of the typical accessions clearly discriminated the three diploid species. T.monococcum and T. boeoticum had rather-similar AFLP fingerprints while T. urartu had a very different fingerprint. The clustering of most atypical accessions was not consistent with the species they were assigned to, but intermediate between T. boeoticum and T. monococcum. Only four susceptible T. monococcum accessions were morphologically and moleculary similar to the typical T. monococcum accessions. Results confirmed that T. boeoticum and T. monococcum are closely related but indicate a clear difference in host-status for the wheat leaf rust fungus in these two species.

Near-isogenic lines (NILs) differing with regard to disease QTLs provide valuable material for a more detailed study into the genetic basis of quantitative resistance. Previously obtained information on QTLs that show an effect on leaf rust (Puccinia hordei) in barley was used in a marker-assisted backcross programme. The genome origin in backcross plants was controlled through AFLP marker analysis and graphical genotyping. Plants obtained after the third generation of backcrossing sufficiently resembled the recurrent parent. For one QTL, BC3S1 plants were evaluated in a disease test and genotyped. NILs containing the desired QTL in homozygous condition in a recipient background were finally obtained. A disease test and verification of the marker genotype confirmed the identity of the NILs. Simultaneous with the backcross programme a simulation study on efficiency of marker-assisted backcrossing was performed.

The infection process of most rust fungi start with spore germination, directional growth of the germ tube towards a stoma, differentiation of an appressorium over the stoma, and penetration into the substomatal cavity. In the South American wild barley Hordeum chilense Roem. & Schult., wide variation occurs in the degree to which several rust fungal species are able to form appressoria over the stomata. Apparently, features of the plant may hamper early stages of the infection process. Such an early defence is called avoidance. In order to find out how germ tube growth is directed towards stomata, and whether the cuticular wax layer plays a role in this orientated growth and in appressorium differentiation, several orientation and differentiation parameters of Puccinia hordei germ tubes were measured on H. chilense leaves with and without the wax layer. Orientated growth of the germ tubes started upon contact with the epidermal cell junctions. The growth of lateral branches of the germ tube over the first epidermal cell junction that it meets, may help the germ tube to grow along the transverse axis of the leaf. No evidence was found of attraction of the germ tube to stomata. Removal of the cuticular wax layer did not result in loss of germ tube orientation. This suggests that the leaf wax layer has no role in the guidance of germ tubes. On high avoidance accessions, removal of the wax layer allowed appressoria to develop over stomata that would otherwise be overgrown. No effect of the cell widths in stomatal complexes was found on the chance that stomata were overgrown. This suggests that the overgrowth of stomata on H. chilense leaves by P. hordei germ tubes is mainly caused by the wax covering of the stomatal apparatus.

Diploid wheat, Triticum monococcum s.l., is a host for the wheat leaf rust fungus, Puccinia triticina. Some accessions have been reported to show a high degree of prehaustorial resistance. This is non-hypersensitivity resistance, which acts before the formation of haustoria by the pathogen. To assess the frequency of prehaustorial resistance 598 accessions of diploid wheat were inoculated with the wheat leaf rust isolate Felix. Most T. monococcum s.s. accessions (84%) were resistant whereas all T. urartu and all but three T. boeoticum accessions were susceptible. Histological components analysis revealed that a high percentage of prehaustorial resistance to P. triticina was found in only three T. monococcum accessions. No haustoria were observed in such infection units confirming the prehaustorial nature of the resistance. Prehaustorial abortion of certain infection units in an accession always coincided with posthaustorial abortion of the other infection units.

Assessments of epidemiological parameters (e.g. infection efficiency, latent period, spore production) are required for the prediction of epidemic progress, for the estimation of components of partial resistance for different host plant cultivars and for the estimation of fitness components of pathogen isolates. A critical evaluation of methods of measurement of monocyclic parameters is presented, suggesting improvements by using more accurate techniques and taking into account different biotic and abiotic factors. Examples of influence of lesion density and age, of meteorological factors and of nutrient status of the host on cereal powdery mildew and rust epidemiological parameters are presented. The use of the ratio of mature lesions rather than the latent period for the estimation of the development rate of the fungus is suggested to allow comparison of a large number of individuals. It is feasible to assess the pathogen biomass by the sterol contents. The need to verify by field experiments epidemiological parameters assessed under controlled conditions is pointed out. Finally, the way to include these monocyclic parameters in epidemiological and forecasting models is discussed using the case of wheat brown rust epidemics.

With standardised near isogenic line (NIL) differentials co-operators were able to present the first comprehensive virulence survey of the European wheat leaf rust population (1996-1999). The work included pathotype identification of 2608 isolates and field tests of NILs. Lr9 and Lr19 were very effective all over Europe. Lr24, Lr25, and Lr28 were also effective, but in some countries and locations substantial virulence frequencies were observed. In addition, the genes Lr12, Lr13, Lr22a, Lr34, Lr35 and Lr37 were effective at the adult plant stage, but locally less so. In general, the indoor seedling tests and adult plant field tests showed good agreement. Virulence to Lr1, Lr2a, Lr24, Lr25, Lr28 and Lr29 tended to increase in the period, for the other Lr-genes the virulence frequency remained more or less stable. Among the 105 pathotypes identified none was clearly predominant in Europe.

Nine accessions of three cucurbit species, ten of eight legume species, three of lettuce (Lactuca sativa) and 34 of 14 Solanaceae species were inoculated with a Dutch isolate of the tomato powdery mildew fungus (Oidium lycopersici) to determine its host range. Macroscopically, no fungal growth was visible on sweet pepper (Capsicum annuum), lettuce, petunia (Petunia spp.) and most legume species (Lupinus albus, L. luteus, L. mutabilis, Phaseolus vulgaris, Vicia faba, Vigna radiata, V. unguiculata). Trace infection was occasionally observed on melon (Cucumis melo), cucumber (Cucumis sativus), courgette (Cucurbita pepo), pea (Pisum sativum) and Solanum dulcamara. Eggplant (Solanum melongena), the cultivated potato (Solanum tuberosum) and three wild potato species (Solanum albicans, S. acaule and S. mochiquense) were more heavily infected in comparison with melon, cucumber, courgette, pea and S. dulcamara, but the fungus could not be maintained on these hosts. All seven tobacco (Nicotiana tabacum) accessions were as susceptible to O. lycopersici as tomato (Lycopersicon esculentum cv Moneymaker), suggesting that tobacco is an alternative host. This host range of the tomato powdery mildew differs from that reported in some other countries, which also varied among each other, suggesting that the causal agent of tomato powdery mildew in the Netherlands differ from that in those countries. Histological observations on 36 accessions showed that the defense to O. lycopersici was associated with a posthaustorial hypersensitive response.

Using AFLP markers, a linkage map was constructed based on a recombinant inbred population of barley derived from a cross between a leaf rust susceptible line, L94, and a partially resistant line, 116-5. The constructed map showed a similar marker distribution pattern as the L94 × Vada map. However, it contained more large gaps, and for some chromosome regions no markers were identified. These regions are most likely derived from L94 because 116-5 was selected from the progeny of a cross of L94 × cv. Cebada Capa. Five QTLs for partial resistance to isolate 1.2.1. were mapped on the L94 × 116-5 map. Three QTLs were effective in the seedling stage, jointly contributing 42% to the total phenotypic variance. Three QTLs were effective in the adult plant stage, collectively explaining 35% of the phenotypic variance. Evidence for two additional linked minor-effect QTLs effective in the adult plant stage was also uncovered. The major-effect QTL, Rphq3, was the only one that was effective in both developmental stages. Moreover, Rphq3, was also identified in the L94 × Vada population, being effective to two rust isolates. The other QTLs were detected in either of the two populations, providing evidence for the existence of many loci for partial resistance to leaf rust on the barley genome. To date, 13 QTLs for partial resistance have been mapped, therefore, a strategy of accumulating many resistance genes in a single cultivar, resulting in a high level of partial resistance, is feasible.

Macroscopical and histological research is useful to characterize quantitative resistance. Analysis of histological components of the resistance to the flax rust fungus (Melampsora lini) was carried out on seedlings of seven flax (Linum usitatissimum) accessions: five fibre flax accessions with various levels of quantitative resistance and two near-isogenic lines of linseed with gene K or NI. Flax seedlings were inoculated with uredospores. Collected leaves of the flax accessions were stained with Chlorazol Black E. It was found that in the pathosystem flax - M. lini, a lower number of pustules (lower infection frequency) on quantitatively resistant accessions did not result from mechanisms acting before stomatal penetration by the fungus. Reduced infection frequency was associated with increased rates of early abortion of the rust fungus but rates of this abortion were not large enough to explain the low infection frequency on the quantitatively resistant accessions Afganistan and Solido. In both of the ...