Two new hosts of anamorphic Erysiphe quercicola: Cinnamomum camphora and Murraya paniculata

Based on collections of powdery mildews (Erysiphales) in Taiwan and combined molecular and morphological analyses, camphor tree (Cinnamomum camphora) and orange jasmine (Murraya paniculata) are recognized as new hosts of the anamorph of the powdery mildew Erysiphe quercicola. The anamorphic powdery mildew on C. camphora has been known as Pseudoidium cinnamomi, but its relationship to a teleomorph was unknown. For M. paniculata as substrate of powdery mildew, only an anamorphic Cystotheca species has been named. Morphological investigation of the fungus on this host shows that the specimens from Taiwan belong to another genus because of the lack of fibrosin bodies. Analysis of internal transcribed spacer sequences indicates that the anamorphic powdery mildews on camphor and orange jasmine belong to a clade representing E. quercicola, with the teleomorph found only on oak species (Quercus, Fagaceae), but with its anamorph reported from a broad host range, particularly in the tropics.     

Molecular phylogenetic analyses reveal a close relationship between powdery mildew fungi on some tropical trees and Erysiphe alphitoides, an oak powdery mildew

To investigate the phylogenetic relationships among the powdery mildew fungi of some economically important tropical trees belonging to Oidium subgenus Pseudoidium, we conducted molecular phylogenetic analyses using 30 DNA sequences of the rDNA internal transcribed spacer (ITS) regions and 26 sequences of the domains D1 and D2 of the 28S rDNA obtained from the powdery mildews on Hevea brasiliensis (para rubber tree), Anacardium occidentale (cashew), Bixa orellana, Citrus spp., Mangifera indica (mango), and Acacia spp. The results indicate that the powdery mildew fungi isolated from these tropical trees are closely related to one another. These powdery mildews are also closely related to E. alphitoides (including Erysiphe sp. on Quercus phillyraeoides). Because of the obligate biotrophic nature of the powdery mildew fungi, the relationship between powdery mildews and their host plants is conservative. However, the present study suggests that a particular powdery mildew species has expanded its host ranges on a wide range of the tropical trees. This article also suggests that a powdery mildew fungus distributed in temperate regions of the Northern Hemisphere expanded its host ranges onto tropical plants and may be a good example of how geographical and host range expansion has occurred in the Erysiphales.     

Molecular mapping of a recessive powdery mildew resistance gene in spelt wheat cultivar Hubel

Wheat powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is one of the most important wheat diseases worldwide. The basis for wheat powdery mildew resistance breeding consists of screening diversified host genetic resources with a range of races of the powdery mildew pathogen. Spelt wheat (Triticum aestivum ssp. spelta 2n = 6x = 42, AABBDD) is a close relative of common wheat (T. aestivum ssp. aestivum) and contains several known disease resistance genes, including Pm1d, Yr5, and Lr65. Here, we report the identification and mapping of a powdery mildew resistance gene in spelt wheat cultivar Hubel, which was introduced to China from Europe and is resistant to Chinese Bgt isolate E09 at the seedling stage. Genetic analysis of a recombinant inbred line population derived from a cross of Hubel and a susceptible early maturing mutant line indicated that Hubel possessed a recessive powdery mildew resistance gene (temporarily designated MlHubel). Markers linked to MlHubel were identified using bulked segregant analysis, simple sequence repeat, and expressed sequence tag-derived sequence tagged site methods. The linked markers were physically located on wheat chromosome 2D. Comparative genomic analysis indicated that the genetic interval covering MlHubel in wheat is highly colinear with the corresponding regions on Brachypodium distachyon chromosome 5 and Oryza sativa chromosome 4. Accordingly, the genetic map of MlHubel was established in comparison with B. distachyon 5L and O. sativa 4L, with the closest marker Xgwm265 being 0.4 cM from MlHubel. The identification of the recessive powdery mildew gene in spelt wheat suggests the potential of this accession along with its closely linked markers in breeding for resistance to powdery mildew.     

Host Phenology and Geography as Drivers of Differentiation in Generalist Fungal Mycoparasites

The question as to why parasites remain generalist or become specialist is a key unresolved question in evolutionary biology. Ampelomyces spp., intracellular mycoparasites of powdery mildew fungi, which are themselves plant pathogens, are a useful model for studies of this issue. Ampelomyces is used for the biological control of mildew. Differences in mycohost phenology promote temporal isolation between sympatric Ampelomyces mycoparasites. Apple powdery mildew (APM) causes spring epidemics, whereas other powdery mildew species on plants other than apple cause epidemics later in the season. This has resulted in genetic differentiation between APM and non-APM strains. It is unclear whether there is genetic differentiation between non-APM Ampelomyces lineages due to their specialization on different mycohosts. We used microsatellites to address this question and found no significant differentiation between non-APM Ampelomyces strains from different mycohosts or host plants, but strong differentiation between APM and non-APM strains. A geographical structure was revealed in both groups, with differences between European countries, demonstrating restricted dispersal at the continent scale and a high resolution for our markers. We found footprints of recombination in both groups, possibly more frequent in the APM cluster. Overall, Ampelomyces thus appears to be one of the rare genuine generalist pathogenic fungi able to parasitize multiple hosts in natural populations. It is therefore an excellent model for studying the evolution of pathogens towards a generalist rather than host-specific strategy, particularly in light of the tritrophic interaction between Ampelomyces mycoparasites, their powdery mildew fungal hosts and the mildew host plants.     

Interspecific and intraspecific diversity in oak powdery mildews in Europe: coevolution history and adaptation to their hosts

Quercus has been reported as the genus with the largest number of attacking powdery mildews. In Europe, oak powdery mildew was rarely reported before 1907, when severe outbreaks were observed. These epidemics were attributed to the newly described species Erysiphe alphitoides, presumed to be of exotic origin. After the burst of interest following the emergence of the disease, research on this topic remained very limited. Interest in research was recently reactivated in response to the availability of molecular tools. This review summarizes current knowledge on the diversity of oak powdery mildews in Europe and their possible evolutionary relationships with European oaks. The most striking results are the evidence of cryptic diversity (detection in France of a lineage closely related to Erysiphe quercicola, previously thought to only have an Asian distribution), large host range (similarity of E. alphitoides and E. quercicola with powdery mildews of tropical plants) but also local adaptation to Quercus robur. These recent findings highlight the complexity of the history of oak powdery mildew in Europe and point to the question of host specialization and host jumps in the evolution of powdery mildew fungi.     

Genetic and comparative genomics mapping reveals that a powdery mildew resistance gene Ml3D232 originating from wild emmer co-segregates with an NBS-LRR analog in common wheat (Triticum aestivum L.)

Powdery mildew caused by Blumeria graminis f. sp. tritici is one of the most important wheat diseases worldwide and breeding for resistance using diversified disease resistance genes is the most promising approach to prevent outbreaks of powdery mildew. A powdery mildew resistance gene, originating from wild emmer wheat (Triticum turgidum var. dicoccoides) accessions collected from Israel, has been transferred into the hexaploid wheat line 3D232 through crossing and backcrossing. Inoculation results with 21 B. graminis f. sp. tritici races indicated that 3D232 is resistant to all of the powdery mildew isolates tested. Genetic analyses of 3D232 using an F₂ segregating population and F₃ families indicated that a single dominant gene, Ml3D232, confers resistance in the host seedling stage. By applying molecular markers and bulked segregant analysis (BSA), we have identified polymorphic simple sequence repeats (SSR), expressed sequence tags (EST) and derived sequence tagged site (STS) markers to determine that the Ml3D232 is located on chromosome 5BL bin 0.59–0.76. Comparative genetic analyses using mapped EST markers and genome sequences of rice and Brachypodium established co-linearity of the Ml3D232 genomic region with a 1.4 Mb genomic region on Brachypodium distachyon chromosome 4, and a 1.2 Mb contig located on the Oryza sativa chromosome 9. Our comparative approach enabled us to develop new EST–STS markers and to delimit the genomic region carrying Ml3D232 to a 0.8 cM segment that is collinear with a 558 kb region on B. distachyon. Eight EST markers, including an NBS-LRR analog, co-segregated with Ml3D232 to provide a target site for fine genetic mapping, chromosome landing and map-based cloning of the powdery mildew resistance gene. This newly developed common wheat germplasm provides broad-spectrum resistance to powdery mildew and a valuable resource for wheat breeding programs.     

Leveillula buddlejae sp. nov., a new species with an asexual morph resembling phylogenetically basal Phyllactinia species

An asexual morph of a powdery mildew was found on Buddleja asiatica (Scrophulariaceae) in Nepal. The morphological traits of this powdery mildew strongly resemble those of Phyllactinia species (including Ovulariopsis) belonging to a phylogenetically basal group closely related to Leveillula species, but phylogenetic examinations and analyses clearly showed that this fungus pertains to Leveillula. Within the latter genus, it seems to be allied to L. duriaei, but clearly differs from this species in having conidiophores arising from superficial hyphae and narrower conidia with different length/width ratios. Therefore, the powdery mildew on B. asiatica is described as Leveillula buddlejae sp. nov.     

Molecular detection of a gene effective against powdery mildew in the wheat cultivar Liangxing 66

Since it was commercialized in 2008, Liangxing 66 is one of the most widely grown cultivars of wheat (Triticum aestivum L.) in winter and facultative wheat-producing regions in northern China. This cultivar displays broad-spectrum resistance to isolates of powdery mildew. To identify the powdery mildew resistance gene in Liangxing 66, genetic analysis and molecular mapping were conducted using the F₂ populations and F_2:3 families derived from the reciprocal crosses of Liangxing 66 and the susceptible cultivar Jingshuang 16. A single dominant gene, tentatively designated PmLX66, conferred resistance in Liangxing 66 to the powdery mildew isolate E09. The results of molecular mapping indicated that this gene was located on the short arm of chromosome 5D and flanked by SCAR203 and Xcfd81 at genetic distances of 0.4 and 2.8?cM, respectively, which is similar to the position of locus Pm2. However, PmLX66 and Pm2 showed different reactions to five of the 42 isolates of powdery mildew tested. Together, these results indicated that PmLX66 was most likely an allele of Pm2. Based on its superior yield and agronomic performance, in combination with powdery mildew resistance, Liangxing 66 is useful as a promising parent for control of powdery mildew and for the development of new disease-resistant cultivars.     

Additional rDNA ITS sequences and its phylogenetic consequences for the genus Leveillula with emphasis on conidium morphology

Taxonomy of the genus Leveillula has long been considered as a challenge in powdery mildew systematics. The rDNA diversity has recently been used for phylogenetic analysis of several specimens of the genus Leveillula. In the present study, additional rDNA ITS sequences are provided and a new phylogenetic analysis is carried out aiming at a better understanding of the genetic diversity in the genus Leveillula. New analyses confirmed that L. taurica is unique in the genus, as it exhibits an intraspecific gene sequence diversity considerably higher than in other species. In several cases L. taurica s. lat. on a certain host plant species has a sequence different from L. taurica on other host plants. Moreover, DNA data indicated different lineages among L. taurica specimens which were hardly distinguishable by morphology. More than one genotype occurring on a single host is sometimes possible. According to these results, several races such as Leveillula on Artemisia, Acroptilon, Onobrychis, are molecularly well characterized. While there is enough molecular evidence to delimit such races as independent taxa, clear morphological delimitations between these new and already published taxa are very difficult or even impossible. However, ecological features, and above all, host specificity for biotrophic fungi such as powdery mildew, would be a good criterion to discriminate cryptic taxa along with rDNA sequences. In fact, many collections of Leveillula strains on different hosts show their own type of conidial morphology, which is usually consistent for a strain on a single host species. Hence, we have proposed to describe new species for Leveillula on some host plants such as Artemisia, Acroptilon, Echinops and Onobrychis.     

ENHANCED DISEASE RESISTANCE4 Associates with CLATHRIN HEAVY CHAIN2 and Modulates Plant Immunity by Regulating Relocation of EDR1 in Arabidopsis

Obligate biotrophs, such as the powdery mildew pathogens, deliver effectors to the host cell and obtain nutrients from the infection site. The interface between the plant host and the biotrophic pathogen thus represents a major battleground for plant-pathogen interactions. Increasing evidence shows that cellular trafficking plays an important role in plant immunity. Here, we report that Arabidopsis thaliana ENHANCED DISEASE RESISTANCE4 (EDR4) plays a negative role in resistance to powdery mildew and that the enhanced disease resistance in edr4 mutants requires salicylic acid signaling. EDR4 mainly localizes to the plasma membrane and endosomal compartments. Genetic analyses show that EDR4 and EDR1 function in the same genetic pathway. EDR1 and EDR4 accumulate at the penetration site of powdery mildew infection, and EDR4 physically interacts with EDR1, recruiting EDR1 to the fungal penetration site. In addition, EDR4 interacts with CLATHRIN HEAVY CHAIN2 (CHC2), and edr4 mutants show reduced endocytosis rates. Taken together, our data indicate that EDR4 associates with CHC2 and modulates plant immunity by regulating the relocation of EDR1 in Arabidopsis.     

Water Stress-Induced Changes in the Morphology of the Powdery Mildew, Leveillula taurica (Lèv.) Arn.

Water stress in Capsiucm annuum L., induced by means of polyethylene glycol influenced the length and branching frequency of conidiophores and conidial dimensions of the powdery mildew, Leveillula taurica (Lèv.) Arn. Conidiophore branching frequency and length as well as conidial length and width decreased with decreasing relative water content. The availability of water to host plants seems to have a direct effect on the growth vigour and development of the powdery mildew possibly is suggested that water stress may be imposed on host plants by the dry harmattan season resulting in reduced growth vigour of the powdery mildew. Morphology of powdery mildew developing under low water potential may be altered.     

Quantitative trait loci mapping of adult-plant resistance to powdery mildew in Chinese wheat cultivar Lumai 21

Powdery mildew, caused by Blumeria graminis f. sp. tritici, is a major fungal disease in common wheat (Triticum aestivum L.) worldwide. The Chinese winter wheat cultivar Lumai 21 has shown good and stable adult plant resistance for 19 years. The aim of this study was to map quantitative trait loci (QTLs) for resistance to powdery mildew in a population of 200 F₃ lines from the cross Lumai 21/Jingshuang 16. The population was tested for powdery mildew reaction in Beijing and Anyang in the 2005–2006 and 2006–2007 cropping seasons, providing data for 4 environments. A total of 1,375 simple sequence repeat (SSR) markers were screened for associations with powdery mildew reactions, initially in bulked segregant analysis. Based on the mean disease values averaged across environments, broad-sense heritabilities of maximum disease severity and area under the disease progress curve were 0.96 and 0.77, respectively. Three QTLs for adult plant resistance were detected by inclusive composite interval mapping. These were designated QPm.caas-2BS, QPm.caas-2BL and QPm.caas-2DL, respectively, and explained from 5.4 to 20.6% of the phenotypic variance across environments. QPm.caas-2BS and QPm.caas-2DL were likely new adult plant resistance QTLs flanked by SSR markers Xbarc98–Xbarc1147 and Xwmc18–Xcfd233, respectively. These markers could be useful for improving wheat powdery mildew resistance in breeding programs.     

Adult plant and seedling resistance to powdery mildew in a Triticum aestivum × Triticum militinae hybrid line

In the progeny of a cross between the common wheat cultivar Tähti and Triticum militinae, a member of the timopheevii group of tetraploid wheats, several hybrid lines were selected that are characterized by improved seedling and adult plant resistance (APR) to powdery mildew. An F₂ single-seed descendant mapping population segregating for seedling resistance and APR to powdery mildew was analysed for the identification of quantitative trait loci (QTL). The main QTL responsible for APR was detected on the long arm of chromosome 4A tightly linked to the Xgwm160 locus on a T. militinae translocation explaining up to 54% of phenotypic variance. The same translocation influenced seedling resistance to powdery mildew upon inoculation of plants with a synthetic population of Blumeria graminis DC. f. sp. tritici, and explained 28–33% of the phenotypic variance.     

Coevolution between a Family of Parasite Virulence Effectors and a Class of LINE-1 Retrotransposons

Parasites are able to evolve rapidly and overcome host defense mechanisms, but the molecular basis of this adaptation is poorly understood. Powdery mildew fungi (Erysiphales, Ascomycota) are obligate biotrophic parasites infecting nearly 10,000 plant genera. They obtain their nutrients from host plants through specialized feeding structures known as haustoria. We previously identified the AVR _k1 powdery mildew-specific gene family encoding effectors that contribute to the successful establishment of haustoria. Here, we report the extensive proliferation of the AVR _k1 gene family throughout the genome of B. graminis, with sequences diverging in formae speciales adapted to infect different hosts. Also, importantly, we have discovered that the effectors have coevolved with a particular family of LINE-1 retrotransposons, named TE1a. The coevolution of these two entities indicates a mutual benefit to the association, which could ultimately contribute to parasite adaptation and success. We propose that the association would benefit 1) the powdery mildew fungus, by providing a mechanism for amplifying and diversifying effectors and 2) the associated retrotransposons, by providing a basis for their maintenance through selection in the fungal genome.     

Four powdery mildew species with catenate conidia infect Galium: molecular and morphological evidence

The Erysiphaceae are a group of obligately biotrophic fungi that cause powdery mildew disease of angiosperms. Due to their inability to be cultured on artificial media, the taxonomy of the Erysiphaceae has generally been based on the morphological characteristics of fresh and herbarium specimens. Thus, several morphological species with wide host ranges have long been maintained in this family, even though they clearly consist of several biological species. Erysiphe galii has been known as a powdery mildew of Galium spp. Recently, the former E. galii var. galii has been reassessed as Neoerysiphe galii and E. galii var. riedliana as Golovinomyces riedlianus, along with a taxonomic revision of the generic concept of the Erysiphaceae. The present study was conducted to evaluate the validity of the taxonomic revision of the two varieties of E. galii. During the course of this study, we found that the Galium powdery mildews consist of at least four different species, viz. Neoerysiphe galii, Golovinomyces orontii, G. riedlianus, and an unknown species collected in Argentina. The latter species is described as a new species, Golovinomyces calceolariae. The three species belonging to Golovinomyces are morphologically very similar to each other, i.e. the discrimination between them is rather difficult. The morphological differences of the three Golovinomyces species of Galium are discussed.     

PMR5, an acetylation protein at the intersection of pectin biosynthesis and defense against fungal pathogens

Powdery mildew (Golovinomyces cichoracearum), one of the most prolific obligate biotrophic fungal pathogens worldwide, infects its host by penetrating the plant cell wall without activating the plant's innate immune system. The Arabidopsis mutant powdery mildew resistant 5 (pmr5) carries a mutation in a putative pectin acetyltransferase gene that confers enhanced resistance to powdery mildew. Here, we show that heterologously expressed PMR5 protein transfers acetyl groups from [¹⁴C]‐acetyl‐CoA to oligogalacturonides. Through site‐directed mutagenesis, we show that three amino acids within a highly conserved esterase domain in putative PMR5 orthologs are necessary for PMR5 function. A suppressor screen of mutagenized pmr5 seed selecting for increased powdery mildew susceptibility identified two previously characterized genes affecting the acetylation of plant cell wall polysaccharides, RWA2 and TBR. The rwa2 and tbr mutants also suppress powdery mildew disease resistance in pmr6, a mutant defective in a putative pectate lyase gene. Cell wall analysis of pmr5 and pmr6, and their rwa2 and tbr suppressor mutants, demonstrates minor shifts in cellulose and pectin composition. In direct contrast to their increased powdery mildew resistance, both pmr5 and pmr6 plants are highly susceptibile to multiple strains of the generalist necrotroph Botrytis cinerea, and have decreased camalexin production upon infection with B. cinerea. These results illustrate that cell wall composition is intimately connected to fungal disease resistance and outline a potential route for engineering powdery mildew resistance into susceptible crop species.     

Morphological and phylogenetic comparisons amongst powdery mildews on Catalpa in the UK

Three species of powdery mildew, Erysiphe elevata, E. catalpae, and Neoerysiphe galeopsidis were identified on Catalpa species in England in 2004. A new disease record, N. galeopsidis was the first Catalpa mildew to appear (in June), but it was later out-competed by E. elevata that caused the most serious damage. Both mildews also attacked C. speciosa, C. ×erubescens and a new host, ×Chitalpa tashkentensis, a Chilopsis × Catalpa hybrid. No powdery mildew was detected on C. bungei, C. ovata, or C. fargesii. Identifications of the pathogens using morphological data were fully supported by DNA analysis yielding characteristic rDNA ITS sequences. The sequences placed E. catalpae within the E. aquilegiae clade. The sequences for E. elevata from southern England and France closely matched those from Hungary and North America, confirming the recent spread of this pathogen from the USA. It eventually overran N. galeopsidis and its sudden appearance in the UK could be due to greater aggressiveness and to the production of more ascomata especially during autumns with delayed leaf fall as in 2001. A further species, Oidium hiratae (i.e. Podosphaera sp.), though described from a 1978 UK collection on C. bignonioides, was not detected in the field.     

Translocation of carbon in powdery mildewed barley

This paper compares translocation in healthy and powdery mildew (Erysiphe graminis f. sp. hordei, race CR3) infected barley (Hordeum vulgare, variety Manchuria). The sink-like properties of the powdery mildew infection were used to determine what effect imposing a sink in the midst of normal source tissue (mature primary leaf) had on the translocation process. The pattern of translocation was determined by monitoring the movement of ¹⁴C which was photosynthetically incorporated from ¹⁴C either by the primary or second leaf. In the healthy primary leaf of barley, ¹⁴C fixed in the tip section of the blade was preferentially translocated to the root, whereas ¹⁴C fixed in the basal section was primarily translocated to the shoot. When a sporulating powdery mildew infection was present in the mid-section of the primary leaf, ¹⁴C fixed in that section or in the acropetal healthy tip section readily accumulated in the infection area. Labeled carbon fixed in the healthy basal section was translocated into the other parts of the plant with only a small fraction moving acropetally into the infected mid-section. The ¹⁴C fixed by the second leaf was translocated to the root and younger shoot with very little entering the primary leaf. The presence of the mildew infection did not alter this pattern.     

Pm50: a new powdery mildew resistance gene in common wheat derived from cultivated emmer

Fungal diseases of wheat, including powdery mildew, cause significant crop, yield and quality losses throughout the world. Knowledge of the genetic basis of powdery mildew resistance will greatly support future efforts to develop and cultivate resistant cultivars. Studies were conducted on cultivated emmer-derived wheat line K2 to identify genes involved in powdery mildew resistance at the seedling and adult plant growth stages using a BC₁ doubled haploid population derived from a cross between K2 and susceptible cultivar Audace. A single gene was located distal to microsatellite marker Xgwm294 on the long arm of chromosome 2A. Quantitative trait loci (QTL) analysis indicated that the gene was also effective at the adult plant stage, explaining up to 79.0 % of the variation in the progeny. Comparison of genetic maps indicated that the resistance gene in K2 was different from Pm4, the only other formally named resistance gene located on chromosome 2AL, and PmHNK54, a gene derived from Chinese germplasm. The new gene was designated Pm50.