中国长春瓜类白粉菌Podosphaera xanthii形态学和分子系统学研究

2009年长春农博园瓜类白粉病大发生,而且大量闭囊产生在葫芦科的四个属（葫芦属、黄瓜属、栝楼属、南瓜属）植物上。对病害发生的严重度和病情指数进行了调查,结果表明园中栽培的所有品种均为感病品种。形态学观察显示分子孢子串生,具纤维体,闭囊壳只含有一个子囊,内含8个子囊孢子,子囊顶端孔较宽（15.6－28.8μm）。有性和无性形态学特征表明该菌应属于Podosphaera xanthii。基于rDNA ITS序列的分子系统学分析表明该菌与GenBank中已报道的该种在其他地区瓜类上的序列以及在其他寄主上的序列聚     

First report of powdery mildew caused by Podosphaera xanthii on Luffa acutangula in Odisha State,India

Abstract

Ridge gourd (Luffa acutangula) is an herbaceous perennial twining vine cultivated globally as vegetable and medicinal plant. During October to January in 2014 and 2015, 40% powdery mildew disease incidence was observed in different areas of Odisha state, India. The pathogenicity experiments confirmed the powdery mildew disease symptoms on artificially inoculated L. acutangula seedlings. Causal organism was identified as Podosphaera xanthii on the basis of morphological and molecular studies. This is the first report of powdery mildew disease on L. acutangula caused by P. xanthii.     

Identification and validation of powdery mildew (Podosphaera xanthii)-resistant loci in recombinant inbred lines of cucumber (Cucumis sativus L.)

Powdery mildew limits cucumber production worldwide. Most resistant cucumber cultivars become susceptible to powdery mildew at low temperatures. Resistance within a wide temperature range is therefore desirable for cucumber production. We constructed a cucumber genetic linkage map based on a population of 111 recombinant inbred lines derived from a cross between CS-PMR1, with strong and temperature-independent resistance, and Santou, with moderate and temperature-dependent resistance. The map spans 693.0 cM and consists of 296 markers segregating into seven linkage groups; the markers include 289 simple sequence repeats (SSRs), six sequence characterized amplified regions, and one inter simple sequence repeat. Due to the presence of 150 common SSR markers, we were able to compare our map with previously published maps obtained by using populations derived from inter- or intra-variety crosses. We also evaluated powdery mildew resistance of the recombinant inbred lines and identified seven quantitative trait loci (QTL) contributed by CS-PMR1 and two QTL contributed by Santou. Four QTL (pm3.1, pm5.1, pm5.2 and pm5.3) were successfully validated by using populations derived from residual heterozygous lines. Some of the QTL identified in our study are in good agreement with previously published results obtained with materials of different origin. The markers reported here would be useful for introducing high and temperature-independent resistance by accumulation of QTL from CS-PMR1 and Santou.     

Molecular phylogenetic status of korean strain of Podosphaera xanthii, a causal pathogen of powdery mildew on Japanese thistle (Cirsium japonicum) in Korea

Powdery mildew diseases are sensitive to climate change and spread can be favored by increased temperature and low moisture. During 2011 to 2012, a powdery mildew disease by a Podosphaera species was observed on the leaves of Japanese thistle (Cirsium japonicum) in Korea. The initial sign of this disease included scattered superficial white mycelia on leaves. As the disease progressed, abundant necrotic black spots exhibiting chasmothecia were formed on the leaves. rDNA ITS and 28S homologies of the fungus (EML-CSPW1) showed 100% identity values with those regions from many strains of P. xanthii (syn. P. fusca) via NCBI BLASTN search.     

Seasonal variations in the occurrence of Golovinomyces orontii and Podosphaera xanthii,causal agents of cucurbit powdery mildew in Northern Italy

Powdery mildew, caused by Golovinomyces orontii and Podosphaera xanthii, is a widespread disease that causes important losses in cucurbit production. To determine the aetiology and the epidemiology of cucurbit powdery mildew disease in the North of Italy, observations on the occurrence of the main disease‐causing fungal species were conducted during the 2010, 2011 and 2012 growing seasons. Samples of infected leaves of zucchini, melon and pumpkin plants, either from field or greenhouse crops, were collected every 15–18 days from May to September/October. To identify the fungal species, both morphological observations based on the asexual stage and molecular identifications by a Multiplex‐PCR reaction with species‐specific primers were performed. Climatic parameters of temperature and relative humidity were also monitored. Pearson's correlation coefficient and Principal Component Analysis showed a negative significant correlation between the two species, and a peculiar epidemiological behaviour was also observed: the earlier infections were caused by G. orontii, which was the predominant species till the end of June–middle of July. At this time, this species progressively decreased in frequency and was replaced by P. xanthii that became the main species infecting cucurbits till the end of the growing season. As the two species have different ecological requirements, these seasonal variations in the cucurbit powdery mildew species composition could possibly be explained by the influence of temperature and relative humidity on the pathogen epidemiology during the growing season but also by the different overwintering strategies adopted by the two species.     

A haustorial‐expressed lytic polysaccharide monooxygenase from the cucurbit powdery mildew pathogen Podosphaera xanthii contributes to the suppression of chitin‐triggered immunity

Podosphaera xanthii is the main causal agent of cucurbit powdery mildew and a limiting factor of crop productivity. The lifestyle of this fungus is determined by the development of specialized parasitic structures inside epidermal cells, termed haustoria, that are responsible for the acquisition of nutrients and the release of effectors. A typical function of fungal effectors is the manipulation of host immunity, for example the suppression of pathogen‐associated molecular pattern (PAMP)‐triggered immunity (PTI). Chitin is a major component of fungal cell walls, and chitin oligosaccharides are well‐known PAMP elicitors. In this work, we examined the role of PHEC27213, the most highly expressed, haustorium‐specific effector candidate of P. xanthii. According to different computational predictions, the protein folding of PHEC27213 was similar to that of lytic polysaccharide monooxygenases (LPMOs) and included a conserved histidine brace; however, PHEC27213 had low sequence similarity with LPMO proteins and displayed a putative chitin‐binding domain that was different from the canonical carbohydrate‐binding module. Binding and enzymatic assays demonstrated that PHEC27213 was able to bind and catalyse colloidal chitin, as well as chitooligosaccharides, acting as an LPMO. Furthermore, RNAi silencing experiments showed the potential of this protein to prevent the activation of chitin‐triggered immunity. Moreover, proteins with similar features were found in other haustorium‐forming fungal pathogens. Our results suggest that this protein is a new fungal LPMO that catalyses chitooligosaccharides, thus contributing to the suppression of plant immunity during haustorium development. To our knowledge, this is the first mechanism identified in the haustorium to suppress chitin signalling.     

Eradication of apple powdery mildew (Podosphaera leucotricha) with dormant season sprays of surface-active agents

selection of surfactants (5%, w/v, a.i.) representative of the three main ionic types eradicated Podosphaera leucotricha from buds on dormant apple trees, cvs Cox's Orange Pippin, Jonathan and Michelin. The pathogen was apparently killed without undue damage to buds, although a phytotoxic response in the form of staining of vascular tissue and some crop reductions were observed. Treatments applied with hand lances were equally effective at reducing infections of vegetative buds on terminals, on non-flowering spurs and flower buds. Indication of a positive correlation between the eradication of mildew from Cox's Orange Pippin trees and an increase in the incidence of apple canker (Nectria galligena) was found at Long Ashton, but this effect was negligible at sites in the West Midlands region. Efficacy of active formulations was reduced by rain soon after application or by a reduction in the concentration of applied material. Applications with a mist-blower were effective on the cv. Cox's Orange Pippin, but less so on the cv. Jonathan, while on both cultivars efficacy of a spray containing 5 % a.i. decreased in the order 2240:1120:560 1/ha. Anti-mildew activity was not associated with any particular chemical or ionic type of surfactant, and there was an indication that eradicant activity of alkyl phenol ethoxylates was related to their HLB value. The ecological and economic implications of the use of surfactants as fungicides are discussed.     

Powdery mildew (Podosphaera xanthii) resistance in melon is categorized into two types based on inhibition of the infection processes

Physiological races of powdery mildew (Podosphaera xanthii) cause different symptoms in eight melon lines. Infection by races 1, 2, and 5 was examined in different melon lines. After a compatible reaction, conidia germination, haustorium initiation from the germ tube, germ tube branching, and sporulation occurred within 12, 24, 48, and 120 h, respectively, and the conidia matured within 240 h. In contrast, type i and ii inhibition were identified through incompatible reactions. The germ tube and haustorium were initiated from conidia, but no germ tube branching occurred in the lines with type i resistance within 48-240 h. In type ii resistance, germ tube branching was observed within 120 h, but no sporulation was observed within 240 h. The number of fluorescing epidermal cells was higher within 24 h in type i, and within 48-120 h in type ii resistance lines than in susceptible lines. Callose accumulation around the haustorium was detected in type ii resistance lines within 48-120 h. This suggests that the rapid hypersensitive response (HR) within 24 h has an important role in the type i response, while HR and callose accumulation in the type ii response occur slowly between 48 and 120 h. Of the resistant lines, PMR 45 and WMR 29 showed a type i incompatible response; the PI 414723 response was entirely type ii; and PMR 5, PI 124112, and MR-1 showed different responses depending on the race. Therefore, the two types of incompatible responses were intermixed in the same germplasm.     

Multilocus phylogenetic analyses within Blumeria graminis, a powdery mildew fungus of cereals

Blumeria graminis, a powdery mildew fungus, is an important plant pathogen that causes serious damage to a variety of cereal crops. In spite of the importance of the pathogen, information on phylogenetic structure within B. graminis is scarce. In this study we conducted phylogenetic analyses of B. graminis based on the DNA sequences of four different DNA regions (ITS, 28S rDNA, chitin synthase 1, and beta-tubulin). The analyses revealed that the protein-coding regions have higher amounts of phylogenetic signals than rDNA regions and are useful for phylogenetic analyses of B. graminis. The present phylogenetic analyses revealed nine distinct groups in the B. graminis isolates used in this study, a result which was commonly supported by all trees constructed from the four DNA regions. Isolates from a single host genus belonged to a single group except for isolates from Lolium and Bromus, in which the isolates were split into two and three groups, respectively. Isolates from Agropyron, Secale and Triticum formed a distinct clade (Triticum clade) with identical or similar DNA sequences. The Hordeum clade was a sister of the Triticum clade, and Poa and Avena clades were distantly related to the Triticum and Hordeum clades. This phylogenetic relationship of B. graminis is well concordant with the level of reproductive isolation between formae speciales and also with phylogeny inferred from a cytological study. Shimodaira-Hasegawa and Templeton tests using sequences of four different DNA regions significantly rejected the tree topology of plants. Therefore, possibility of co-speciation between B. graminis and its host plants was obscure in this study.     

The knock‐down of the expression of MdMLO19 reduces susceptibility to powdery mildew (Podosphaera leucotricha) in apple (Malus domestica)

Varieties resistant to powdery mildew (PM; caused by Podosphaera leucotricha) are a major component of sustainable apple production. Resistance can be achieved by knocking‐out susceptibility S‐genes to be singled out among members of the MLO (Mildew Locus O) gene family. Candidates are MLO S‐genes of phylogenetic clade V up‐regulated upon PM inoculation, such as MdMLO11 and 19 (clade V) and MdMLO18 (clade VII). We report the knock‐down through RNA interference of MdMLO11 and 19, as well as the complementation of resistance with MdMLO18 in the Arabidopsis thaliana triple mlo mutant Atmlo2/6/12. The knock‐down of MdMLO19 reduced PM disease severity by 75%, whereas the knock‐down of MdMLO11, alone or in combination with MdMLO19, did not result in any reduction or additional reduction of susceptibility compared with MdMLO19 alone. The test in A. thaliana excluded a role for MdMLO18 in PM susceptibility. Cell wall appositions (papillae) were present in both PM‐resistant and PM‐susceptible plants, but were larger in resistant lines. No obvious negative phenotype was observed in plants with mlo genes knocked down. Apparently, MdMLO19 plays the pivotal role in apple PM susceptibility and its knock‐down induces a very significant level of resistance.     

Abscisic acid negatively regulates post-penetration resistance of Arabidopsis to the biotrophic powdery mildew fungus

Pytohormone abscisic acid(ABA) plays important roles in defense responses.Nonetheless,how ABA regulates plant resistance to biotrophic fungi remains largely unknown.Arabidopsis ABA-deficient mutants,aba2-1 and aba3-1,displayed enhanced resistance to the biotrophic powdery mildew fungus Golovinomyces cichoracearum.Moreover,exogenously administered ABA increased the susceptibility of Arabidopsis to G.cichoracearum.Arabidopsis ABA perception components mutants,abil-1 and abi2-1,also displayed similar phenotypes to ABA-deficient mutants in resistance to G.cichoracearum.However,the resistance to G.cichoracearum is not changed in downstream ABA signaling transduction mutants,abi3-1,abi4-1,and abi5-1.Microscopic examination revealed that hyphal growth and conidiophore production of G.cichoracearum were compromised in the ABA deficient mutants,even though pre-penetration and penetration growth of the fungus were not affected.In addition,salicylic acid(SA) and MPK3 are found to be involved in ABA-regulated resistance to G.cichoracearum.Our work demonstrates that ABA negatively regulates post-penetration resistance of Arabidopsis to powdery mildew fungus G.cichoracearum,probably through antagonizing the function of SA.     

Changes in specific activities of peroxidase, chitinase and phenylalanine ammonia-lyase and phenolic content in cucumber leaves inoculated with Podosphaera fusca, the causal agent of powdery mildew

In this investigation, the effects of powdery mildew disease [caused by Podosphaera fusca (syn. Sphaerotheca fuliginea)] on specific activities of several defense-related enzymes and phenolic content were studied in cucumber leaves. Spore suspension of the fungus was sprayed on cucumber (cv. Super Dominus) plants in greenhouse and leaves from both inoculated and non-inoculated control plants were sampled at 0, 24, 48, 72 and 144 hours after inoculation (HAI). Spore germination and tissue colonization of P. fusca were microscopically studied on the inoculated surface of leaf samples. Further, Phenolic content (PHE) and specific activities of peroxidase (POX), chitinase (CHI) and phenylalanine ammonia-lyase (PAL) were spectrophotometrically measured in leaf extracts. Time-course of disease progress on the leaf surface showed that maximum spore germination occurred within 24 HAI and host penetration and disease development process began during 24-48 HAI. Evaluation of enzyme activities showed that POX specific activity in inoculated plants significantly increased at 72 HAI onwards and reached 2.5 times of that of control at 144 HAI. CHI specific activity showed a transient reduction in inoculated plants between 48-72 HAI and thereafter increased significantly in relation to control. PAL specific activity in inoculated plants was not significantly different from that of control. PHE in inoculated plants showed a significant increase compared to control at 48 HAI and thereafter. Comparison of time-course of disease progress with changes in enzyme activities indicated that POX activity had an increasing trend during disease progress whereas CHI activity showed a transient decrease at the early stages and then increased during the later stages of infection: PAL activity did not show any changes during the infection and PHE increased at the early stages of infection process and remained constant at rest of the time.     

Sawadaea nankinensis comb. nov.: a powdery mildew fungus of Acer buergerianum

The anamorphic state of a powdery mildew on trident maple (Acer buergerianum, Aceraceae), belonging to Sawadaea, has been observed since 1980 in Tokyo and other areas of Japan. Since the autumn of 2003, this fungus has begun to produce chasmothecia in various areas of Japan, which were consistent with Erysiphe nankinensis (= Uncinula nankinensis), but apparently contradictory to the characteristics of the anamorph. Based on a molecular phylogenetic analysis using DNA separately extracted from the anamorph and teleomorph of the fungus on A. buergerianum, it could be demonstrated that sequences of this fungus are sister to Sawadaea. As the anamorph belongs to Oidium subgen. Octagoidium and because of the phylogenetic position within the Sawadaea clade, the new combination Sawadaea nankinensis is proposed for this species. The genus Sawadaea is emended to comprise species with consistently unbranched appendages.     

The isoelectric point of proteins influences their translocation to the extrahaustorial matrix of the barley powdery mildew fungus

Many biotrophic fungal plant pathogens develop feeding structures, haustoria, inside living plant cells, which are essential for their success. Extrahaustorial membranes (EHMs) surround haustoria and delimit the extrahaustorial matrices (EHMxs). Little is known about transport mechanisms across EHMs and what properties proteins and nutrients need in order to cross these membranes. To investigate this further, we expressed fluorescent proteins in the cytosol of infected barley leaf epidermal cells after particle bombardment and investigated properties that influenced their localisation in the powdery mildew EHMx. We showed that this translocation is favoured by a neutral isoelectric point (pI) between 6.0 and 8.4. However, for proteins larger than 50 kDa, pI alone does not explain their localisation, hinting towards a more complex interplay between pI, size, and sequence properties. We discuss the possibility that an EHM translocon is involved in protein uptake into the EHMx.     

The tomato Arp2/3 complex is required for resistance to the powdery mildew fungus Oidium neolycopersici

The actin‐related protein 2/3 complex (Arp2/3 complex), a key regulator of actin cytoskeletal dynamics, has been linked to multiple cellular processes, including those associated with response to stress. Herein, the Solanum habrochaites ARPC3 gene, encoding a subunit protein of the Arp2/3 complex, was identified and characterized. ShARPC3 encodes a 174‐amino acid protein possessing a conserved P21‐Arc domain. Silencing of ShARPC3 resulted in enhanced susceptibility to the powdery mildew pathogen Oidium neolycopersici (On‐Lz), demonstrating a role for ShARPC3 in defence signalling. Interestingly, a loss of ShARPC3 coincided with enhanced susceptibility to On‐Lz, a process that we hypothesize is the result of a block in the activity of SA‐mediated defence signalling. Conversely, overexpression of ShARPC3 in Arabidopsis thaliana, followed by inoculation with On‐Lz, showed enhanced resistance, including the rapid induction of hypersensitive cell death and the generation of reactive oxygen. Heterologous expression of ShARPC3 in the arc18 mutant of Saccharomyces cerevisiae (i.e., ?arc18) resulted in complementation of stress‐induced phenotypes, including high‐temperature tolerance. Taken together, these data support a role for ShARPC3 in tomato through positive regulation of plant immunity in response to O. neolycopersici pathogenesis.     

The conserved oligomeric Golgi complex is involved in penetration resistance of barley to the barley powdery mildew fungus

Membrane trafficking is vital to plant development and adaptation to the environment. It is suggested that post‐Golgi vesicles and multivesicular bodies are essential for plant defence against directly penetrating fungal parasites at the cell wall. However, the actual plant proteins involved in membrane transport for defence are largely unidentified. We applied a candidate gene approach and single cell transient‐induced gene silencing for the identification of membrane trafficking proteins of barley involved in the response to the fungal pathogen Blumeria graminis f.sp. hordei. This revealed potential components of vesicle tethering complexes [putative exocyst subunit HvEXO70F‐like and subunits of the conserved oligomeric Golgi (COG) complex] and Golgi membrane trafficking (COPIγ coatomer and HvYPT1‐like RAB GTPase) as essential for resistance to fungal penetration into the host cell.     

Functional conservation of wheat and rice Mlo orthologs in defense modulation to the powdery mildew fungus

Homologs of barley Mlo are found in syntenic positions in all three genomes of hexaploid bread wheat, Triticum aestivum, and in rice, Oryza sativa. Candidate wheat orthologs, designated TaMlo-A1, TaMlo-B1, and TaMlo-D1, encode three distinct but highly related proteins that are 88% identical to barley MLO and appear to originate from the three diploid ancestral genomes of wheat. TaMlo-B1 and the rice ortholog, OsMlo2, are able to complement powdery mildew-resistant barley mlo mutants at the single-cell level. Overexpression of TaMlo-B1 or barley Mlo leads to super-susceptibility to the appropriate powdery mildew formae speciales in both wild-type barley and wheat. Surprisingly, overexpression of either Mlo or TaMlo-B1 also mediates enhanced fungal development to tested inappropriate formae speciales. These results underline a regulatory role for MLO and its wheat and rice orthologs in a basal defense mechanism that can interfere with forma specialis resistance to powdery mildews.