Appearance of race Pfs:5 of spinach downy mildew fungus, Peronospora farinosa f. sp. spinaciae, in Japan

The races for the causal agent of spinach downy mildew Peronospora farinosa f. sp. spinaciae were identified by inoculation of race-differential cultivars. One isolate was identified as Pfs:5s and the others belonged to a new race. This is the first report of race Pfs:5 and another new race in Japan.     

First report of spinach downy mildew caused by race Pfs:8 of <Emphasis Type="Italic">Peronospora farinosa</Emphasis> f. sp. <Emphasis Type="Italic">spinaciae</Emphasis> in Japan

The race of field isolates of Peronospora farinosa f. sp. spinaciae (Pfs), causal agent of spinach downy mildew, were identified using race-differential cultivars. One isolate was similar to race Pfs:6. Three isolates were identified as race Pfs:8, the first time the race has been reported in Japan as far as we know. The differential reaction caused by the other two isolates did not match any known to be caused by races Pfs:1 through Pfs:11; thus, this strain appears to a new pathogenic strain in Japan.     

Recent races of spinach downy mildew pathogen <Emphasis Type="Italic">Peronospora farinosa</Emphasis> f. sp. <Emphasis Type="Italic">spinaciae</Emphasis> in Japan

Isolates of spinach downy mildew fungus, Peronospora farinosa f. sp. spinaciae, collected at seven sites in Japan from December 2013 to March 2016, were identified as belonging to races 8, 10, 12 and 13, and one was unauthorized. Races 10, 12, 13 and the unauthorized were first found in Japan.     

Characterization of a resistance locus (Pfs-1) to the spinach downy mildew pathogen (Peronospora farinosa f. sp. spinaciae) and development of a molecular marker linked to Pfs-1

Downy mildew is a destructive disease of spinach worldwide. There have been 10 races described since 1824, six of which have been identified in the past 10 years. Race identification is based on qualitative disease reactions on a set of diverse host differentials which include open-pollinated cultivars, contemporary hybrid cultivars, and older hybrid cultivars that are no longer produced. The development of a set of near-isogenic open-pollinated spinach lines (NILs), having different resistance loci in a susceptible and otherwise common genetic background, would facilitate identification of races of the downy mildew pathogen, provide a tool to better understand the genetics of resistance, and expedite the development of molecular markers linked to these disease resistance loci. To achieve this objective, the spinach cv. Viroflay, susceptible to race 6 of Peronospora farinosa f. sp. spinaciae, was used as the recurrent susceptible parent in crosses with the hybrid spinach cv. Lion, resistant to race 6. Resistant F(1) progeny were subsequently backcrossed to Viroflay four times with selection for race 6 resistance each time. Analysis of the segregation data showed that resistance was controlled by a single dominant gene, and the resistance locus was designated Pfs-1. By bulk segregant analysis, an amplified fragment length polymorphism (AFLP) marker (E-ACT/M-CTG) linked to Pfs-1 was identified and used to develop a co-dominant Sequence characterized amplified region (SCAR) marker. This SCAR marker, designated Dm-1, was closely linked ( approximately 1.7 cM) to the Pfs-1 locus and could discriminate among spinach genotypes that were homozygous resistant (Pfs-1Pfs-1), heterozygous resistant (Pfs-1pfs-1), or homozygous susceptible (pfs-1pfs-1) to race 6 within the original mapping population. Evaluation of a wide range of commercial spinach lines outside of the mapping population indicated that Dm-1 could effectively identify Pfs-1 resistant genotypes; the Dm-1 marker correctly predicted the disease resistance phenotype in 120 out of 123 lines tested. In addition, the NIL containing the Pfs-1 locus (Pfs-1Pfs-1) was resistant to multiple races of the downy mildew pathogen indicating Pfs-1 locus may contain a cluster of resistance genes.     

Spinach: better management of downy mildew and white rust through genomics

Spinach (Spinacia oleracea) has become an increasingly important vegetable crop in many parts of the world. Significant changes in production practices, particularly in the U.S. and E.U., have occurred in the past 10–15 years as a result of increased product demand. These changes likely increased the incidence and severity of downy mildew, caused by Peronospora farinosa f. sp. spinaciae. Recently, progress has been made to define the genetics of resistance to this pathogen and the closely related white rust pathogen, Albugo occidentalis. In this paper, we outline the genetic and genomic resources currently available for spinach, draw parallels between spinach diseases and more thoroughly characterized pathosystems, and describe efforts currently underway to develop new genetic and genomic tools to better understand downy mildew and white rust of spinach. Presently, many crucial tools and resources required to define the molecular underpinnings of disease are unavailable for either spinach or its pathogens. New resources and information for spinach genomics would provide a jumpstart for ongoing efforts to define (and deploy) genetic resistance against downy mildew and white rust.     

Characterization of resistance mechanisms activated by Trichoderma harzianum T39 and benzothiadiazole to downy mildew in different grapevine cultivars

Downy mildew, caused by Plasmopara viticola, is one of the most destructive diseases of grapevine and is controlled with intense application of chemical fungicides. Treatment with Trichoderma harzianum T39 (T39) or benzothiadiazole‐7‐carbothioic acid S‐methyl ester (BTH) has been previously shown to activate grapevine resistance to downy mildew and reduce disease symptoms in the Pinot noir cultivar. However, enhancement of plant resistance can be affected by several factors, including plant genotype. In order to further extend the use of resistance inducers against downy mildew, the physiological and molecular properties of T39‐ and BTH‐activated resistance in different cultivars of table and wine grapes were characterized under greenhouse conditions. T39 treatment reduced downy mildew symptoms, but the degree of efficacy differed significantly among grapevine cultivars. However, efficacy of BTH‐activated resistance was consistently high in the different cultivars. Expression profiles of defence‐related genes differed among cultivars in response to resistance inducers and to pathogen inoculation. T39 treatment enhanced the expression of defence‐related genes in the responsive cultivars, before and after P. viticola inoculation. A positive correlation between the efficacy of T39 and the expression level of defence‐related genes was found in Primitivo and Pinot noir plants, while different genes or more complex processes were probably activated in Sugraone and Negroamaro. The data reported here suggest that the use of a responsive cultivar is particularly important to maximize the efficacy of resistance inducers and new natural inducers should be explored for the less responsive cultivars.     

A comparison of two pathosystems: Downy mildew on Spinacia oleracea and on Chenopodium album

The development of downy mildew (Peronospora farinosa) on a cultivated spinach crop (Spinacia oleracea) was compared to that on lambsquarters (Chenopodium album), a naturally occurring weed. Experiments were carried out on small spinach plots, in which epidemics could develop from point sources (PS) or area sources (AS).Two lambsquarter populations were studied, in which downy mildew epidemics arose from oospore infections. The downy mildew epidemics, on spinach and lambtquarters, were related to the environmental factors: temperature, vapour pressure deficit (VPD) and rainfall. It is concluded that the two different formae speciales ofP. farinosa, one on spinach and one on lambsquarters, react in the same way to weather changes. The epidemiological patterns were almost similar.Samenvatting De ontwikkeling van valse meeldauw (Peronospora farinosa) op het cultuurgewas spinazie (Spinacia oleracea) werd vergeleken met die op het zich natuurlijk ontwikkelende onkruid melganzevoet (Chenopodium album). Daarvoor werden experimenten uitgevoerd op spinazieperceeltjes, waarop in enkele gevallen puntbroninfecties (PS), en in andere gevallen oppervlaktebrondinfecties (AS) waren aangebracht. Er werd gevarieerd in inoculatietijdstip.Bij melganzevoet werden twee populaties bestudeerd, waarin de ziekte via oösporeinfectie op natuurlijke wijze was ontstaan.Het geheel werd in verband gebracht met de omgevingsfactoren: temperatuur, dampdruk-deficiet (VPD) en regenval. Geconcludeerd kan worden, dat deP. farinosa formae speciales zowel op spinazie als op melganzevoet op gelijke wijze reageerden op veranderingen in het weer. Het verloop van de epidemieeën was sterk vergelijkbaar.     

First report of downy mildew of carnation caused by Peronospora dianthicola in Japan

Yellowish lesions with downy, gray growth developed on the leaves of carnations (Dianthus caryophyllus) in a greenhouse in Hokkaido Prefecture, Japan, in March 2011. We identified the fungus on diseased plants as Peronospora dianthicola on the basis of the morphologies of the conidia, conidiophores, and oospores. To confirm pathogenicity of the fungus, we inoculated healthy plants with a conidial suspension of the fungus, and the plants developed the same signs and symptoms as the naturally infected plants. We then reidentified P. dianthicola from the diseased lesions. This is the first report of downy mildew of carnation in Japan.     

Variation for virulence among Scandinavian isolates of Peronospora viciae f.sp. pisi (pea downy mildew) and responses of pea genotypes

Variation for virulence among Scandinavian isolates of Peronospora viciae f.sp. pisi (downy mildew) on different pea genotypes was investigated. Variation for virulence was found within and between mass-conidial isolates which were originally obtained from oospore populations. Virulence towards pea cultivars that have not been commerically cultivated in Scandinavia was observed. The specific resistance of the pea cultivars Puget, Cobri, Gastro, Starcovert and Starnain was confirmed. One pea breeding line showed a stable partial resistance to different isolates of the fungus and it was also highly resistant to race 8'from The Netherlands which is considered to be virulent on most pea genotypes carrying known specific resistance factors.     

Activity of the New Fungicide Benthiavalicarb Against Plasmopara viticola and its Efficacy in Controlling Downy Mildew in Grapevines

Benthiavalicarb is a new fungicide active against Oomycetes fungal plant pathogens. The present study shows that benthiavalicarb is effective for controlling the Oomycete fungal pathogen Plasmopara viticola, which causes downy mildew in grapevines. The fungicide did not affect zoospore discharge from sporangia of P. viticola, but strongly inhibited zoospore encystment, cystospore germination in vitro and mycelial growth, together with sporangial production in vivo. Benthiavalicarb showed strong prophylactic and local activity in intact plants or detached leaves and low translaminar activity. The compound was not translocated from leaf to leaf in either a acropetal or basipetal direction. Benthiavalicarb applied at 1, 3 and 6 days post-inoculation protected grapevine plants against downy mildew and inhibited sporulation of the pathogen. Similar results were obtained on leaf disks if benthiavalicarb was applied up to 96 h post-inoculation. Benthiavalicarb diminished the sporulation of P. viticola when applied to established disease in the tissue. Benthiavalicarb remained active on leaves for a period up to 28 days. Two foliar applications of benthiavalicarb, 2 weeks apart, to field-grown grapevines inhibited downy mildew development and were as effective as the standard metalaxyl-Cu treatment in controlling the disease. A formulated mixture of benthiavalicarb + Folpet was similar or superior in performance to metalaxyl-Cu and the new strobilurin trifloxystrobin in controlling downy mildew. The effectiveness of benthiavalicarb makes it well suited for integration into a control programme against downy mildew disease in vineyards, and as a component to delay resistance buildup.     

Induction of β-1,3-glucanase in Seedlings of Pearl Millet in Response to Infection by Sclerospora graminicola

Differential resistance of pearl millet cultivars to downy mildew disease was correlated with the levels of -1,3-glucanase in their seeds. Higher activity of the enzyme in highly resistant cultivars and lower activity in the highly susceptible ones suggested the possible use of -1,3-glucanase as a biochemical marker for screening pearl millet cultivars for downy mildew disease. Inoculation of seedlings with the downy mildew pathogen Sclerospora graminicola resulted in increased enzyme levels in resistant cultivars. Mesocotyl and shoot regions of seedlings recorded higher levels of enzyme than the root. Isoelectric focusing revealed four basic isoforms with pI 9.6, 9.0, 8.9 and 8.2 and two acidic isoforms with pI 4.9 and 6.2 of -1,3-glucanase in pearl millet. The pI 9.6 isoform was a major isoform of the enzyme in the pearl millet seedlings with a probable developmental function. Isoforms pI 6.2 and pI 8.2 appeared to be involved in resistance and pI 4.9 isoform seemed to be involved in pathogenesis of pearl millet-downy mildew.     

Microscopical studies of the effect of metalaxyl on the interaction between sunflower,Helianthus annuus L. and downy mildew,Plasmopara halstedii

The mode of action of the fungicide metalaxyl againstPlasmopara halstedii, the causal agent of sunflower downy mildew, was studied following its application before, during and after artificial contamination of seedlings. The development of the fungus within the treated seedlings was examined microscopically and compared to that occuring in untreated genetically susceptible or resistant genotypes. Hypersensitive-like reactions and necrotic zones leading to the inhibition of fungus growth within the hypocotyl were observed for the three modes of application. This suggests that sunflower defence mechanisms activated by genetical resistance are also involved in the control of downy mildew by metalaxyl.     

A new pathotype of<Emphasis Type="Italic">Pseudoperonospora cubensis</Emphasis> causing downy mildew in cucurbits in Israel

Downy mildew, incited by the comycetePseudoperonospora cubensis (Berk.et Curt.) Rost., was recorded in Israel during the years 1979–2001 on cucumber (Cucumis sativus) and melon (Cucumis melo) and classified as pathotype 3. In July 2002 severe outbreaks of downy mildew occurred on pumpkin (Cucurbita moschata) and summer squash (Cucurbita pepo subsp.pepo). Host range pathogenicity studies revealed high compatibility of the new population with cucumber, melon, pumpkin and summer squash but low compatibility with watermelon. This new population was therefore designated as pathotype 6. The possible origin of this new pathotype in Israel is discussed. http://www.phytoparasitica.org posting Oct. 9, 2003.     

弱光胁迫下黄瓜霜霉病抗性评价与分析

为明确弱光胁迫下黄瓜霜霉病田间发病规律及遗传特性,筛选抗霜霉病黄瓜品种,以15份黄瓜自交系为试材,在弱光条件下通过子叶接菌诱导寄主植株发病,同时结合田间发病率及病情指数调查进行复合抗性鉴定,进而筛选抗病组合,并对其分离世代进行抗性鉴定与遗传特征分析。结果表明,供试15份材料中共筛选出4个典型品系HB1、HB2、HB3和HB4,其病情指数分别是14.17、28.71、63.33和78.33,分别属于高抗、抗病、感病和高感病类型。这4个亲本的F1代及分离世代群体中霜霉病抗性与亲本抗性均存在正相关关系,抗性亲本的后代表现出较强的抗性;F2代群体病株分离现象比较明显,呈偏态分布,有明显的主基因+多基因存在特征,表明弱光胁迫下黄瓜霜霉病受主基因+多基因控制,主基因效应值较大。经田间抗病性鉴定分析,初步筛选出组合HB12、HB13和HB21为抗霜霉病组合,可用于后续抗病品种的选育。     

Monitoring of QoI fungicide resistance in Plasmopara viticola populations in Japan

BACKGROUND: The increasing occurrence of QoI fungicide resistance in Plasmopara viticola (Berk. & MA Curtis) Berl. & DeToni populations is becoming a serious problem in the control of grapevine downy mildew. In Japan, the existence of QoI‐fungicide‐resistant P. viticola was reported in 2009. RESULTS: The QoI fungicide resistance in P. viticola samples collected from vineyards in Japan in 2008 and 2009 was monitored. Resistant P. viticola were detected in the regions where QoI fungicides have been introduced in accordance with the pest management programme, whereas in Hokkaido vineyards, where QoI fungicides have not yet been introduced, QoI‐fungicide‐resistant P. viticola were not found. CONCLUSION: Japan comprises thousands of islands and is physically isolated from other countries by the sea. Monitoring the emergence, incidence and distribution of QoI fungicide resistance in P. viticola populations in Japan is necessary to improve pest management strategies for downy mildew disease in Japanese vineyards. Copyright © 2010 Society of Chemical Industry     

Resistance to acylalanines in Pseudoperonospora cubensis1

Resistance to acylalanine fungicides in Pseudoperonospora cubensis, the downy mildew of cucurbits, is reviewed. Insensitive strains of the fungus predominated during the second year of metalaxyl application in cucumber glasshouses in Greece and Israel. Resistance was detected either on detached cucumber leaves floated on metalaxyl solutions or on potted plants treated (sprayed or drenched) with this fungicide. Metalaxyl-resistant strains of P. cubensis showed good pathogenicity and fitness, and competed favorably with the sensitive strains in the absence of the fungicide. A synergism between these two biotypes resulted in an increased virulence of the resistant forms on metalaxyl-treated plants. Metalaxyl-resistant strains of the pathogen exhibited cross resistance to other acylalanine fungicides. Strains resistant to acylalanines and to the chemically unrelated oomycete-fungicides fosetyl-Al and propamocarb were isolated in Israel, as well. In Greece, however, protective sprays with fosetyl-Al, cymoxanil and mancozeb gave good control of cucumber downy mildew when the pathogen was resistant to metalaxyl. P. cubensis is the first case of fungus which developed resistance to acylalanines in the field.     

Application timing of herbicides,glyphosate and atrazine,sway respective epidemics of foliar pathogens in herbicide-tolerant rapeseed

Rapeseed (Brassica napus) production in Australia relies heavily on triazine-or glyphosate-tolerant cultivars. For 14 triazine-tolerant cultivars, disease development of Neopseudocercosporella capsellae (white leaf spot), Alternaria brassicae and A. japonica (Alternaria leaf spot), and Hyaloperonospora brassicae (downy mildew) were all dependent upon herbicide application timing (p < 0.001), with significant differences between cultivars (p < 0.001) and a significant interaction (p < 0.001) between herbicide application timing and cultivars. Atrazine applied preinfection by N. capsellae, A. brassicae, or A. japonica enhanced disease incidence, severity, and leaf collapse, while atrazine application postinfection for these same pathogens reduced all three disease parameters. However, for H. brassicae, application of atrazine after, and especially prior to, infection resulted in lower disease incidence, severity, and leaf collapse. Application of glyphosate on five glyphosate-tolerant cultivars for N. capsellae resulted in significant differences (p < 0.05) between glyphosate application treatments, and between host cultivars in terms of incidence and consequent leaf collapse. For A. brassicae, A. japonica, and H. brassicae, glyphosate resulted in significant differences (p < 0.001) across application timings between cultivars, and a significant interaction (p < 0.001) between herbicide application timings and cultivars. Glyphosate applied on glyphosate-resistant rapeseed after, and especially prior to, attack by H. brassicae, reduced downy mildew. These are the first studies to highlight how the timing of application of triazine or glyphosate in relation to pathogen infection is critical to the susceptibility of rapeseed to white leaf spot, Alternaria leaf spot, and downy mildew. This new understanding offers fresh possibilities for improved management of these diseases in herbicide-tolerant rapeseed crops.     

Race 3, a new race of<Emphasis Type="Italic"> Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">lactucae </Emphasis>determined by a differential system with commercial cultivars

 Pathogenic variation among 26 Japanese isolates of Fusarium oxysporum f. sp. lactucae (FOL) was tested using 21 lettuce cultivars to select commercial lettuce cultivars as race differential indicators. Cultivar Costa Rica No. 4 was resistant to race 1 but susceptible to race 2, consistent with the conventional standard differential line VP1010. Cultivar Banchu Red Fire was susceptible to race 1 but resistant to race 2, which showed an opposite type of reaction as another differential line VP1013. Cultivar Patriot was susceptible to both races. The resistance reactions of the three cultivars under field conditions were identical with that observed in the seedlings. Thus cv. Costa Rica No. 4 and cv. Banchu Red Fire can be used as differential hosts to identify pathogenic races of FOL. This differential system showed that all FOL isolates obtained from diseased butterhead lettuce in Fukuoka, Japan were new races (i.e., pathogenic to three cultivars). We propose that the new race be designated race 3. Isolates of FOL, the pathogen of Fusarium wilt in lettuce, obtained from California showed the same reaction as that of race 1. Furthermore, the Japanese isolate SB1-1 (race 1) and California isolate HL-2 belonged to the same vegetative compatibility group. Our results suggest that both of the fungi are the same forma specialis. Received: March 25, 2002 / Accepted: August 26, 2002     

Monitoring physiological races of<Emphasis Type="Italic">Podosphaera xanthii</Emphasis> (syn.<Emphasis Type="Italic">Sphaerotheca fuliginea</Emphasis>), the causal agent of powdery mildew in cucurbits: Factors affecting race identification and the importance for research and commerce

Identification of the physiological races ofPodosphaera xanthii (syn.Sphaerotheca fuliginea), the causal agent of powdery mildew in cucurbits, is based upon the differing responses of various melon cultigens to the pathogen. Eight races of the pathogen have been identified to date in the USA, Africa, Europe and around the Mediterranean Sea, and four new races were reported from greenhouse melons in the major growing area of Japan. Plant responses to powdery mildew may be affected by environmental factors such as light intensity, temperature and humidity, as well as by age and nutritional status of the plants. The same factors affect the accuracy and reliability of race identification. In an attempt to overcome those obstacles, the genetic diversity ofP. xanthii was studied using molecular markers. Unfortunately, no correlation was found between DNA polymorphism and the race of the pathogen as identified by biological tests. The usefulness of race identification as a guide for the grower in selecting appropriate cultivars is limited because changes or shifts in the pathogen population are common. Such changes may be found among growing seasons, geographic regions and hosts, and also within a single greenhouse during a single season. On the other hand, race identification is important for basic research and is especially important for the commercial seed industry, which requires accuracy in declaring the type and level of resistance to powdery mildew in its products. http://www.phytoparasitica.org posting March 2, 2004. Contribution from the Agricultural Research Organization. No. 501/04.     

Downy mildew of busy lizzie caused by Plasmopara obducens in Japan

In June 2010 and March 2012, busy lizzie (Impatiens sultanii) plants in a glasshouse in Yamagata Prefecture and in nurseries in a plastic house in Shimane Prefecture, Japan were observed with yellowing of the adaxial surface of leaves or cotyledons and a white downy growth on the abaxial surface of leaves or cotyledons. We identified the causal pathogen as Plasmopara obducens based on the morphologies and rDNA-large subunit sequences. This new disease was named “downy mildew of busy lizzie” in Japan.