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Sample records for plasmopara viticola oospores

  1. Pinus pinaster Knot: A Source of Polyphenols against Plasmopara viticola.

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    Gabaston, Julien; Richard, Tristan; Cluzet, Stéphanie; Palos Pinto, Antonio; Dufour, Marie-Cécile; Corio-Costet, Marie-France; Mérillon, Jean-Michel

    2017-09-29

    Pine knot extract from Pinus pinaster byproducts was characterized by UHPLC-DAD-MS and NMR. Fourteen polyphenols divided into four classes were identified as follows: lignans (nortrachelogenin, pinoresinol, matairesinol, isolariciresinol, secoisolariciresinol), flavonoids (pinocembrin, pinobanksin, dihydrokaempferol, taxifolin), stilbenes (pinosylvin, pinosylvin monomethyl ether, pterostilbene), and phenolic acids (caffeic acid, ferulic acid). The antifungal potential of pine knot extract, as well as the main compounds, was tested in vitro against Plasmopara viticola. The ethanolic extract showed a strong antimildew activity. In addition, pinosylvins and pinocembrin demonstrated significant inhibition of zoospore mobility and mildew development. These findings strongly suggest that pine knot is a potential biomass that could be used as a natural antifungal product.

  2. Phenotypic and histochemical traits of the interaction between Plasmopara viticola and resistant or susceptible grapevine varieties

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    Toffolatti Silvia

    2012-08-01

    Full Text Available Abstract Background Grapevine downy mildew, caused by Plasmopara viticola, is a very serious disease affecting mainly Vitis vinifera cultivated varieties around the world. Breeding for resistance through the crossing with less susceptible species is one of the possible means to reduce the disease incidence and the application of fungicides. The hybrid Bianca and some of its siblings are considered very promising but their resistance level can vary depending on the pathogen strain. Moreover, virulent strains characterized by high fitness can represent a potential threat to the hybrid cultivation. Results The host response and the pathogen virulence were quantitatively assessed by artificially inoculating cv Chardonnay, cv Bianca and their siblings with P. viticola isolates derived from single germinating oospores collected in various Italian viticultural areas. The host phenotypes were classified as susceptible, intermediate and resistant, according to the Area Under the Disease Progress Curve caused by the inoculated strain. Host responses in cv Bianca and its siblings significantly varied depending on the P. viticola isolates, which in turn differed in their virulence levels. The fitness of the most virulent strain did not significantly vary on the different hybrids including Bianca in comparison with the susceptible cv Chardonnay, suggesting that no costs are associated with virulence. Among the individual fitness components, only sporangia production was significantly reduced in cv Bianca and in some hybrids. Comparative histological analysis revealed differences between susceptible and resistant plants in the pathogen diffusion and cytology from 48 h after inoculation onwards. Defence mechanisms included callose depositions in the infected stomata, increase in peroxidase activity, synthesis of phenolic compounds and flavonoids and the necrosis of stomata and cells immediately surrounding the point of invasion and determined alterations in

  3. Stilbenes from Vitis vinifera L. waste: a sustainable tool for controlling Plasmopara viticola.

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    Gabaston, Julien; Cantos-Villar, Emma; Biais, Benoit; Waffo-Teguo, Pierre; Renouf, Elodie; Corio Costet, Marie France; Richard, Tristan; Merillon, Jean-Michel

    2017-03-13

    Stilbene-enriched extracts from Vitis vinifera waste (cane, wood and root) were characterized by UHPLC-MS. Eleven stilbenes were identified and quantified as follows: ampelopsin A, E-piceatannol, pallidol, E-resveratrol, hopeaphenol, isohopeaphenol, E-ε-viniferin, E-miyabenol C, E-ω-viniferin, r2-viniferin and r-viniferin. The fungicide concentration inhibiting 50% of growth of Plasmopara viticola sporulation (IC50) was determined for the extracts and also for the main compounds isolated. r-Viniferin followed by hopeaphenol and r2-viniferin showed low IC50 and thus high efficacy against Plasmopara viticola. Regarding stilbene extracts, wood extract followed by root extract showed the highest antifungal activities. These data suggest that stilbene complex mixtures from Vitis vinifera waste could be used as a cheap source of bioactive stilbenes for the development of natural fungicides.

  4. Revisiting Vitis vinifera subtilase gene family: a possible role in grapevine resistance against Plasmopara viticola

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    Joana Figueiredo

    2016-11-01

    Full Text Available Subtilisin-like proteases, also known as subtilases, are a very diverse family of serine peptidases present in many organisms. In grapevine, there are hints of the involvement of subtilases in defence mechanisms, but their role is not yet understood. The first characterization of the subtilase gene family was performed in 2014. However, simultaneously, the grapevine genome was re-annotated and several sequences were re-annotated or retrieved. We have performed a re-characterization of this family in grapevine and identified 82 genes coding for 97 putative proteins, as result of alternative splicing. All the subtilases identified present the characteristic S8 peptidase domain and the majority of them also have a pro-domain I9 inhibitor, a protease-associated (PA domain and a signal peptide for targeting to the secretory pathway. Phylogenetic studies revealed six subtilase groups denominated VvSBT1 to VvSBT6. As several evidences have highlighted the participation of plant subtilases in response to biotic stimulus, we have investigated subtilase participation in grapevine resistance to Plasmopara viticola, the causative agent of downy mildew. Fourteen grapevine subtilases presenting either high homology to P69C from tomato, SBT3.3 from Arabidopsis thaliana or located near the Resistance to Plasmopara viticola (RPV locus were selected. Expression studies were conducted in the grapevine-P. viticola pathosystem with resistant and susceptible cultivars. Our results may indicate that some of grapevine subtilisins are potentially participating in the defence response against this biotrophic oomycete.

  5. Solar radiation affects grapevine susceptibility to Plasmopara Viticola Radiação solar e susceptibilidade da videira à Plasmopara Viticola

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    Anna Dalla Marta

    2008-12-01

    Full Text Available Solar radiation plays an important role in the development of some fungal diseases due to its direct action on the microorganisms and also its indirect effect on the production of specific plant compounds. This experiment examined the effect of two light environments (100% and 35% of full strength on the polyphenolic content of grapevine leaves and quantified their relation to resistance to downy mildew (Plasmopara viticola. Leaf epidermal polyphenolic contents were non-destructively measured during the growing season 2006 using the Dualex chlorophyll fluorescence-based portable leaf-clip. The experimental design consisted of six parcels of 30 vines and measurements were performed on the 12 central vines. The leaves were inoculated with a sporangia suspension containing 50,000 sporangia of P. viticola per mL and the disease severity was assessed after the appearance of symptoms. Leaves maintained at 100% sun had high polyphenolic content and significantly lower disease severity compared to leaves under shading nets. These results indicate an inverse relationship between produced polyphenolics and downy mildew severity.A radiação solar exerce papel importante no desenvolvimento de algumas doenças fúngicas, devido à sua ação direta sobre os microorganismos e também devido ao seu efeito indireto sobre a produção de compostos específicos nas plantas. Este experimento examinou o efeito de dois ambientes de luz (100% e 35% da intensidade máxima sobre o conteúdo de polifenóis nas folhas da videira e quantificou sua relação com a resistência ao míldio (Plasmopara viticola. As concentrações de polifenóis na epiderme das folhas foram quantificadas de forma não destrutiva, durante a estação de crescimento de 2006, usando o equipamento portátil Dualex, baseado na fluorescência da clorofila. O delineamento experimental consistiu de seis parcelas de 30 videiras e as medidas foram feitas nas 12 plantas centrais. As folhas foram

  6. Impact of Plasmopara viticola infection of Merlot and Cabernet Sauvignon grapes on wine composition and flavor.

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    Pons, Alexandre; Mouakka, Nadia; Deliere, Laurent; Crachereau, Jean Christophe; Davidou, Ludivine; Sauris, Pierre; Guilbault, Pascal; Darriet, Philippe

    2018-01-15

    This work reports the identification of volatile compounds involved in the particular and atypical flavor detected in Vitis vinifera red Merlot and Cabernet Sauvignon wines made with grapes infected and wilted by brown rot (Plasmopara viticola). Must made from withered grapes had green aromas while red wines were marked by intense odor reminiscent of green, herbaceous notes but also figs and cooked fruit. Thanks to GC-O and GC-MS analysis, cooked fruit notes were identified as 3-methyl-2,4-nonanedione, γ-nonalactone and γ-decalactone, whereas herbaceous and green aromas were identified as (Z)-1,5-octadien-3-one and 3-isobutyl-2-methoxypyrazine. We show that the organoleptic impact of P. viticola is more pronounced in Merlot wines compared to Cabernet Sauvignon ones. The highest levels of 3-methyl-2,4-nonanedione (75.3ng/L) were found in old Merlot wines made with 20% infected berries, suggesting the incidence of berry quality on the ability of a wine to age. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. SOYBEAN AND CASEIN HYDROLYSATES INDUCE GRAPEVINE IMMUNE RESPONSES AND RESISTANCE AGAINST PLASMOPARA VITICOLA

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    Nihed eLachhab

    2014-12-01

    Full Text Available Plasmopara viticola, the causal agent of grapevine downy mildew, is one of the most devastating grape pathogen in Europe and North America. Although phytochemicals are used to control pathogen infections, the appearance of resistant strains and the concern for possible adverse effects on environment and human health are increasing the search for alternative strategies. In the present investigation, we successfully tested two protein hydrolysates from soybean (soy and casein (cas to trigger grapevine resistance against P. viticola. On Vitis vinifera cv. Marselan plants, the application of soy and cas reduced the infected leaf surface by 76 and 63%, as compared to the control, respectively. Since both hydrolysates might trigger the plant immunity, we investigated their ability to elicit grapevine defence responses. On grapevine cell suspensions, a different free cytosolic calcium signature was recorded for each hydrolysate, whereas a similar transient phosphorylation of two MAP kinases of 45 and 49 kDa was observed. These signalling events were followed by transcriptome reprogramming, including the up-regulation of defence genes encoding pathogenesis-related (PR proteins and the stilbene synthase enzyme responsible for the biosynthesis of resveratrol, the main grapevine phytoalexin. Liquid chromatography analyses confirmed the production of resveratrol and its dimer metabolites, δ- and ε-viniferins. Overall, soy effects were more pronounced as compared to the cas one. Both hydrolysates proved to act as elicitors to enhance grapevine immunity against pathogen attack.

  8. Identification of Biomarkers for Defense Response to Plasmopara viticola in a Resistant Grape Variety

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    Giulia Chitarrini

    2017-09-01

    Full Text Available Downy mildew (Plasmopara viticola is one of the most destructive diseases of the cultivated species Vitis vinifera. The use of resistant varieties, originally derived from backcrosses of North American Vitis spp., is a promising solution to reduce disease damage in the vineyards. To shed light on the type and the timing of pathogen-triggered resistance, this work aimed at discovering biomarkers for the defense response in the resistant variety Bianca, using leaf discs after inoculation with a suspension of P. viticola. We investigated primary and secondary metabolism at 12, 24, 48, and 96 h post-inoculation (hpi. We used methods of identification and quantification for lipids (LC-MS/MS, phenols (LC-MS/MS, primary compounds (GC-MS, and semi-quantification for volatile compounds (GC-MS. We were able to identify and quantify or semi-quantify 176 metabolites, among which 53 were modulated in response to pathogen infection. The earliest changes occurred in primary metabolism at 24–48 hpi and involved lipid compounds, specifically unsaturated fatty acid and ceramide; amino acids, in particular proline; and some acids and sugars. At 48 hpi, we also found changes in volatile compounds and accumulation of benzaldehyde, a promoter of salicylic acid-mediated defense. Secondary metabolism was strongly induced only at later stages. The classes of compounds that increased at 96 hpi included phenylpropanoids, flavonols, stilbenes, and stilbenoids. Among stilbenoids we found an accumulation of ampelopsin H + vaticanol C, pallidol, ampelopsin D + quadrangularin A, Z-miyabenol C, and α-viniferin in inoculated samples. Some of these compounds are known as phytoalexins, while others are novel biomarkers for the defense response in Bianca. This work highlighted some important aspects of the host response to P. viticola in a commercial variety under controlled conditions, providing biomarkers for a better understanding of the mechanism of plant defense and a

  9. Observations on the Behaviour of Different Populations of Plasmopara viticola Resistant to QoI Fungicides in Italian Vineyards

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    M.L. Gullino

    2004-12-01

    Full Text Available Grapevine downy mildew, caused by Plasmopara viticola, is probably the most damaging fungal disease of grapevine world-wide. Among the fungicides recently developed for downy mildew control is the QoI class of fungicides, which inhibits mitochondrial respiration. Since 1999, selected P. viticola populations in northern Italy have been monitored for resistance to QoI fungicides. Detached leaf discs and whole potted plants were used under controlled conditions to test the sampled populations. QoI-resistant populations of P. viticola were found in all the vineyards sampled in 2001 and 2002 in Trentino Alto Adige and Friuli Venezia Giulia, where failure in QoI control was reported. Many of the populations had minimum inhibition concentration (MIC values 3– 30 times higher than those of sensitive reference populations. Populations of P. viticola sampled from vineyards in Piedmont, where no QoI fungicides had previously been used, showed MIC values equal to, or lower than those of the reference populations. Most of the P. viticola populations collected in Trentino Alto Adige in 2001 showed high virulence in leaf disc test and were not controlled by QoI fungicides, applied both at field and double field rates in the whole plant test. Most of these populations retained their virulence in the subsequent leaf disc test in water.

  10. Extracts of Inula viscosa Control Downy Mildew of Grapes Caused by Plasmopara viticola.

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    Cohen, Yigal; Wang, Wenqiao; Ben-Daniel, Bat-Hen; Ben-Daniel, Yigal

    2006-04-01

    ABSTRACT The oily paste extracts of Inula viscosa leaves made with organic solvents were effective in controlling downy mildew caused by Plasmopara viticola in detached leaf tissues of grapes in growth chambers. Thin-layer chromatography of such extracts revealed the presence of 11 Rf regions of which four contained highly effective compounds against the disease. Two major inhibitory compounds, each comprising 10.6% of the total paste weight, were identified as tomentosin and costic acid. An emulsified concentrate formulation of the oily paste extracts provided excellent activity against the disease in the field. The effective dose (concentration) required for 90% control of the disease in treated shoots in the field was below 0.125% (paste in water). No seasonal fluctuations were observed in the control efficacy of six extracts made from I. viscosa leaves harvested during the period of May to July. In whole vines, treated and artificially inoculated, the paste concentration required for 90% control of the disease ranged between 0.30 to 0.37%, whereas in naturally infected vines it was 0.58%. It appears that I. viscosa is a valuable source for fungicidal preparations against downy mildew of grapes. This is the first report on the control of a disease with I. viscosa extracts under field conditions.

  11. Inheritance of downy mildew (Plasmopara viticola) and anthracnose (Sphaceloma ampelinum) resistance in grapevines.

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    Poolsawat, O; Mahanil, S; Laosuwan, P; Wongkaew, S; Tharapreuksapong, A; Reisch, B I; Tantasawat, P A

    2013-12-13

    Downy mildew (Plasmopara viticola) and anthracnose (Sphaceloma ampelinum) are two of the major diseases of most grapevine (Vitis vinifera L.) cultivars grown in Thailand. Therefore, breeding grapevines for improved downy mildew and anthracnose resistance is crucial. Factorial crosses were made between three downy mildew and/or anthracnose resistant lines ('NY88.0517.01', 'NY65.0550.04', and 'NY65.0551.05'; male parents) and two or three susceptible cultivars of V. vinifera ('Black Queen', 'Carolina Black Rose', and/or 'Italia'; female parents). F1 hybrid seedlings were evaluated for downy mildew and anthracnose resistance using a detached/excised leaf assay. For both diseases, the general combining ability (GCA) variance among male parents was significant, while the variance of GCA among females and the specific combining ability (SCA) variance were not significant, indicating the prevalence of additive over non-additive gene actions. The estimated narrow sense heritabilities of downy mildew and anthracnose resistance were 55.6 and 79.2%, respectively, suggesting that downy mildew/anthracnose resistance gene(s) were highly heritable. The 'Carolina Black Rose x NY65.0550.04' cross combination is recommended for future use.

  12. Response of grapevine leaves to Plasmopara viticola infection by means of measurement of reflectance and fluorescence signals

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    David Šebela

    2012-01-01

    Full Text Available Response of grapevine leaf tissue naturally infected by Plasmopara viticola in field was measured by means of chlorophyll fluorescence and reflectance signals. Three susceptible grapevine varieties (Cabernet Sauvignon, Pinot Blanc and Pinot Gris were used in this study. Since the infection impairs photosynthetic activity, distribution of FV/FM parameter (maximum quantum yield of Photosystem II over the leaf was effective to discriminate healthy and naturally infected leaf tissue. FV/FM was reduced ~ 25% in all infected leaf parts. Infected leaf spots expressed significantly altered chlorophyll fluorescence induction kinetics expressing much slower electron transport rate both on donor and acceptor site of PSII. Vegetation reflectance indices followed the variations in pigment content after the fungal infection. R750/R700 (R2 = 0.877 and CRI (carotenoid reflectance index; R2 = 0.735 were the most potent to follow changes in chlorophylls and carotenoids contents, respectively. Infected leaf tissue exhibited decrease in chlorophyll a (~50 % as well as carotenoids (~70%. We conclude that combination of chlorophyll fluorescence and reflectance measurements can be used as an effective non-invasive tool for an early detection of Plasmopara viticola in field as well as for estimation of the level of infection.

  13. cDNA-AFLP analysis of plant and pathogen genes expressed in grapevine infected with Plasmopara viticola

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    Kortekamp Andreas

    2008-03-01

    Full Text Available Abstract Background The oomycete Plasmopara viticola (Berk. and Curt. Berl. and de Toni causes downy mildew in grapevine (Vitis vinifera L.. This pathogen is strictly biotrophic, thus completely dependent on living host cells for its survival. The molecular basis of compatibility and disease development in this system is poorly understood. We have carried out a large-scale cDNA-AFLP analysis to identify grapevine and P. viticola genes associated with the infection process. Results We carried out cDNA-AFLP analysis on artificially infected leaves of the susceptible cultivar Riesling at the oil spot stage, on water-treated leaves and on a sample of pure sporangia as controls. Selective amplifications with 128 primer combinations allowed the visualization of about 7000 transcript-derived fragments (TDFs in infected leaves, 1196 of which (17% were differentially expressed. We sequenced 984 fragments, 804 of which were identified as grapevine transcripts after homology searching, while 96 were homologous to sequences in Phytophthora spp. databases and were attributed to P. viticola. There were 82 orphan TDFs. Many grapevine genes spanning almost all functional categories were downregulated during infection, especially genes involved in photosynthesis. Grapevine genes homologous to known resistance genes also tended to be repressed, as were several resistance gene analogs and carbonic anhydrase (recently implicated in pathogen resistance. In contrast, genes encoding cytoskeletal components, enzymes of the phenylpropanoid and beta-oxidation pathways, and pathogenesis related proteins were primarily upregulated during infection. The majority of P. viticola transcripts expressed in planta showed homology to genes of unknown function or to genomic Phytophthora sequences, but genes related to metabolism, energy production, transport and signal transduction were also identified. Conclusion This study provides the first global catalogue of grapevine and P

  14. Development and application of loop-mediated isothermal amplification (LAMP) for detection of Plasmopara viticola

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    Kong, X.; Qin, W.; Xiaoqing, X.; Kong, F.; Schoen, C.D.; Feng, J.; Wang, Z.; Zhang, H.

    2016-01-01

    A rapid LAMP (loop-mediated isothermal amplification) detection method was developed on the basis of the ITS sequence of P. viticola, the major causal agent of grape downy mildew. Among the 38 fungal and oomycete species tested, DNA isolated exclusively from P. viticola resulted in a specific

  15. Separation and identification of highly fluorescent compounds derived from trans-resveratrol in the leaves of Vitis vinifera infected by Plasmopara viticola.

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    Tříska, Jan; Vrchotová, Naděžda; Olejníčková, Julie; Jílek, Rudolf; Sotolář, Radek

    2012-03-06

    A method for identification of highly fluorescent compounds in vine leaves infected by Plasmopara viticola was developed using reversed phase liquid chromatography with simultaneous diode array and fluorometric detection. Fluorescent compounds were extracted from leaves with a methanol-water mixture (70:30). Separation by HPLC was performed using a C(18) column and gradient elution with water-acetonitrile mixtures (20-80% of acetonitrile). The main unknown fluorescent compound was identified by line spectral comparison with a standard obtained by UV photoisomerization of trans-resveratrol glucoside, and its structure was confirmed by liquid chromatography-mass spectrometry. Identification and structural elucidation of the fluorescent compound in the leaves of Vitis vinifera allows early detection of Plasmopara viticola invasion.

  16. Effect of dose rate and mixtures of fungicides on selection for QoI resistance in populations of Plasmopara viticola.

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    Genet, Jean-Luc; Jaworska, Grazyna; Deparis, Francine

    2006-02-01

    Resistance to QoI fungicides (strobilurins, famoxadone and fenamidone) in populations of Plasmopara viticola (Berk & Curt) Berlese & de Toni developed soon after their introduction in France and Italy. Current resistance management strategies include limitation of the number of applications, use of mixtures and alternation of fungicides with different modes of action. The selection pressure resulting from QoI fungicides applied alone or in mixtures with non-QoI fungicides was investigated in whole plant experiments under controlled conditions. QoI-resistant populations of P. viticola gradually reverted to full sensitivity following consecutive transfers to untreated plants, suggesting that resistant phenotypes were less competitive than sensitive ones. When cycled on QoI-treated plants, reduction in sensitivity was greater for the QoI fungicide which had greater intrinsic activity on P. viticola. Sensitivity decreased at each subsequent cycle, resulting in almost full resistance after four generations. Mixture experiments indicated that selection pressure was affected most by the dose of the QoI fungicide and the nature of the partner fungicide. Folpet delayed selection pressure most effectively when it was associated with famoxadone or azoxystrobin. Mancozeb was least effective at reducing the rate of selection compared with the QoI alone, and fosetyl-aluminium was intermediate. Higher rates of selection were recorded when the dose of the QoI fungicide, solo or in a mixture, was increased from 1 to 4 microg ml(-1). Increasing the dose of the non-QoI partner fungicide in the mixture from 10 to 30 microg ml(-1) resulted in reduced selection pressure. These results suggest that the choice of the fungicide partner and its dosage in the mixture can significantly affect the success of QoI resistance management strategies under practical conditions.

  17. Resistance to Plasmopara viticola in a grapevine segregating population is associated with stilbenoid accumulation and with specific host transcriptional responses

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    Delledonne Massimo

    2011-08-01

    Full Text Available Abstract Background Downy mildew, caused by the oomycete Plasmopara viticola, is a serious disease in Vitis vinifera, the most commonly cultivated grapevine species. Several wild Vitis species have instead been found to be resistant to this pathogen and have been used as a source to introgress resistance into a V. vinifera background. Stilbenoids represent the major phytoalexins in grapevine, and their toxicity is closely related to the specific compound. The aim of this study was to assess the resistance response to P. viticola of the Merzling × Teroldego cross by profiling the stilbenoid content of the leaves of an entire population and the transcriptome of resistant and susceptible individuals following infection. Results A three-year analysis of the population's response to artificial inoculation showed that individuals were distributed in nine classes ranging from total resistance to total susceptibility. In addition, quantitative metabolite profiling of stilbenoids in the population, carried out using HPLC-DAD-MS, identified three distinct groups differing according to the concentrations present and the complexity of their profiles. The high producers were characterized by the presence of trans-resveratrol, trans-piceid, trans-pterostilbene and up to thirteen different viniferins, nine of them new in grapevine. Accumulation of these compounds is consistent with a resistant phenotype and suggests that they may contribute to the resistance response. A preliminary transcriptional study using cDNA-AFLP selected a set of genes modulated by the oomycete in a resistant genotype. The expression of this set of genes in resistant and susceptible genotypes of the progeny population was then assessed by comparative microarray analysis. A group of 57 genes was found to be exclusively modulated in the resistant genotype suggesting that they are involved in the grapevine-P. viticola incompatible interaction. Functional annotation of these transcripts

  18. Ação de quitosana sobre o desenvolvimento de Plasmopara viticola e Elsinoe ampelina, in vitro e em videiras cv. Isabel Chitosan action on Plasmopara viticola and Elsinoe ampelina development in vitro and in grapevines cv. Isabel

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    Aline José Maia

    2010-09-01

    Full Text Available Este trabalho teve como objetivos avaliar o efeito da quitosana no controle dos fungos Plasmopara viticola e Elsinoe ampelina, agentes causais do míldio e da antracnose da videira, respectivamente. As concentrações de 0, 20, 40, 80 e 160 mg L-1 de quitosana foram utilizadas nos seguintes experimentos: testes de crescimento micelial, de germinação de esporos e ensaio em condições de campo. Para os dois últimos ensaios, adicionou-se tratamentos padrões com mancozeb e calda bordalesa. Verificou-se redução no crescimento micelial de E. ampelina sendo que a maior concentração de quitosana (160 mg L-1 reduziu em 57% o desenvolvimento do fungo, 192 horas após incubação. Nos testes de germinação, a dose de 160 mg L-1 de quitosana reduziu a germinação de esporos de E. ampelina em aproximadamente 98% e de P. viticola em 60%, não diferindo dos tratamentos com calda bordalesa e mancozeb. Nos ensaios a campo as maiores doses de quitosana (80 e 160 mg L-1 apresentaram um decréscimo na severidade de antracnose entre 93 e 81%. Para o míldio, a concentração de 160 mg L-1 apresentou um decréscimo de aproximadamente 81%. Baseando-se nestes resultados, pode-se concluir que a quitosana tem um grande potencial no controle do míldio e da antracnose da videira.This work aimed to evaluate the effect of chitosan on the control of fungi Elsinoe ampelina and Plasmopara viticola, the causal agents of downy mildew and anthracnose in grapevines, respectively. The concentrations of 0, 20, 40, 80 and 160 mg L-1 chitosan were used in the following trials: mycelial growth, spore germination, and experiment in field conditions. For the last two trials standard treatments with mancozeb and bordeax mixture were added. There was a reduction in E. ampelina mycelial growth and the highest chitosan concentration (160 mg L-1 decreased by 57% the fungus development at 192 hours after incubation. In the germination tests, the concentration of 160 mg L-1 chitosan

  19. Impact de l’endomycorhization sur la résistance naturelle et induite de la vigne à Plasmopara viticola (agent causal du mildiou)

    OpenAIRE

    ZERBIB, Jérémie

    2015-01-01

    Plasmopara viticola est l’oomycète responsable du mildiou chez la vigne cultivée (Vitis vinifera L.). Les viticulteurs ont souvent recours à une lutte chimique (fongicide) massive pour contrôler cette maladie. Cependant, l’utilisation systématique de produits phytopharmaceutiques implique aussi bien un coût économique pour le viticulteur et des répercussions avérées sur l’environnement et la santé humaine. Dans la cadre du plan EcoPhyto, qui encourage une forte diminution de l’utilisation de ...

  20. Response of phytohormones and correlation of SAR signal pathway genes to the different resistance levels of grapevine against Plasmopara viticola infection.

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    Liu, Shao-Li; Wu, Jiao; Zhang, Pei; Hasi, Gerile; Huang, Yu; Lu, Jiang; Zhang, Ya-Li

    2016-10-01

    Phytohormones play an important role in the process of disease resistance in plants. Here, we investigated which among salicylic acid, jasmonic acid, and abscisic acid performs a key role in plant defense after Plasmopara viticola infection in grapevine. We used grapevines possessing different resistance levels against P. viticola infection to study the relationship between the expression of key genes in the related resistance signaling pathways and the level of resistance. We performed high-performance liquid chromatography-mass spectrometry to estimate the phytohormone contents in grape leaves at different time points after the infection. Furthermore, we performed quantitative analyses of key genes such as EDS1, PAD4, ICS2, PAL, NPR1, TGA1, and PR1 in the systemic acquired resistance pathway by quantitative reverse transcription-polymerase chain reaction. The results showed an increased variation in the SA content, which was maintained at high levels, after P. viticola infection in plant species exhibiting stronger resistance to the pathogen; this finding highlights the importance of SA in plant defense mechanisms. Moreover, EDS1 and PAD4 expression did not show a positive correlation with disease resistance in grape; however, higher expression of other genes that were analyzed was observed in highly resistant grape varieties. Our results provide insights into the role of phytohormone regulation in the induction and maintenance of plant defense response to pathogens.

  1. Incidence of downy mildew Plasmopara viticola (Berk. et Curtis ex. de Bary Berl. et de Toni in Terceira island, Azores

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    Matej Zorman

    2008-12-01

    Full Text Available Downy mildew is one of the most economically important disease affecting vine production in Terceira island, Azores. By its incidence, frequency and quantity of pathogens, supported with favourable temperate humid climate conditions, this disease is difficult to control in terms of sustainable integrated plant protection policy. The objectives of INTERFRUTA project were to determine the incidence of the disease, to better understand it and to adopt an adequate forecasting model with necessary adjustments to predict the disease appearance and its in-time integrated plant protection control measures. Plasmopara viticola was directly observed in two weeks time periods on seven representative vineyards at different altitudes. Three of the monitored plots were established during study inside one plantation. Incidence was registered as result of visual symptoms in percentage, where 10% of plants in each vineyard were observed. Seasonal evolution of the disease was studied in one of the wine producing area.O Míldio é uma das doenças que mais afecta a vinha, na ilha Terceira, Açores. É uma doença fúngica de difícil controlo, pela sua incidência e frequência de aparecimento. Os objectivos do projecto INTERFRUTA II foram determinar a incidência desta doença para melhorar o conhecimento sobre esta e adoptar e validar um modelo matemático adequado à previsão do aparecimento da doença de modo a permitir a aplicação de medidas de protecção integrada. A evolução do míldio foi acompanhada em sete parcelas de vinha, da área vitivinícola dos Biscoitos, através da observação visual de folhas. Numa das parcelas, três dos talhões não foram tratados para servirem de controlo. Em cada parcela foram observadas 10% das plantas, tendo sido registada a percentagem de plantas com sintomas visuais da presença de míldio.

  2. Proteome of Plasmopara viticola-infected Vitis vinifera provides insights into grapevine Rpv1/Rpv3 pyramided resistance to downy mildew.

    Science.gov (United States)

    Nascimento-Gavioli, Maria Carolina Andrade; Agapito-Tenfen, Sarah Zanon; Nodari, Rubens Onofre; Welter, Leocir José; Sanchez Mora, Fernando David; Saifert, Luciano; da Silva, Aparecido Lima; Guerra, Miguel Pedro

    2017-01-16

    Grapevine is one of the major fruit crops worldwide and requires phytochemical use due to susceptibility to numerous pests, including downy mildew. The pyramiding of previous identified QTL resistance regions allows selection of genotypes with combined resistance loci in order to build up sustainable resistance. This study investigates resistance response of pyramided plants containing Rpv1 and Rpv3 loci to Plasmopara viticola infection process. Phenotypic characterization showed complete resistance and lack of necrotic hypersensitive response spots. Principal Component Analysis revealed infected 96hpi (hours post-inoculation) samples with the most distant proteomes of the entire dataset, followed by the proteome of infected 48hpi samples. Quantitative and qualitative protein differences observed using 2-DE gels coupled to nanoHPLC-ESI-MS/MS analysis showed a lack of transient breakdown in defense responses (biphasic modulation) accompanying the onset of disease. Forty-one proteins were identified, which were mainly included into functional categories of redox and energy metabolism. l-ascorbate degradation pathway was the major altered pathway and suggests up-regulation of anti-oxidant metabolism in response to apoplastic oxidative burst after infection. Overall, these data provide new insights into molecular basis of this incompatible interaction and suggests several targets that could potentially be exploited to develop new protection strategies against this pathogen. This study provide new insights into the molecular basis of incompatible interaction between Plasmopara viticola and pyramided Rpv1/Rpv3 grapevine and suggests several targets that could potentially be exploited to develop new protection strategies against this pathogen. This is the first proteomic characterization of resistant grapevine available in the literature and it presents contrasting proteomic profiles of that of susceptible plants. The resistance against downy mildew in grapevine has been

  3. The sulfated laminarin triggers a stress transcriptome before priming the SA- and ROS-dependent defenses during grapevine's induced resistance against Plasmopara viticola.

    Directory of Open Access Journals (Sweden)

    Adrien Gauthier

    Full Text Available Grapevine (Vitis vinifera is susceptible to many pathogens which cause significant losses to viticulture worldwide. Chemical control is available, but agro-ecological concerns have raised interest in alternative methods, especially in triggering plant immunity by elicitor treatments. The β-glucan laminarin (Lam and its sulfated derivative (PS3 have been previously demonstrated to induce resistance in grapevine against downy mildew (Plasmopara viticola. However, if Lam elicits classical grapevine defenses such as oxidative burst, pathogenesis-related (PR-proteins and phytoalexin production, PS3 triggered grapevine resistance via a poorly understood priming phenomenon. The aim of this study was to identify the molecular mechanisms of the PS3-induced resistance. For this purpose we studied i the signaling events and transcriptome reprogramming triggered by PS3 treatment on uninfected grapevine, ii grapevine immune responses primed by PS3 during P. viticola infection. Our results showed that i PS3 was unable to elicit reactive oxygen species (ROS production, cytosolic Ca(2+ concentration variations, mitogen-activated protein kinase (MAPK activation but triggered a long lasting plasma membrane depolarization in grapevine cells, ii PS3 and Lam shared a common stress-responsive transcriptome profile that partly overlapped the salicylate- (SA and jasmonate-(JA-dependent ones. After P. viticola inoculation, PS3 specifically primed the SA- and ROS-dependent defense pathways leading to grapevine induced resistance against this biotroph. Interestingly pharmacological approaches suggested that the plasma membrane depolarization and the downstream ROS production are key events of the PS3-induced resistance.

  4. The transcription factor VvWRKY33 is involved in the regulation of grapevine (Vitis vinifera) defense against the oomycete pathogen Plasmopara viticola.

    Science.gov (United States)

    Merz, Patrick R; Moser, Tina; Höll, Janine; Kortekamp, Andreas; Buchholz, Günther; Zyprian, Eva; Bogs, Jochen

    2015-03-01

    Grapevine (Vitis vinifera ssp. vinifera) is one of the most important fruit species; however, it is highly susceptible to various pathogens, which can cause severe crop losses in viticulture. It has been shown that several WRKY class transcription factors (TFs) are part of the signal transduction cascade, which leads to the activation of plant defense reactions against various pathogens. In the present investigation, a full-length cDNA was isolated from V. vinifera leaf tissue encoding a predicted protein, designated VvWRKY33, which shows the characteristics of group I WRKY protein family. VvWRKY33 induction correlates with the expression of VvPR10.1 (pathogenesis-related 10.1) gene in the leaves of the resistant cultivar 'Regent' after infection with Plasmopara viticola, whereas in the susceptible cultivar 'Lemberger' VvWRKY33 and VvPR10.1 are not induced. Corresponding expression of the TF and VvPR10.1 was even obtained in uninfected ripening berries. In planta, analysis of VvWRKY33 has been performed by ectopic expression of VvWRKY33 in grapevine leaves of greenhouse plants mediated via Agrobacterium tumefaciens transformation. In consequence, VvWRKY33 strongly increases resistance to P. viticola in the susceptible cultivar 'Shiraz' and reduces pathogen sporulation of about 50-70%, indicating a functional role for resistance in grapevine. Complementation of the resistance-deficient Arabidopsis thaliana Columbia-0 (Col-0) mutant line wrky33-1 by constitutive expression of VvWRKY33 restores resistance against Botrytis cinerea to wild-type level and in some complemented mutant lines even exceeds the resistance level of the parental line Col-0. Our results support the involvement of VvWRKY33 in the defense reaction of grapevine against different pathogens. © 2014 Scandinavian Plant Physiology Society.

  5. 叶片湿润时间对葡萄霜霉病发生状况的影响%Effect of Different Humidity Time on Occurrence of Plasmopara viticola in Grape

    Institute of Scientific and Technical Information of China (English)

    沙月霞; 王国珍; 樊仲庆

    2011-01-01

    The experiments set up different time of leaves infected by fungal suspension, different humidity time of leaves inoculated by Plasmorara viticola and different humidity condition in laboratory to explore the effect of different thumidity time on occurrence extent of Plasmopara viticola on grape leaves. Results showed that grape downy mildew would take place when grape leaves were infected by fungal suspension beyond 0. 5 h and the severity of disease became worse with time enlongation. The humid condition of grape downy mildew was beyond RH 76. 00% and disease was worse with higher humid. The key condition for Plasmorara viticola infection and disease apparition was keeping humidity beyond 2 -3 h. The shortest incubation period of Plasmorara viticola was 2~3 d at 20 °C. The highest apparition period of disease was 5 - 7 d after wetting grape blade back.%从不同菌悬液与叶片接触时间、接菌后叶片不同保湿时间和不同保湿条件3方面,研究叶片湿润时间对葡萄霜霉病发生状况的影响.结果表明,葡萄霜霉病菌悬液与叶片接触0.5h以上即可导致霜霉病发生,接触时间越长发病越重;接菌后在RH=76%保湿条件下,葡萄霜霉病即发生,湿度越高发病越重;霜霉病菌完成侵入到显症需要的叶片湿润条件是叶片湿润2~3 h以上;20℃条件下,霜霉病的潜伏期最短2~3 d,而病斑出现的高峰期是使叶背湿润后的5~7 d.

  6. Caractérisation de polyphénols stilbéniques et de dérivés induits ou constitutifs de la vigne impliqués dans sa défense contre l'agent pathogène du mildiou de la vigne, Plasmopara viticola (Berk. and Curt.)

    OpenAIRE

    Jean-Denis, Julien Bernard; R. Tabacchi

    2006-01-01

    Le Mildiou de la vigne, causé par un oomycète parasite obligatoire, Plasmopara viticola, est une des maladies les plus redoutées par les viticulteurs. Son apparition est occasionnée par des éclaboussures de pluie et de terre sur les tissues de la vigne après les fortes précipitations véhiculant les zoospores infectieuses présentes à la surface du sol. Certaines variétés comme le Chasselas (V. vinifera) sont sensibles au mildiou et laissent P. viticola germer et se développer. D’autres variété...

  7. Morphological variability of oospores of Chara baueri A. Braun (Characeae

    Directory of Open Access Journals (Sweden)

    Andrzej Hutorowicz

    2011-01-01

    Full Text Available The shape and size of present-day specimens of Chara baueri are described. C. baueri is believed to have practically died out in Europe. Oospores of specimens of C. baueri collected in Kazachstan were subjected to morpholgical analysis. Data recorded included oospore length, oospore diameter, the ratio of length to diameter, the number of lateral convolutions, ridge width, the width of the fossules at the equator, and the diameter of the basal pore. Variation in oospore length and the number of convolutions was higher than had been previously reported in the literature, whereas variation in oospore width was identical to previous reports.

  8. Production, survival and infectivity of oospores of Phytophthora infestans

    NARCIS (Netherlands)

    Turkensteen, L.J.; Flier, W.G.; Mulder, A.; Wanningen, R.

    2000-01-01

    The formation of oospores of Phytophthora infestans was studied in tomato and potato crops and volunteer plants under field conditions, and in laboratory tests with leaf discs of potato cultivars differing in their level of race-nonspecific resistance. Oospores were readily detected in

  9. Sorosphaera viticola, a plasmodiophorid parasite of grapevine

    OpenAIRE

    Neuhauser, Sigrid; Huber, Lars; Kirchmair, Martin

    2009-01-01

    Sorosphaera viticola is a soil-borne, endophytic parasite of grapevine. It is classified within the plasmodiophorids, an enigmatic group of obligate biotrophic parasites of higher plants. Sorosphaera viticola has been found abundantly in the roots of Vitis spp. in Germany and Canada. This may indicate a global distribution of this root parasite. But its biphasic life-cycle, its soil-borne nature and its co-occurrence with other soil-borne pathogens make an assessment of the disease pattern or...

  10. Sorosphaera viticola, a plasmodiophorid parasite of grapevine

    Directory of Open Access Journals (Sweden)

    S. Neuhauser

    2009-05-01

    Full Text Available >Sorosphaera viticola is a soil-borne, endophytic parasite of grapevine. It is classifi ed within the plasmodiophorids, an enigmatic group of obligate biotrophic parasites of higher plants. Sorosphaera viticola has been found abundantly in the roots of Vitis spp. in Germany and Canada. This may indicate a global distribution of this root parasite. But its biphasic life-cycle, its soil-borne nature and its co-occurrence with other soil-borne pathogens make an assessment of the disease pattern or a possible yield reduction of this fungus diffi cult.

  11. Formation, production and viability of oospores of Phytophthora infestans from potato and Solanum demissum in the Toluca Valley, central Mexico

    NARCIS (Netherlands)

    Flier, W.G.; Grünwald, N.J.; Fry, W.E.; Turkensteen, L.J.

    2001-01-01

    Aspects of the ecology of oospores of Phytophthora infestans were studied in the highlands of central Mexico. From an investigation of a random sample of strains, it was found that isolates differed in their average capability to form oospores when engaged in compatible pairings. Most crosses produc

  12. Metabolic Fingerprint of PS3-Induced Resistance of Grapevine Leaves against Plasmopara viticola Revealed Differences in Elicitor-Triggered Defenses.

    Science.gov (United States)

    Adrian, Marielle; Lucio, Marianna; Roullier-Gall, Chloé; Héloir, Marie-Claire; Trouvelot, Sophie; Daire, Xavier; Kanawati, Basem; Lemaître-Guillier, Christelle; Poinssot, Benoît; Gougeon, Régis; Schmitt-Kopplin, Philippe

    2017-01-01

    Induction of plant resistance against pathogens by defense elicitors constitutes an attractive strategy to reduce the use of fungicides in crop protection. However, all elicitors do not systematically confer protection against pathogens. Elicitor-induced resistance (IR) thus merits to be further characterized in order to understand what makes an elicitor efficient. In this study, the oligosaccharidic defense elicitors H13 and PS3, respectively, ineffective and effective to trigger resistance of grapevine leaves against downy mildew, were used to compare their effect on the global leaf metabolism. Ultra high resolution mass spectrometry (FT-ICR-MS) analysis allowed us to obtain and compare the specific metabolic fingerprint induced by each elicitor and to characterize the associated metabolic pathways. Moreover, erythritol phosphate was identified as a putative marker of elicitor-IR.

  13. Specific adjustments in grapevine leaf proteome discriminating resistant and susceptible grapevine genotypes to Plasmopara viticola

    DEFF Research Database (Denmark)

    Figueiredo, Andreia; Martins, Joana; Sebastiana, Mónica

    2017-01-01

    differentiating the resistant genotype. Lipid associated signalling events, particularly related to jasmonates appear also to play a major role in the establishment of resistance. The findings from this study contribute to a better understanding of genotype-specific differences that account for a successful...

  14. Enzymatic Activity of the Mycelium Compared with Oospore Development During Infection of Pea Roots by Aphanomyces euteiches.

    Science.gov (United States)

    Kjøller, R; Rosendahl, S

    1998-09-01

    ABSTRACT To describe the disease cycle of the root pathogen Aphanomyces euteiches, enzymatic activity in the mycelium was compared with the development of oospores in pea roots. Plants were inoculated with two zoospore concentrations to achieve different disease levels. Hyphae were stained for fungal alkaline phosphatase activity in the roots. Additionally, enzyme activity was measured after electrophoresis of an A. euteiches-specific glucose-6-phosphate isozyme. Development of oospores in the roots was measured after staining the oospores with trypan blue. In plants inoculated with the higher zoospore concentration, the enzymatic activity of the pathogen mycelium peaked 10 to 14 days after inoculation, when oospore formation was initiated. Oospore formation was associated with a gradual increase in disease symptoms. At the last harvest, plants inoculated with the higher zoospore concentration had died. In these plants, oospores were found in 90% of the root length, while the enzymatic activity of the mycelium was low. This suggests that the pathogen mycelium is only active on living plants and does not grow saprophytically on dead plant material.

  15. Plasmopara halstedii virus causes hypovirulence in Plasmopara halstedii, the downy mildew pathogen of the sunflower.

    Science.gov (United States)

    Grasse, Wolfgang; Zipper, Reinhard; Totska, Maria; Spring, Otmar

    2013-08-01

    Plasmopara halstedii virus (PhV) is an isometric virus recently found in the oomycete Plasmopara halstedii. The fully sequenced virus genome consists of two ss(+)RNA strands encoding for the virus polymerase and the coat protein, respectively. Most of previously screened field isolates of P. halstedii were found to harbor PhV, but effects of PhV on the pathogenicity and aggressiveness of the oomycete have not been investigated yet. To assess the influence of PhV on the infectivity of P. halstedii, virus-free isolates of the oomycete were searched for, cultivated on sunflower and used for single zoospore infection. Four genetically homogenous strains belonging to three different pathotypes (710, 730, 750) were established. Subcultures of each strain were successfully infected with PhV. This afforded pairs of isogenic strains with and without virus and allowed assessment of the pathogenicity (susceptibility to specific sunflower genotypes) and aggressiveness (intensity of infection, time scale and density of sporulation) in cultivation of sunflower. While no significant difference was found in the pathogenicity of P. halstedii strains with and without virus towards sunflower seedlings of different resistance (pathotype differentials), the aggressiveness of the oomycete was diminished by PhV. Compared to the virus-free strains, the time required for the first sporulation (latent period) increased by about 1 day post inoculation. Progression of the pathogen from the hypocotyl into the epicotyl of sunflower (systemic infection) was reduced by about one third in the presence of virus. In the virus containing strains, the average density of sporangia produced per cm² cotyledon reached only 75% of the virus-free controls. In summary, the presence of PhV leads to hypovirulence effects by weakening the aggressiveness of P. halstedii.

  16. Enzymatic Activity of the Mycelium Compared with Oospore Development During Infection of Pea Roots by Aphanomyces euteiches

    DEFF Research Database (Denmark)

    Kjøller, Rasmus; Rosendahl, Søren

    1998-01-01

    To describe the disease cycle of the root pathogen Aphanomyces euteiches, enzymatic activity in the mycelium was compared with the development of oospores in pea roots. Plants were inoculated with two zoospore concentrations to achieve different disease levels. Hyphae were stained for fungal alka....... In these plants, oospores were found in 90% of the root length, while the enzymatic activity of the mycelium was low. This suggests that the pathogen mycelium is only active on living plants and does not grow saprophytically on dead plant material.......To describe the disease cycle of the root pathogen Aphanomyces euteiches, enzymatic activity in the mycelium was compared with the development of oospores in pea roots. Plants were inoculated with two zoospore concentrations to achieve different disease levels. Hyphae were stained for fungal...... concentration, the enzymatic activity of the pathogen mycelium peaked 10 to 14 days after inoculation, when oospore formation was initiated. Oospore formation was associated with a gradual increase in disease symptoms. At the last harvest, plants inoculated with the higher zoospore concentration had died...

  17. Sensitivity to copper in Xanthomonas campestris pv. viticola

    OpenAIRE

    2009-01-01

    ABSTRACT Bacterial canker, caused by Xanthomonas campestris pv. viticola, affects grapevines in the irrigated areas of the São Francisco river valley, in the states of Pernambuco and Bahia. Several practices for disease management have been adopted including copper sprays.This is the only available chemical control method and most frequently used in the areas affected by the disease. The objective of this work was to determine the sensitivity to copper of strains of X. campestris pv. vi...

  18. I romanismi nella terminologia viticola dell'Istria slovena

    OpenAIRE

    Rada Cossutta

    1995-01-01

    Il  presente saggio trae spunto da un attento riesame del materiale che ho raccolto ed analizzato per la mia tesi di dottorato di ricerca dal titolo Poljedelska in vi­ nogradniska terminologija v govorih Slovenske Istre (La terminologia agricola e viticola nelle parlate dell'lstria slovena), tesi che ho discusso presso la Facoltà di Filosofia di Lubiana il 14 gennaio 1994, relatore il prof. Tine Logar, correlatore la prof. Alenka Šivic-Dular. L'opera si inserisce nel filone di ricerche lessic...

  19. Meio semi-seletivo para isolamento de Xanthomonas campestris pv. viticola

    OpenAIRE

    2006-01-01

    O cancro bacteriano causado por Xanthomonas campestris pv. viticola é a fitobacteriose mais importante da videira no Submédio São Francisco. O isolamento de X. campestris pv. viticola de tecidos vegetais infectados é dificultado pela presença de contaminantes bacterianos, entre os quais Microbacterium barkeri. Objetivando-se a formulação de meio de cultura semi-seletivo, 22 isolados de X. campestris pv. viticola foram testados com relação a 30 antibióticos. O meio semi-seletivo NYDAM (extrato...

  20. Meio semi-seletivo para isolamento de Xanthomonas campestris pv. viticola

    Directory of Open Access Journals (Sweden)

    Peixoto Ana Rosa

    2006-01-01

    Full Text Available O cancro bacteriano causado por Xanthomonas campestris pv. viticola é a fitobacteriose mais importante da videira no Submédio São Francisco. O isolamento de X. campestris pv. viticola de tecidos vegetais infectados é dificultado pela presença de contaminantes bacterianos, entre os quais Microbacterium barkeri. Objetivando-se a formulação de meio de cultura semi-seletivo, 22 isolados de X. campestris pv. viticola foram testados com relação a 30 antibióticos. O meio semi-seletivo NYDAM (extrato de carne 3, peptona 5, glicose 10, extrato de levedura 5, ágar 18 e ampicilina 0,1 em g L-1 inibiu M. barkeri e bactérias fitopatogênicas podendo ser utilizado para isolar X. campestris pv. viticola de hospedeiros com infecção natural em campo.

  1. I romanismi nella terminologia viticola dell'Istria slovena

    Directory of Open Access Journals (Sweden)

    Rada Cossutta

    1995-12-01

    Full Text Available Il  presente saggio trae spunto da un attento riesame del materiale che ho raccolto ed analizzato per la mia tesi di dottorato di ricerca dal titolo Poljedelska in vi­ nogradniska terminologija v govorih Slovenske Istre (La terminologia agricola e viticola nelle parlate dell'lstria slovena, tesi che ho discusso presso la Facoltà di Filosofia di Lubiana il 14 gennaio 1994, relatore il prof. Tine Logar, correlatore la prof. Alenka Šivic-Dular. L'opera si inserisce nel filone di ricerche lessicali orientate a ri­ scrivere la storia linguistica di questa regione così complessa per le innumerevoli vicende storiche e per i molteplici contatti linguistici tuttora fertili.

  2. Reconstruction of the charophyte community of Lake Shinji by oospore collection

    Directory of Open Access Journals (Sweden)

    Komuro T.

    2016-01-01

    Full Text Available Submerged aquatic vegetation (SAV aids in maintaining a clear stable state in shallow lakes. However, charophytes are more effective in increasing transparency compared to angiosperms. Lake Shinji was more transparent prior to the beginning of herbicide use for rice weed control in the mid-1950s, because its bottom was covered by SAV up to 3 m depth. Although Chara braunii C.C. Gmelin and Nitella hyaline (De Candolle C. Agardh were recorded in the 1960s, there are no reports on SAV in the 1950s. Therefore, in the present study, we aimed to show that the SAV of Lake Shinji was mostly composed of charophytes prior to the 1950s, by conducting a seed analysis. We obtained charophyte oospores from the sediment, but seeds of angiosperms were not identified. In addition to C. braunii that was previously recorded in Lake Shinji, we also found two newly identified species, Chara corallina Willdenow  and Chara fibrosa C. Agardh ex Bruzelius. Overall, this study indicates that seed analysis is helpful in reconstructing the former flora of Lake Shinji.

  3. ‘Moldova’, a table grape varity highly resistant to Plasmopara viticola%高抗霜霉病鲜食葡萄品种--摩尔多瓦

    Institute of Scientific and Technical Information of China (English)

    赵胜建; 郭紫娟; 马爱红

    2006-01-01

    1997年引入摩尔多瓦(Moldova)葡萄品种进行试栽观察.该品种果穗圆锥形,平均穗重650g;果粒短椭圆形,平均果粒重8.0~9.0g;果皮蓝黑色,着色易,且均匀一致.肉质中等,无香味,品质上,可溶性固形物含量达16.0%,含酸量0.54%,耐贮运.高抗葡萄霜霉病,极丰产.适宜小棚架栽培,中短梢修剪.

  4. Genetic signature of a range expansion and leap-frog event after the recent invasion of Europe by the grapevine downy mildew pathogen Plasmopara viticola

    NARCIS (Netherlands)

    Fontaine, Michael C.; Austerlitz, Frederic; Giraud, Tatiana; Labbe, Frederic; Papura, Daciana; Richard-Cervera, Sylvie; Delmotte, Francois

    2013-01-01

    Biologic invasions can have important ecological, economic and social consequences, particularly when they involve the introduction and spread of plant invasive pathogens, as they can threaten natural ecosystems and jeopardize the production of human food. Examples include the grapevine downy mildew

  5. The Loricrin-Like Protein (LLP) of Phytophthora infestans Is Required for Oospore Formation and Plant Infection

    Science.gov (United States)

    Guo, Ting; Wang, Xiao-Wen; Shan, Kun; Sun, Wenxian; Guo, Li-Yun

    2017-01-01

    Loricrin-like protein (LLP) is characterized by a high content of glycine residues and is a major component of plant cell wall. Here, we identified a Phytophthora infestans ortholog of plant LLP, named PiLLP. In P. infestans, PiLLP is strongly expressed in asexual and sexual developmental stages, including in sporangia, zoospores and germinating cysts, and during oospore formation, as well as in the early stages of infection and during hydrogen peroxide stress. Compared with the wild type, the PiLLP-silenced transformants were defective in oospore formation, had slower colony expansion rates, produced less sporangia with lower germination and zoospore-release rates, and were more sensitive to hydrogen peroxide. Moreover, Nile red staining, and PiLLP-red fluorescent protein fusions indicated that PiLLP is involved in oogonia formation. The silenced transformants also had severely diminished virulence levels that could be partially restored with diphenyleneiodium treatments. The analysis of catalase activity showed a decrease of catalase activity in silenced transformants. Thus, PiLLP is important for sexual and asexual reproduction, and is required for oxidative stress tolerance and plant infection. PMID:28232841

  6. Influence of epiphytic bacteria from grapevine leaves on Phomopsis viticola Sacc.

    Directory of Open Access Journals (Sweden)

    Ewa Król

    2013-12-01

    Full Text Available Out of 282 isolates of epiphytic bacteria population originating from grapevine leaves 15% inhibited growth of Phomopsis viticola on potato-dextrose agar. Protective activity of 16 isolates, representing different morphological groups was tested on one-year old of grapevine stems fragments. Majority of bacterial isolates which inhibited the fungus growth in-vitro provided no protection of stems fragments against P. viticola. The efficiency of protection activity of bacterial isolates tested decreased with time. The most effective isolates of bacteria in protection of grapevine stemswere identified as Bacillus sp. and Pseudomonas fluorescens.

  7. The nucleotide sequence and genome organization of Plasmopara halstedii virus

    Directory of Open Access Journals (Sweden)

    Göpfert Jens C

    2011-03-01

    Full Text Available Abstract Background Only very few viruses of Oomycetes have been studied in detail. Isometric virions were found in different isolates of the oomycete Plasmopara halstedii, the downy mildew pathogen of sunflower. However, complete nucleotide sequences and data on the genome organization were lacking. Methods Viral RNA of different P. halstedii isolates was subjected to nucleotide sequencing and analysis of the viral genome. The N-terminal sequence of the viral coat protein was determined using Top-Down MALDI-TOF analysis. Results The complete nucleotide sequences of both single-stranded RNA segments (RNA1 and RNA2 were established. RNA1 consisted of 2793 nucleotides (nt exclusive its 3' poly(A tract and a single open-reading frame (ORF1 of 2745 nt. ORF1 was framed by a 5' untranslated region (5' UTR of 18 nt and a 3' untranslated region (3' UTR of 30 nt. ORF1 contained motifs of RNA-dependent RNA polymerases (RdRp and showed similarities to RdRp of Scleropthora macrospora virus A (SmV A and viruses within the Nodaviridae family. RNA2 consisted of 1526 nt exclusive its 3' poly(A tract and a second ORF (ORF2 of 1128 nt. ORF2 coded for the single viral coat protein (CP and was framed by a 5' UTR of 164 nt and a 3' UTR of 234 nt. The deduced amino acid sequence of ORF2 was verified by nano-LC-ESI-MS/MS experiments. Top-Down MALDI-TOF analysis revealed the N-terminal sequence of the CP. The N-terminal sequence represented a region within ORF2 suggesting a proteolytic processing of the CP in vivo. The CP showed similarities to CP of SmV A and viruses within the Tombusviridae family. Fragments of RNA1 (ca. 1.9 kb and RNA2 (ca. 1.4 kb were used to analyze the nucleotide sequence variation of virions in different P. halstedii isolates. Viral sequence variation was 0.3% or less regardless of their host's pathotypes, the geographical origin and the sensitivity towards the fungicide metalaxyl. Conclusions The results showed the presence of a single and new

  8. The nucleotide sequence and genome organization of Plasmopara halstedii virus

    Science.gov (United States)

    2011-01-01

    Background Only very few viruses of Oomycetes have been studied in detail. Isometric virions were found in different isolates of the oomycete Plasmopara halstedii, the downy mildew pathogen of sunflower. However, complete nucleotide sequences and data on the genome organization were lacking. Methods Viral RNA of different P. halstedii isolates was subjected to nucleotide sequencing and analysis of the viral genome. The N-terminal sequence of the viral coat protein was determined using Top-Down MALDI-TOF analysis. Results The complete nucleotide sequences of both single-stranded RNA segments (RNA1 and RNA2) were established. RNA1 consisted of 2793 nucleotides (nt) exclusive its 3' poly(A) tract and a single open-reading frame (ORF1) of 2745 nt. ORF1 was framed by a 5' untranslated region (5' UTR) of 18 nt and a 3' untranslated region (3' UTR) of 30 nt. ORF1 contained motifs of RNA-dependent RNA polymerases (RdRp) and showed similarities to RdRp of Scleropthora macrospora virus A (SmV A) and viruses within the Nodaviridae family. RNA2 consisted of 1526 nt exclusive its 3' poly(A) tract and a second ORF (ORF2) of 1128 nt. ORF2 coded for the single viral coat protein (CP) and was framed by a 5' UTR of 164 nt and a 3' UTR of 234 nt. The deduced amino acid sequence of ORF2 was verified by nano-LC-ESI-MS/MS experiments. Top-Down MALDI-TOF analysis revealed the N-terminal sequence of the CP. The N-terminal sequence represented a region within ORF2 suggesting a proteolytic processing of the CP in vivo. The CP showed similarities to CP of SmV A and viruses within the Tombusviridae family. Fragments of RNA1 (ca. 1.9 kb) and RNA2 (ca. 1.4 kb) were used to analyze the nucleotide sequence variation of virions in different P. halstedii isolates. Viral sequence variation was 0.3% or less regardless of their host's pathotypes, the geographical origin and the sensitivity towards the fungicide metalaxyl. Conclusions The results showed the presence of a single and new virus type in

  9. Limpeza clonal de mudas de videira infectadas por Xanthomonas campestris pv. viticola

    OpenAIRE

    2013-01-01

    O cancro bacteriano da videira é causado por Xanthomonas campestris pv. viticola (Xcv). Visando à limpeza clonal de mudas de 'Red Globe', foram estudados: tamanho ideal de ápices e gemas axilares para cultivo em meio de Galzy modificado (MGM); efeito da termoterapia (38ºC/30 dias); e ação de antibióticos na eliminação de Xcv em videiras infectadas. Os percentuais de contaminação por Xcv e de regeneração foram analisados, e as plantas obtidas foram indexadas em meio ágar nutritivo-dextrose-ext...

  10. Identificação de potenciais plantas hospedeiras alternativas de Xanthomonas campestris pv. Viticola.

    OpenAIRE

    2014-01-01

    Este estudo teve como objetivo identificar possíveis hospedeiras alternativas de Xanthomonas campestris pv. viticola (Xcv), visando a fornecer subsídios para o manejo do cancro bacteriano da videira. Vinte e seis espécies vegetais foram inoculadas artificialmente com o isolado Xcv3 e mantidas em condições de casa de vegetação, sendo avaliada a evolução sintomatológica da doença, como manchas necróticas angulares e lesões nas nervuras. O Xcv3 foi reisolado a partir de cada hospedeiro alternati...

  11. Diagnose molecular do cancro bacteriano da videira causado por Xanthomonas campestris pv. viticola

    OpenAIRE

    2009-01-01

    A bactéria Xanthomonas campestris pv. viticola (Nayudu) Dye, agente do cancro bacteriano da videira, foi relatada pela primeira vez nas áreas irrigadas do Submédio do Vale São Francisco em Petrolina, Pernambuco, em 1998. A doença também foi identificada em Juazeiro, Bahia, e posteriormente no Piauí, no Ceará, Roraima e Goiás. O uso de material propagativo livre do patógeno tornou-se uma preocupação, uma vez que a ocorrência da bacteriose ainda esta restrita a essas regiões no p...

  12. Sobrevivência de Xanthomonas campestris pv. viticola em tecido infectado de videira.

    OpenAIRE

    2012-01-01

    Tecidos de plantas infectados constituem uma importante fonte de inóculo primário para fitobacterioses. O objetivo deste trabalho foi investigar a sobrevivência de Xanthomonas campestris pv. viticola (Xcv), agente do cancro bacteriano da videira, em tecidos infectados na superfície do solo e durante a compostagem de restos de poda. Mudas de videira 'Festival' foram inoculadas com mutante resistente à rifampicina Xcv2Rif e mantidas em casa de vegetação até apresentarem alta severidade da doenç...

  13. Chemotaxis and oospore formation in Phytophthora sojae are controlled by G-protein-coupled receptors with a phosphatidylinositol phosphate kinase domain.

    Science.gov (United States)

    Yang, X; Zhao, W; Hua, C; Zheng, X; Jing, M; Li, D; Govers, F; Meijer, H J G; Wang, Y

    2013-04-01

    G-protein-coupled receptors (GPCRs) are key cellular components that mediate extracellular signals into intracellular responses. Genome mining revealed that Phytophthora spp. have over 60 GPCR genes among which a prominent class of 12 encoding novel proteins with an N-terminal GPCR domain fused to a C-terminal phosphatidylinositol phosphate kinase (PIPK) domain. This study focuses on two GPCR-PIPKs (GKs) in Phytophthora sojae. PsGK4 and PsGK5 are differentially expressed during the life cycle with the highest expression in cysts and during cyst germination, and at late infection stages. In P. sojae transformants that constitutively express RFP-tagged PsGK4 and PsGK5, the fusion proteins in hyphae reside in small, rapidly moving vesicular-like structures. Functional analysis using gene silencing showed that PsGK4-silenced transformants displayed higher levels of encystment and a reduced cyst germination rate when compared with the recipient strain. Moreover, GK4 deficiency (or reduction) resulted in severe defects in zoospore chemotaxis towards isoflavones and soybean roots. In contrast, PsGK5-silenced transformants exhibited no obvious defects in asexual development but oospore production was severely impaired. Both, PsGK4- and PsGK5-silenced transformants showed reduced pathogenicity. These results point to involvement of GKs in zoospore behaviour, chemotaxis and oospore development, and suggest that PsGK4 and PsGK5 each head independent signalling pathways.

  14. Limpeza clonal de mudas de videira infectadas por Xanthomonas campestris pv. viticola Clonal cleaning of grapevine plants infected by Xanthomonas campestris pv. viticola

    Directory of Open Access Journals (Sweden)

    Adriano Márcio Freire Silva

    2013-03-01

    Full Text Available O cancro bacteriano da videira é causado por Xanthomonas campestris pv. viticola (Xcv. Visando à limpeza clonal de mudas de 'Red Globe', foram estudados: tamanho ideal de ápices e gemas axilares para cultivo em meio de Galzy modificado (MGM; efeito da termoterapia (38ºC/30 dias; e ação de antibióticos na eliminação de Xcv em videiras infectadas. Os percentuais de contaminação por Xcv e de regeneração foram analisados, e as plantas obtidas foram indexadas em meio ágar nutritivo-dextrose-extrato de levedura-ampicilina (NYDAM, seguindo-se teste de patogenicidade. O cultivo de explantes com 3 mm possibilitou a obtenção de plantas livres da bactéria, com regeneração 14,3 vezes maior que explantes com 1 mm. A termoterapia de mudas infectadas, associada ao cultivo in vitro, não eliminou o patógeno. O cultivo de explantes com 10 mm, durante 40 dias em MGM + cefotaxima (300 mg L-1, proporcionou limpeza clonal das mudas. A indexação de plantas de videira regeneradas in vitro, quanto à infecção por Xcv utilizando NYDAM, seguida de teste de patogenicidade, é uma alternativa econômica e eficiente para produção de mudas de alta qualidade fitossanitária.Bacterial canker is caused by Xanthomonas campestris pv. viticola (Xcv. In order to eliminate Xcv from 'Red Globe' plants it was studied: optimal size of meristem tips and axillary buds for cultivation in modified Galzy's medium (MGM; effects of thermotherapy (38ºC/30 days; and action of antibiotics in the elimination of Xcv in infected grapevines. The percentages of contamination by Xcv and regeneration were analyzed and plants obtained were indexed using the semi-selective culture medium nutrient agar-dextrose-yeast extract-ampicilin (NYDAM followed by a pathogenicity test. The cultivation of 3 mm explants permitted to obtain plants free of bacteria with regeneration 14.3 times higher than 1 mm explants. The thermotherapy of infected plants associated to the in vitro culture

  15. Expressed Sequence Tags from the oomycete Plasmopara halstedii, an obligate parasite of the sunflower

    Directory of Open Access Journals (Sweden)

    Mouzeyar Said

    2007-12-01

    Full Text Available Abstract Background Sunflower downy mildew is a major disease caused by the obligatory biotrophic oomycete Plasmopara halstedii. Little is known about the molecular mechanisms underlying its pathogenicity. In this study we used a genomics approach to gain a first insight into the transcriptome of P. halstedii. Results To identify genes from the obligatory biotrophic oomycete Plasmopara halstedii that are expressed during infection in sunflower (Helianthus annuus L. we employed the suppression subtraction hybridization (SSH method from sunflower seedlings infected by P. halstedii. Using this method and random sequencing of clones, a total of 602 expressed sequence tags (ESTs corresponding to 230 unique sequence sets were identified. To determine the origin of the unisequences, PCR primers were designed to amplify these gene fragments from genomic DNA isolated either from P. halstedii sporangia or from Helianthus annuus. Only 145 nonredundant ESTs which correspond to a total of 373 ESTs (67.7% proved to be derived from P. halstedii genes and that are expressed during infection in sunflower. A set of 87 nonredundant sequences were identified as showing matches to sequences deposited in public databases. Nevertheless, about 7% of the ESTs seem to be unique to P. halstedii without any homolog in any public database. Conclusion A summary of the assignment of nonredundant ESTs to functional categories as well as their relative abundance is listed and discussed. Annotation of the ESTs revealed a number of genes that could function in virulence. We provide a first glimpse into the gene content of P. halstedii. These resources should accelerate research on this important pathogen.

  16. Produção e caracterização de anticorpos policlonais contra Xanthomonas campestris pv. viticola Production and characterization of polyclonal antibodies against Xanthomonas campestris pv. viticola

    Directory of Open Access Journals (Sweden)

    João Sebastião de Paula Araujo

    2005-03-01

    Full Text Available O objetivo deste trabalho foi a produção de anticorpos policlonais contra Xanthomonas campestris pv. viticola e sua caracterização pelo método Elisa indireto. Os resultados apontaram a qualidade dos anticorpos policlonais produzidos, os quais mostraram-se altamente reativos e específicos para o patovar com potencial para ser empregado no diagnóstico da doença e em programas de certificação.The objective of this work was to produce polyclonal antibodies against Xanthomonas campestris pv. viticola and characterize these antibodies through Elisa serological indirect method. Results indicate that polyclonal antibodies produced were highly reactive against bacterial cells, showing specificity at the pathovar level and potential to be used for diagnosis and certification purposes.

  17. Produção e caracterização de anticorpos policlonais contra Xanthomonas campestris pv. viticola.

    OpenAIRE

    2005-01-01

    O objetivo deste trabalho foi a produção de anticorpos policlonais contra Xanthomonas campestris pv. viticola e sua caracterização pelo método Elisa indireto. Os resultados apontaram a qualidade dos anticorpos policlonais produzidos, os quais mostraram-se altamente reativos e específicos para o patovar com potencial para ser empregado no diagnóstico da doença e em programas de certificação.

  18. The Resistance of Six Grape Varieties to Uncinula necator and Plasmopara viticola%6个葡萄品种对白粉病和霜霉病的抗性

    Institute of Scientific and Technical Information of China (English)

    刘会宁; 吴广宇

    2010-01-01

    采取田间自然鉴定法,鉴定了6个欧亚种葡萄(Vitis vinifera)品种对白粉病和霜霉病的抗性.结果表明,品种间的白粉病和霜霉病的抗性存在差异,9307为高感品种;凤凰51号、研选5号和维多利亚均为感病品种;矢富罗莎为抗病品种;早紫对白粉病表现为高抗,而对霜霉病表现为抗病.

  19. ANALYSIS OF TWO STANDARTS TO EVALUATE THE RESISTANCE OF PLASMOPARA VITICOLA%两种葡萄霜霉病抗性鉴定标准的分析比较

    Institute of Scientific and Technical Information of China (English)

    刘延琳; 贺普超

    1998-01-01

    本研究比较了葡萄霜霉病抗性鉴定中反应型与严重度两种鉴定标准.结果表明,两种抗性鉴定标准呈极显著相关,均可以反映葡萄植株对霜霉病的抗病性.反应型标准简便、准确、稳定,是定性的分级方法,而严重度标准可以反映植株抗病性的微小差异.

  20. Limpeza clonal de mudas de videira infectadas por Xanthomonas campestris pv. viticola

    Directory of Open Access Journals (Sweden)

    Adriano Márcio Freire Silva

    2013-03-01

    Full Text Available O cancro bacteriano da videira é causado por Xanthomonas campestris pv. viticola (Xcv. Visando à limpeza clonal de mudas de 'Red Globe', foram estudados: tamanho ideal de ápices e gemas axilares para cultivo em meio de Galzy modificado (MGM; efeito da termoterapia (38ºC/30 dias; e ação de antibióticos na eliminação de Xcv em videiras infectadas. Os percentuais de contaminação por Xcv e de regeneração foram analisados, e as plantas obtidas foram indexadas em meio ágar nutritivo-dextrose-extrato de levedura-ampicilina (NYDAM, seguindo-se teste de patogenicidade. O cultivo de explantes com 3 mm possibilitou a obtenção de plantas livres da bactéria, com regeneração 14,3 vezes maior que explantes com 1 mm. A termoterapia de mudas infectadas, associada ao cultivo in vitro, não eliminou o patógeno. O cultivo de explantes com 10 mm, durante 40 dias em MGM + cefotaxima (300 mg L-1, proporcionou limpeza clonal das mudas. A indexação de plantas de videira regeneradas in vitro, quanto à infecção por Xcv utilizando NYDAM, seguida de teste de patogenicidade, é uma alternativa econômica e eficiente para produção de mudas de alta qualidade fitossanitária.

  1. Identificação de potenciais plantas hospedeiras alternativas de Xanthomonas campestris pv. viticola

    Directory of Open Access Journals (Sweden)

    Morgana Mateus Santos

    2014-04-01

    Full Text Available Este estudo teve como objetivo identificar possíveis hospedeiras alternativas de Xanthomonas campestris pv. viticola (Xcv, visando a fornecer subsídios para o manejo do cancro bacteriano da videira. Vinte e seis espécies vegetais foram inoculadas artificialmente com o isolado Xcv3 e mantidas em condições de casa de vegetação, sendo avaliada a evolução sintomatológica da doença, como manchas necróticas angulares e lesões nas nervuras. O Xcv3 foi reisolado a partir de cada hospedeiro alternativo com sintomas, sendo identificado por PCR (Polymerase Chain Reaction, com iniciadores específicos. As espécies inoculadas que apresentaram os sintomas típicos da doença foram Glycine sp., Senna obtusifolia, Desmodium discolor, Amaranthus deflexus, Azadirachta indica, Solanum lycopersicum e Vigna unguiculata. As espécies da família Poaceae, Bidens pilosa, Emilia fosbergii, Praxelis pauciflora, Macroptilium lathyroides e Portulaca oleracea não apresentaram sintomas durante o período da avaliação.

  2. Presencia de Phomopsis viticola (Saccardo Saccardo, agente causal de excoriosis, en viñedos de Córdoba, Argentina Presence of Phomopsis viticola (Saccardo Saccardo, causal agent of excoriosis, in vineyards of Córdoba, Argentina

    Directory of Open Access Journals (Sweden)

    C.I. Cragnolini

    2009-12-01

    Full Text Available Desde 2001 se observan en viñedos de Córdoba, Argentina, hojas de Vitis vinifera y V. labrusca con manchas cloróticas y centro necrótico. Las plantas presentan nervaduras, pecíolos, raquis y entrenudos con puntos necróticos que al confluir agrietan el tejido cortical. Las bayas pueden marchitarse y desprenderse. En invierno los sarmientos toman color blanquecino con manchas negras, y en ellos y en órganos secos aparecen picnidios con cirros blanco-crema con conidios alfa y beta. El objetivo fue confirmar la presencia de Phomopsis viticola (excoriosis y cuantificar su incidencia. Se realizaron aislamientos a partir de pecíolos, brotes, zarcillos, bayas y sarmientos que produjeron micelio claro con picnidios y conidios alfa (7,5m±1,5 x 2,8m±2 y beta (19m±1 x 1,2m±0,5 o sólo alfa, típicos de P. viticola. La identificación fue confirmada mediante secuenciación molecular. En brotes de las variedades Malbec y Pinot noir inoculados con 3 10(4 conidios/mL, las hojas mostraron lesiones y picnidios 8 días después, y las bayas cirros amarillos a los 15-18 días. P. viticola (Sacc. Sacc. fue reaislado de los cirros y las colonias produjeron picnidios y conidios característicos 10-20 días después. Se detectó excoriosis en 80% de los viñedos evaluados, con mayor incidencia en viñedos añosos de las variedades Pinot noir e Isabella.Leaves with chlorotic spots and necrotic centers have been observed in Vitis vinifera and V. labrusca in vineyards of Córdoba, Argentina, since 2001. Plants exhibit veins, petioles, rachises, and internodes with necrotic spots that coalesce to form cracks in the cortical tissue. Grape berries can shrivel and fall off. Winter canes became whitish with black spots; pycnidia appear in canes and in dry organs, exuding whitish-cream cirri with alpha and beta conidia. The aim of this work was to confirm the presence of Phomopsis viticola (excoriosis and quantify its incidence. Isolates obtained from petioles

  3. Produção e caracterização de anticorpos policlonais contra Xanthomonas campestris pv. viticola Production and characterization of polyclonal antibodies against Xanthomonas campestris pv. viticola

    OpenAIRE

    2005-01-01

    O objetivo deste trabalho foi a produção de anticorpos policlonais contra Xanthomonas campestris pv. viticola e sua caracterização pelo método Elisa indireto. Os resultados apontaram a qualidade dos anticorpos policlonais produzidos, os quais mostraram-se altamente reativos e específicos para o patovar com potencial para ser empregado no diagnóstico da doença e em programas de certificação.The objective of this work was to produce polyclonal antibodies against Xanthomonas campestris pv. vitic...

  4. Sobrevivência de Xanthomonas campestris pv. viticola em tecido infectado de videira

    Directory of Open Access Journals (Sweden)

    Adriano Márcio Freire Silva

    2012-09-01

    Full Text Available Tecidos de plantas infectados constituem uma importante fonte de inóculo primário para fitobacterioses. O objetivo deste trabalho foi investigar a sobrevivência de Xanthomonas campestris pv. viticola (Xcv, agente do cancro bacteriano da videira, em tecidos infectados na superfície do solo e durante a compostagem de restos de poda. Mudas de videira 'Festival' foram inoculadas com mutante resistente à rifampicina Xcv2Rif e mantidas em casa de vegetação até apresentarem alta severidade da doença. No primeiro experimento, ramos fragmentados e folhas inteiras destas plantas foram acondicionados em bolsas de malha plástica e colocados na superfície de microparcelas. No segundo experimento, ramos e folhas fragmentados foram acondicionados em bolsas plásticas e depositados no interior de pilhas de compostagem de restos de poda de videira. A sobrevivência de Xcv2Rif em tecidos infectados foi monitorada a intervalos de 8 e 10 dias a partir do início do primeiro e segundo experimentos, respectivamente. No primeiro experimento, foi também avaliada a decomposição de tecidos, e no segundo, as curvas de temperatura das pilhas, conteúdo de fenóis e microbiota antagonista a Xcv2Rif. O patógeno sobreviveu em altas populações até 80 dias em tecidos infectados na superfície do solo. A compostagem eliminou Xcv2Rif de restos culturais em 10 dias, devido às altas temperaturas, liberação de compostos fenólicos e antagonismo microbiano. Concluindo, em tecidos infectados de videira Xcv sobrevive na superfície do solo por, pelo menos, 80 dias mas é eliminada pela compostagem em 10 dias.

  5. Impact of major gene resistance management for sunflower on fitness of Plasmopara halstedii (downy mildew populations

    Directory of Open Access Journals (Sweden)

    Tourvieille de Labrouhe Denis

    2010-01-01

    Full Text Available Changes in virulence of Plasmopara halstedii populations under different major gene (Pl management strategies were studied over 5 years continuous cropping of one sunflower hybrid under netting cages. Strategies were monoculture of forms of the hybrid with 1 gene or with combinations of 2 genes, alternation of different genes, and mixtures of several different forms of the hybrid. Monoculture with single resistance genes led to loss of efficient resistance after 3 years, with high levels of disease and increased variability of the pathogen, whatever the Pl gene used. Combinations of genes, alternation and mixtures gave longer term control of downy mildew. In particular, combinations of resistance genes coming from both female and male parents of the hybrid (such that even impurities had a resistance gene gave the best control and least variation in pathogen virulence. Results are discussed with the object of durable control of downy mildew by all methods available.

  6. Foreword: Special issue on fungal grapevine diseases

    Science.gov (United States)

    An impressively large proportion of fungicides applied in European, North American and Australian agriculture has been used to manage grapevine powdery mildew (Erysiphe necator), grapevine downy mildew (Plasmopara viticola), and botrytis bunch rot (Botrytis cinerea). These fungal and oomycetous plan...

  7. Identification of a novel Plasmopara halstedii elicitor protein combining de novo peptide sequencing algorithms and RACE-PCR

    Directory of Open Access Journals (Sweden)

    Madlung Johannes

    2010-05-01

    Full Text Available Abstract Background Often high-quality MS/MS spectra of tryptic peptides do not match to any database entry because of only partially sequenced genomes and therefore, protein identification requires de novo peptide sequencing. To achieve protein identification of the economically important but still unsequenced plant pathogenic oomycete Plasmopara halstedii, we first evaluated the performance of three different de novo peptide sequencing algorithms applied to a protein digests of standard proteins using a quadrupole TOF (QStar Pulsar i. Results The performance order of the algorithms was PEAKS online > PepNovo > CompNovo. In summary, PEAKS online correctly predicted 45% of measured peptides for a protein test data set. All three de novo peptide sequencing algorithms were used to identify MS/MS spectra of tryptic peptides of an unknown 57 kDa protein of P. halstedii. We found ten de novo sequenced peptides that showed homology to a Phytophthora infestans protein, a closely related organism of P. halstedii. Employing a second complementary approach, verification of peptide prediction and protein identification was performed by creation of degenerate primers for RACE-PCR and led to an ORF of 1,589 bp for a hypothetical phosphoenolpyruvate carboxykinase. Conclusions Our study demonstrated that identification of proteins within minute amounts of sample material improved significantly by combining sensitive LC-MS methods with different de novo peptide sequencing algorithms. In addition, this is the first study that verified protein prediction from MS data by also employing a second complementary approach, in which RACE-PCR led to identification of a novel elicitor protein in P. halstedii.

  8. Efeito do tratamento de bacelos de videira 'Red Globe' no controle do cancro bacteriano causado por Xanthomonas campestris pv. viticola

    Directory of Open Access Journals (Sweden)

    Carine Rosa Naue

    2014-12-01

    Full Text Available A disseminação de Xanthomonas campestris pv. viticola (Xcv, agente do cancro bacteriano da videira, ocorre, dentre outras formas, por meio de mudas e bacelos infectados. Foi estudada a obtenção de material propagativo livre do patógeno, testando a eficiência do tratamento de bacelos com termoterapia, bactericidas e sanitizantes. Os isolados de Xcv foram testados quanto à patogenicidade e realizado o teste de sensibilidade in vitro aos produtos, em diferentes concentrações. A erradicação de Xcv em bacelos de videira foi testada em experimentos com termoterapia (50ºC por 30 e 40 min; 53ºC por 5 e 10 min; bactericidas [oxitetraciclina+sulfato de cobre (150+2.000; 165+2.200; 180+2.400 e 195+2.600 mg L-1 de H2O e oxitetraciclina (600; 700; 800 e 900 mg L-1]; e sanitizantes [cloreto de dodecildimetil amônio (600; 1.200; 1.800; 2.400 e 3.000 µL L-1; hipoclorito de sódio (5.000; 10.000; 20.000; 30.000 e 40.000 µL L-1 e cloreto de benzalcônio (125; 167;250; 334 e 500 µL L-1]. Foram avaliados período de incubação, incidência e severidade da doença. O bactericida oxitetraciclina e os sanitizantes cloreto de dodecildimetil amônio e hipoclorito de sódio proporcionaram os maiores halos de inibição de Xcv in vitro. No entanto, apesar dos diversos tratamentos testados, não foi possível recomendar tratamento termoterápico ou produto que erradicasse Xcv de bacelos infectados. Porém, ficou confirmada a grande importância destes na disseminação do agente do cancro bacteriano da videira.

  9. Development of a molecular method for detection and identification of Xanthomonas campestris pv. viticola Desenvolvimento de um método molecular para detecção e identificação de Xanthomonas campestris pv. viticola

    Directory of Open Access Journals (Sweden)

    Loiselene Carvalho da Trindade

    2007-03-01

    Full Text Available In order to develop a molecular method for detection and identification of Xanthomonas campestris pv. viticola (Xcv the causal agent of grapevine bacterial canker, primers were designed based on the partial sequence of the hrpB gene. Primer pairs Xcv1F/Xcv3R and RST2/Xcv3R, which amplified 243- and 340-bp fragments, respectively, were tested for specificity and sensitivity in detecting DNA from Xcv. Amplification was positive with DNA from 44 Xcv strains and with DNA from four strains of X. campestris pv. mangiferaeindicae and five strains of X. axonopodis pv. passiflorae, with both primer pairs. However, the enzymatic digestion of PCR products could differentiate Xcv strains from the others. None of the primer pairs amplified DNA from grapevine, from 20 strains of nonpathogenic bacteria from grape leaves and 10 strains from six representative genera of plant pathogenic bacteria. Sensitivity of primers Xcv1F/Xcv3R and RST2/Xcv3R was 10 pg and 1 pg of purified Xcv DNA, respectively. Detection limit of primers RST2/Xcv3R was 10(4 CFU/ml, but this limit could be lowered to 10² CFU/ml with a second round of amplification using the internal primer Xcv1F. Presence of Xcv in tissues of grapevine petioles previously inoculated with Xcv could not be detected by PCR using macerated extract added directly in the reaction. However, amplification was positive with the introduction of an agar plating step prior to PCR. Xcv could be detected in 1 µl of the plate wash and from a cell suspension obtained from a single colony. Bacterium identity was confirmed by RFLP analysis of the RST2/Xcv3R amplification products digested with Hae III.Com o objetivo de desenvolver um método molecular para detecção e identificação de Xanthomonas campestris pv. viticola (Xcv, agente causal do cancro bacteriano da videira, oligonucleotídeos (primers foram desenhados com base na seqüência parcial do gene hrpB. As combinações de primers Xcv1F/Xcv3R e RST2/Xcv3R que

  10. Variabilité de l’agressivité de Plasmopara halstedii, étude de sa stabilité en test de laboratoire

    Directory of Open Access Journals (Sweden)

    Tourvieille De Labrouhe Denis

    2009-01-01

    Full Text Available Evolution of pathogenicity in Plasmopara halstedii was analysed by successive artificial infections of two strains with different levels of aggressiveness on two sunflower genotypes not carrying major gene resistance but which show different levels of partial resistance. Symptoms, in particular of plant size and sporulation density, confirmed the differences between the two lines observed in the field. The parasite populations studied showed considerable adaptation to infection conditions: the percentage of plants showing severe symptoms increased by 5% at each infection cycle, for both pathogen strains and both sunflower genotypes. These first laboratory results need to be confirmed in the field to judge the possible durability of quantitative resistances which could be introduced in sunflower breeding programmes.

  11. Molecular cloning and sequence analysis of a PVGOX gene encoding glucose oxidase in Penicillium viticola F1 strain and it's expression quantitation.

    Science.gov (United States)

    Khan, Ibrar; Qayyum, Sadia; Ahmed, Shehzad; Niaz, Zeeshan; Fatima, Nighat; Chi, Zhen-Ming

    2016-11-05

    The PVGOX gene (accession number: KT452630) was isolated from genomic DNA of the marine fungi Penicillium viticola F1 by Genome Walking and their expression analysis was done by Fluorescent RT-PCR. An open reading frame of 1806bp encoding a 601 amino acid protein (isoelectric point: 5.01) with a calculated molecular weight of 65,535.4 was characterized. The deduced protein showed 75%, 71%, 69% and 64% identity to those deduced from the glucose oxidase (GOX) genes from different fungal strains including; Talaromyces variabilis, Beauveria bassiana, Aspergillus terreus, and Aspergillus niger, respectively. The promoter of the gene (intronless) had two TATA boxes around the base pair number -88 and -94 and as well as a CAAT box at -100. However, the terminator of the PVGOX gene does not contain any polyadenylation site (AATAAA). The protein deduced from the PVGOX gene had a signal peptide containing 17 amino acids, three cysteine residues and six potential N-linked glycosylation sites, among them, -N-K-T-Y- at 41 amino acid, -N-R-S-L- at 113 amino acid, -N-G-T-I- at 192 amino acid, -N-T-T-A at 215 amino acid, -N-F-T-E at 373 amino acid and -N-V-T-A- at 408 amino acid were the most possible N-glycosylation sites. Furthermore, the relative transcription level of the PVGOX gene was also stimulated in the presence of 4% (w/v) of calcium carbonate and 0.5 % (v/v) of CSL in the production medium compared with that of the PVGOX gene when the fungal strain F1 was grown in the absence of calcium carbonate and CSL in the production medium, suggesting that under the optimal conditions, the expression of the PVGOX gene responsible for gluconic acid biosynthesis was enhanced, leading to increased gluconic acid production. Therefore, the highly glycosylated oxidase enzyme produced by P. viticola F1 strain might be a good producer in the fermentation process for the industrial level production of gluconic acid.

  12. Transcriptomic analysis of the interaction between Helianthus annuus and its obligate parasite Plasmopara halstedii shows single nucleotide polymorphisms in CRN sequences

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    Gouzy Jérôme

    2011-10-01

    Full Text Available Abstract Background Downy mildew in sunflowers (Helianthus annuus L. is caused by the oomycete Plasmopara halstedii (Farl. Berlese et de Toni. Despite efforts by the international community to breed mildew-resistant varieties, downy mildew remains a major threat to the sunflower crop. Very few genomic, genetic and molecular resources are currently available to study this pathogen. Using a 454 sequencing method, expressed sequence tags (EST during the interaction between H. annuus and P. halstedii have been generated and a search was performed for sites in putative effectors to show polymorphisms between the different races of P. halstedii. Results A 454 pyrosequencing run of two infected sunflower samples (inbred lines XRQ and PSC8 infected with race 710 of P. halstedii, which exhibit incompatible and compatible interactions, respectively generated 113,720 and 172,107 useable reads. From these reads, 44,948 contigs and singletons have been produced. A bioinformatic portal, HP, was specifically created for in-depth analysis of these clusters. Using in silico filtering, 405 clusters were defined as being specific to oomycetes, and 172 were defined as non-specific oomycete clusters. A subset of these two categories was checked using PCR amplification, and 86% of the tested clusters were validated. Twenty putative RXLR and CRN effectors were detected using PSI-BLAST. Using corresponding sequences from four races (100, 304, 703 and 710, 22 SNPs were detected, providing new information on pathogen polymorphisms. Conclusions This study identified a large number of genes that are expressed during H. annuus/P. halstedii compatible or incompatible interactions. It also reveals, for the first time, that an infection mechanism exists in P. halstedii similar to that in other oomycetes associated with the presence of putative RXLR and CRN effectors. SNPs discovered in CRN effector sequences were used to determine the genetic distances between the four races

  13. Reação de clones de videira a Xanthomonas campestris pv. viticola, baseada nos componentes epidemiológicos do cancro bacteriano

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    Nascimento Ana Rosa Peixoto

    2006-01-01

    Full Text Available O cancro bacteriano, causado por Xanthomonas campestris pv. viticola (Xcv, é a doença bacteriana mais importante da videira na região do Submédio São Francisco. A reação de 20 clones de videira, sendo 13 de copa e sete de porta-enxerto, foi avaliada quanto à resistência ao patógeno, em casa de vegetação. As plantas foram inoculadas com a suspensão do isolado Xcv1 (A570=108UFC mL-1, incubadas em casa de vegetação e observadas diariamente quanto aos componentes epidemiológicos do cancro bacteriano: período de incubação, incidência de folhas com sintomas, incidência de folhas com cancro, severidade da doença, taxa de progresso da incidência da doença, área abaixo da curva de progresso da severidade da doença. Todos os clones foram suscetíveis ao patógeno, embora diferindo significativamente entre si (P=0,05 para a maioria das variáveis analisadas. Em geral, 'Brasil' apresentou os maiores níveis de doença para todas as variáveis testadas, enquanto'Isabel' e 'Paulsen 1103' destacaram-se ao propiciarem os maiores valores de período de incubação e os menores valores de incidência de folhas com sintomas, com cancros, severidade da doença, taxa de progresso de incidência da doença e área abaixo da curva de progresso da severidade da doença, indicando grande potencial de utilização em programas de melhoramento genético e manejo integrado. As correlações significativas (P=0,05 verificadas entre as variáveis estudadas indicam que qualquer delas pode ser utilizada em pesquisas envolvendo reação de clones ao cancro bacteriano da videira. Quando considerado o conjunto dos componentes epidemiológicos, o agrupamento pelo método UPGMA (agrupamento aos pares pela média aritmética não ponderada permitiu a separação dos clones de copa e porta-enxerto em três grupos de similaridade cada.

  14. Detecção e variabilidade de Plasmopara halstedii no Brasil e avaliação da resistência de genótipos de girassol ao míldio Detection and variability of Plasmopara halstedii in Brazil and resistance of sunflower genotypes to downy mildew

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    Regina Maria Villas Bôas de Campos Leite

    2007-12-01

    Full Text Available Este trabalho foi conduzido com o objetivo de identificar a raça fisiológica de Plasmopara halstedii que ocorreu em plantas de girassol coletadas no campo experimental da Embrapa Soja, Londrina, PR, em 1998, 2001 e 2002 e avaliar a reação de genótipos de girassol ao míldio. Plântulas de girassol das diferenciadoras de raças e das cultivares foram inoculadas com suspensão de zoosporângios do patógeno e foram plantadas em caixas contendo areia autoclavada. As plântulas foram mantidas em câmara climatizada, com temperatura controlada em 21ºC, por 11 dias. Em seguida, as plantas foram aspergidas intensamente com água destilada, cobertas com saco plástico e mantidas no escuro, a 18ºC. No dia seguinte, foi observada a presença de esporulação nos cotilédones. As plantas que apresentaram esporulação foram consideradas suscetíveis e as sem esporulação foram resistentes. O resultado indicou tratar-se da raça 330 (antiga raça 7 americana, nas três ocasiões. Os genótipos de girassol Embrapa 122, BRS 191 e as cultivares de girassol ornamental BRS Capri M, BRS Encanto M, BRS Oásis, BRS Paixão M, BRS Pesqueiro M, BRS Refúgio M, BRS Saudade M e BRS Saudade U e seus respectivos parentais foram suscetíveis a P. halstedii raça 330. Os genótipos AGROBEL 910, AGROBEL 920, AGROBEL 960, AGROBEL 965, C11, EXP38, M734, M742 e RUMBOSOL 91 foram resistentes à raça 330 do patógeno e podem ser indicados aos agricultores para uso em regiões de risco de ocorrência da doença.This research was carried out for identifying the physiological race of Plasmopara halstedii occurring in sunflower at the experimental field of Embrapa Soybean, Londrina, PR, Brazil, in 1998, 2001 and 2002 by evaluating the reaction of sunflower genotypes inoculated with downy mildew pathogen. Sunflower seedlings of the differentials set to identify races and of the cultivars were inoculated by immersion in zoosporangia suspension and were grown in autoclaved

  15. REPCO 501452 : Replacement of copper fungicides in organic production of grapevine and apple in Europe : publishable final activity report

    NARCIS (Netherlands)

    Köhl, J.

    2007-01-01

    Final Activity Report: The objective of REPCO was to contribute to the replacement of copper fungicides in organic agriculture by new measures for control of downy mildew (Plasmopara viticola) in grapevine and scab (Venturia inaequalis) in apple. Both major European organic crops strongly depend on

  16. Contributions of EU-project REPCO to apple scab control

    NARCIS (Netherlands)

    Köhl, J.; Heijne, B.; Hockenhull, J.; Lindhard Pedersen, H.; Trapman, M.; Eiben, U.; Tamm, L.

    2006-01-01

    The objective of REPCO is to contribute to the replacement of copper fungicides in organic agriculture by new measures for control of downy mildew (Plasmopara viticola) in grapevine and scab (Venturia inaequalis) in apple. Both major European organic crops strongly depend on copper fungicides. Permi

  17. Macro- and microscopic leaf characteristics of six grapevine genotypes (Vitis spp.) with different susceptibilities to grapevine downy mildew

    OpenAIRE

    Boso Alonso, Susana; Alonso-Villaverde Iglesias, Virginia; Santiago Blanco, José Luis; Gago Montaña, Pilar; Dürrenberger, M.; Düggelin, M.; Kassemeyer, H. H.; Martínez Rodríguez, María del Carmen

    2010-01-01

    This work reports the leaf morphology of six grapevine genotypes, five belonging to Vitis vinifera and one to Vitis riparia. Earlier studies on these genotypes showed different levels of susceptibility to grapevine downy mildew (Plasmopara viticola). The aim of this work was to detect differences between the leaf morphology of these cultivars at the macro- and microscopic levels, and to characterize morphological traits which could be associated with susceptibility and resistance to downy ...

  18. Effects of Some Plant Essential Oils Against Botrytis cinerea and Tetranychus urticae on Grapevine

    OpenAIRE

    DOĞU, Duygu MERMER; ZOBAR, Damla

    2014-01-01

    The cultivation of grapes in the world in terms of the first-ranked Turkey as well as the appropriate climatic zone, has potential for rich gene and ancient viticulture culture. Grape growers, starting from production until it reaches the consumer is faced with various problems in the process. Plant protection faced in the vineyards of the manufacturers in terms of the most important causes of losses; Downy Mildew (Plasmopara viticola), Powdery Mildew (Uncinula necator), Gray mold (Botrytis c...

  19. REPCO 501452 : Replacement of copper fungicides in organic production of grapevine and apple in Europe : publishable final activity report

    OpenAIRE

    Köhl, J

    2007-01-01

    Final Activity Report: The objective of REPCO was to contribute to the replacement of copper fungicides in organic agriculture by new measures for control of downy mildew (Plasmopara viticola) in grapevine and scab (Venturia inaequalis) in apple. Both major European organic crops strongly depend on copper fungicides. Permitted amounts will be reduced stepwise in the years ahead (Council Regulation (EEC) 2092/91, Annex II) to avoid environmental risks. In European countries where copper fungic...

  20. Components of variance and heritability of resistance to important fungal diseases agents in grapevine

    Directory of Open Access Journals (Sweden)

    Nikolić Dragan

    2006-01-01

    Full Text Available In four interspecies crossing combinations of grapevine (Seedling 108 x Muscat Hamburg, Muscat Hamburg x Seedling 108, S.V.I8315 x Muscat Hamburg and Muscat Hamburg x S.V.I2375 during three years period, resistance to important fungal diseases agents (Plasmopara viticola and Botrytis cinerea were examined. Based on results of analysis of variance, for investigated characteristics, components of variance, coefficients of genetic and phenotypic variation and coefficient of heritability in a broader sense were calculated. It was established that for both characteristics and in all crossing combinations, genetic variance took the biggest part in total variability. The lowest coefficients of genetic and phenotypic variation were established for both properties in crossing combination Seedling 108 x Muscat Hamburg. The highest coefficients of genetic and phenotypic variation were determined for leaf resistance to Plasmopara viticola in crossing combination Muscat Hamburg x S.V.I2375, and for bunch resistance to Botrytis cinerea in crossing combination Muscat Hamburg x Seedling 108. Considering all investigated crossing combinations, coefficient of heritability for leaf resistance to Plasmopara viticola was from 87.23% to 94.88%, and for bunch resistance to Botrytis cinerea from 88.04% to 93.32%. .

  1. Generation of Antifungal Stilbenes Using the Enzymatic Secretome of Botrytis cinerea.

    Science.gov (United States)

    Gindro, Katia; Schnee, Sylvain; Righi, Davide; Marcourt, Laurence; Nejad Ebrahimi, Samad; Codina, Josep Massana; Voinesco, Francine; Michellod, Emilie; Wolfender, Jean-Luc; Queiroz, Emerson Ferreira

    2017-04-28

    The protein secretome of Botrytis cinerea was used to perform the biotransformation of resveratrol, pterostilbene, and a mixture of both. Metabolite profiling by UHPLC-HRMS revealed the presence of compounds with unusual molecular formula, suggesting the existence of new products. To isolate these products, the reactions were scaled-up, and 21 analogues were isolated and fully characterized by NMR and HRESIMS analyses. The reaction with pterostilbene afforded five new compounds, while the reaction with a mixture of pterostilbene and resveratrol afforded seven unusual stilbene dimers. The antifungal properties of these compounds were evaluated using in vitro bioassays against Plasmopara viticola. The cytological effects of the isolated antifungal compounds on the ultrastructure of P. viticola were also evaluated.

  2. The Influence of the Climatic Factors on the Main Vineyard Diseases When Organic Products Are Used in Fetească Regală Variety

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    MIHAI - OROIAN C-tin

    2010-12-01

    Full Text Available This paper presents the main results of a study undertaken during 2005 - 2007 in Blaj Center vineyard in whichwe followed the evolution of pathogens Plasmopara viticola, Uncinula Necator and Botryitis cinerea in wine grapevariety Fetească Royal, in conjunction with climatic and treatment options without organic synthesis. There was a verybig influence of climatic conditions, in particular precipitation and relative air humidity that have an uneven distributionduring the growing season, which causes fungal diseases in each year of research. Research conducted during the threeexperimental years shows that the largest share in total degree assault had gray mold which met favorable conditions inthis area.

  3. The role of oospores in the epidemiology of potato late blight

    NARCIS (Netherlands)

    Kessel, G.J.T.; Andersson, B.; Widmark, A.K.; Yuen, J.E.; Evenhuis, A.; Turkensteen, L.J.; Lehtinen, A.; Nielsen, B.; Ravnskov, S.; Hansen, J.G.; Hermansen, A.; Brurberg, M.B.; Nordskog, B.

    2009-01-01

    Potato late blight (Phytophthora infestans) is a plant disease feared globally by farmers and the potato industry. P. infestans is a heterothallic oomycete with two mating types. Until recently the pathogen was limited to surviving between seasons as living mycelia in its host plant in most parts of

  4. Resistance induced component of management of diseases of grapevine

    Directory of Open Access Journals (Sweden)

    Jusciélio Barbosa

    2009-08-01

    Full Text Available The use of induced resistance presents as a viable alternative in the management of diseases of the vine. Accordingly, the objective of this study was to evaluate the efficiency of Agro Mos® and potassium phosphite in controlling diseases of grapevine under field conditions in the Valley San Francisco. O test was conducted under field conditions in the experimental area IFSertão Pernambucano, Petrolina, PE, using the cultivar Petit Sirah. The experimental design was in randomized blocks, composed of five treatments and five replicates: T1 - control; T2 - Cabrio Top® - CT (2kg ha-1; T3 - Agro Mos® - AM (3mL L-1; T4 - Fosfito de potássio - FP (4mL L-1; T5 - Agro Mos® - AM (3mL L-1 interleaved with the fungicide Cabrio Top® - CT (2kg ha-1. Each plot consisted of eight plants. Data were subjected to analysis of variance and averages compared by Tukey test at 5%. Conditions in which the experiment was developed, the use of potassium phosphite and Agro-Mos® promoted a significant reduction in the incidence of Plasmopara viticola and Uncinula necator.Key-words: resistance induced, Plasmopara viticola, Uncinula necator.

  5. Inhibitory effects of macrotetrolides from Streptomyces spp. on zoosporogenesis and motility of peronosporomycete zoospores are likely linked with enhanced ATPase activity in mitochondria

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    Md Tofazzal Islam

    2016-11-01

    Full Text Available The release of zoospores from sporangia and motility of the released zoospores are critical in the disease cycle of the Peronosporomycetes that cause devastating diseases in plants, fishes, animals and humans. Disruption of any of these asexual life stages eliminates the possibility of pathogenesis. In the course of screening novel bioactive secondary metabolites, we found that extracts of some strains of marine Streptomyces spp. rapidly impaired motility and caused subsequent lysis of zoospores of the grapevine downy mildew pathogen Plasmopara viticola at 10 g/ml. We tested a number of secondary metabolites previously isolated from these strains and found that macrotetrolide antibiotics such as nonactin, monactin, dinactin and trinactin, and nactic acids such as (+-nonactic acid, (+-homonactic acid, nonactic acid methyl ester, homonactic acid methyl ester, bonactin and feigrisolide C impaired motility and caused subsequent lysis of P. viticola zoospores in a dose- and time-dependent manners with dinactin being the most active compound (MIC 0.3 g/ml. A cation channel-forming compound, gramicidin, and a carrier of monovalent cations, nigericin also showed similar biological activities. Among all 12 compounds tested, gramicidin most potently arrested the motility of zoospores at concentrations starting from 0.1 g/ml. All macrotetrolide antibiotics also displayed similar motility impairing activities against P. viticola, Phytophthora capsici and Aphanomyces cochlioides zoospores indicating non-specific biological effects of these compounds towards peronosporomyctes. Furthermore, macrotetrolide antibiotics and gramicidin also markedly suppressed the release of zoospores from sporangia of P. viticola in a dose-dependent manner. As macrotetrolide antibiotics and gramicidin are known as enhancers of mitochondrial ATPase activity, inhibition of zoosporogenesis and motility of zoospores by these compounds are likely linked with hydrolysis of ATP

  6. Downy mildew resistance induced by Trichoderma harzianum T39 in susceptible grapevines partially mimics transcriptional changes of resistant genotypes

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    Perazzolli Michele

    2012-11-01

    Full Text Available Abstract Background Downy mildew, caused by Plasmopara viticola, is one of the most severe diseases of grapevine and is commonly controlled by fungicide treatments. The beneficial microorganism Trichoderma harzianum T39 (T39 can induce resistance to downy mildew, although the molecular events associated with this process have not yet been elucidated in grapevine. A next generation RNA sequencing (RNA-Seq approach was used to study global transcriptional changes associated with resistance induced by T39 in Vitis vinifera Pinot Noir leaves. The long-term aim was to develop strategies to optimize the use of this agent for downy mildew control. Results More than 14.8 million paired-end reads were obtained for each biological replicate of T39-treated and control leaf samples collected before and 24 h after P. viticola inoculation. RNA-Seq analysis resulted in the identification of 7,024 differentially expressed genes, highlighting the complex transcriptional reprogramming of grapevine leaves during resistance induction and in response to pathogen inoculation. Our data show that T39 has a dual effect: it directly modulates genes related to the microbial recognition machinery, and it enhances the expression of defence-related processes after pathogen inoculation. Whereas several genes were commonly affected by P. viticola in control and T39-treated plants, opposing modulation of genes related to responses to stress and protein metabolism was found. T39-induced resistance partially inhibited some disease-related processes and specifically activated defence responses after P. viticola inoculation, causing a significant reduction of downy mildew symptoms. Conclusions The global transcriptional analysis revealed that defence processes known to be implicated in the reaction of resistant genotypes to downy mildew were partially activated by T39-induced resistance in susceptible grapevines. Genes identified in this work are an important source of markers

  7. Breakdown of resistance to grapevine downy mildew upon limited deployment of a resistant variety

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    Bellin Diana

    2010-07-01

    Full Text Available Abstract Background Natural disease resistance is a cost-effective and environmentally friendly way of controlling plant disease. Breeding programmes need to make sure that the resistance deployed is effective and durable. Grapevine downy mildew, caused by the Oomycete Plasmopara viticola, affects viticulture and it is controlled with pesticides. Downy mildew resistant grapevine varieties are a promising strategy to control the disease, but their use is currently restricted to very limited acreages. The arising of resistance-breaking isolates under such restricted deployment of resistant varieties would provide valuable information to design breeding strategies for the deployment of resistance genes over large acreages whilst reducing the risks of the resistance being defeated. The observation of heavy downy mildew symptoms on a plant of the resistant variety Bianca, whose resistance is conferred by a major gene, provided us with a putative example of emergence of a resistance-breaking isolate in the interaction between grapevine and P. viticola. Results In this paper we describe the emergence of a P. viticola isolate (isolate SL that specifically overcomes Rpv3, the major resistance gene carried by Bianca at chromosome 18. We show that isolate SL has the same behaviour as two P. viticola isolates avirulent on Bianca (isolates SC and SU when inoculated on susceptible plants or on resistant plants carrying resistances derived from other sources, suggesting there is no fitness cost associated to the virulence. Molecular analysis shows that all three isolates are genetically closely related. Conclusions Our results are the first description of a resistance-breaking isolate in the grapevine/P. viticola interaction, and show that, despite the reduced genetic variability of P. viticola in Europe compared to its basin of origin and the restricted use of natural resistance in European viticulture, resistance-breaking isolates overcoming monogenic

  8. 新型杀菌剂苯噻菌胺%A New Fungicide,Benthiavalicarb

    Institute of Scientific and Technical Information of China (English)

    陈启辉

    2004-01-01

    苯噻菌胺(benthiavalicarb-isopropyl)是一种新型杀菌剂,对多种作物的卵菌纲病菌都有很好的活性,以较低的剂量(25~75g a.i./hm2)能够有效地控制马铃薯和番茄的晚疫病(Phytophthora infestans)、葡萄霜霉病(Plasmopara viticola)、瓜类霜霉病(Pseudoperonospora cubensis)、寄生霜霉病(Peronospora parasitica).苯噻菌胺具有较好的毒理性和环保性,对许多蔬菜和水果无毒害.同时介绍了该品种的理化性质、毒性、作用机理、合成方法等.

  9. Toward the Identification of Two Glycoproteins Involved in the Stomatal Deregulation of Downy Mildew-Infected Grapevine Leaves.

    Science.gov (United States)

    Guillier, Christelle; Gamm, Magdalena; Lucchi, Géraldine; Truntzer, Caroline; Pecqueur, Delphine; Ducoroy, Patrick; Adrian, Marielle; Héloir, Marie-Claire

    2015-11-01

    Stomata remain abnormally opened and unresponsive to abscisic acid in grapevine leaves infected by downy mildew. This deregulation occurs from 3 days postinoculation and increases concomitantly with leaf colonization by the pathogen. Using epidermal peels, we demonstrated that the active compound involved in this deregulation is located in the apoplast. Biochemical assays showed that the active compound present in the apoplastic fluids isolated from Plasmopara viticola-infected grapevine leaves (IAF) is a CysCys bridge-independent, thermostable and glycosylated protein. Fractionation guided assays based on chromatography coupled to stomatal response and proteomic analysis allowed the identification of both plant and pathogen proteins in the active fraction obtained from IAF. Further in silico analysis and discriminant filtrations based on the comparison between predictions and experimental indications lead to the identification of two Vitis vinifera proteins as candidates for the observed stomatal deregulation.

  10. DINÂMICA DE INÓCULOS E DOENÇAS EM VIDEIRA SOB SISTEMA CONVENCIONAL E PROTEGIDO

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    DIÓGENES DA CRUZ BATISTA

    2015-01-01

    Full Text Available The effect of the covered and conventional cultivation of grapevine (cv. Superior Seedless on the aerial dispersion of pathogen inoculum and dynamic of diseases was investigated in two-year (2009 and 2010 and period similar to the year. The experiment was conducted at the experimental station of Embrapa semiarid. The traps were maintained at 0.5 meters above the canopy of the grapevine, and the samples taken every 24 hours with the exchange of slides 9:00 a.m. Greater dispersion of pathogens (Plasmopara viticola, Phakopsora euvitis, Alternaria alternata and intensity of diseases (downy mildew, rust, and bacterial canker were detected in grapevines under conventional cultivation. The aerial dispersal of the fungus Lasiodiplodia theobromae was similar under grapevine covered and uncovered.

  11. Genetic dissection of a TIR-NB-LRR locus from the wild North American grapevine species Muscadinia rotundifolia identifies paralogous genes conferring resistance to major fungal and oomycete pathogens in cultivated grapevine.

    Science.gov (United States)

    Feechan, Angela; Anderson, Claire; Torregrosa, Laurent; Jermakow, Angelica; Mestre, Pere; Wiedemann-Merdinoglu, Sabine; Merdinoglu, Didier; Walker, Amanda R; Cadle-Davidson, Lance; Reisch, Bruce; Aubourg, Sebastien; Bentahar, Nadia; Shrestha, Bipna; Bouquet, Alain; Adam-Blondon, Anne-Françoise; Thomas, Mark R; Dry, Ian B

    2013-11-01

    The most economically important diseases of grapevine cultivation worldwide are caused by the fungal pathogen powdery mildew (Erysiphe necator syn. Uncinula necator) and the oomycete pathogen downy mildew (Plasmopara viticola). Currently, grapegrowers rely heavily on the use of agrochemicals to minimize the potentially devastating impact of these pathogens on grape yield and quality. The wild North American grapevine species Muscadinia rotundifolia was recognized as early as 1889 to be resistant to both powdery and downy mildew. We have now mapped resistance to these two mildew pathogens in M. rotundifolia to a single locus on chromosome 12 that contains a family of seven TIR-NB-LRR genes. We further demonstrate that two highly homologous (86% amino acid identity) members of this gene family confer strong resistance to these unrelated pathogens following genetic transformation into susceptible Vitis vinifera winegrape cultivars. These two genes, designated resistance to Uncinula necator (MrRUN1) and resistance to Plasmopara viticola (MrRPV1) are the first resistance genes to be cloned from a grapevine species. Both MrRUN1 and MrRPV1 were found to confer resistance to multiple powdery and downy mildew isolates from France, North America and Australia; however, a single powdery mildew isolate collected from the south-eastern region of North America, to which M. rotundifolia is native, was capable of breaking MrRUN1-mediated resistance. Comparisons of gene organization and coding sequences between M. rotundifolia and the cultivated grapevine V. vinifera at the MrRUN1/MrRPV1 locus revealed a high level of synteny, suggesting that the TIR-NB-LRR genes at this locus share a common ancestor. © 2013 The Authors The Plant Journal © 2013 John Wiley & Sons Ltd.

  12. Prediction of biological sensors appearance with ARIMA models as a tool for Integrated Pest Management protocols

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    María Fernández-González

    2015-12-01

    Full Text Available [b]Introduction and objectives.[/b] Powdery mildew caused by [i]Uncinula necator[/i] and Downy mildew produced by [i]Plasmopara viticola[/i] are the most common diseases in the North-West Spain vineyards. Knowledge of airborne spore concentrations could be a useful tool in the Integrated Pest Management protocols in order to reduce the number of pesticide treatments, applied only when there is a real risk of infection. [b]Material and methods. [/b]The study was carried out in a vineyard of the D. O. Ribeiro, in the North-West Spain, during the grapevine active period 2004–2012. A Hirts-type volumetric spore-trap was used for the aerobiological monitoring. [b]Results.[/b] During the study period the annual total [i]U. necator[/i] spores amount ranged from the 578 spores registered in 2007 to the 4,145 spores sampled during 2008. The highest annual total [i]P. viticola[/i] spores quantity was observed in 2010 (1,548 spores and the lowest in 2005 (210 spores. In order to forecast the concentration of fungal spores, ARIMA models were elaborated. [b]Conclusions[/b]. The most accurate models were an ARIMA (3.1.3 for [i]U. necator[/i] and (1.0.3 for [i]P. viticola[/i]. The possibility to forecast the spore presence 72 hours in advance open an important horizon for optimizing the organization of the harvest processes in the vineyard.

  13. General and species-specific transcriptional responses to downy mildew infection in a susceptible (Vitis vinifera and a resistant (V. riparia grapevine species

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    Lovato Arianna

    2010-02-01

    Full Text Available Abstract Background Downy mildew is a destructive grapevine disease caused by Plasmopara viticola (Berk. and Curt. Berl. and de Toni, which can only be controlled by intensive fungicide treatments. Natural sources of resistance from wild grapevine (Vitis species are used in conventional breeding approaches, but the signals and effectors involved in resistance in this important crop species are not well understood. Results Early transcriptional changes associated with P. viticola infection in susceptible V. vinifera and resistant V. riparia plants were analyzed using the Combimatrix microarray platform. Transcript levels were measured 12 and 24 h post-inoculation, reflecting the time points immediately preceding the onset of resistance in V. riparia, as determined by microscopic analysis. Our data indicate that resistance in V. riparia is induced after infection, and is not based on differences in basal gene expression between the two species. The strong and rapid transcriptional reprogramming involves the induction of pathogenesis-related proteins and enzymes required for the synthesis of phenylpropanoid-derived compounds, many of which are also induced, albeit to a lesser extent, in V. vinifera. More interestingly, resistance in V. riparia also involves the specific modulation of numerous transcripts encoding components of signal transduction cascades, hypersensitive reaction markers and genes involved in jasmonate biosynthesis. The limited transcriptional modulation in V. vinifera represents a weak attempted defense response rather than the activation of compatibility-specific pathways. Conclusions Several candidate resistance genes were identified that could be exploited in future biotechnological approaches to increase disease resistance in susceptible grapevine species. Measurements of jasmonic acid and methyl jasmonate in infected leaves suggest that this hormone may also be involved in V. riparia resistance to P. viticola.

  14. General and species-specific transcriptional responses to downy mildew infection in a susceptible (Vitis vinifera) and a resistant (V. riparia) grapevine species

    Science.gov (United States)

    2010-01-01

    Background Downy mildew is a destructive grapevine disease caused by Plasmopara viticola (Berk. and Curt.) Berl. and de Toni, which can only be controlled by intensive fungicide treatments. Natural sources of resistance from wild grapevine (Vitis) species are used in conventional breeding approaches, but the signals and effectors involved in resistance in this important crop species are not well understood. Results Early transcriptional changes associated with P. viticola infection in susceptible V. vinifera and resistant V. riparia plants were analyzed using the Combimatrix microarray platform. Transcript levels were measured 12 and 24 h post-inoculation, reflecting the time points immediately preceding the onset of resistance in V. riparia, as determined by microscopic analysis. Our data indicate that resistance in V. riparia is induced after infection, and is not based on differences in basal gene expression between the two species. The strong and rapid transcriptional reprogramming involves the induction of pathogenesis-related proteins and enzymes required for the synthesis of phenylpropanoid-derived compounds, many of which are also induced, albeit to a lesser extent, in V. vinifera. More interestingly, resistance in V. riparia also involves the specific modulation of numerous transcripts encoding components of signal transduction cascades, hypersensitive reaction markers and genes involved in jasmonate biosynthesis. The limited transcriptional modulation in V. vinifera represents a weak attempted defense response rather than the activation of compatibility-specific pathways. Conclusions Several candidate resistance genes were identified that could be exploited in future biotechnological approaches to increase disease resistance in susceptible grapevine species. Measurements of jasmonic acid and methyl jasmonate in infected leaves suggest that this hormone may also be involved in V. riparia resistance to P. viticola. PMID:20167053

  15. Microscopic observations on oospore germination of Phytophthora sojae%大豆疫病卵孢子萌发的显微观察

    Institute of Scientific and Technical Information of China (English)

    罗加风; 黄国明

    2001-01-01

    @@ 大豆疫病Phytophthora sojae Kaufmann&Gerdemann作为典型的土传真菌病害,随病土远距离传播的卵孢子是病菌扩散的主要来源.国内外许多专家都认为该病不能种传,周肇蕙和严进发现:通过人工接种大豆疫霉可以得到有卵孢子的病种子,认为该病可以种传[1~2],但种子中卵孢子的活性测定仍未解决.

  16. Análise da diversidade genética por AFLP e identificação de marcadores associados à resistência a doenças em videira Genetic diversity and identification of AFLP markers associated with diseases resistance in grapevine

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    Paulo Ricardo Dias de Oliveira

    2005-12-01

    Full Text Available Com objetivo de estudar diversidade genética e identificar marcadores associados à resistência ao míldio (Plasmopara viticola e ao oídio (Uncinula necator, foram analisadas as cultivares de videira A 1976, CG 87746, CNPUV 154-27, Crimson Seedless, Gota de Ouro, Itália, Seyve Villard 12327 e Seyve Villard 12375. As análises de polimorfismo de comprimento de fragmento amplificado (AFLP seguiram as etapas de digestão, ligação, pré-amplificação e amplificação. Efetuou-se a separação dos fragmentos amplificados em gel de 7% de poliacrilamida desnaturante (uréia 7M e coloração com nitrato de prata. A similaridade foi estimada com base no coeficiente de Jaccard, e a agregação, por UPGMA. Foi gerada uma matriz de similaridade com bom ajustamento da agregação (r = 0,84. Os agrupamentos obtidos corresponderam à origem e classificação botânica das cultivares. Foram identificadas 15 marcas dissimilares associadas à resistência, sendo oito para míldio e sete para oídio.The grapevine cultivars A 1976, CG 87746, CNPUV 154-27, Crimson Seedless, Gota de Ouro, Itália, Seyve Villard 12327 and Seyve Villard 12375 were analyzed to establish the genetic relationship and to identify markers associated with resistance to downy mildew (Plasmopara viticola and powdery mildew (Uncinula necator. The amplified fragment lenght polymorphism (AFLP analysis comprise the steps of digestion, ligation, preamplification and amplification. The amplified fragments were separeted on a 7% denaturing polyacrylamide gel (7M urea and silver stained. The similarity was estimated using Jaccard's coefficient and the clustering was estimated by UPGMA. A similarity matrix was generated with a good fit for aggregation (r = 0,84. The clusters corresponded to origin and botany classification of the cultivars. Fifteen markers were associated with resistance, eight for downy mildew and seven for powdery mildew.

  17. Extrato de alho e óleo vegetal no controle do míldio da videira

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    Carla Daiane Leite

    2011-06-01

    Full Text Available O objetivo deste trabalho foi verificar o efeito do extrato de alho e do óleo vegetal no controle do míldio da videira cv. Isabel (Vitis labrusca. A severidade da doença, expressa pela área abaixo da curva de progresso da doença (AACPD, e a germinação de esporângios de seu agente causal Plasmopara viticola foram as variáveis avaliadas. Os tratamentos consistiram em 0; 5; 10; 15; 20; 25 ou 30 mL L-1 de extrato de alho adicionados de 2,5 mL L-1 óleo vegetal, calda bordalesa (1:1:100 e testemunha (sem tratamento. No teste de germinação, utilizou-se mancozebe (2 g L-1 como tratamento-padrão. Em condições de campo, observou-se redução da severidade do míldio com o óleo vegetal, sendo que o extrato de alho, a partir de 20 mL L-1, potencializou tal ação biocida. A germinação dos esporângios de P. viticola variou em função do tempo de exposição ao extrato de alho, não apresentando boa eficiência quando comparada ao tratamento com calda bordalesa e mancozebe. O óleo vegetal não influenciou na germinação dos esporângios desse patógeno.

  18. Cytochrome b gene structure and consequences for resistance to Qo inhibitor fungicides in plant pathogens.

    Science.gov (United States)

    Grasso, Valeria; Palermo, Simona; Sierotzki, Helge; Garibaldi, Angelo; Gisi, Ulrich

    2006-06-01

    The cytochrome b (cyt b) gene structure was characterized for different agronomically important plant pathogens, such as Puccinia recondita f sp tritici (Erikss) CO Johnston, P graminis f sp tritici Erikss and Hennings, P striiformis f sp tritici Erikss, P coronata f sp avenae P Syd & Syd, P hordei GH Otth, P recondita f sp secalis Roberge, P sorghi Schwein, P horiana Henn, Uromyces appendiculatus (Pers) Unger, Phakopsora pachyrhizi Syd & P Syd, Hemileia vastatrix Berk & Broome, Alternaria solani Sorauer, A alternata (Fr) Keissl and Plasmopara viticola (Berk & Curt) Berlese & de Toni. The sequenced fragment included the two hot spot regions in which mutations conferring resistance to QoI fungicides may occur. The cyt b gene structure of these pathogens was compared with that of other species from public databases, including the strobilurin-producing fungus Mycena galopoda (Pers) P Kumm, Saccharomyces cerevisiae Meyer ex Hansen, Venturia inaequalis (Cooke) Winter and Mycosphaerella fijiensis Morelet. In all rust species, as well as in A solani, resistance to QoI fungicides caused by the mutation G143A has never been reported. A type I intron was observed directly after the codon for glycine at position 143 in these species. This intron was absent in pathogens such as A alternata, Blumeria graminis (DC) Speer, Pyricularia grisea Sacc, Mycosphaerella graminicola (Fuckel) J Schröt, M fijiensis, V inaequalis and P viticola, in which resistance to QoI fungicides has occurred and the glycine is replaced by alanine at position 143 in the resistant genotype. The present authors predict that a nucleotide substitution in codon 143 would prevent splicing of the intron, leading to a deficient cytochrome b, which is lethal. As a consequence, the evolution of resistance to QoI fungicides based on G143A is not likely to evolve in pathogens carrying an intron directly after this codon.

  19. Historical introgression of the downy mildew resistance gene Rpv12 from the Asian species Vitis amurensis into grapevine varieties.

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    Silvia Venuti

    Full Text Available The Amur grape (Vitis amurensis Rupr. thrives naturally in cool climates of Northeast Asia. Resistance against the introduced pathogen Plasmopara viticola is common among wild ecotypes that were propagated from Manchuria into Chinese vineyards or collected by Soviet botanists in Siberia, and used for the introgression of resistance into wine grapes (Vitis vinifera L.. A QTL analysis revealed a dominant gene Rpv12 that explained 79% of the phenotypic variance for downy mildew resistance and was inherited independently of other resistance genes. A Mendelian component of resistance-a hypersensitive response in leaves challenged with P. viticola-was mapped in an interval of 0.2 cM containing an array of coiled-coil NB-LRR genes on chromosome 14. We sequenced 10-kb genic regions in the Rpv12(+ haplotype and identified polymorphisms in 12 varieties of V. vinifera using next-generation sequencing. The combination of two SNPs in single-copy genes flanking the NB-LRR cluster distinguished the resistant haplotype from all others found in 200 accessions of V. vinifera, V. amurensis, and V. amurensis x V. vinifera crosses. The Rpv12(+ haplotype is shared by 15 varieties, the most ancestral of which are the century-old 'Zarja severa' and 'Michurinets'. Before this knowledge, the chromosome segment around Rpv12(+ became introgressed, shortened, and pyramided with another downy mildew resistance gene from North American grapevines (Rpv3 only by phenotypic selection. Rpv12(+ has an additive effect with Rpv3(+ to protect vines against natural infections, and confers foliar resistance to strains that are virulent on Rpv3(+ plants.

  20. Stepwise flow diagram for the development of formulations of non spore-forming bacteria against foliar pathogens: The case of Lysobacter capsici AZ78.

    Science.gov (United States)

    Segarra, Guillem; Puopolo, Gerardo; Giovannini, Oscar; Pertot, Ilaria

    2015-12-20

    The formulation is a significant step in biopesticide development and is an efficient way to obtain consistency in terms of biological control under field conditions. Nonetheless, there is still a lack of information regarding the processes needed to achieve efficient formulation of non spore-forming bacterial biological control agents. In response to this, we propose a flow diagram made up of six steps including selection of growth parameters, checking of minimum shelf life, selection of protective additives, checking that the additives have no adverse effects, validation of the additive mix under field conditions and choosing whether to use additives as co-formulants or tank mix additives. This diagram is intended to provide guidance and decision-making criteria for the formulation of non spore-forming bacterial biological control agents against foliar pathogens. The diagram was then validated by designing an efficient formulation for a Gram-negative bacterium, Lysobacter capsici AZ78, to control grapevine downy mildew caused by Plasmopara viticola. A harvest of 10(10)L. capsici AZ78cellsml(-1) was obtained in a bench top fermenter. The viability of cells decreased by only one order of magnitude after one year of storage at 4°C. The use of a combination of corn steep liquor, lignosulfonate, and polyethyleneglycol in the formulation improved the survival of L. capsici AZ78 cells living on grapevine leaves under field conditions by one order of magnitude. Furthermore, the use of these additives also guaranteed a reduction of 71% in P. viticola attacks. In conclusion, this work presents a straightforward stepwise flow diagram to help researchers develop formulations for biological control agents that are easy to prepare, stable, not phytotoxic and able to protect the microorganims under field conditions.

  1. Protein kinase C is likely to be involved in zoosporogenesis and maintenance of flagellar motility in the peronosporomycete zoospores.

    Science.gov (United States)

    Islam, Md Tofazzal; von Tiedemann, Andreas; Laatsch, Hartmut

    2011-08-01

    The motility of zoospores is critical in the disease cycles of Peronosporomycetes that cause devastating diseases in plants, fishes, vertebrates, and microbes. In the course of screening for secondary metabolites, we found that ethyl acetate extracts of a marine Streptomyces sp. strain B5136 rapidly impaired the motility of zoospores of the grapevine downy mildew pathogen Plasmopara viticola at 0.1 μg/ml. The active principle in the extracts was identified as staurosporine, a known broad-spectrum inhibitor of protein kinases, including protein kinase C (PKC). In the presence of staurosporine (2 nM), zoospores moved very slowly in their axis or spun in tight circles, instead of displaying straight swimming in a helical fashion. Compounds such as K-252a, K-252b, and K-252c structurally related to staurosporine also impaired the motility of zoospores in a similar manner but at varying doses. Among the 22 known kinase inhibitors tested, the PKC inhibitor chelerythrine was the most potent to arrest the motility of zoospores at concentrations starting from 5 nM. Inhibitors that targeted kinase pathways other than PKC pathways did not practically show any activity in impairing zoospore motility. Interestingly, both staurosporine (5 nM) and chelerythrine (10 nM) also inhibited the release of zoospores from the P. viticola sporangia in a dose-dependent manner. In addition, staurosporine completely suppressed downy mildew disease in grapevine leaves at 2 μM, suggesting the potential of small-molecule PKC inhibitors for the control of peronosporomycete phytopathogens. Taken together, these results suggest that PKC is likely to be a key signaling mediator associated with zoosporogenesis and the maintenance of flagellar motility in peronosporomycete zoospores.

  2. Overexpression of a stress-responsive U-box protein gene VaPUB affects the accumulation of resistance related proteins in Vitis vinifera 'Thompson Seedless'.

    Science.gov (United States)

    Jiao, Li; Zhang, Yali; Lu, Jiang

    2017-03-01

    Many U-box proteins have been identified and characterized as important factors against environmental stresses such as chilling, heat, salinity and pathogen attack in plant. Our previous research reported the cloning of a novel U-box protein gene VaPUB from Vitis amurensis 'Zuoshanyi' grape and suggested a function of it in related to cold stress in the model plant Arabidopsis system. In this study, the role of VaPUB in response to biotic and abiotic stress was further analyzed in the homologous grapevine system by studying the transcript regulation and the protein accumulation in VaPUB transgenic vines. The expression analysis assay shown that VaPUB was significantly up-regulated 6 h after cold treatment and as early as 2 h post inoculation with Plasmopara viticola, a pathogen causing downy mildew disease in grapevine. Over-expressing VaPUB in V. Vinifera 'Thompson Seedless' affected the microstructure of leaves. The proteome assay shown that the accumulation of pathogenesis-related protein PR10 and many proteins involved in carbon and energy metabolism, oxidation reaction and protein metabolism were significantly altered in transgenic vines. In comparison with wild type plants, the expression level of PR10 family genes was significantly decreased in VaPUB transgenic vines under P. viticola treatment or cold stress. Results from this study showed that the U-box protein gene PUB quickly responded to both biotic stress and abiotic stress and significantly influenced the accumulation of resistance related proteins in grapevine. Copyright © 2016. Published by Elsevier Masson SAS.

  3. Pea broth enhances the biocontrol efficacy of Lysobacter capsici AZ78 by triggering cell motility associated with biogenesis of type IV pilus

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    Selena Tomada

    2016-07-01

    Full Text Available Bacterial cells can display different types of motility, due to the presence of external appendages such as flagella and type IV pili. To date, little information on the mechanisms involved in the motility of the Lysobacter species has been available. Recently, L. capsici AZ78, a biocontrol agent of phytopathogenic oomycetes, showed the ability to move on jellified pea broth. Pea broth medium improved also the biocontrol activity of L. capsici AZ78 against Plasmopara viticola under greenhouse conditions. Noteworthy, the quantity of pea residues remaining on grapevine leaves fostered cell motility in L. capsici AZ78. Based on these results, this unusual motility related to the composition of the growth medium was investigated in bacterial strains belonging to several Lysobacter species. The six L. capsici strains tested developed dendrite-like colonies when grown on jellified pea broth, while the development of dendrite-like colonies was not recorded in the media commonly used in motility assays. To determine the presence of genes responsible for biogenesis of the flagellum and type IV pili, the genome of L. capsici AZ78 was mined. Genes encoding structural components an d regulatory factors of type IV pili were upregulated in L. capsici AZ78 cells grown on the above-mentioned medium, as compared with the other tested media. These results provide new insight into the motility mechanism of L. capsici members and the role of type IV pili and pea compounds on the epiphytic fitness and biocontrol features of L. capsici AZ78.

  4. Over-expression of VvWRKY1 in grapevines induces expression of jasmonic acid pathway-related genes and confers higher tolerance to the downy mildew.

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    Chloé Marchive

    Full Text Available Most WRKY transcription factors activate expression of defence genes in a salicylic acid- and/or jasmonic acid-dependent signalling pathway. We previously identified a WRKY gene, VvWRKY1, which is able to enhance tolerance to fungal pathogens when it is overexpressed in tobacco. The present work analyzes the effects of VvWRKY1 overexpression in grapevine. Microarray analysis showed that genes encoding defence-related proteins were up-regulated in the leaves of transgenic 35S::VvWRKY1 grapevines. Quantitative RT-PCR analysis confirmed that three genes putatively involved in jasmonic acid signalling pathway were overexpressed in the transgenic grapes. The ability of VvWRKY1 to trans-activate the promoters of these genes was demonstrated by transient expression in grape protoplasts. The resistance to the causal agent of downy mildew, Plasmopara viticola, was enhanced in the transgenic plants. These results show that VvWRKY1 can increase resistance of grapevine against the downy mildew through transcriptional reprogramming leading to activation of the jasmonic acid signalling pathway.

  5. Genome size determination in peronosporales (Oomycota) by Feulgen image analysis.

    Science.gov (United States)

    Voglmayr, H; Greilhuber, J

    1998-12-01

    Genome size was determined, by nuclear Feulgen staining and image analysis, in 46 accessions of 31 species of Peronosporales (Oomycota), including important plant pathogens such as Bremia lactucae, Plasmopara viticola, Pseudoperonospora cubensis, and Pseudoperonospora humuli. The 1C DNA contents ranged from 0.046 (45. 6 Mb) to 0.163 pg (159.9 Mb). This is 0.041- to 0.144-fold that of Glycine max (soybean, 1C = 1.134 pg), which was used as an internal standard for genome size determination. The linearity of Feulgen absorbance photometry method over this range was demonstrated by calibration of Aspergillus species (1C = 31-38 Mb) against Glycine, which revealed differences of less than 6% compared to the published CHEF data. The low coefficients of variation (usually between 5 and 10%), repeatability of the results, and compatibility with CHEF data prove the resolution power of Feulgen image analysis. The applicability and limitations of Feulgen photometry are discussed in relation to other methods of genome size determination (CHEF gel electrophoresis, reassociation kinetics, genomic reconstruction) that have been previously applied to Oomycota. Copyright 1998 Academic Press.

  6. Arbuscular mycorrhizal symbiosis stimulates key genes of the phenylpropanoid biosynthesis and stilbenoid production in grapevine leaves in response to downy mildew and grey mould infection.

    Science.gov (United States)

    Bruisson, Sébastien; Maillot, Pascale; Schellenbaum, Paul; Walter, Bernard; Gindro, Katia; Deglène-Benbrahim, Laurence

    2016-11-01

    Grapevine (Vitis spp) is susceptible to serious fungal diseases usually controlled by chemical treatments. Arbuscular mycorrhizal fungi (AMF) are obligate plant symbionts which can stimulate plant defences. We investigated the effect of mycorrhization on grapevine stilbenoid defences. Vitis vinifera cvs Chasselas, Pinot noir and the interspecific hybrid Divico, on the rootstock 41B, were mycorrhized with Rhizophagus irregularis before leaf infection by Plasmopara viticola or Botrytis cinerea. Gene expression analysis showed an up-regulation of PAL, STS, and ROMT, involved in the stilbenoid biosynthesis pathway, in plant leaves, 48 h after pathogen inoculation. This defense response could be potentiated under AMF colonization, with an intensity level depending on the gene, the plant cultivar and/or the pathogen. We also showed that higher amounts of active forms of stilbenoids (i.e trans-form of resveratrol, ε- and δ-viniferins and pterostilbene) were produced in mycorrhized plants of the three genotypes in comparison with non-mycorrhized ones, 10 days post-inoculation with either pathogen. These results support the hypothesis that AMF root colonization enhances defence reactions against a biotrophic and a necrotrophic pathogen, in the aerial parts of grapevine. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Pesticides' influence on wine fermentation.

    Science.gov (United States)

    Caboni, Pierluigi; Cabras, Paolo

    2010-01-01

    Wine quality strongly depends on the grape quality. To obtain high-quality wines, it is necessary to process healthy grapes at the correct ripeness stage and for this reason the farmer has to be especially careful in the prevention of parasite attacks on the grapevine. The most common fungal diseases affecting grape quality are downy and powdery mildew (Plasmopara viticola and Uncinula necator), and gray mold (Botrytis cinerea). On the other hand, the most dangerous insects are the grape moth (Lobesia botrana), vine mealybug (Planococcus ficus), and the citrus mealybug (Planococcus citri). Farmers fight grape diseases and insects applying pesticides that can be found at harvest time on grapes. The persistence of pesticides depends on the chemical characteristic of the active ingredients as well as on photodegradation, thermodegradation, codistillation, and enzymatic degradation. The pesticide residues on grapes can be transferred to the must and this can influence the selection and development of yeast strains. Moreover, yeasts can also influence the levels of the pesticides in the wine by reducing or adsorbing them on lees. During the fermentative process, yeasts can cause the disappearance of pesticide residues by degradation or absorption at the end of the fermentation when yeasts are deposited as lees. In this chapter, we reviewed the effect of commonly used herbicides, insecticides, and fungicides on yeasts. We also studied the effect of alcoholic and malolactic fermentation on pesticide residues.

  8. Pea Broth Enhances the Biocontrol Efficacy of Lysobacter capsici AZ78 by Triggering Cell Motility Associated with Biogenesis of Type IV Pilus.

    Science.gov (United States)

    Tomada, Selena; Puopolo, Gerardo; Perazzolli, Michele; Musetti, Rita; Loi, Nazia; Pertot, Ilaria

    2016-01-01

    Bacterial cells can display different types of motility, due to the presence of external appendages such as flagella and type IV pili. To date, little information on the mechanisms involved in the motility of the Lysobacter species has been available. Recently, L. capsici AZ78, a biocontrol agent of phytopathogenic oomycetes, showed the ability to move on jellified pea broth. Pea broth medium improved also the biocontrol activity of L. capsici AZ78 against Plasmopara viticola under greenhouse conditions. Noteworthy, the quantity of pea residues remaining on grapevine leaves fostered cell motility in L. capsici AZ78. Based on these results, this unusual motility related to the composition of the growth medium was investigated in bacterial strains belonging to several Lysobacter species. The six L. capsici strains tested developed dendrite-like colonies when grown on jellified pea broth, while the development of dendrite-like colonies was not recorded in the media commonly used in motility assays. To determine the presence of genes responsible for biogenesis of the flagellum and type IV pili, the genome of L. capsici AZ78 was mined. Genes encoding structural components and regulatory factors of type IV pili were upregulated in L. capsici AZ78 cells grown on the above-mentioned medium, as compared with the other tested media. These results provide new insight into the motility mechanism of L. capsici members and the role of type IV pili and pea compounds on the epiphytic fitness and biocontrol features of L. capsici AZ78.

  9. Association of RGA-SSCP markers with resistance to downy mildew and anthracnose in grapevines.

    Science.gov (United States)

    Tantasawat, P A; Poolsawat, O; Prajongjai, T; Chaowiset, W; Tharapreuksapong, A

    2012-07-02

    Downy mildew (Plasmopara viticola) and anthracnose (Sphaceloma ampelinum) are two major diseases that severely affect most grapevine (Vitis vinifera) cultivars grown commercially in Thailand. Progress of conventional breeding programs of grapevine for improved resistance to these diseases can be speeded up by selection of molecular markers associated with resistance traits. We evaluated the association between 13 resistance gene analog (RGA)-single-strand conformation polymorphism (SSCP) markers with resistance to downy mildew and anthracnose in 71 segregating progenies of seven cross combinations between susceptible cultivars and resistant lines. F(1) hybrids from each cross were assessed for resistance to downy mildew and anthracnose (isolates Nk4-1 and Rc2-1) under laboratory conditions. Association of resistance traits with RGA-SSCP markers was evaluated using simple linear regression analysis. Three RGA-SSCP markers were found to be significantly correlated with anthracnose resistance, whereas significant correlation with downy mildew resistance was observed for only one RGA-SSCP marker. These results demonstrate the usefulness of RGA-SSCP markers. Four candidate markers with significant associations to resistance to these two major diseases of grapevine were identified. However, these putative associations between markers and resistance need to be verified with larger segregating populations before they can be used for marker-assisted selection.

  10. Antimicrobial Activity of Resveratrol Analogues

    Directory of Open Access Journals (Sweden)

    Malik Chalal

    2014-06-01

    Full Text Available Stilbenes, especially resveratrol and its derivatives, have become famous for their positive effects on a wide range of medical disorders, as indicated by a huge number of published studies. A less investigated area of research is their antimicrobial properties. A series of 13 trans-resveratrol analogues was synthesized via Wittig or Heck reactions, and their antimicrobial activity assessed on two different grapevine pathogens responsible for severe diseases in the vineyard. The entire series, together with resveratrol, was first evaluated on the zoospore mobility and sporulation level of Plasmopara viticola (the oomycete responsible for downy mildew. Stilbenes displayed a spectrum of activity ranging from low to high. Six of them, including the most active ones, were subsequently tested on the development of Botrytis cinerea (fungus responsible for grey mold. The results obtained allowed us to identify the most active stilbenes against both grapevine pathogens, to compare the antimicrobial activity of the evaluated series of stilbenes, and to discuss the relationship between their chemical structure (number and position of methoxy and hydroxy groups and antimicrobial activity.

  11. Genome analyses of the sunflower pathogen Plasmopara halstedii provide insights into effector evolution in downy mildews and Phytophthora

    NARCIS (Netherlands)

    Sharma, Rahul; Xia, Xiaojuan; Cano, Liliana M; Evangelisti, Edouard; Kemen, Eric; Judelson, Howard; Oome, Stan; Sambles, Christine; van den Hoogen, D Johan; Kitner, Miloslav; Klein, Joël; Meijer, Harold J G; Spring, Otmar; Win, Joe; Zipper, Reinhard; Bode, Helge B; Govers, Francine; Kamoun, Sophien; Schornack, Sebastian; Studholme, David J; Van den Ackerveken, Guido; Thines, Marco

    2015-01-01

    BACKGROUND: Downy mildews are the most speciose group of oomycetes and affect crops of great economic importance. So far, there is only a single deeply-sequenced downy mildew genome available, from Hyaloperonospora arabidopsidis. Further genomic resources for downy mildews are required to study thei

  12. Characterization of Vitis vinifera NPR1 homologs involved in the regulation of Pathogenesis-Related gene expression

    Science.gov (United States)

    Le Henanff, Gaëlle; Heitz, Thierry; Mestre, Pere; Mutterer, Jerôme; Walter, Bernard; Chong, Julie

    2009-01-01

    Background Grapevine protection against diseases needs alternative strategies to the use of phytochemicals, implying a thorough knowledge of innate defense mechanisms. However, signalling pathways and regulatory elements leading to induction of defense responses have yet to be characterized in this species. In order to study defense response signalling to pathogens in Vitis vinifera, we took advantage of its recently completed genome sequence to characterize two putative orthologs of NPR1, a key player in salicylic acid (SA)-mediated resistance to biotrophic pathogens in Arabidopsis thaliana. Results Two cDNAs named VvNPR1.1 and VvNPR1.2 were isolated from Vitis vinifera cv Chardonnay, encoding proteins showing 55% and 40% identity to Arabidopsis NPR1 respectively. Constitutive expression of VvNPR1.1 and VvNPR1.2 monitored in leaves of V. vinifera cv Chardonnay was found to be enhanced by treatment with benzothiadiazole, a SA analog. In contrast, VvNPR1.1 and VvNPR1.2 transcript levels were not affected during infection of resistant Vitis riparia or susceptible V. vinifera with Plasmopara viticola, the causal agent of downy mildew, suggesting regulation of VvNPR1 activity at the protein level. VvNPR1.1-GFP and VvNPR1.2-GFP fusion proteins were transiently expressed by agroinfiltration in Nicotiana benthamiana leaves, where they localized predominantly to the nucleus. In this system, VvNPR1.1 and VvNPR1.2 expression was sufficient to trigger the accumulation of acidic SA-dependent Pathogenesis-Related proteins PR1 and PR2, but not of basic chitinases (PR3) in the absence of pathogen infection. Interestingly, when VvNPR1.1 or AtNPR1 were transiently overexpressed in Vitis vinifera leaves, the induction of grapevine PR1 was significantly enhanced in response to P. viticola. Conclusion In conclusion, our data identified grapevine homologs of NPR1, and their functional analysis showed that VvNPR1.1 and VvNPR1.2 likely control the expression of SA-dependent defense genes

  13. 致病疫霉在中国云南的马铃薯田间形成卵孢子%FORMATION OF OOSPORES OF PHYTOPHTHORA INFESTANS IN NATURAL POTATO FIELD IN YUNNAN PROVINCE OF CHINA

    Institute of Scientific and Technical Information of China (English)

    赵志坚; 李成云; 王淑芬; 隋启君

    2001-01-01

    @@ 由致病疫霉Phytophthora infestans (Mont.)de Bary引起的马铃薯和番茄晚疫病是世界性的作物病害,每年均造成巨大的经济损失和社会影响.致病疫霉是异宗配合的卵菌,有两个已知交配型A1和A2,两个相对交配型互作时可进行有性生殖产生卵孢子(Gallegly & Galindo,1958).过去许多年一直认为致病疫霉在除墨西哥以外的国家中只存在A1交配型,通过产生孢子囊进行无性繁殖.近年来,由于致病疫霉新致病群体的产生以及全球范围的迁移和替代,A2交配型菌株先后在欧洲、美洲、亚洲和非洲的许多国家被发现.

  14. Climatic conditions for grapevine cultivation in Boa Vista, Roraima, Brazil = Aptidão climática para o cultivo da videira em Boa Vista, Roraima

    Directory of Open Access Journals (Sweden)

    Marco Antônio Fonseca Conceição

    2014-12-01

    Full Text Available Grapevine plants have been grown in different tropical regions of Brazil. In the state of Roraima, this culture was introduced commercially in 2005 in Boa Vista. Unlike temperate regions, tropical regions present thermal conditions for grape production throughout the year. To evaluate the climatic characteristics on different periods of the year, it can be used different indices. The objective of this study was to characterize climate suitability of Boa Vista, Roraima, Brazil, for grapevine cultivation at different months of the year. For climate classification it was used the Geoviticulture Multicriteria Climatic Classification System (CCM, which is composed by three indices: Heliothermal (IH, Cool Night (IF, and Drought (IS. It was also used the Zuluaga Index (IZ, to assess the risk of incidence of fungal diseases on the vine, especially in relation to the incidence of downy mildew (Plasmopara viticola, one of the main grapevine diseases in humid regions. Based on the evaluated indices, it was found that the most favorable period for grape production goes from October to March because of the lower values of rainfall. During the other months, it can be employed branch pruning without fruit production, as occurs in other tropical locations of Brazil. =A videira tem sido cultivada em diferentes regiões tropicais do Brasil. No estado de Roraima, ela foi implantada comercialmente em 2005 em Boa Vista. Ao contrário das regiões de clima temperado, as regiões tropicais apresentam condições térmicas para a produção de uvas durante o ano inteiro. Para se avaliar as características climáticas dos diferentes períodos do ano pode-se recorrer a diversos índices. Objetivou-se, com o presente trabalho, avaliar a aptidão climática de Boa Vista, Roraima, para o cultivo da videira, considerando-se diferentes períodos de produção ao longo do ano. Na caracterização climática, foi utilizado o Sistema de Classificação Climática Multicrit

  15. Influência das condições climáticas sobre a eficácia de fungicidas empregados para o controle do míldio em Vitis vinifera Effect of environmental conditions on efficacy of fungicides to downy mildew control in Vitis vinifera

    Directory of Open Access Journals (Sweden)

    Ana Beatriz Costa Czermainski

    2004-02-01

    Full Text Available Um dos fatores a serem manejados para a máxima produtividade de um vinhedo é o míldio, doença causada por Plasmopara viticola, que afeta regiões produtoras de uva do mundo inteiro. No Brasil, são necessárias diversas aplicações de fungicida para seu controle. Diferentes princípios ativos são encontrados no mercado. Com base em resultados de ensaios de fungicidas para controle do míldio em Vitis vinifera conduzidos em 1995 e 1996, foi analisada a influência dos fatores climáticos temperatura do ar, precipitação e umidade relativa do ar sobre a eficácia dos produtos disponíveis. Os experimentos foram instalados na Embrapa Uva e Vinho, sobre a cv. Tannat, conduzida no sistema espaldeira. A análise dos fatores ambientais apontou períodos de infecções primárias e secundárias. A maior ocorrência de chuvas e alta umidade relativa do ar foram determinantes da maior severidade do míldio no ano de 1996. A análise da incidência e do índice de intensidade de doença em folhas e em cachos, indicou maior eficácia dos fungicidas cymoxanil + mancozeb, metalaxil + mancozeb e dithianona, nos dois anos. Os princípios ativos cúpricos - oxicloreto de cobre, isolado ou combinado com mancozeb, e sulfato de cobre - não foram eficazes para controlar o míldio nos cachos sob condições climáticas favoráveis à doença.One of the factors to be managed for maximum vineyard productivity is the downy mildew, disease caused by Plasmopara viticola that affects grape production in many parts of the world. In Brazil, several fungicide applications for its control are necessary. Different active ingredients are available on the market. The influence of climatic factors (air temperature, rainfall and relative humidity on fungicide efficacy in Vitis vinifera was analyzed using results of fungicide trials for downy mildew control as a base. The experiments were carried out at Embrapa Uva e Vinho with the cv. Tannat conducted on vertical trellis

  16. Laboratory Co-toxicity of Cyprodinil and Carbamic Acid Complex against Pathogenic Fungi on Five Kinds of Crops%嘧菌环胺和缬霉威复配对5种作物病原菌的室内联合毒力

    Institute of Scientific and Technical Information of China (English)

    刘润强; 曹前辉; 王清峻; 陈锡岭

    2016-01-01

    The synergistic activity of cyprodinil and carbamic acid complex against Fusarium oxysporum f. sp. Fragariae,Colletotrichum orbiculare (Berk. & Ment. ) Arx,Plasmopara viticola(Berk. et Curtis) Berl. et de Toni, Pythium aphanidermatum ( Edson) Fitzpatrick and Botrytis cinerea Pers,the pathogenic fungi on 5 kinds of crops,strawberry,watermelon,grape,tobacco,and rose,respectively,was studied based on the mycelial growth method in the laboratory. The bioassay results showed that the EC50 values of cyprodinil against 5 fungi ranged from 0. 19 mg/L to 1. 92 mg/L,and the EC50 values of carbamic acid against 5 fungi ranged from 0. 23 mg/L to 1. 02 mg/L. When the ratio of cyprodinil and carbamic acid was 15:1,the co-toxicity cofficient(CTC) against mycelial growth of Fusarium oxysporum f. sp. Fragariae was maximum ( 217 . 70 );When the ratio of the mixture was 8:1 , the CTC against mycelial growth of Colletotrichum orbiculare (Berk.& Ment. ) Arx was maximum(203. 59); When the ratio of the mixture was 3:1,the CTC against mycelial growth of Pythium aphanidermatum ( Edson) Fitzpatrick was maximum (196. 29);When the ratio of the mixture was 1:6,the CTC against mycelial growth of Plasmopara viticola (Berk. et Curtis) Berl. et de Toni was maximum(201. 98);When the ratio of the mixture was 9:1, the CTC against mycelial growth of Botrytis cinerea Pers was maximum ( 202 . 56 ) . The results suggested that cyprodinil and carbamic acid had significant synergistic activity against 5 different fungi when combined by appropriate ratio.%采用室内菌丝生长速率法,以草莓枯萎病菌、西瓜炭疽病菌、烟草猝倒病菌、葡萄霜霉病菌和月季灰霉病菌为作用目标,研究嘧菌环胺和缬霉威混配对5种病菌的联合毒力,为嘧菌环胺和缬霉威混配使用提供科学依据.结果显示,嘧菌环胺对5种病菌的有效中质量浓度(EC50)为0.19~1.92 mg/L,缬霉威对5种病菌的EC50为0.23~1.02 mg/L;当嘧菌环胺和缬霉威的混配比例为15

  17. Phenolic Compounds and Bioactivity of Healthy and Infected Grapevine Leaf Extracts from Red Varieties Merlot and Vranac (Vitis vinifera L.).

    Science.gov (United States)

    Anđelković, Marko; Radovanović, Blaga; Anđelković, Ana Milenkovic; Radovanović, Vladimir

    2015-09-01

    We investigated the phenolic composition, radical scavenging activity, and antimicrobial activity of grapevine leaf extracts from two red grape varieties, Vranac and Merlot (Vitis vinifera L.). The extracts were prepared from healthy grapevine leaves and those infected by Plasmopara viticola (downy mildew). The phenolic composition of the grapevine leaf extracts was determined using spectrophotometric assays and reverse-phase high performance liquid chromatography (RP-HPLC). The radical scavenging activity of grapevine leaf extracts was determined by the 2,2-diphenyl-1-picrylhydrazyl assay, and their antimicrobial activity was determined by microwell dilution tests. The total phenolic content was higher in healthy grapevine leaf extracts than in infected grapevine leaf extracts. The RP-HPLC analysis detected significant amounts of flavonols, phenolic acids, and flavan-3-ols, and small amounts of stilbenes in the grapevine leaf extracts. Compared with the infected grapevine leaf extracts, the healthy grapevine leaf extracts were richer in flavonols, phenolic acids, and flavan-3-ols, but had lower stilbenes contents. All extracts showed strong free radical scavenging activity, which was strongly correlated with the total phenolic content (R(2) = 0.978). The extracts showed a stronger antimicrobial activity towards Gram-positive bacterial strains than towards Gram-negative bacterial strains and yeast. The phenolic compounds in grapevine leaves were responsible for their strong radical scavenging and antimicrobial activities. Together, these results demonstrate that grapevine leaves have high nutritional value and can be used as a fresh food and to prepare extracts that can be used as additives in food and medicines.

  18. Mandipropamid targets the cellulose synthase-like PiCesA3 to inhibit cell wall biosynthesis in the oomycete plant pathogen, Phytophthora infestans.

    Science.gov (United States)

    Blum, Mathias; Boehler, Martine; Randall, Eva; Young, Vanessa; Csukai, Michael; Kraus, Sabrina; Moulin, Florence; Scalliet, Gabriel; Avrova, Anna O; Whisson, Stephen C; Fonne-Pfister, Raymonde

    2010-03-01

    Oomycete plant pathogens cause a wide variety of economically and environmentally important plant diseases. Mandipropamid (MPD) is a carboxylic acid amide (CAA) effective against downy mildews, such as Plasmopara viticola on grapes and potato late blight caused by Phytophthora infestans. Historically, the identification of the mode of action of oomycete-specific control agents has been problematic. Here, we describe how a combination of biochemical and genetic techniques has been utilized to identify the molecular target of MPD in P. infestans. Phytophthora infestans germinating cysts treated with MPD produced swelling symptoms typical of cell wall synthesis inhibitors, and these effects were reversible after washing with H(2)O. Uptake studies with (14)C-labelled MPD showed that this oomycete control agent acts on the cell wall and does not enter the cell. Furthermore, (14)C glucose incorporation into cellulose was perturbed in the presence of MPD which, taken together, suggests that the inhibition of cellulose synthesis is the primary effect of MPD. Laboratory mutants, insensitive to MPD, were raised by ethyl methane sulphonate (EMS) mutagenesis, and gene sequence analysis of cellulose synthase genes in these mutants revealed two point mutations in the PiCesA3 gene, known to be involved in cellulose synthesis. Both mutations in the PiCesA3 gene result in a change to the same amino acid (glycine-1105) in the protein. The transformation and expression of a mutated PiCesA3 allele was carried out in a sensitive wild-type isolate to demonstrate that the mutations in PiCesA3 were responsible for the MPD insensitivity phenotype.

  19. Effect of climate change on infection of grapevine by downy and powdery mildew under controlled environment.

    Science.gov (United States)

    Pugliese, M; Gullino, M L; Garibaldi, A

    2011-01-01

    Plant responses to elevated CO2 and temperature have been much studied in recent years, but effects of climate change on pathological responses are largerly unknown. The pathosystems grapevine (Vitis vinifera) - downy mildew (Plasmopara viticola) and powdery mildew (Erysiphe necatrix) were chosen as models to assess the potential impact of increased CO2 and temperature on disease incidence and severity under controlled environment. Grapevine potted plants were grown in phytotrons under 4 different simulated climatic conditions: (1) standard temperature (ranging from 18 degrees to 22 degrees C) and standard CO2 concentration (450 ppm); (2) standard temperature and elevated CO2 concentration (800 ppm); (3) elevated temperature (ranging from 22 degrees to 26 degrees C, 4 degrees C higher than standard) and standard CO2 concentration; (4) elevated temperature and CO2 concentration. Each plant was inoculated with a spore suspension containing 5x10(5) cfu/ml. Disease index and physiological parameters (chlorophyll content, fluorescence, assimilation rate) were assessed. Results showed an increase of the chlorophyll content with higher temperatures and CO2 concentration, to which consequently corresponded an higher fluorescence index. Disease incidence of downy mildew increased when both CO2 and temperatures were higher, while an increase in CO2 did not influenced powdery mildew incidence, probably due to the increased photosynthetic activity of plants under such conditions. Considering that the rising concentrations of CO2 and other greenhouse gases will lead to an increase in global temperature and longer seasons, we can assume that this will allow more time for pathogens evolution and could increase pathogen survival, indirectly affecting downy and powdery mildews of grapevine.

  20. Effects of potential food sources on biological and demographic parameters of the predatory mites Kampimodromus aberrans, Typhlodromus pyri and Amblyseius andersoni.

    Science.gov (United States)

    Lorenzon, Mauro; Pozzebon, Alberto; Duso, Carlo

    2012-11-01

    Kampimodromus aberrans, Typhlodromus pyri and Amblyseius andersoni are generalist predatory mites important in controlling tetranychid and eriophyoid mites in European vineyards. They can persist by exploiting various non-prey foods when their main prey is absent or scarce. A comparative analysis of the effects of various prey and non-prey foods on the life history of these predators is lacking. In the laboratory, predatory mites were reared on herbivorous mites (Panonychus ulmi, Eotetranychus carpini and Colomerus vitis), a potential alternative prey (Tydeus caudatus) and two non-prey foods, i.e. the pollen of Typha latifolia and the mycelium of Grape downy mildew (GDM) Plasmopara viticola. Developmental times, survival, sex ratio and fecundity as well as life table parameters were estimated. Kampimodromus aberrans developed faster on E. carpini, C. vitis or pollen than on P. ulmi and laid more eggs on pollen than on prey. Low numbers of this predator developed on GDM infected leaves. Tydeus caudatus was not suitable as prey for any of the three predatory mites. Kampimodromus aberrans showed the highest intrinsic rate of population increase when fed on pollen. Developmental times of T. pyri on prey or pollen were similar but fecundity was higher on pollen than on P. ulmi. Typhlodromus pyri had higher intrinsic rates of population increase on C. vitis and pollen than on P. ulmi; E. carpini showed intermediate values whereas GDM resulted in the lowest r ( m ) values. Development of A. andersoni females was faster on pollen and C. vitis than on P. ulmi and GDM. Fecundity was higher on pollen and mites compared to GDM. Life table parameters of A. andersoni did not differ when predators were fed with prey or pollen while GDM led to a lower r ( m ) value. On a specific diet A. andersoni exhibited faster development and higher fecundity than T. pyri and K. aberrans. These findings improve knowledge on factors affecting the potential of predatory mites in controlling

  1. Estudos dos constituintes químicos e atividade antibacteriana do óleo essencial de Lippia gracilis a Xanthomonas campestris pv. viticola "in vitro"

    Directory of Open Access Journals (Sweden)

    Morgana Mateus Santos

    2014-09-01

    Full Text Available Este trabalho foi realizado com o objetivo de estudar a eficácia de óleos essenciais de Lippia gracilis, no crescimento in vitro de Xcv. Foram utilizados oito óleos extraídos de L. gracilis, obtidos de diferentes manejos de cultivo, em três dosagens diferentes (200; 250 e 300 µL.L-1, testemunha com o oxicloreto de cobre (controle padrão e a testemunha absoluta. A suscetibilidade do isolado Xcv3 aos óleos testados foi avaliada na primeira etapa pela presença e ausência do crescimento bacteriano e, na segunda etapa, pela porcentagem de inibição do crescimento bacteriano. A análise da composição química dos óleos essenciais extraídos de L. gracilis, mostrou, o carvacrol (73,9 a 77% como composto majoritário e o timol (4,9 a 10,3%. Os óleos referentes aos tratamentos 01, 06 e 07 foram os que proporcionaram maiores porcentagens de inibição no crescimento bacteriano de Xcv: 94,75%, 96,50% e 94,02%, respectivamente, sendo superiores ao oxicloreto de cobre (49,6%.

  2. Estudos dos constituintes químicos e atividade antibacteriana do óleo essencial de Lippia gracilis a Xanthomonas campestris pv. viticola in vitro.

    OpenAIRE

    2015-01-01

    Este trabalho foi realizado com o objetivo de estudar a eficácia de óleos essenciais de Lippia gracilis, no crescimento in vitro de Xcv. Foram utilizados oito óleos extraídos de L. gracilis, obtidos de diferentes manejos de cultivo, em três dosagens diferentes (200; 250 e 300 µL.L-1), testemunha com o oxicloreto de cobre (controle padrão) e a testemunha absoluta. A suscetibilidade do isolado Xcv3 aos óleos testados foi avaliada na primeira etapa pela presença e ausência do crescimento bacteri...

  3. 76 FR 60025 - Product Cancellation Order for Certain Pesticide Registrations

    Science.gov (United States)

    2011-09-28

    ... action under docket identification (ID) number EPA-HQ-OPP-2009-1017. Publicly available docket materials...% Permectrin Pour-On Permethrin Insecticide. 069592-00002 Laginex AS Lagenidium giganteum, mycelium or oospores 069592-00003 Technical Laginex......... Lagenidium giganteum, mycelium or oospores 070506-00202...

  4. Identification of Molecular Genetic Markers Tightly Linked to Downy Mildew Resistant Genes in Grape%与葡萄抗霜霉病基因紧密连锁的 分子遗传标记

    Institute of Scientific and Technical Information of China (English)

    罗素兰; 贺普超; 周鹏; 郑学勤

    2001-01-01

    Bulk segregant analysis (BSA), randomly amplified polymorphic DNA(RAPD) and sequence characterized amplified region (SCAR) methods were used to tag the downy mildew-resistant genes of grape with molecular markers. Parents and their 60 individuals of an F1 progeny resulting from a cross 88-110 between 83-4-96 (Vitis quinquangularis, downy mildew-resistant) and Muscat Rose (V. vinifera, downy mildew-susceptible), three F2 progenies resulting from three crosses of self-cross and inter-cross of 88-110 F1 seedlings, as well as another interspecific F1 hybrids of 88-84 cross [Xun-3 (V. quinquangularis, downy mildew-resistant)×Ugni Blanc (V. vinifera, downy mildew-susceptible)] and the wild grapes native to China were used for the study. Among 280 Operon primers, 160 gave distinct band patterns. One RAPD marker OPO06-1500 was tightly linked to a major gene resistant to Plasmopara viticola (RPv-1). Based on Mapmaker software analysis, the map distance between RPv-1 and OPO06-1500 was 1.7cM. Marker OPO06-1500 was cloned and sequenced. According to the sequence, two specific primers were designed to amplify all plant materials. RAPD marker was converted into SCAR marker (SCO06-1500). One distinct single band only in resistant plants was amplified, whose size was the same as that of the RAPD marker. The SCAR marker’s popularity was confirmed, and it could be used for the identification of hybrid resistant to P. viticola and will be potentially useful in the development of new resistant grape cultivars.%以种间杂交组合88-110[83-4-96(毛葡萄,抗霜霉病)×粉红玫瑰(欧洲葡萄,感霜霉病)]的F1代(60个单株)及其自交或互交所得的3个F2代为试材,应用BSA、RAPD和SCAR方法研究了葡萄抗霜霉病基因的分子标记。共筛选了280个Operon引物,其中160个引物扩增出了清晰的DNA条带,发现了RAPD标记OPO06-1500与葡萄抗霜霉病主效基因(RPv-1)紧密连锁,经Mapmarker软件连锁分析,OPO06

  5. Características físico-químicas e produtivas da videira 'BRS Clara' sob cultivo protegido

    Directory of Open Access Journals (Sweden)

    Larissa Abgariani Colombo

    2011-09-01

    Full Text Available Atualmente, a preferência por uvas sem sementes vem aumentando nos mercados interno e externo, sendo uma alternativa a produção de uvas sob cultivo protegido. No entanto, o custo de produção de uvas finas para mesa tem sido afetado pela intensa necessidade de controle de doenças fúngicas, como o míldio (Plasmopara viticola. O trabalho teve como objetivo avaliar a produção e as características físicoquímicas dos frutos da videira 'BRS Clara' sobre os porta-enxertos 'IAC 572 Jales' e 'IAC 766 Campinas', sob diferentes tipos de cultivo protegido. O experimento foi realizado no município de Marialva-PR, durante duas safras regulares (set.-dez.2007, ago-dez.2008. O delineamento experimental foi o de blocos ao acaso, com quatro repetições, em arranjo fatorial 7 x 2 (sete tipos de cultivo protegido e dois porta-enxertos. Os tratamentos foram os seguintes tipos de cultivo: a. tela plástica sem fungicidas para míldio; b. tela plástica com fungicidas para míldio (padrão de controle da região; c. cobertura plástica sem fungicidas para míldio; d. cobertura plástica e 50% de redução do padrão de fungicidas para míldio; e. cobertura plástica e 75% de redução do padrão de fungicidas para míldio; f. cobertura plástica com fosfito e cobre, e g. cobertura plástica sem fungicidas. Verificou-se que o cultivo protegido não alterou as características produtivas da videira 'BRS Clara', como número de cachos e produção por planta; os porta-enxertos 'IAC 766' e 'IAC 572' são indicados para a produção da uva 'BRS Clara', e a utilização da cobertura plástica permite a redução do número de aplicações de fungicidas para míldio no cultivo da uva 'BRS Clara'.

  6. 葡萄病程相关蛋白1基因的克隆和表达分析%Gene Cloning and Expression Analysis of Pathogenesis-Related Protein 1 in Vitis vinifera L

    Institute of Scientific and Technical Information of China (English)

    侯丽霞; 高超; 车永梅; 赵方贵; 刘新

    2012-01-01

    Using homology cloning method, the full-length cDNA of pathogenesis-related protein 1 (PR1) named VvPR-1 was cloned from leaves of Vitis vinifera cultivar 'Zuoyouhong' tissue culture seedling. Bioin-formatic analysis indicated that VvPR-1 consisted of 486 nucleotides encoding 161 amino acid with molecular weight 17.5 kDa, isoelectric point 8.69, VvPR-1 possessed six conserved cysteine and four conserved allergen V5/Tpx-1 related domain. The VvPR-1 was highiy homologous to PR1 in other plants. Real-time PCR analysis showed that expression level of VvPR-1 was the highest in leaves, VvPRl was significantly induced by Plasmo-para viticola inoculation, salt stress and osmotic stress, signaling molecules such as salicylic acid (SA), jas-monate (JA), abscisic acid (ABA) as well as nitric oxide (NO), hydrogen peroxide (H2O2), hydrogen sulfide (H2S) also had inductive effects on VvPR-1 expression. It suggested that VvPRl was concerned with various bi-otic stresses and abiotic stresses.%以葡萄品种‘左优红’,组培苗叶片为材料,利用同源克隆法获得其病程相关蛋白1基因VvPR1的cDNA全长序列.扩增片段大小为486bp,编码161个氨基酸,分子量17.5 kDa,等电点PI=8.69,含有6个保守半胱氨酸,4个allergen V5/Tpx-1 related 保守结构域.VvPR1与多种植物PR1高度同源.实时定量PCR检测结果表明VvPRI在葡萄叶片中相对表达量最高;霜霉病菌、低温、盐和干旱胁迫均可显著诱导其表达;水杨酸、脱落酸、茉莉酸、一氧化氮、过氧化氢和硫化氢等亦可诱导其大量表达,据此推测,VvPR1参与了多种生物胁迫和非生物胁迫过程.

  7. Immobilization of copper by biochar in Cu-enriched agricultural soils depends on interactions with soil organic carbon

    Science.gov (United States)

    Mlinkov, Slađana; Zehetner, Franz; Rosner, Franz; Dersch, Georg; Soja, Gerhard

    2017-04-01

    The appearance of downy mildew (Plasmopara viticola) in European vineyards of the 19th century was the starting point for the search of effective fungicides to avoid severe yield losses. Copper has been found as an important ingredient for several fungicides that have been used in agriculture and horticulture. For organic viticulture, several diseases can only be antagonized with Cu-containing fungicides as the application of organic fungicides is not permitted. This long-lasting dependence on Cu-fungicides has led to a gradual Cu enrichment of vineyard soils in traditional wine-growing areas, locally exceeding 300 mg/kg. Although these concentrations do not affect the vines or wine quality, they may impair soil microbiological functions in the top soil layer or the root growth of green cover plants. Therefore, measures are demanded that reduce the bioavailability of copper, thereby reducing the ecotoxicological effects. The use of biochar and compost as soil amendment has been suggested as a strategy to immobilize Cu and reduce the exchangeable fractions. In our study we have tested the hypothesis that biochar immobilizes the bioavailability of Cu for soil cover crops and reduces soil pore water concentrations. This study had the objective to test the interactions of compost and biochar with respect to Cu immobilization in vineyard soils. A Cu-enriched vineyard soil (250 mg Cu kg-1) was analyzed both in greenhouse and field experiments. In both experiments, soil with or without biochar and/or compost and mixtures of the two components were used. In the greenhouse experiments, was used as test plant Lolium multiflorum for Cu uptake; in the field, Lolium perenne and Trifolium repens were analyzed. Greenhouse experiment: Soil pore water concentrations showed clearer differences in Cu concentration than Lolium multiflorum shoots. Compost increased dissolved organic carbon (DOC) and Cu in soil pore water and biochar reduced it significantly. The mixtures of compost and

  8. Soil microbial toxicity assessment of a copper-based fungicide in two contrasting soils

    Science.gov (United States)

    Dober, Melanie; Deltedesco, Evi; Jöchlinger, Lisa; Schneider, Martin; Gorfer, Markus; Bruckner, Alexander; Zechmeister-Boltenstern, Sophie; Soja, Gerhard; Zehetner, Franz; Keiblinger, Katharina Maria

    2016-04-01

    The infestation with the fungus downy mildew (Plasmopara viticola) causes dramatic losses in wine production. Copper (Cu) based fungicides have been used in viticulture since the end of the 19th century, and until today both conventional and organic viticulture strongly rely on Cu to prevent and reduce fungal diseases. Consequently, Cu has built up in many vineyard soils and it is still unclear how this affects soil functioning. The aim of the present study is the evaluation of the soil microbial toxicity of Cu contamination. Two contrasting agricultural soils, an acidic sandy soil and a calcareous loamy soil, were sampled to conduct an eco-toxicological greenhouse pot experiment. The soils were spiked with a commonly used fungicide based on copper hydroxid in seven concentrations (0, 50, 100, 200, 500, 1500 and 5000 mg Cu kg-1 soil) and Lucerne (Medicago sativa L. cultivar. Plato) was grown in the pots for 3 months. Sampling was conducted at the beginning and at the end of the study period to test copper's soil microbial toxicity in total microbial biomass and basal respiration, as well as enzyme activities, such as exoglucanase, β-glucosidase, exochitinase, phosphatase, protease, phenol-, peroxidase and urease. Additionally, DOC, TN, Cmic, Nmic, NO3 and NH4 were determined to provide further insight into the carbon and nitrogen cycle. Microbial community structure was analysed by phospholipid fatty acids (PLFAs), and ergosterol as a fungal biomarker. In addition, molecular tools were applied by extracting soil DNA and performing real time quantitative polymerase chain reaction (qPCR) and a metagenomic approach using 16S and ITS amplification and sequencing with MiSeq platform for the second sampling. Hydrolytic extracellular enzymes were not clearly affected by rising Cu concentrations, while a trend of increasing activity of oxidative enzymes (phenol- and peroxidase) was observed. Microbial respiration rate as well as the amount of Cmic and Nmic decreased with

  9. Biochar and compost as amendments in copper-enriched vineyard soils - stabilization or mobilization of copper?

    Science.gov (United States)

    Soja, Gerhard; Fristak, Vladimir; Wimmer, Bernhard; Bell, Stephen; Chamier Glisczinski, Julia; Pardeller, Georg; Dersch, Georg; Rosner, Franz; Wenzel, Walter; Zehetner, Franz

    2016-04-01

    Copper is an important ingredient for several fungicides that have been used in agriculture. For organic viticulture, several diseases as e.g. downy mildew (Plasmopara viticola) can only be antagonized with Cu-containing fungicides. This long-lasting dependence on Cu-fungicides has led to a gradual Cu enrichment of vineyard soils in traditional wine-growing areas, occasionally exceeding 300 mg/kg. Although these concentrations do not affect the vines or wine quality, they may impair soil microbiological functions in the top soil layer or the root growth of green cover plants. Therefore measures are demanded that reduce the bioavailability of copper, thereby reducing the ecotoxicological effects. The use of biochar and compost as soil amendment has been suggested as a strategy to immobilize Cu and reduce the exchangeable fractions. This study consisted of lab and greenhouse experiments that were designed to test the sorption and desorption behavior of copper in vineyard soils with or without biochar and/or compost as soil amendment. Slightly acidic soils (pHalkaline soils. The analyses of leachate waters of microlysimeter experiments showed that the biochar effects were more evident for a reduction of the ionic form Cu2+ than for total soluble copper, even in alkaline soils. Biochar modified with citric or tartaric acid did not significantly decrease the solubility of copper based on total dissolved concentrations although CEC was higher than in unmodified biochar. Treatments consisting of compost only or that had an equal amount of compost and biochar rather had a mobilizing effect on biochar. Sorption experiments with different DOC concentrations and biochar, however, showed a positive effect on copper sorption. Apparently in vineyard soils the predisposition to form organic-Cu-complexes may outbalance the binding possibilities of these complexes to biochar, occasionally resulting in enhanced mobilization. Presumably immobilization of copper with biochar would

  10. 果树真菌病害拮抗细菌的筛选、鉴定及拮抗机制初探%Screening and identification of antagonistic bacteria against fungal pathogens in fruit trees and their antifungal mechanisms

    Institute of Scientific and Technical Information of China (English)

    于晓丽; 亓超; 王培松; 李宝燕; 王英姿

    2016-01-01

    [目的]分离获得对果树真菌病害[苹果轮纹病(Botryospuaeria dothidea)、苹果斑点落叶病(Alternaria alternaria)、葡萄炭疽病(Colletotrichum gloeosporioides)、葡萄白腐病(Coniella diplodiella)和葡萄霜霉病(Plasmopara viticola)]具有明显拮抗作用的菌株,初步研究其作用机制,为果树真菌病害的生物防治提供潜在的材料菌株.[方法]从不同植物上分离内生菌,采用平板对峙法、菌落生长速率法及叶盘法等试验筛选拮抗菌;根据菌株的形态、生理生化特征、16SrDNA序列分析、gyrA基因序列分析等对其进行鉴定;通过测定拮抗菌发酵液对温度、pH值的敏感度分析拮抗物质的稳定性;通过测定拮抗菌发酵液对菌丝的影响,初步分析其拮抗机制.[结果]从不同植物根、茎、叶中共分离得到58株细菌,其中编号为YTB1407的细菌菌株对上述5种植物病原真菌有较好的拮抗作用,经鉴定该菌株为解淀粉芽孢杆菌,且该菌株发酵液的拮抗活性在温度为20~100℃、pH为2.0~10.0,比较稳定.经该菌株发酵液处理后的病原菌菌丝畸形.[结论]该拮抗菌株为Bacillus amyloliquefaciens;温度和pH值对该菌株拮抗物质活性影响不大;该拮抗菌株可以通过影响病原菌菌丝正常生长发挥其拮抗作用.

  11. Reproductive strategies of field Phytophthora infestans populations on potato and tomato in some regions of Russia

    Directory of Open Access Journals (Sweden)

    Smirnov A.N.

    2009-01-01

    Full Text Available The occurrence of zoosporangia and oospores is analyzed for 88 field populations of Phytophthora infestans (Mont. de Bary in Moscow region and other regions of Russia from 1997 to 2006. For estimation of zoosporangia and oospore frequencies in the populations, the indexes IZ (for zoosporangia and IO (for oospores are suggested. The combinations of these indexes enabled a reveal of 4 strategies for P. infestans reproduction, which were found in the field populations of the pathogen. These strategies were more numerous in P. infestans field populations on tomato than on potato (4 versus 2 in spite of the fact that potato populations in Moscow region strongly predominated. It was connected with rareness or absence of oospores in potato and their presence in tomato, especially in fruit of many populations. The knowledge about strategies of reproduction can be applied for verification of efficiency of new methods for crop protection against late blight, and for investigations of P. infestans populations.

  12. Evaluation of Risk of Resistance in Plant Pathogenous Fungi to Fungicides%植物病原菌对杀菌剂抗性风险评估

    Institute of Scientific and Technical Information of China (English)

    王文桥; 马志强; 张小风; 张文吉

    2001-01-01

    植物病原菌对杀菌剂的抗性风险由基本风险和治理风险组成。杀菌剂使用之前或之初可根据人工诱变、药剂选择或驯化实验、田间野生敏感菌株敏感性变异、抗药菌株的生物及遗传特征、杀菌剂作用方式等进行基本抗药风险预测;杀菌剂使用数年之后可根据人工诱变、药剂选择或驯化、田间药效与抗药性发生、抗药菌株的生物及遗传特征、杀菌剂作用方式与使用对策等已有资料进行抗药风险评估。目前已有4种方法用于抗药风险评估。由杀菌剂与病害共同决定的基本抗药风险可分成低、中和高度。基本抗性风险高的药剂合理使用可延缓田间抗药性发生,中度基本抗性风险药剂不合理使用也可引发田间抗药性发生和药效明显降低。%Risk of resistance in plant pathogenous fungi to fungicides is composed of inherent risk and management risk. At the beginning of the application of target fungicide, inherent risk of resistance can be predicted by artificial induction or fungicide selection, sensitivity variation of wild-type sensitive strains, biological and genetic characteristics of resistant strains and mode of action of target fungicide. Several years after application of target fungicide, risk of resistance can be assessed according to the known experimental data, such as artificial induction or fungicide selection, efficacy appearance and resistance occurrence in field, biological and genetic characteristics of resistant strains, mode of action and application strategies of target fungicide. Currently, 4 methods have been adopted to evaluate risk of resistance. Inherent risk of resistance caused by diseases and fungicides can be divided into low, intermediate, and high level. High risk of resistance to phenylamide fungicides and lower risk of resistance to cymoxanil have been assessed in Phytophthora infestans and Plasmopara viticola. Low or intermediate risk of

  13. Potato late blight epidemics and population structure of Phytophthora infestans.

    NARCIS (Netherlands)

    Zwankhuizen, M.J.

    1998-01-01

    Potato late blight is caused by the fungus Phytophthora infestans . To study the relative importance of oospores in the epidemiology, and to estimate the relative impact of various infection sources, late blight epidemics in Southern Flevoland (The Netherlands) were studied using epidemiological and

  14. Phytophthora porri in leek: epidemiology and resistance.

    NARCIS (Netherlands)

    Smilde, W.D.

    1996-01-01

    In winter, Phytophthora porri is an important pathogen of leek ( Allium porrum L.) in the Netherlands. The fungus survives the crop-free period in summer by oospores in soil, and infects the leaves in autumn. Field studies indicated that dispersal by rain splash is crucial for initiation of an epid

  15. Real-time PCR and spore trap-based detection of the downy mildew pathogen, Peronospora effusa

    Science.gov (United States)

    Peronospora effusa is an obligate pathogen and the causal agent of downy mildew on spinach. The pathogen can be dispersed by splashing rain and wind, and may overwinter as oospores. Outbreaks of downy mildew on spinach are common in the cool climate of central coastal California, including the Sal...

  16. 76 FR 47579 - Notice of Receipt of Requests To Voluntarily Cancel Certain Pesticide Registrations

    Science.gov (United States)

    2011-08-05

    ... information whose disclosure is restricted by statute. Certain other material, such as copyrighted material... available docket materials are available either in the electronic docket at http://www.regulations.gov , or.... 069592-00002 Laginex AS Lagenidium giganteum, mycelium or oospores. 069592-00003 Technical...

  17. Oospora populations of Peronospora viciae: quantification, germinability andsurvival.

    NARCIS (Netherlands)

    Gaag, van der D.J.

    1997-01-01

    Peronospora viciae causes downy mildew on pea, field and broad bean. Two pathogens can be distinguished, P. viciae f.sp. pisi on pea (Pisum sativum) and P. viciae f. sp. fabae on field and broad bean (Vicia faba). These pathogens form resting spores, called oospores, in their host plant tissue. The

  18. Biologische beheersstrategie van Valse Meeldauw in zonnebloem

    NARCIS (Netherlands)

    Duyvesteijn, R.G.E.; Breeuwsma, S.J.; Helm, van der F.P.M.; Lans, van der A.M.; Boer, de M.

    2010-01-01

    Zonnebloemen zijn een belangrijk onderdeel van het biologische boeket. Echter de teelt van zonnebloemen wordt bedreigd door valse meeldauw. De Latijnse naam van valse meeldauw is Plasmopara halstedii. Deze oömyceet infecteert de zonnebloem systemisch en dit leidt tot grote verliezen. In dit project

  19. Development and dissection of diagnostic SNP markers for the downy mildew resistance genes PlArg and Pl8 and maker-assisted gene pyramiding in sunflower (Helianthus annuus L.)

    Science.gov (United States)

    Downy mildew, which is caused by fungus Plasmopara halstedii (Farl.) Berlese & de Toni, is one of the most important diseases that affect sunflower production globally. Two downy mildew resistance genes, PlArg and Pl8, were discovered in the late 1980s. Over two decades, PlArg is still effective aga...

  20. Pl17 is a novel gene independent of known downy mildew resistance genes in the cultivated sunflower (Helianthus annuus L.)

    Science.gov (United States)

    Downy mildew (DM), caused by Plasmopara halstedii (Farl.) Berl. et de Toni, is one of the serious sunflower diseases in the world due to its high virulence and the variability of the pathogen. DM resistance in the USDA inbred line, HA 458, has been shown to be effective against all virulent races of...

  1. Effect of sequential fermentations and grape cultivars on volatile compounds and sensory profiles of Danish wines

    DEFF Research Database (Denmark)

    Liu, Jing; Arneborg, Nils; Toldam-Andersen, Torben

    2017-01-01

    : The different non-Saccharomyces strains and cool-climate grape cultivars produced wines with a distinctive aromatic profile. A total of 67 volatile compounds were identified, including 43 esters, 14 alcohols, five acids, two ketones, a C13-norisoprenoid, a lactone and a sulfur compound. The use of M. viticola...

  2. Reassessment of Phomopsis species on grapevines

    NARCIS (Netherlands)

    Niekerk, van J.M.; Groenewald, J.Z.; Farr, D.F.; Fourie, P.H.; Halleen, F.; Crous, P.W.

    2005-01-01

    Ten species of Phomopsis have previously been identified from grapevines. Of these, P. viticola, the causal agent of Phomopsis cane and leaf spot, and P. vitimegaspora, the causal agent of swelling arm of grapevines, have been confirmed as severe pathogens of this host. Earlier taxonomic treatments

  3. Effect of phosphorus and the arbuscular mycorrhizal fungus Glomus intraradices on disease severity of root rot of peas (Pisum sativum) caused by Aphanomyces euteiches

    DEFF Research Database (Denmark)

    Bødker, Lars; Kjøller, Rasmus; Rosendahl, Søren

    1998-01-01

    The effects of inorganic phosphate levels and the presence of arbuscular mycorrhiza on disease severity of Aphanomyces euteiches in pea roots were studied. Disease severity on roots and epicotyl as well as the oospore number within infected root tissue were correlated with the phosphorus (P) level...... in the growth medium. The arbuscular mycorrhizal fungus Glomus intraradices increased P uptake and the P concentration in the plant but reduced disease development in peas. Polyacrylamide gel electrophoresis followed by densitometry of glucose-6-phosphate dehydrogenase specific to A.euteiches was used...

  4. Stage-specific gene expression during sexual development in Phytophthora infestans

    DEFF Research Database (Denmark)

    Fabritius, Anna-Liisa; Cvitanich, Cristina; Judelson, Howard S.

    2002-01-01

    Eight genes that are upregulated during sexual development in the heterothallic oomycete, Phytophthora infestans, were identified by suppression subtractive hybridization. Two genes showed very low but detectable expression in vegetative hyphae and became induced about 40- to >100-fold early...... in mating, before gametangial initials appeared. The remaining six loci were not induced until later in mating, coincident with the formation of gametangia and oospores, with induction levels ranging from 60- to >100-fold. Five genes were single copy, and three were members of families. Sequence analysis...

  5. Saprolegnia brachydanis, a new oomycete isolated from zebra fish.

    Science.gov (United States)

    Ke, Xiaoli; Wang, Jianguo; Gu, Zemao; Li, Ming; Gong, Xiaoning

    2009-02-01

    Saprolegnia brachydanis is described from zebra fish (Brachydanio rerio) in Wuhan, Hubei Province, China. The species is illustrated and compared with other species of the genus. The distinctive characteristics of S. brachydanis are the production of glomerulate oogonia wrapped around by predominantly monoclinous antheridia which can be up to eight in one oogonium. The oogonial stalks are short, straight, or curved and the antheridia, twisted, can enwind one or more oogonia. The oospores cannot mature or easily abort. Morphological features of the oomycete and the ITS sequence of its rDNA as well as the comparison with related species are discussed in this article.

  6. Fungal trunk pathogens associated with wood decay of pistachio trees in Iran

    Directory of Open Access Journals (Sweden)

    Hamid Mohammadi

    2015-06-01

    Full Text Available Over the growing seasons of 2011–2013, various pistachio (Pistacia vera L. cv. Fandoghi, and wild pistachio (P. atlantica Desf. subsp. mutica trees were inspected in Iran to determine the aetiology of trunk diseases with specific reference to species of Phaeoacremonium and Botryosphaeriaceae spp. Samples were collected from branches of trees exhibiting yellowing, defoliation, canker and dieback, as well as wood discoloration in cross sections. Fungal trunk pathogens were identified using morphological and cultural characteristics as well as comparisons of DNA sequence data of the ITS and TEF-1α (for Botryosphaeriaceae species and β-tubulin gene (for Phaeoacremonium species regions. Phaeoacremonium parasiticum was the dominant species followed by Phaeoacremonium aleophilum, Botryosphaeria dothidea, Neofusicoccum parvum, Phaeoacremoniumcinereum, Phaeoacremonium viticola and Dothiorella viticola. Pathogenicity tests were undertaken to determine the role of these species on pistachio under field conditions. Neofusicoccum parvum and Pm. aleophilum caused the longest and smallest lesions respectively. This study represents the first report on the occurrence and pathogenicity of Phaeoacremonium species on P. vera cv. Fandoghi. This also represents the first report of Pleurostomophora sp. on pistachio and Pm. parasiticum and D. viticola on wild pistachio.

  7. Antifungal activity against plant pathogens of metabolites from the endophytic fungus Cladosporium cladosporioides.

    Science.gov (United States)

    Wang, Xiaoning; Radwan, Mohamed M; Taráwneh, Amer H; Gao, Jiangtao; Wedge, David E; Rosa, Luiz H; Cutler, Horace G; Cutler, Stephen J

    2013-05-15

    Bioassay-guided fractionation of Cladosporium cladosporioides (Fresen.) de Vries extracts led to the isolation of four compounds, including cladosporin, 1; isocladosporin, 2; 5'-hydroxyasperentin, 3; and cladosporin-8-methyl ether, 4. An additional compound, 5',6-diacetylcladosporin, 5, was synthesized by acetylation of compound 3. Compounds 1-5 were evaluated for antifungal activity against plant pathogens. Phomopsis viticola was the most sensitive fungus to the tested compounds. At 30 μM, compound 1 exhibited 92.7, 90.1, 95.4, and 79.9% growth inhibition against Colletotrichum acutatum , Colletotrichum fragariae , Colletotrichum gloeosporioides , and P. viticola, respectively. Compound 2 showed 50.4, 60.2, and 83.0% growth inhibition at 30 μM against Co. fragariae, Co. gloeosporioides, and P. viticola, respectively. Compounds 3 and 4 were isolated for the first time from Cl. cladosporioides. Moreover, the identification of essential structural features of the cladosporin nuclei has also been evaluated. These structures provide new templates for the potential treatment and management of plant diseases.

  8. Yeast diversity on grapes in two German wine growing regions.

    Science.gov (United States)

    Brysch-Herzberg, Michael; Seidel, Martin

    2015-12-01

    The yeast diversity on wine grapes in Germany, one of the most northern wine growing regions of the world, was investigated by means of a culture dependent approach. All yeast isolates were identified by sequence analysis of the D1/D2 domain of the 26S rDNA and the ITS region. Besides Hanseniaspora uvarum and Metschnikowia pulcherrima, which are well known to be abundant on grapes, Metschnikowia viticola, Rhodosporidium babjevae, and Curvibasidium pallidicorallinum, as well as two potentially new species related to Sporidiobolus pararoseus and Filobasidium floriforme, turned out to be typical members of the grape yeast community. We found M. viticola in about half of the grape samples in high abundance. Our data strongly suggest that M. viticola is one of the most important fermenting yeast species on grapes in the temperate climate of Germany. The frequent occurrence of Cu. pallidicorallinum and strains related to F. floriforme is a new finding. The current investigation provides information on the distribution of recently described yeast species, some of which are known from a very few strains up to now. Interestingly yeasts known for their role in the wine making process, such as Saccharomyces cerevisiae, Saccharomyces bayanus ssp. uvarum, Torulaspora delbrueckii, and Zygosaccharomyces bailii, were not found in the grape samples.

  9. Pathogenic variation and sexual reproduction in Swedish populations of Bremia lactucae.

    Science.gov (United States)

    Gustafsson, M; Liljeroth, E; Gustafsson, I

    1985-09-01

    The host-pathogen interaction between lettuce (Lactuca sativa) and downy mildew (Bremia lactucae) is mainly differential and the resistance so far utilized in the host is vertical. As in many other obligate parasites, the introduction of cultivars with new vertical resistance has exerted a strong selection pressure on the pathogen resulting in significant changes in virulence frequencies and in the establishment of races with new combinations of virulence. Genetic diversity in pathogen populations may arise through mutation and gene flow, and new virulence genotypes may then be established through parasexuality and sexual recombination. In Swedish populations of Bremia lactucae, the pattern of variation in the parasite agrees well with that which might be expected in a diploid, outcrossing organism with frequent sexual reproduction. This is supported by: two or more isolates, different in virulence and mating type, may occur together on the same lettuce leaf; zygotes (oospores) are formed in all populations investigated and the frequency varies from 22% to 98%; oospores germinate rather frequently under suitable conditions. To breed for resistance in dynamic host-pathogen systems such as this one is difficult and the program should preferably be based on race-non-specific resistance.

  10. Phytophthora aquimorbida sp. nov. and Phytophthora taxon 'aquatilis' recovered from irrigation reservoirs and a stream in Virginia, USA.

    Science.gov (United States)

    Hong, Chuanxue; Richardson, Patricia A; Hao, Wei; Ghimire, Sita R; Kong, Ping; Moorman, Gary W; Lea-Cox, John D; Ross, David S

    2012-01-01

    Two distinct subgroups (L2 and A(-2)) were recovered from irrigation reservoirs and a stream in Virginia, USA. After molecular, morphological and physiological examinations, the L2 subgroup was named Phytophthora aquimorbida and the A(-2) designated as Phytophthora taxon 'aquatilis'. Both taxa are homothallic. P. aquimorbida is characterized by its noncaducous and nonpapillate sporangia, catenulate and radiating hyphal swellings and thick-walled plerotic oospores formed in globose oogonia mostly in the absence of an antheridium. P. taxon 'aquatilis' produces plerotic oospores in globose oogonia mostly with a paragynous antheridium. It has semi-papillate, caducous sporangia with variable pedicels, but it does not have hyphal swelling. Analyses of ITS, CO1, β-tubulin and NADH1 sequences revealed that P. aquimorbida is closely related to P. hydropathica, P. irrigata and P. parsiana, and P. taxon 'aquatilis' is related to P. multivesiculata. The optimum temperature for culture growth is 30 and 20 C for P. aquimorbida and P. taxon 'aquatilis' respectively. Both taxa were pathogenic to rhododendron plants and caused root discoloration, pale leaves, wilting, tip necrosis and dieback. Their plant biosecurity risk also is discussed.

  11. Emerging oomycete threats to plants and animals.

    Science.gov (United States)

    Derevnina, Lida; Petre, Benjamin; Kellner, Ronny; Dagdas, Yasin F; Sarowar, Mohammad Nasif; Giannakopoulou, Artemis; De la Concepcion, Juan Carlos; Chaparro-Garcia, Angela; Pennington, Helen G; van West, Pieter; Kamoun, Sophien

    2016-12-05

    Oomycetes, or water moulds, are fungal-like organisms phylogenetically related to algae. They cause devastating diseases in both plants and animals. Here, we describe seven oomycete species that are emerging or re-emerging threats to agriculture, horticulture, aquaculture and natural ecosystems. They include the plant pathogens Phytophthora infestans, Phytophthora palmivora, Phytophthora ramorum, Plasmopara obducens, and the animal pathogens Aphanomyces invadans, Saprolegnia parasitica and Halioticida noduliformans For each species, we describe its pathology, importance and impact, discuss why it is an emerging threat and briefly review current research activities.This article is part of the themed issue 'Tackling emerging fungal threats to animal health, food security and ecosystem resilience'.

  12. Dissemination of Botryosphaeriaceae conidia in vineyards in the semiarid Mediterranean climate of the Valparaíso Region of Chile

    Directory of Open Access Journals (Sweden)

    Diego VALENCIA

    2015-09-01

    Full Text Available The dispersal of Botryosphaeriaceae conidia was studied in two vineyards in the Valparaiso Region of Chile, where semiarid Mediterranean climate conditions prevail. The objective of this study was to record trappings of Botryosphaeriaceae conidia over one year on a weekly basis and to associate these records with weather and agrochemical application data. Two Chardonnay vineyards in Casablanca and two Cabernet Sauvignon vineyards in Panquehue were selected. In each vineyard at each weekly sampling, five microscope slides covered with petroleum jelly (Vaseline were installed near grapevines affected by Botryosphaeria cankers. In addition, Botryosphaeriaceae species associated with cankers in these vineyards were identified. Seasonal peaks of Botryosphaeriaceae conidia were detected among the different grape producing areas. In Casablanca, peaks of conidium dispersal were observed in winter 2013 and in autumn and winter 2014. Peaks of large numbers of conidia were mainly associated with precipitation events equal to or greater than 0.2 mm. The species observed in the spore traps were preliminarily identified as Diplodia seriata, Spencermartinsia viticola and Neofusicoccum sp., and isolations from adjacent plants confirmed the presence of D. seriata, S. viticola and N. australe in these vineyards. In Panquehue, peaks of conidium dispersal were observed in winter 2013, with a lower relative abundance of spores detected compared with Casablanca. Conidia dispersal recommenced in autumn 2014 but remained low later in winter. In this location, D. seriata was the main species detected, followed by S. viticola. No Neofusicoccum species was detected in this area. No association was found between conidium dispersal and the volume of water (≤ 1000 L ha-1 used in the application of agrochemical products.

  13. IN VITRO ANTIFUNGAL ACTIVITY OF ESSENTIAL OILS ON GROWTH OF PHYTOPATHOGENIC FUNGI

    Directory of Open Access Journals (Sweden)

    Jasenka Ćosić

    2010-12-01

    Full Text Available Eleven essential oils (clove, rosemary, cinnamon leaf, sage, scots pine, neroli, peppermint, aniseed, caraway, lavander, common thyme were tested for in vitro antifungal activity on twelve plant pathogenic fungi (Fusarium graminearum, F. verticillioides, F. subglutinans, F. oxysporum, F. avenaceum, Diaporthe helianthi, Diaporthe phaseolorum var. caulivora, Phomopsis longicolla, P. viticola, Helminthosporium sativum, Colletotrichum coccodes, Thanatephorus cucumeris. The results indicated that all oils except scots pine and neroli had antifungal activity against some or all tested fungi. The best antifungal activity had common thyme, cinnamon leaf, clove and aniseed oils. When compared to control, scots pine, neroli and sage oils stimulated mycelium growth of some investigated fungi.

  14. Saprolegnia oliviae sp. nov. isolated from an Argentine river (Tierra del Fuego Province, Argentina).

    Science.gov (United States)

    Steciow, Mónica Mirta

    2003-02-28

    Saprolegnia oliviae sp. nov. is described from litter (floating dead twigs, leaves and roots) in the Olivia River, Ushuaia Department, Tierra del Fuego Province (Argentina). The new species is illustrated and compared with other species of the genus. Distinguishing characteristics of S. oliviae are the production of smooth oogonia (with some lateral or terminal projections) and the absence of antheridial branches on the majority of the oogonia, but when present, they are mostly diclinous, at times oogonia are supplied with androgynous and monoclinous antheridial branches. The oogonial stalks are predominantly short and straight or long and bent, curved or many times coiled; oospores are distinctive subcentric, (1-) 15-50 (-70) per oogonium. Morphological details of the new species and its comparison with other described species are discussed here.

  15. Pythium campanulatum sp. nov., isolated from the rhizosphere of maize, its taxonomy, ITS region of rDNA, and comparison with related species.

    Science.gov (United States)

    Mathew, Ritta; Singh, Kusum Kumari; Paul, Bernard

    2003-09-12

    Pythium campanulatum sp. nov. was isolated from some soil samples taken in the rhizosphere of maize (Zea mays) in north-eastern India. This species is characterized by the absence of zoospores and sporangia, antheridial branches wrapping around the oogonia leaving one to two campanulate antheridial cells after fertilization, and aplerotic oospores. The ITS region of its rDNA is comprised of 922 bases. This oomycete is closely related to Pythium orthogonon, Pythium nunn and Pythium toruloides. However, it has its own characteristic features and is completely devoid of zoospores. Taxonomic description of this new species and its comparison with related oomycetes, together with the sequence of the PCR-amplified internal transcribed region (spacers ITS1, ITS2, and the gene 5.8S) of its rDNA are given here.

  16. PsMPK7, a stress-associated mitogen-activated protein kinase (MAPK) in Phytophthora sojae, is required for stress tolerance, reactive oxygenated species detoxification, cyst germination, sexual reproduction and infection of soybean.

    Science.gov (United States)

    Gao, Jian; Cao, Mingna; Ye, Wenwu; Li, Haiyang; Kong, Liang; Zheng, Xiaobo; Wang, Yuanchao

    2015-01-01

    The sensing of stress signals and their transduction into appropriate responses are crucial for the adaptation, survival and infection of phytopathogenic fungi and oomycetes. Amongst evolutionarily conserved pathways, mitogen-activated protein kinase (MAPK) cascades function as key signal transducers that use phosphorylation to convey information. In this study, we identified a gene, designated PsMPK7, one of 14 predicted genes encoding MAPKs in Phytophthora sojae. PsMPK7 was highly transcribed in each tested stage, but was up-regulated in the zoospore, cyst and cyst germination stages. Silencing of PsMPK7 affected the growth of germinated cysts, oospore production and the pathogenicity of soybean. PsMPK7 transcription was induced by stresses from sorbitol, NaCl and hydrogen peroxide. Transformants in which PsMPK7 expression was silenced (PsMPK7-silenced) were significantly more sensitive to osmotic and oxidative stress. Aniline blue and diaminobenzidine staining revealed that the silenced lines did not suppress the host reactive oxygen species (ROS) burst, indicating that either the inoculated plants activated stronger defence responses to the transformants and/or the PsMPK7-silenced transformants failed to overcome plant defences. In addition, extracellular secretion of laccase decreased in the silenced lines. Overall, our results indicate that the PsMPK7 gene encodes a stress-associated MAPK in P. sojae that is important not only for responses to various stresses, but also for ROS detoxification, cyst germination, sexual oospore production and infection of soybean.

  17. Botryosphaeriaceae as potential pathogens of prunus species in South Africa, with descriptions of Diplodia africana and Lasiodiplodia plurivora sp. nov.

    Science.gov (United States)

    Damm, Ulrike; Crous, Pedro W; Fourie, Paul H

    2007-01-01

    Botryosphaeriaceae are common dieback and canker pathogens of woody host plants, including stone fruit trees. In the present study the diversity of members of the Botryosphaeriaceae isolated from symptomatic wood of Prunus species (plum, peach, nectarine and apricot) was determined in stone fruit-growing areas in South Africa. Morphological and cultural characteristics as well as DNA sequence data (5.8S rDNA, ITS-1, ITS-2 and EF-1a) were used to identify known members and describe novel members of Botryosphaeriaceae. From the total number of wood samples collected (258) 67 isolates of Botryosphaeriaceae were obtained, from which eight species were identified. All species were associated with wood necrosis. Diplodia seriata (= "Botryosphaeria" obtusa) was dominant, and present on all four Prunus species sampled, followed by Neofusicoccum vitifusiforme and N. australe. First reports from Prunus spp. include N. vitifusiforme, Dothiorella viticola and Diplodia pinea. This is also the first report of D. mutila from South Africa. Two species are newly described, namely Lasiodiplodia plurivora sp. nov. from P. salicina and Diplodia africana sp. nov. from P. persica. All species, except Dothiorella viticola, caused lesions on green nectarine and/or plum shoots in a detached shoot pathogenicity assay.

  18. APPLICATION OF BIOFUNGICIDES CALLED GUAPSIN AND TRIHODERMIN DURING THE PERIOD OF PRODUCTION OF GRAFTED PLANTING MATERIAL AT THE STAGE OF STRATIFICATION OF GRAFTED GRAPES CUTTINGS

    Directory of Open Access Journals (Sweden)

    Stranishevskaya E. P.

    2016-09-01

    Full Text Available The article gives the results of the three years of research to identify the main species composition of fungal pathogens when stratified open and closed methods. Biological efficiency of Guapsin, 0,2%, and Trichodermin, 0,5% is shown. During the period of stratification, using an open method "on the water", the following pathogens have evolved: Cladosporium herbarum (18,6%, Trichotecium roseum (16,3%, Phomopsis viticola (13,9%, Alternaria spp. (13,4%, Penicillium sp. ( 9,1%, Aspergillus sp. (5,6%, Pythium sp. (4,3%, Gonatobotrys flava (4,3%, Mycelia sterilia (2,8%, Botrytis cinerea (2,2%. The studies found that the close method of stratification in the peat developed the following specific composition of pathogens: Penicillium spp. (21,1%, Phomopsis viticola (16,4%, Botrytis cinerea (15,3%, Trichotecium roseum (11,1%, Alternaria spp. (8,2%, Mycelia sterilia (1,0%. During the period of stratification, biological efficiency Guapsin, 0.2%, and Trichodermin, 0,5%, was 31,9-88,0% and 28,2-86,3%, respectively

  19. Penicillium daejeonium sp. nov., a new species isolated from a grape and schisandra fruit in Korea.

    Science.gov (United States)

    Sang, Hyunkyu; An, Tae-Jin; Kim, Chang Sun; Choi, Young Phil; Deng, Jian-Xin; Paul, Narayan Chandra; Sung, Gi-Ho; Yu, Seung Hun

    2013-08-01

    Two isolates of monoverticillate Penicillium species were collected from a grape and schisandra fruit in Korea. Multigene phylogenetic analyses with the nuclear ribosomal internal transcribed spacer (ITS) region and genes encoding β-tubulin (benA) and calmodulin (cmd), as well as morphological analyses revealed that the two isolates are members of the P. sclerotiorum complex in Penicillium subgenus Aspergilloides, but different from species of the P. sclerotiorum complex. The isolates are closely related to P. cainii, P. jacksonii, and P. viticola in terms of their multigene phylogeny, but their colony and conidiophore morphologies differ from those of closely related species. The name P. daejeonium is proposed for this unclassified new species belonging to the P. sclerotiorum complex in subgenus Aspergilloides.

  20. Manganese Oxidation in Petri Disease Fungi as a Novel Taxonomic Character

    Directory of Open Access Journals (Sweden)

    B.E. Overton

    2006-04-01

    Full Text Available Manganese (Mn oxidation was evaluated for Petri disease fungi by using potato dextrose agar (PDA amended with Mn sulfate. Phaeomoniella chlamydospora showed reduced growth on PDA amended with 3000 ppm Mn sulfate and exhibited no growth at higher concentrations. All Phaeoacremonium spp. evaluated were manganese tolerant and grew on PDA amended with as much as 6000 ppm Mn sulfate. Phaeoacremonium viticola and P. angustius s.s. oxidized manganese in vitro. Stereum hirsutum and Fomitiporia punctata, known white-rot basidiomycetes, were used as controls. Stereum hirsutum oxidized Mn in vitro and grew on PDA amended with 6000 ppm Mn sulfate. Fomitiporia punctata had reduced growth at 300 ppm and no growth at higher concentrations of Mn sulfate with no clear evidence of Mn oxidation.

  1. The sedimentary sequence from the Lake Ķūži outcrop, central Latvia: implications for late glacial stratigraphy

    Directory of Open Access Journals (Sweden)

    Tiiu Koff

    2011-06-01

    Full Text Available Sediment samples from an outcrop in the near-shore area of Lake Ķūži (Vidzeme Heights, Central Latvia were investigated using palaeobotanical (pollen and macrofossil analysis and lithological (grain-size analysis methods and accelerator mass spectrometry 14C dating. A dark, organic-rich sediment layer was found below 1.7 m of sandy layers approximately 30 cm above the present lake level. Radiocarbon dating of a wood sample from the lowermost layer (11 050 ± 60 14C BP, 13 107–12 721 cal BP shows that the layer is of late glacial age. The composition of the pollen spectra is characterized by Betula nana, Cyperaceae pollen and spores of Equisetum, confirming that the lowermost sediments were formed during the late glacial. Fossils of obligate aquatic organisms in the upper layer, which include oospores of Characeae and seeds of Potamogeton, indicate an open water environment. Pollen of Myriophyllum and Potamogeton and non-pollen palynomorphs, such as algal Botryococcus and Pediastrum cf. boryanum, confirm this conclusion. The pollen assemblage from the greyish loam layer following this lacustrine phase shows a pattern characteristic of the Younger Dryas vegetation before the start of the real expansion of birch forests at the beginning of the Holocene.

  2. Identification of Phytophthora sojae genes involved in asexual sporogenesis

    Indian Academy of Sciences (India)

    Ziying Wang; Xhaoxia Wang; Jie Shen; Guangyue Wang; Xiaoxi Zhu; Hongxia Lu

    2009-08-01

    To explore the molecular mechanisms involved in asexual spore development in Phytophthora sojae, the zoospores of strain PS26 were treated with ultraviolet (UV) irradiation. After selection, a mutant progeny, termed PS26-U03, was obtained and demonstrated to exhibit no oospore production. A suppression subtractive hybridization (SSH) approach was developed to investigate differences in gene expression between PS26 and PS26-U03 during asexual sporogenesis. Of the 126 sequences chosen for examination, 39 putative unigenes were identified that exhibit high expression in PS26. These sequences are predicted to encode proteins involved in metabolism, cell cycle, protein biosynthesis, cell signalling, cell defence, and transcription regulation. Seven clones were selected for temporal expression analysis using RT-PCR based on the results of the dot-blot screens. Three of the selected genes, developmental protein DG1037 (UB88), glycoside hydrolase (UB149) and a hypothetical protein (UB145), were expressed only in PS26, whereas the transcripts of phosphatidylinositol-4-phosphate 5-kinase (UB36), FAD-dependent pyridine nucleotide-disulphide oxidoreductase (UB226) and sugar transporter (UB256) were expressed at very low levels in PS26-U03 but at high levels in PS26.

  3. Occurrence and characterization of a Phytophthora sp. pathogenic to asparagus (Asparagus officinalis) in Michigan.

    Science.gov (United States)

    Saude, C; Hurtado-Gonzales, O P; Lamour, K H; Hausbeck, M K

    2008-10-01

    A homothallic Phytophthora sp. was recovered from asparagus (Asparagus officinalis) spears, storage roots, crowns, and stems in northwest and central Michigan in 2004 and 2005. Isolates (n = 131) produced ovoid, nonpapillate, noncaducous sporangia 45 microm long x 26 microm wide and amphigynous oospores of 25 to 30 microm diameter. Mycelial growth was optimum at 25 degrees C with no growth at 5 and 30 degrees C. All isolates were sensitive to 100 ppm mefenoxam. Pathogenicity studies confirmed the ability of the isolates to infect asparagus as well as cucurbits. Amplified fragment length polymorphism analysis of 99 isolates revealed identical fingerprints, with 12 clearly resolved fragments present and no clearly resolved polymorphic fragments, suggesting a single clonal lineage. The internal transcribed spacer regions of representative isolates were homologous with a Phytophthora sp. isolated from diseased asparagus in France and a Phytophthora sp. from agave in Australia. Phylogenetic analysis supports the conclusion that the Phytophthora sp. isolated from asparagus in Michigan is a distinct species, and has been named Phytophthora asparagi.

  4. Green fluorescent protein (GFP) as a vital marker for studying the interaction of Phytophthora sojae and soybean

    Institute of Scientific and Technical Information of China (English)

    CHEN XiaoRen; CHENG BaoPing; WANG XinLe; DONG SuoMeng; WANG YongLin; ZHENG XiaoBo; WANG YuanChao

    2009-01-01

    Transgenic Phytophthora sojae strains that produce green fluorescent protein (GFP) were obtained after stable DNA integration using the Hsp70 promoter and the Ham34 terminator of Bremia lactucae.The expression of GFP during different developmental stages of P.sojae was observed using fluorescent microscopy.Based on this reporter system,the histopathologic events caused by the pathogen in soybean leaves,hypocotyls and roots were monitored.Meanwhile,the difference in resistance between different soybean cultivars against P.sojae was analyzed microscopically in roots.The results indicate that GFP can be stably expressed in zoosporangia,zoospores,cysts,hyphae and oospores of .sojae.Using the GFP marker,the infecting pathogens in leaves,hypocotyls and roots of host could be distinctly visualized.The germ tube length of cysts germinating on the roots of resistant cultivar Nannong 8848 was longer than that on the roots of susceptible culUvar Hefeng 35.These results show for the first time that this eukaryotic reporter can be used in P.sojae as a stable and vital marker,allowing the study of genetics of this hemibiotrophic pathogen.

  5. Disentangling Peronospora on Papaver: phylogenetics, taxonomy, nomenclature and host range of downy mildew of opium poppy (Papaver somniferum and related species.

    Directory of Open Access Journals (Sweden)

    Hermann Voglmayr

    Full Text Available Based on sequence data from ITS rDNA, cox1 and cox2, six Peronospora species are recognised as phylogenetically distinct on various Papaver species. The host ranges of the four already described species P. arborescens, P. argemones, P. cristata and P. meconopsidis are clarified. Based on sequence data and morphology, two new species, P. apula and P. somniferi, are described from Papaver apulum and P. somniferum, respectively. The second Peronospora species parasitizing Papaver somniferum, that was only recently recorded as Peronospora cristata from Tasmania, is shown to represent a distinct taxon, P. meconopsidis, originally described from Meconopsis cambrica. It is shown that P. meconopsidis on Papaver somniferum is also present and widespread in Europe and Asia, but has been overlooked due to confusion with P. somniferi and due to less prominent, localized disease symptoms. Oospores are reported for the first time for P. meconopsidis from Asian collections on Papaver somniferum. Morphological descriptions, illustrations and a key are provided for all described Peronospora species on Papaver. cox1 and cox2 sequence data are confirmed as equally good barcoding loci for reliable Peronospora species identification, whereas ITS rDNA does sometimes not resolve species boundaries. Molecular phylogenetic data reveal high host specificity of Peronospora on Papaver, which has the important phytopathological implication that wild Papaver spp. cannot play any role as primary inoculum source for downy mildew epidemics in cultivated opium poppy crops.

  6. Aproximación a las comunidades de carófitos que existieron en la Albufera de Valencia a partir del estudio de las oósporas del sedimento

    Directory of Open Access Journals (Sweden)

    Soulié-Märsche, Ingeborg

    2009-12-01

    Full Text Available Charophyte oospores and their features (size, shape, colour, outer wall ornamentation, etc. are able to persist in seed banks of aquatic ecosystems. Thus, these characteristics can be used for taxonomic species identification, thereby providing knowledge about the specific composition of the charophyte community in a habitat that currently lacks, or has greatly altered, submerged vegetation. Several years ago, l’Albufera de València lost the charophyte meadows that covered the lake bottom in the past, which had never been studied in detail. Several sediment cores were extracted from the lagoon at three sites and the oospores and gyrogonites were isolated and characterized. The oospore/gyrogonite densities in the sediment layers were estimated. Ten different taxa have been identified: six belonging to genus Chara [Ch. vulgaris, Ch. Hispida var. major, Ch. fragilis (Ch. globularis, Ch. Hispida var. baltica, Ch. aspera, Ch. tomentosa]; one Nitella species (N. hyalina, two Tolypella species (T. glomerata and T. hispanica and one Lamprothamnium species (L. papulosum. The latter, which indicates the period when water salinity was high in the lagoon, was found in the deepest sediment layers. An increase in the oospore abundance and diversity was observed in line with sediment depth. This indicates, on the one hand, that the lagoon went through different ecological stages (with important changes in salinity, mainly and, on the other hand, the eutrophication process that the lagoon has been undergoing for at least the last 45 years, has progressively affected the abundance and diversity of the charophyte community.

    Las oósporas de carófitos y sus características (tamaño, forma, color, ornamentación de la pared, etc. pueden persistir en los bancos de semillas del sedimento de ecosistemas acuáticos, y pueden ser utilizadas para identificar los diferentes táxones y obtener de este modo una

  7. A high-temperature tolerant species in clade 9 of the genus Phytophthora: P. hydrogena sp. nov.

    Science.gov (United States)

    Yang, Xiao; Gallegly, Mannon E; Hong, Chuanxue

    2014-01-01

    A previously unknown Phytophthora species was isolated from irrigation water in Virginia, USA. This novel species produces abundant noncaducous and nonpapillate sporangia in soil water extract solution. It sometimes produces chlamydospores and hyphal swellings in aged cultures and in Petri's solution. This species has optimum vegetative growth at 30 C and grows well at 35 C. The lowest and highest temperatures for growth are 5 and 40 C. All isolates examined in this study are compatibility type A1 and produce mostly plerotic oospores when paired with an A2 mating-type tester of P. cinnamomi. Sequence analyses of the rDNA internal transcribed spacer (ITS) regions and the mitochondrially encoded cytochrome c oxidase 1 (cox 1) gene placed this species in clade 9 of the genus Phytophthora. These characteristics support the description of this taxon as a new species for which we propose the name P. hydrogena sp. nov. Further phylogenetic and physiological investigations of clade 9 species revealed a high-temperature tolerant cluster including P. hydrogena, P. aquimorbida, P. hydropathica, P. irrigata, P. chrysanthemi, P. insolita, P. polonica and P. parsiana. These species all grow well at 35 C. The monophyly of the species in this heat-tolerant cluster except P. insolita and P. polonica is highly supported by the maximum-likelihood analyses of the ITS and cox 1 sequences.

  8. The importin α subunit PsIMPA1 mediates the oxidative stress response and is required for the pathogenicity of Phytophthora sojae.

    Science.gov (United States)

    Yang, Xinyu; Ding, Fa; Zhang, Lei; Sheng, Yuting; Zheng, Xiaobo; Wang, Yuanchao

    2015-09-01

    The sensing of extracellular signals and their transduction into an appropriate response are crucial for the survival and virulence of plant pathogens. Eukaryotic plant pathogens must overcome the obstacles posed by nuclear membranes to manipulate gene expression to adapt to the host challenge. A highly sophisticated mechanism is the use of importins to transport proteins into the nucleus. In this study, we identified a conserved importin α gene, PsIMPA1, in Phytophthora sojae that was differentially expressed during the life cycle of this soybean pathogen. PsIMPA1 expression was lowest in zoospores and cysts but relatively consistent during the other life cycle stages, except for a slight increase at 6h post infection. Silenced mutants Psimpa1 had a decreased growth rate, an aberrant mycelial morphology, and a severely impaired ability to form oospores and sporangia. In addition, the Psimpa1 mutants exhibited reduced pathogenicity compared to the wild type. 3,3-Diaminobenzidine (DAB) staining and in vitro hydrogen peroxide tolerance assays showed that the scavenging of reactive oxygen species by these mutants was significantly impaired. Taken together, these results indicate that PsIMPA1 regulates multiple processes during the life cycle of P. sojae.

  9. Genome Sequencing and Mapping Reveal Loss of Heterozygosity as a Mechanism for Rapid Adaptation in the Vegetable Pathogen Phytophthora capsici

    Energy Technology Data Exchange (ETDEWEB)

    Lamour, Kurt H.; Mudge, Joann; Gobena, Daniel; Hurtado-Gonzales, Oscar P.; Schmutz, Jeremy; Kuo, Alan; Miller, Neil A.; Rice, Brandon J.; Raffaele, Sylvain; Cano, Liliana M.; Bharti, Arvind K.; Donahoo, Ryan S.; Finely, Sabra; Huitema, Edgar; Hulvey, Jon; Platt, Darren; Salamov, Asaf; Savidor, Alon; Sharma, Rahul; Stam, Remco; Sotrey, Dylan; Thines, Marco; Win, Joe; Haas, Brian J.; Dinwiddie, Darrell L.; Jenkins, Jerry; Knight, James R.; Affourtit, Jason P.; Han, Cliff S.; Chertkov, Olga; Lindquist, Erika A.; Detter, Chris; Grigoriev, Igor V.; Kamoun, Sophien; Kingsmore, Stephen F.

    2012-02-07

    The oomycete vegetable pathogen Phytophthora capsici has shown remarkable adaptation to fungicides and new hosts. Like other members of this destructive genus, P. capsici has an explosive epidemiology, rapidly producing massive numbers of asexual spores on infected hosts. In addition, P. capsici can remain dormant for years as sexually recombined oospores, making it difficult to produce crops at infested sites, and allowing outcrossing populations to maintain significant genetic variation. Genome sequencing, development of a high-density genetic map, and integrative genomic or genetic characterization of P. capsici field isolates and intercross progeny revealed significant mitotic loss of heterozygosity (LOH) in diverse isolates. LOH was detected in clonally propagated field isolates and sexual progeny, cumulatively affecting >30percent of the genome. LOH altered genotypes for more than 11,000 single-nucleotide variant sites and showed a strong association with changes in mating type and pathogenicity. Overall, it appears that LOH may provide a rapid mechanism for fixing alleles and may be an important component of adaptability for P. capsici.

  10. Phytophthora inundata sp. nov., a part heterothallic pathogen of trees and shrubs in wet or flooded soils.

    Science.gov (United States)

    Brasier, Clive M; Sanchez-Hernandez, Esperanza; Kirk, Susan A

    2003-04-01

    A Phytophthora pathogen of trees and shrubs previously designated Phytophthora sp. O-group is formally named as P. inundata sp. nov. P. inundata falls within the P. gonapodyides-P. megasperma major ITS Clade 6, its present nearest known relative being P. humicola. It has non-papillate sporangia, fairly large oogonia (average ca 40 microns) with thick walled oospores, amphigynous antheridia, a distinctive colony type, a high optimum temperature for growth of 28-30 degrees C, fast growth at the optimum, and a high upper temperature limit for growth of ca 35-37 degrees. A study of the breeding system of eight P. inundata isolates showed them to be classically heterothallic with A1 and A2 compatibility types. However some P. inundata A1 x A2 combinations failed to mate even though the same isolates mated successfully with P. drechsleri testers. Others were 'silent' A1s or A2s, unable to produce their own gametangia but able to induce gametangial formation in the opposite sexual compatibility type of another species. This indicates a partial breakdown of the sexual mechanism in the species. Two isolates (one A1 and one A2) were unpredictably and chimaerically self-fertile, suggesting A1 + A2 chromosomal heteroploidy. The association of P. inundata with ponds and rivers and with root and collar roots of trees and shrubs after flooding is discussed.

  11. Transformation of Pythium aphanidermatum to geneticin resistance.

    Science.gov (United States)

    Weiland, John J

    2003-03-01

    Conditions for the production of protoplasts and gene transfer in Pythium aphanidermatum were investigated. Efficient protoplast generation was possible after culture of mycelium in potato dextrose broth followed by digestion with 0.5% (w/v) each of cellulase and beta- d-glucanase. Plasmid pHAMT35N/SK encoding the nptII gene under control of the Ham34 promoter from the oomycete Bremia lactucae was used to define electroporation parameters for gene transfer. A square-wave electroporation pulse of 2500 V/cm at 50 microF capacitance reproducibly produced transformants, albeit at low efficiency (0.1-0.4 transformants from approximately 10(5) regenerable protoplasts per microgram of DNA). Thirty-two independent transformants exhibited wild-type growth on potato dextrose agar amended with geneticin at 50 microg/ml, a concentration that near completely inhibited the growth of untransformed P. aphanidermatum. Southern blot analysis indicated that transforming DNA was integrated into the oomycete genome and that the DNA was stably inherited through sporogenesis. Growth on geneticin-free media, the ability to form zoospores or oospores, and the ability to cause disease in sugarbeet seedlings in the laboratory were indistinguishable between a subset of the transformed isolates and the progenitor isolate 898B. Co-electroporation of pHAMT35N/SK with plasmid pACT-GUS encoding the Escherichia coli gusA gene controlled by oomycete transcriptional promoter and terminator sequences or with pEGFP encoding enhanced green fluorescent protein under the control of the immediate early promoter from the mammalian cytomegalovirus produced, respectively, stable beta-glucuronidase and transient expression of blue-green fluorescence. Application of the technique to studies on the biochemical basis for pathogenesis in this agriculturally important group of fungi is discussed.

  12. Molecular detection of Peronospora variabilis in quinoa seed and phylogeny of the quinoa downy mildew pathogen in South America and the United States.

    Science.gov (United States)

    Testen, Anna L; del Mar Jiménez-Gasco, María; Ochoa, José B; Backman, Paul A

    2014-04-01

    Quinoa (Chenopodium quinoa) is an important export of the Andean region, and its key disease is quinoa downy mildew, caused by Peronospora variabilis. P. variabilis oospores can be seedborne and rapid methods to detect seedborne P. variabilis have not been developed. In this research, a polymerase chain reaction (PCR)-based detection method was developed to detect seedborne P. variabilis and a sequencing-based method was used to validate the PCR-based method. P. variabilis was detected in 31 of 33 quinoa seed lots using the PCR-based method and in 32 of 33 quinoa seed lots using the sequencing-based method. Thirty-one of the quinoa seed lots tested in this study were sold for human consumption, with seed originating from six different countries. Internal transcribed spacer (ITS) and cytochrome c oxidase subunit 2 (COX2) phylogenies were examined to determine whether geographical differences occurred in P. variabilis populations originating from Ecuador, Bolivia, and the United States. No geographical differences were observed in the ITS-derived phylogeny but the COX2 phylogeny indicated that geographical differences existed between U.S. and South American samples. Both ITS and COX2 phylogenies supported the existence of a Peronospora sp., distinct from P. variabilis, that causes systemic-like downy mildew symptoms on quinoa in Ecuador. The results of these studies allow for a better understanding of P. variabilis populations in South America and identified a new causal agent for quinoa downy mildew. The PCR-based seed detection method allows for the development of P. variabilis-free quinoa seed, which may prove important for management of quinoa downy mildew.

  13. Antifungal compounds from turmeric and nutmeg with activity against plant pathogens.

    Science.gov (United States)

    Radwan, Mohamed M; Tabanca, Nurhayat; Wedge, David E; Tarawneh, Amer H; Cutler, Stephen J

    2014-12-01

    The antifungal activity of twenty-two common spices was evaluated against plant pathogens using direct-bioautography coupled Colletotrichum bioassays. Turmeric, nutmeg, ginger, clove, oregano, cinnamon, anise, fennel, basil, black cumin, and black pepper showed antifungal activity against the plant pathogens Colletotrichum acutatum, Colletotrichum fragariae, and Colletotrichum gloeosporioides. Among the active extracts, turmeric and nutmeg were the most active and were chosen for further investigation. The bioassay-guided fractionation led to the isolation of three compounds from turmeric (1-3) and three compounds from nutmeg (4-6). Their chemical structures were elucidated by spectroscopic analysis including HR-MS, 1D, and 2D NMR as curcumin (1), demethoxycurcumin (2) and bisdemethoxy-curcumin (3), erythro-(7R,8R)-Δ(8')-4,7-dihydroxy-3,3',5'-trimethoxy-8-O-4'-neolignan (4), erythro-(7R,8R)-Δ8'-7-acetoxy-3,4,3',5'-tetra-methoxy-8-O-4'-neolignan (5), and 5-hydroxy-eugenol (6). The isolated compounds were subsequently evaluated using a 96-well microbioassay against plant pathogens. At 30 μM, compounds 2 and 3 possessed the most antifungal activity against Phomopsis obscurans and Phomopsis viticola, respectively.

  14. Fungi inhabiting healthy grapevine canes (Vitis spp. in some nurseries

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    Ewa Król

    2012-12-01

    Full Text Available The purpose of this study, conducted in the years 2000 - 2002, was to identify fungi species colonizing apparently healthy canes and to investigate whether canes storage modify the quantitative and qualitative composition of these fungi. The plant material was collected from 5 commercial plantations growing in various regions of Poland, taking into consideration 8 cultivars which were the most frequently cultivated. From each plantation and cultivar 20 apparently healthy canes were randomly sampled in two terms: before storage - November/December (term I and 3-4 months after storage - February/March (term II. The results showed that from asymptomatic canes 2746 isolates of fungi belonging to 23 species were obtained, but the majority of them origined from canes analysed after storage. It was found that P. viticola is able to live latently within grapevine tissue in Polish conditions because isolates of this fungus from visually healty canes the all studied plantations and terms were obtained. Among the other fungi species inhabiting grapevine canes Alternaria alternata and Fusarium spp. dominated. Moreover, both in term I and term II Botrytis cinerea, Phoma spp., Epicoccum purpurascens and Cladosporium cladosporioides were frequently isolated, whereas fungi from the genus Acremonium only in the term I. Each time isolates of Trichoderma spp. and Gliocladium spp. were also obtained. Inhabitation of grapevine canes by various fungi species shown in the present experiment indicate the danger of pathogens spread with propagation material on the new plantations.

  15. Fungal communities living in the wood of different cultivars of young Vitis vinifera plants

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    L. Casieri

    2009-05-01

    Full Text Available The fungal communities associated with fi ve Vitis vinifera cultivars grown in Switzerland (‘Humagne’, ‘Chasselas’, ‘Arvine’, ‘Gamaret’ and ‘Gamay’ were examined. Of the 703 fungal isolates obtained in pure culture, 66 operational taxonomic units (OTUs were defi ned. The results show that: the great majority of the fungi isolated in this study were ascomycetes, with a high proportion of Sordariomycetes (mainly Hypocreales, Sordariales and Diaporthales; different fungal OTUs were associated with different cultivars; graft and rootstock contributed equally to the fungal community composition; Esca- or Petri-related species occurred sporadically in the different cultivars, with some of them occupying specifi c tissues or parts of the plant (e.g.: Botryosphaeriaceous species, Phaeomoniella chlamydospora and Phomopsis viticola; almost 25% of OTUs occurred in different plant parts in most cultivars, which suggests an easy spread outwards from the infected material (graft or rootstock, which might be explained by the fungal propagules being transported through the xylem vessels.

  16. Development and dissection of diagnostic SNP markers for the downy mildew resistance genes Pl Arg and Pl 8 and maker-assisted gene pyramiding in sunflower (Helianthus annuus L.).

    Science.gov (United States)

    Qi, L L; Talukder, Z I; Hulke, B S; Foley, M E

    2017-02-03

    Diagnostic DNA markers are an invaluable resource in breeding programs for successful introgression and pyramiding of disease resistance genes. Resistance to downy mildew (DM) disease in sunflower is mediated by Pl genes which are known to be effective against the causal fungus, Plasmopara halstedii. Two DM resistance genes, Pl Arg and Pl 8 , are highly effective against P. halstedii races in the USA, and have been previously mapped to the sunflower linkage groups (LGs) 1 and 13, respectively, using simple sequence repeat (SSR) markers. In this study, we developed high-density single nucleotide polymorphism (SNP) maps encompassing the Pl arg and Pl 8 genes and identified diagnostic SNP markers closely linked to these genes. The specificity of the diagnostic markers was validated in a highly diverse panel of 548 sunflower lines. Dissection of a large marker cluster co-segregated with Pl Arg revealed that the closest SNP markers NSA_007595 and NSA_001835 delimited Pl Arg to an interval of 2.83 Mb on the LG1 physical map. The SNP markers SFW01497 and SFW06597 delimited Pl 8 to an interval of 2.85 Mb on the LG13 physical map. We also developed sunflower lines with homozygous, three gene pyramids carrying Pl Arg , Pl 8 , and the sunflower rust resistance gene R 12 using the linked SNP markers from a segregating F2 population of RHA 340 (carrying Pl 8 )/RHA 464 (carrying Pl Arg and R 12 ). The high-throughput diagnostic SNP markers developed in this study will facilitate marker-assisted selection breeding, and the pyramided sunflower lines will provide durable resistance to downy mildew and rust diseases.

  17. Kajian Penggunaan Pupuk Hayati untuk Mengendalikan Penyakit Akar Gada (Plasmodiophora brassicae pada Tanaman Sawi Daging

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    Diding Rachmawati

    2016-03-01

    Full Text Available Pada budidaya tanaman sawi daging (pakcoi  dijumpai berbagai masalah  serius  yang menghambat upaya peningkatan produksi dan kualitas hasil. Salah satu kendala utama adalah penyakit tular tanah yang disebabkan oleh cendawan Plasmopara brassicae Wor . Serangan patogen tular tanah dapat menekan produksi tanaman hortikultura secara significan. Berbagai upaya telah dilakukan untuk mengendalikan patogen tular tanah antara lain dengan menggunakan bekterisida sistemik . Salah satu alternatif pengendalian yang paling prospektif adalah dengan menggunakan pupuk hayati yang telah diperkaya dengan mikroorganisme. antara lain bakteri selulotik, Azotobacter sp., Azospirillium sp., Rhizobium sp., Pseudomonas sp., Lactobacillus sp., dan  bakteri pelarut fosfat yang bertujuan untuk memperbaiki struktur tanah dan mengendalikan penyakit tular tanah. Penelitian dilakukan di kebun percobaan Karangploso BPTP Jatim,  pada bulan Januari sampai dengan April 2014, menggunakan rancangan acak kelompok, 4 perlakuan dan 6 ulangan. Perlakuan  terdiri dari  : A = Pupuk hayati dosis 15 kg/ha,   B = Pupuk hayati dosis 30 kg/ha,  C = Pupuk hayati dosis 45 kg/ha, D = Cara petani. Tujuan penelitian adalah untuk mengetahui efektifitas pupuk hayati dalam mengendalikan penyakit akar gada  P.brassicae  pada tanaman sawi daging. Hasil penelitian menunjukkan bahwa pemberian pupuk hayati dosis 45 kg/ha dapat memberikan pertumbuhan yang baik terhadap tinggi tanaman ( 26,50 cm, jumlah daun (21 helai, lebar tajuk (33,25 cm, panjang akar (14,38 cm dan bobot/tanaman (380 g/tanaman. Persentase serangan penyakit akar gada terendah juga ditunjukkan oleh pemberian pupuk hayati dosis 45 kg/ha, yaitu sebesar 1,75 % dan penekanan penyakit sebesar 70,83 %.Kata Kunci : Brassica juncea, pupuk hayati, penyakit bengkak akar

  18. Cancro bacteriano da videira: etiologia, epidemiologia e medidas de controle Bacterial canker of grapevine: etiology, epidemiology and control strategies

    Directory of Open Access Journals (Sweden)

    Ana Rosa Peixoto Nascimento

    2004-02-01

    Full Text Available No início de 1998, o cancro bacteriano da videira, causado pela bactéria Xanthomonas campestris pv. viticola, foi detectado pela primeira vez, no Brasil, em parreirais do Submédio São Francisco, onde a doença vem ocasionando prejuízos nas cultivares suscetíveis Red Globe, Itália, Festival, Brasil, Piratininga, Patrícia, Benitaka e Catalunha. Os sintomas, nas folhas, surgem como pontos necróticos (1 a 2mm de diâmetro com ou sem halos amarelados, algumas vezes coalescendo e causando a morte de extensas áreas do limbo foliar. Nas nervuras e pecíolos, nos ramos e ráquis dos frutos, formam-se manchas escuras alongadas que evoluem para fissuras longitudinais de coloração negra conhecidas como cancros. Descoloração vascular é também observada. As bagas são desuniformes em tamanho e cor podendo apresentar lesões necróticas. A disseminação do patógeno ocorre através de material propagativo infectado, material de colheita (contentores, tesouras de poda e raleio, luvas, tratos culturais (desbrota, poda, raleio de bagas, colheita, ventos e chuvas. Apesar da região apresentar um curto período chuvoso, a disseminação da bactéria é mais eficiente durante essa época. Em condições de umidade e temperatura elevadas, o patógeno sobrevive em restos de cultura. Para o controle da doença, recomenda-se o uso de material propagativo sadio, inspeção no campo, poda drástica de órgãos infectados, eliminação de plantas severamente infectadas, condução da época de poda de produção, desinfestação de veículos, de equipamentos e de materiais para poda, utilização de fungicidas protetores cúpricos e tiocarbamatos, e utilização de quebra-ventos para reduzir a disseminação do patógeno.In the begining of 1998, bacterial canker of grapevine, caused by the bacterium Xanthomonas campestris pv. viticola, was detected for the first time in Brazil in vineyards of the “Submédio São Francisco”. Losses in susceptible

  19. Viability of Botryosphaeriaceae species pathogenic to grapevine after hot water treatment

    Directory of Open Access Journals (Sweden)

    Georgina ELENA

    2015-09-01

    Full Text Available The viability of eight species of Botryosphaeriaceae pathogenic to grapevine was studied after a hot water treatment (HWT in order to evaluate the feasibility of this technique as a potential tool to control these species during the grapevine propagation process. In a first trial (in vitro, mycelial plugs contained in Eppendorf tubes with sterile distilled water were subjected to different combinations of temperature (50–54°C and exposure time (15, 30 and 45 min in a hot water bath. Growth rates of treated mycelia were compared to untreated controls. Significant differences in survival and growth for all factors (species, temperature and time and their 2-way interactions were observed. Fungal survival and growth generally decreased with increasing temperatures and exposure times. Diplodia seriata, Neofusicoccum luteum, N. parvum and Spencermartinsia viticola were the most susceptible species to temperature, while Lasiodiplodia theobromae and N. vitifusiforme were the most tolerant. In a second trial (in planta, the fungi were inoculated into grapevine canes (Richter 110 rootstock. Inoculated canes were incubated at 25°C for 3 weeks to allow the fungi to colonize the wood and then subjected to HWT in the range of 50–53°C for 30 min, and survival of fungi after HWT was assessed. Survival of all species was sharply reduced after HWT of 30 min at 51°C and higher temperatures. At 50°C, Lasiodiplodia theobromae was the most tolerant species whereas N. luteum was the most susceptible. Results obtained in this study demonstrate the feasibility of controlling these pathogens by HWT in the nursery grapevine propagation process.

  20. Molecular Phylogeny, Diversity, and Bioprospecting of Endophytic Fungi Associated with wild Ethnomedicinal North American Plant Echinacea purpurea (Asteraceae).

    Science.gov (United States)

    Carvalho, Camila R; Wedge, David E; Cantrell, Charles L; Silva-Hughes, Alice F; Pan, Zhiqiang; Moraes, Rita M; Madoxx, Victor L; Rosa, Luiz H

    2016-07-01

    The endophytic fungal community associated with the ethnomedicinal plant Echinacea purpurea was investigated as well as its potential for providing antifungal compounds against plant pathogenic fungi. A total of 233 endophytic fungal isolates were obtained and classified into 42 different taxa of 16 genera, of which Alternaria alternata, Colletotrichum dematium, and Stagonosporopsis sp. 2 are the most frequent colonizers. The extracts of 29 endophytic fungi displayed activities against important phytopathogenic fungi. Eight antifungal extracts were selected for chemical analysis. Forty fatty acids were identified by gas chromatography-flame-ionization detection (GC-FID) analysis. The compounds (-)-5-methylmellein and (-)-(3R)-8-hydroxy-6-methoxy-3,5-dimethyl-3,4-dihydroisocoumarin were isolated from Biscogniauxia mediterraneaEPU38CA crude extract. (-)-5-Methylmellein showed weak activity against Phomopsis obscurans, P. viticola, and Fusarium oxysporum, and caused growth stimulation of C. fragariae, C. acutatum, C. gloeosporioides, and Botrytis cinerea. (-)-(3R)-8-Hydroxy-6-methoxy-3,5-dimethyl-3,4-dihydroisocoumarin appeared slightly more active in the microtiter environment than 5-methylmellein. Our results indicate that E. purpurea lives symbiotically with different endophytic fungi, which are able to produce bioactive fatty acids and aromatic compounds active against important phytopathogenic fungi. The detection of the different fatty acids and aromatic compounds produced by the endophytic community associated with wild E. purpurea suggests that it may have intrinsic mutualistic resistance against phytopathogen attacks in its natural environment.

  1. Synthesis and Biological Activity of Substituted Urea and Thiourea Derivatives Containing 1,2,4-Triazole Moieties

    Directory of Open Access Journals (Sweden)

    David E. Wedge

    2013-03-01

    Full Text Available A series of novel thiourea and urea derivatives containing 1,2,4-triazole moieties were synthesized and evaluated for their antifungal and larvicidal activity. Triazole derivatives 3a–e and 4a–e were synthesized by reacting thiocarbohydrazide with thiourea and urea compounds 1a–e and 2a–e, respectively, in a 130–140 °C oil bath. The proposed structures of all the synthesized compounds were confirmed using elemental analysis, UV, IR, 1H-NMR and mass spectroscopy. All compounds were evaluated for antifungal activity against plant pathogens, larvicidal and biting deterrent activity against the mosquito Aedes aegypti L. and in vitro cytotoxicity and anti-inflammatory activity against some human cell lines. Phomopis species were the most sensitive fungi to these compounds. Compounds 1b, 1c, 3a and 4e demonstrated selectively good activity against Phomopis obscurans and only 1b and 4e showed a similar level of activity against P. viticola. Compound 3d, with a LD50 value of 67.9 ppm, followed by 1c (LD50 = 118.8 ppm and 3e (LD50 = 165.6 ppm, showed the highest toxicity against Aedes aegypti larvae. Four of these compounds showed biting deterrent activity greater than solvent control, with the highest activity being seen for 1c, with a proportion not biting (PNB value of 0.75, followed by 1e, 2b and 1a. No cytotoxicity was observed against the tested human cancer cell lines. No anti-inflammatory activity was observed against NF-kB dependent transcription induced by phorbol myristate acetate (PMA in human chondrosarcoma cells.

  2. Fungal Planet description sheets: 320-370.

    Science.gov (United States)

    Crous, P W; Wingfield, M J; Guarro, J; Hernández-Restrepo, M; Sutton, D A; Acharya, K; Barber, P A; Boekhout, T; Dimitrov, R A; Dueñas, M; Dutta, A K; Gené, J; Gouliamova, D E; Groenewald, M; Lombard, L; Morozova, O V; Sarkar, J; Smith, M Th; Stchigel, A M; Wiederhold, N P; Alexandrova, A V; Antelmi, I; Armengol, J; Barnes, I; Cano-Lira, J F; Castañeda Ruiz, R F; Contu, M; Courtecuisse, Pr R; da Silveira, A L; Decock, C A; de Goes, A; Edathodu, J; Ercole, E; Firmino, A C; Fourie, A; Fournier, J; Furtado, E L; Geering, A D W; Gershenzon, J; Giraldo, A; Gramaje, D; Hammerbacher, A; He, X-L; Haryadi, D; Khemmuk, W; Kovalenko, A E; Krawczynski, R; Laich, F; Lechat, C; Lopes, U P; Madrid, H; Malysheva, E F; Marín-Felix, Y; Martín, M P; Mostert, L; Nigro, F; Pereira, O L; Picillo, B; Pinho, D B; Popov, E S; Rodas Peláez, C A; Rooney-Latham, S; Sandoval-Denis, M; Shivas, R G; Silva, V; Stoilova-Disheva, M M; Telleria, M T; Ullah, C; Unsicker, S B; van der Merwe, N A; Vizzini, A; Wagner, H-G; Wong, P T W; Wood, A R; Groenewald, J Z

    2015-06-01

    Novel species of fungi described in the present study include the following from Malaysia: Castanediella eucalypti from Eucalyptus pellita, Codinaea acacia from Acacia mangium, Emarcea eucalyptigena from Eucalyptus brassiana, Myrtapenidiella eucalyptorum from Eucalyptus pellita, Pilidiella eucalyptigena from Eucalyptus brassiana and Strelitziana malaysiana from Acacia mangium. Furthermore, Stachybotrys sansevieriicola is described from Sansevieria ehrenbergii (Tanzania), Phacidium grevilleae from Grevillea robusta (Uganda), Graphium jumulu from Adansonia gregorii and Ophiostoma eucalyptigena from Eucalyptus marginata (Australia), Pleurophoma ossicola from bone and Plectosphaerella populi from Populus nigra (Germany), Colletotrichum neosansevieriae from Sansevieria trifasciata, Elsinoë othonnae from Othonna quinquedentata and Zeloasperisporium cliviae (Zeloasperisporiaceae fam. nov.) from Clivia sp. (South Africa), Neodevriesia pakbiae, Phaeophleospora hymenocallidis and Phaeophleospora hymenocallidicola on leaves of a fern (Thailand), Melanconium elaeidicola from Elaeis guineensis (Indonesia), Hormonema viticola from Vitis vinifera (Canary Islands), Chlorophyllum pseudoglobossum from a grassland (India), Triadelphia disseminata from an immunocompromised patient (Saudi Arabia), Colletotrichum abscissum from Citrus (Brazil), Polyschema sclerotigenum and Phialemonium limoniforme from human patients (USA), Cadophora vitícola from Vitis vinifera (Spain), Entoloma flavovelutinum and Bolbitius aurantiorugosus from soil (Vietnam), Rhizopogon granuloflavus from soil (Cape Verde Islands), Tulasnella eremophila from Euphorbia officinarum subsp. echinus (Morocco), Verrucostoma martinicensis from Danaea elliptica (French West Indies), Metschnikowia colchici from Colchicum autumnale (Bulgaria), Thelebolus microcarpus from soil (Argentina) and Ceratocystis adelpha from Theobroma cacao (Ecuador). Myrmecridium iridis (Myrmecridiales ord. nov., Myrmecridiaceae fam. nov.) is also

  3. Epidemiology of Basil Downy Mildew.

    Science.gov (United States)

    Cohen, Yigal; Ben Naim, Yariv; Falach, Lidan; Rubin, Avia E

    2017-06-09

    Basil downy mildew (BDM) caused by the oomycete Peronospora belbahrii is a destructive disease of sweet basil (Ocimum basilicum) worldwide. It originated in Uganda in the 1930s and recently spread to Europe, the Middle East, Americas, and the Far East. Seed transmission may be responsible for its quick global spread. The pathogen attacks leaf blades, producing chlorotic lesions with ample dark asexual spores on the lower leaf surface. Oospores may form in the mesophyll of infected leaves. The asexual spores germinate on a wet leaf surface within 2 h and penetrate into the epidermis within 4 h. Spore germination and infection occur at a wide range of temperatures from 5 to 28.5°C. Infection intensity depends on the length of dew period, leaf temperature, and inoculum dose. The duration of latent period (from infection to sporulation) extends from 5 to 10 days, depending on temperature and light regime. The shortest is 5 days at 25°C under continuous light. Sporulation requires high humidity but not free leaf wetness. Sporulation occurs at 10 to 26°C. At the optimum temperature of 18°C, the process of sporulation requires 7.5 h at relative humidity ≥ 85%, with 3 h for sporophores emergence from stomata and 4.5 h for spore formation. Sporophores can emerge under light or darkness, but spore formation occurs in the dark only. Limited data are available on spore dispersal. Spores dispersed from sporulating plants contaminate healthy plants within 2 h of exposure. Settled spores may survive on leaf surface of healthy plants for prolonged periods, depending on temperature. Seed transmission of the disease occurs in Europe, but not in Israel or the United States. P. belbahrii in Israel also attacks species belonging to Rosemarinus, Nepeta, Agastache, Micromeria, and Salvia but not Plectranthus (coleus). A Peronospora species that infects coleus does not infect sweet basil. Control of BDM includes chemical, physical, and genetic means. The fungicide mefenoxam was

  4. Oogonial biometry and phylogenetic analyses of the Pythium vexans species group from woody agricultural hosts in South Africa reveal distinct groups within this taxon.

    Science.gov (United States)

    Spies, Christoffel F J; Mazzola, Mark; Botha, Wilhelm J; Van Der Rijst, Marieta; Mostert, Lizel; Mcleod, Adéle

    2011-02-01

    Pythium vexans fits into the internal transcribed spacer (ITS) clade K sensu Lévesque & De Cock (2004). Within clade K, P. vexans forms a distinct clade containing two enigmatic species, Pythium indigoferae and Pythium cucurbitacearum of which no ex-type strains are available. In South Africa, as well as in other regions of the world, P. vexans isolates are known to be heterogeneous in their ITS sequences and may consist of more than one species. This study aimed to investigate the diversity of South African P. vexans isolates, mainly from grapevines, but also citrus and apple using (i) phylogenetic analyses of the ITS, cytochrome c oxidase (cox) I, cox II, and β-tubulin regions and (ii) seven biometric oogonial parameters. Each of the phylogenies clustered P. vexans isolates into a single well-supported clade, distinct from other clade K species. The β-tubulin region was phylogenetically uninformative regarding the P. vexans group. The ITS phylogeny and combined cox I and II phylogenies, although each revealing several P. vexans subclades, were incongruent. One of the most striking incongruences was the presence of one cox subclade that contained two distinct ITS subclades (Ib and IV). Three groups (A-C) were subjectively identified among South African P. vexans isolates using (i) phylogenetic clades (ITS and cox), (ii) univariate analysis of oogonial diameters, and (iii) multivariate analyses of biometric oogonial parameters. Group A is considered to be P. vexans s. str. since it contained the P. vexans CBS reference strain from Van der Plaats-Niterink (1981). This group had significantly smaller oogonial diameters than group B and C isolates. Group B contained the isolates from ITS subclades Ib and IV, which formed a single cox subclade. The ITS subclade IV isolates were all sexually sterile or produced mainly abortive oospores, as opposed to the sexually fertile subclade Ib isolates, and may thus represent a distinct assemblage within group B. Although ITS

  5. 养殖日本鳗鲡腐皮病真菌性病原的分离与鉴定%ISOLATION AND IDENTIFICATION OF FUNGAL PATHOGEN FROM DERMAL ULCERATION DISEASE ON CULTURED ANGUILLA JAPONICA

    Institute of Scientific and Technical Information of China (English)

    曾占壮; 樊海平; 卓玉琛; 吴斌; 张新艳; 陈灵

    2011-01-01

    , the sexual structure could not be produced at those temperatures hereinbefore, even cultured for 4 weeks, but it could be produced at 15℃ when the strain was cultured for more than 3 weeks. Zoosporangium was clavate or spindly and renewed by internal or cymose proliferation, zoospores were diplanetic. Every oogonium could form more than one oospore, which were smooth. Flagella stain with AgNO3 revealed that there were two flagella with almost equal length on the zoospore. According to the characteristics of morphology and life history, such as the modality of the hyphae, the reproductive modes, the modality and the mode of movement of the zoospores, the amount and modality of the oospores, etc, strain Js80122 was identified as Saprolegniaceae, Protoachlya Coker, Protoachlya para-doxa.Epidemiological investigation showed that the epidemic temperature of the dermal ulceration disease was under 20℃, but it was nonoccurrence when the aquatic temperature exceeded to 23℃, and it could not be prevented and controlled by using multifarious disinfectors or antibacterial medicaments in breed aquatics of Japanese eels, contrarily, it would be more serious if the antibacterial medicaments were used in the beginning. Additionally, the fungal hyphae could be easily observed under the microscope even at the initial stage of the disease, when the derma of the diseased eels was unworn and the muscle had not been rotted. Strain Js80122 could cause dermal ulceration disease even the experimental eels were infected with spores alone as already noted. According to the results of epidemiological investigation and this study, it could be confirmed that the primary pathogen of dermal ulceration disease on cultured Japanese eels was not any pathogenic bacteria but fungi, in the concrete, it was Protoachlya paradoxa, the bacteria caused secondary infection, and they brought on nothing more than the worse illness.%针对近年来人工养殖日本鳗鲡(Anguillajaponica)在越冬过程

  6. Molecular detection of Phytophthora infestans based on Ypt1 gene%以Ypt1基因为靶标的致病疫霉菌Phytophthora infestans的分子检测

    Institute of Scientific and Technical Information of China (English)

    孟军

    2012-01-01

    Yptl is a Ras-related gene, in yeast, the gene encoding a protein associated with GTP binding. In this study, over 30 oomycetes Yptl DNA sequences were compared and a pair of primers (Pil/Pi2) was designed for detecting P. Infestans. 144 isolates representing 26 species of Phytophthora and 29 fungal species of plant pathogens were used to test the specificity of the primers. PCR amplification with species-specific (Pi) primers resulted in a product of 369 bp only from isolates of P. Infestans. The detection sensitivity with Pi primers was 100 pg of genomic DNA. Using YptlF/YptlR as first-round amplification primers, followed by a second round using the primer pair Pil/Pi2, a nested PCR procedure was developed, which increased the detection sensitivity 10-fold to 10 pg. For primers Pil/Pi2, one oospore or 3 zoospores could be detected by nested PCR. The results showed that Yptl gene could be a molecular target for detection of P. Infestans. PCR with the Pi primers could also be used to detect P. Infestans from naturally infected potato tissues and diseased soil samples.%三磷酸鸟苷(GTP)结合蛋白基因(Ypt1)是一个与原癌基因Ras(Rat sarcoma)相关的基因,在酵母中,该基因编码一个与Ras相关GTP结合蛋白.为了研究以Ypt1基因为分子靶标的致病疫霉菌(P.infestans)检测技术,比较了30种卵菌Ypt1基因的序列,以该序列为靶标设计了1对针对P.infestans的特异性PCR引物Pi1/Pi2.试验结果表明,在供试的55种不同疫霉菌和真菌的144个菌株中,利用这1对引物只能从P.infestans基因组DNA中分别扩增出1条分子量为369 bp的特异性条带,这1对引物的检测灵敏度为100 pg.以疫霉菌Ypt1通用引物Yph1F/Yph2R结合这1对特异引物进行套式PCR扩增,使引物Pi1/Pi2的检测灵敏度提高了10倍,检测到10 pg的基因组DNA.通过套式PCR,引物Pi1/Pi2对游动孢子和卵孢子的检测灵敏度分别为3个游动孢子和1个卵孢子;以Pi1/Pi2引物,分别采用单

  7. Interception of Phytophthora syringae on Citrus fruits imported from California, USA%进境美国加州脐橙中丁香疫霉Phytophthora syringae截获

    Institute of Scientific and Technical Information of China (English)

    罗加凤; 刘跃庭; 廖芳; 胡晓红; 刘鹏; 黄国明

    2012-01-01

    Several brown rot fruits have been found from Californian fresh orange sample. Three isolates similar to Phytophthora syringae were obtained from these rot fruits. The colonies grew slowly on PDA and V8 juice agar (V8A). They were stellate, tiled and thin on V8A, but on PDA, the margin was chrysanthemum petaloid; mycelia dense, milk-white-colored. Sporangia and catenulate hyphal swellings formed in sterilizing water and soil extract water in 48h. The isolates were homothallic. Oospores were produced abundantly on PDA and V8A mixed with fresh Citrus fruit tissue or leaf of rhododendron. The isolates were wounded-inoculated on fresh Citrus fruits. Typical brown rot symptoms are apparent on inoculated fruits in 7d. DNA of hyphae was amplified with ITS1/ITS4 primers. In comparision with P. Syringae from NCBIGenBank databases, the sequences had 99% identity. All the results of tests showed that the isolates were Phytophthorasyringae.%从产自美国加利福尼亚州的新鲜脐橙样品中发现多个腐烂病果,通过分离培养得到3个疑似丁香疫霉Phytophthora syringae菌株,对3个菌株进行形态学研究、致病性测定和分子序列比对分析.结果表明病菌在V8A培养基上菌落稀疏、平铺,呈星状,菌丝紧贴培养基生长或埋于基质内生长;在PDA培养基上菌落呈菊花花瓣状,菌丝致密,乳白色;游动孢子囊和菌丝膨大体在无菌水和土壤浸出液中黑暗条件下48h后产生;菌株为同宗配合,卵孢子在带有新鲜脐橙果实组织或杜鹃叶片的V8A培养基中大量产生;创伤接种脐橙果实,7d后接种脐橙出现典型的褐腐症状;通用引物ITS 1/ITS4扩增测序,Blastn分析表明序列与GenBank中P.syringae序列相似性为99%.依据上述研究结果,将分离获得的3株菌鉴定为丁香疫霉Phytophthora syringae,系国内首次截获的一种植物检疫性真菌病害.

  8. Temporal variation of Nitella furcata subsp. mucronata var. mucronata f. oligospira (Charophyceae in the Ninféias pond, São Paulo State, southeast Brazil Variação temporal de Nitella furcata subsp. mucronata var. mucronata f. oligospira (Charophyceae no Lago das Ninféias, São Paulo, sudeste do Brasil

    Directory of Open Access Journals (Sweden)

    Norma Catarina Bueno

    2006-03-01

    Full Text Available Temporal and spatial variation of Nitella furcata (Roxburgh ex Bruzelius C. Agardh emend. R.D. Wood subsp. mucronata (A. Braun R.D. Wood var. mucronata f. oligospira (A. Braun R.D. Wood were studied at the Ninféias pond (23°38'18.9"S, 46°37'16.3"W, a tropical, shallow, mesotrophic reservoir located in the Parque Estadual das Fontes do Ipiranga Biological Reserve, Municipality of São Paulo, southeast Brazil. Water samples were collected monthly from October/1996 to October/1997 for turbidity, electric conductivity, pH, alkalinity, dissolved oxygen, nutrients, biomass, and algal phenology analysis. Spatial distribution pattern of algal biomass showed that all four sampling stations were similar among themselves during the whole study period. Regarding the temporal variation, greatest algal total biomass values (98.35-266.06 g.m-2 were measured during the rainy period whereas the smallest ones (48.86-170.56 g.m-2 were detected during the dry period. Algal population density peaked at all four sampling stations from October/1996 to April/1997 (rainy period when the greatest values of air and water temperature, precipitation, solar radiation, turbidity, total nitrogen, and ammonium were measured allowing favorable conditions for the algal growth. Period from May/1997 (late fall to October/1997 (early spring was favorable to the development of reproductive structures (nucules and globules with consequent decrease of algal biomass and population density, clearly suggesting the greatest energy allocation towards the development of sexual reproductive structures. This fact was confirmed by the canonic correspondence analysis that indicated strong correlation between nucules length and width, number of nucules per plant, and oospores length and width with the low values of pH and high ones of dissolved inorganic matter detected during the dry period. Consequently, environmental factors such as increasing of water temperature and nutrients availability