Thermal inactivation of Phytophthora capsici oospores.

Science.gov (United States)

Etxeberria, Aitzol; Mendarte, Sorkunde; Larregla, Santiago

2011-01-01

Phytophthora capsici is a major fungal plant pathogen that causes root and crown rot of pepper crops and its oospores are the most resistant propagules. To evaluate the effect of different temperature regimes and exposure times on the survival of P. capsici oospores. Thermal inactivation treatments simulated field conditions, through the use of different constant and cycling temperature regimes, in moistened sterilized soil (15-53 °C) and sterilized water (45-53 °C). The plasmolysis method evaluated oospore viability. Relationships between oospores viability and exposure time were statistically determined by linear regression. Interpolation was used to calculate the estimated times required to kill a determined percentage of the population. The required time to reduce P. capsici oospores viability decreased with increasing temperatures. Times required to kill 100% of oospores were 199-22-6.6-4.7-1.0 hours at 40-45-47.5-50-53°C respectively in moistened soil and 31-1.0-0.2 hours at 45-50-53 °C in water. Oospores were scarcely affected at temperatures ≤ 35 °C. With 1,680 hours at 15-35 °C, oospores survival in soil ranged from 88 to 36%. The 4 hours-40 °C regime killed 100% of oospores after 28days, while the 5 hours-35°C regime after 70 days killed only 75%. Time required to achieve total oospores death was remarkably shortened in water when compared with moistened soil. The developed models can be used to predict survival values at any exposure time with constant temperatures ranging from 40 to 53 °C in moistened soil and from 45 to 53 °C in water. The weakening of P. capsici oospores under sublethal heating, is a useful observation that can be applied for pathogen control with solarization. Copyright © 2010 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

Temperature and Water Effects on the Viability of Alginate-Formulated Oospores of Pythrium Oligandrum

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A. K. Al-Rawahi

1998-06-01

Full Text Available The low germination rates of oospores hinder the use of Pythium oligandrum as a biocontrol agent of soilborne plant pathogens. To improve the reliability of oospore-inoculum of P. oligandrum for the biological control of root diseases, oospores were formulated in calcium alginate pellets (oospore-pellets. Oospore-pellets of five isolates of P. oligandrum stored at 4°C under dry conditions remained viable (more or equal than 99% germination for each time interval over 12 months regardless of the culture medium used to produce the oospores. In contrast, oospore-pellets stored at 25°C under the same conditions maintained their viability for 3 months but declined to 50%, viability in the following 9 months. Germination of oospore-pellets was at the highest (98% at solute potentials (y8 higher than -l.33 MPa or relative humidities (RH higher than 97% at 25° and 33°C. Germination declined sharply as (y8, decreased from -1.33 to -2.59 MPa and RH decreased from 97 to 94% with no germination occurring at -3.59 MPa and 92% RH. Only 60% germination was found at 16°C in the top range of y, and RH, and germination declined sharply as y, and RH were decreased, with no germination occurring at - l .33 MPa and 95% RH. Mycelial growth exhibited similar ranges of tolerance under the same moisture conditions and temperatures. High degrees of survival during storage accompanied by high percentages of germination under relatively dry conditions suggest that the oospore-pellet inoculum of P. oligandrum can be used for the control of pathogens in soil or a potting mix at high temperatures during a warm season or under greenhouse conditions.

Comparative study of the oospore morphology of two populations of a rare species Chara baueri A. Braun in Cedynia (Poland and Batzlow (Germany

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Andrzej Pukacz

2012-06-01

Full Text Available Morphological features of oospores of Chara baueri A. Braun, one of the rarest charophyte species worldwide, were studied based on 100 oospores collected from a small and temporarily dried mid-field pond near Cedynia, Western Poland. This is the first Polish and fifth presently known locality of this species. For comparison 67 oospores from a German population (similar pond localized near Batzlow, Germany were also measured. So far, data on morphology of C. baueri oospores as well as the species ecology are limited. The only more detailed study of oospores for this species was earlier performed on 15 oospores from Kazakhstan. Largest polar axis (LPA, length, largest equatorial diameter (LED, width, isopolarity index (ISI = LPA/LED × 100, number of ridges, width of fossa, distance from apical pole to LED (AND and anisipolarity index (ANI = AND/LPA × 100 were measured. The comparative analysis revealed that the oospores from Poland are generally bigger and more prolate than the Ger­man ones. The differences for most of studied parameters were statistically significant. The finding is discussed in the context of habitat differentiation of both studied sites. Moreover, the results obtained of oospore measurements for both populations differs from most of the data known so far from the literature.

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

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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.

Genetic modification of European winegrapes with genes from an American wild relative confers resistance to the major diseases powdery and downy mildew

Science.gov (United States)

The two most economically important diseases of grapevine cultivation worldwide are caused by the fungal pathogen powdery mildew (Erysiphe necator syn. Uncinula necator) and the oomycete, downy mildew (Plasmopara viticola). These pathogens, endemic to North America, were introduced into Europe in t...

Grapevine downy mildew control in organic farming.

Science.gov (United States)

La Torre, A; Spera, G; Lolletti, D

2005-01-01

Cupric products at low dose and alternative compounds have been tested to control the downy mildew in an organic vineyard. It has found that copper compounds control downy mildew in a satisfactory way, reducing, at the same time, the dose of copper metal. The alternative products were not satisfactory to control Plasmopara viticola.

Induction of defence mechanisms in grapevine leaves by emodin- and anthraquinone-rich plant extracts and their conferred resistance to downy mildew.

Science.gov (United States)

Godard, Sophie; Slacanin, Ivan; Viret, Olivier; Gindro, Katia

2009-09-01

The ability of two plant extracts, Rheum palmatum root extract (RPRE) and Frangula alnus bark extract (FABE), to protect Vitis vinifera leaves from Plasmopara viticola infection was evaluated. These natural products are toxic to the pathogen and induce defence reactions in a susceptible cultivar of V. vinifera (V. vinifera cv. Chasselas), including stilbenic phytoalexin accumulation, enhanced peroxidase (EC 1.11.1.7) activity, and a hypersensitive reaction. Inhibition of the first stage of biotrophic hyphal development of P. Viticola by the two plant extracts was observed. HPLC-DAD-MS analysis showed that these two natural extracts contain many phenolic compounds belonging to the anthraquinone family, such as rhein, frangulin A, emodin, aloe-emodin, chrysophanol, and physcion. Emodin alone is able to impair P. viticola development and to stimulate viniferins and the accumulation of pterostilbene.

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...

Effectiveness of some microorganisms in the limitation of grapevine cuttings infection by Phomopsis viticola Sacc.

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Ewa Król

2013-12-01

Full Text Available The possibilities of using antagonistic fungi and bacteria in the limitation of grapevine stems infection by Phomopsis viticola Sacc. were studied. Trichodema koningii Oud., T.viride Persoon ex S.F.,T.harzianum Rifai, Gliocladium catenulatum Gilman and Abbott, G.fimbriatum Gilman and Abbott, Bacillus sp., Pseudomonas fluorescens and five unidentified isolates of bacteria i.e.: 22a, 35, 40, 45, 66 were estimated. It was appeared what Trichoderma spp. were the most effective in protection of grapevine stems against the infection by P.viticola. After these antagonistic fungi were used on protected grapevine canes not numerous necrosis were observed and few cultures of pathogen were reisolated from them. Moreover, Trichoderma spp. survived on the grapevine stems during the period of experiment. The abilities of other microorganisms tested to protect grapevine cuttings against P.viticola infection and to exist on the stems were less than Trichoderma spp.

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

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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

Towards optical detection of plasmopara viticola infection in the field

Czech Academy of Sciences Publication Activity Database

Šebela, David; Olejníčková, Julie; Sotolář, R.; Vrchotová, Naděžda; Tříska, Jan

2014-01-01

Roč. 96, č. 2 (2014), s. 309-320 ISSN 1125-4653 R&D Projects: GA MŠk(CZ) ED1.1.00/02.0073 Institutional support: RVO:67179843 Keywords : Downy mildew * grapevine * UV-induced fluorescence * chlorophyll fluorescence * reflectance * transresveratrol * 2,4,6-trihydroxyphenanthrene-2-O-glucoside Subject RIV: EH - Ecology, Behaviour Impact factor: 1.043, year: 2014

Proteomic analysis of grapevine resistance induced by Trichoderma harzianum T39 reveals specific defence pathways activated against downy mildew

Science.gov (United States)

Perazzolli, Michele

2012-01-01

Downy mildew is caused by the oomycete Plasmopara viticola and is one of the most serious diseases of grapevine. The beneficial microorganism Trichoderma harzianum T39 (T39) has previously been shown to induce plant-mediated resistance and to reduce the severity of downy mildew in susceptible grapevines. In order to better understand the cellular processes associated with T39-induced resistance, the proteomic and histochemical changes activated by T39 in grapevine were investigated before and 1 day after P. viticola inoculation. A comprehensive proteomic analysis of T39-induced resistance in grapevine was performed using an eight-plex iTRAQ protocol, resulting in the identification and quantification of a total of 800 proteins. Most of the proteins directly affected by T39 were found to be involved in signal transduction, indicating activation of a complete microbial recognition machinery. Moreover, T39-induced resistance was associated with rapid accumulation of reactive oxygen species and callose at infection sites, as well as changes in abundance of proteins involved in response to stress and redox balance, indicating an active defence response to downy mildew. On the other hand, proteins affected by P. viticola in control plants mainly decreased in abundance, possibly reflecting the establishment of a compatible interaction. Finally, the high-throughput iTRAQ protocol allowed de novo peptide sequencing, which will be used to improve annotation of the Vitis vinifera cv. Pinot Noir proteome. PMID:23105132

Synthesis and fungicidal activity of N-thiazol-4-yl-salicylamides, a new family of anti-oomycete compounds.

Science.gov (United States)

Sulzer-Mosse, Sarah; Cederbaum, Fredrik; Lamberth, Clemens; Berthon, Guillaume; Umarye, Jayant; Grasso, Valeria; Schlereth, Alexandra; Blum, Mathias; Waldmeier, Rita

2015-05-01

A novel class of experimental fungicides has been discovered, which consists of special N-thiazol-4-yl-salicylamides. They originated from amide reversion of lead structures from the patent literature and are highly active against important phytopathogens, such as Phytophthora infestans (potato and tomato late blight), Plasmopara viticola (grapevine downy mildew) and Pythium ultimum (damping-off disease). Structure-activity relationship studies revealed the importance of a phenolic or enolic hydroxy function in the β-position of a carboxamide. An efficient synthesis route has been worked out, which for the first time employs the carbonyldiimidazole-mediated Lossen rearrangement in the field of thiazole carboxylic acids. Copyright © 2015 Elsevier Ltd. All rights reserved.

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.

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

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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

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

Science.gov (United States)

2012-01-01

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 for selecting novel

Diagnostic moléculaire de la présence de mildiou (Plasmopara halstedii dans les semences de tournesol

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Says-Lesage Véronique

2001-05-01

Full Text Available Le mildiou du tournesol, causé par l’oomycète Plasmopara halstedii, est une maladie potentiellement dévastatrice et économiquement importante. Du fait du risque de propagation de la maladie par les semences infectées et de l’observation croissante de l’apparition d’isolats résistants au métalaxyl, il est apparu nécessaire de mettre au point une méthode de détection du parasite dans les graines ou les tissus de la plante. Nous avons donc développé un test moléculaire pour le diagnostic de la présence du mildiou dans les semences. Plusieurs méthodes d’extraction ont été comparées pour améliorer le rendement et la qualité de l’ADN du parasite présent en faible quantité. Des échantillons de graines contaminées entières et décortiquées ont été analysés par PCR, avec des amorces spécifiques de Plasmopara halstedii. L’ADN de l’agent pathogène est toujours détecté, notamment dans les coques. Ces données sont importantes pour le développement d’un kit de diagnostic.

Use of radiation for improving vines regarding their resistance to mildew

International Nuclear Information System (INIS)

Coutinho, M.P.

1977-01-01

Vines (Vitis vinifera) resistant to mildew (Plasmopara viticola) offer real advantages in Europe, and the problems of producing such vines were studied for a long time. At first conventional techniques were used, obtaining plants with high yield and with resistance persisting under widely differing ecological conditions; moreover, pathogenically different biotypes of Plasmopara have never been found. However, various factors - such as the quantitative nature of this resistance, which is controlled by a polygenic system, certain genetic correlations between resistance and poor quality of the grapes and the lack of resistance sources in V. vinifera - suggested that mutagenesis should be included among the methods used for the improvement of vines. Hence shoots and, more particularly, seeds have been undergoing irradiation with X-rays and neutrons since 1966. The exposure of seeds to low radiation doses (about 1000rad), particularly of X-rays, has in most varieties produced a very welcome increase in the germination percentage. Seedlings from irradiated seeds are selected mainly on the basis of the characteristics of the infection spots which develop on the leaves. In V. vinifera, resistance to mildew is reflected not in necroses but in small and usually not very sporulated spots. Using these criteria, the author has already selected - after natural infections and inoculations - plants representing approximately 0.001-0.0025% of the total number of plants screened. Tables illustrate the selection procedure up to 1976

The Top 10 oomycete pathogens in molecular plant pathology.

Science.gov (United States)

Kamoun, Sophien; Furzer, Oliver; Jones, Jonathan D G; Judelson, Howard S; Ali, Gul Shad; Dalio, Ronaldo J D; Roy, Sanjoy Guha; Schena, Leonardo; Zambounis, Antonios; Panabières, Franck; Cahill, David; Ruocco, Michelina; Figueiredo, Andreia; Chen, Xiao-Ren; Hulvey, Jon; Stam, Remco; Lamour, Kurt; Gijzen, Mark; Tyler, Brett M; Grünwald, Niklaus J; Mukhtar, M Shahid; Tomé, Daniel F A; Tör, Mahmut; Van Den Ackerveken, Guido; McDowell, John; Daayf, Fouad; Fry, William E; Lindqvist-Kreuze, Hannele; Meijer, Harold J G; Petre, Benjamin; Ristaino, Jean; Yoshida, Kentaro; Birch, Paul R J; Govers, Francine

2015-05-01

Oomycetes form a deep lineage of eukaryotic organisms that includes a large number of plant pathogens which threaten natural and managed ecosystems. We undertook a survey to query the community for their ranking of plant-pathogenic oomycete species based on scientific and economic importance. In total, we received 263 votes from 62 scientists in 15 countries for a total of 33 species. The Top 10 species and their ranking are: (1) Phytophthora infestans; (2, tied) Hyaloperonospora arabidopsidis; (2, tied) Phytophthora ramorum; (4) Phytophthora sojae; (5) Phytophthora capsici; (6) Plasmopara viticola; (7) Phytophthora cinnamomi; (8, tied) Phytophthora parasitica; (8, tied) Pythium ultimum; and (10) Albugo candida. This article provides an introduction to these 10 taxa and a snapshot of current research. We hope that the list will serve as a benchmark for future trends in oomycete research. © 2014 BSPP AND JOHN WILEY & SONS LTD.

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.

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

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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.

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.

Antimicrobial Activity of Resveratrol Analogues

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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.

Chitosan and Laminarin as Alternatives to Copper for Plasmopara viticola Control: Effect on Grape Amino Acid.

Science.gov (United States)

Garde-Cerdán, T; Mancini, V; Carrasco-Quiroz, M; Servili, A; Gutiérrez-Gamboa, G; Foglia, R; Pérez-Álvarez, E P; Romanazzi, G

2017-08-30

Copper fungicide use is limited by the European regulation; therefore, new strategies have been developed to prevent grapevine downy mildew (GDM). However, there is poor information about their effects on grape amino acid composition. This field trial aimed to evaluate the effect on grape amino acid composition of chitosan and of a mixture of laminarin and Saccharomyces extracts (LamE), applied in different strategies with copper hydroxide. The results showed that all the treatments applied to grapevines decreased the concentration of several amino acids. Moreover, treatments that have mostly decreased these compounds are those with copper hydroxide, especially when applied individually. LamE applied individually or alternately with copper hydroxide had the least negative effect on grape amino acid content. These results provide further information about the negative effects of copper on grape quality, which can be reduced when it is used in strategy with LamE or chitosan in GDM control.

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.

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

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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

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

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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.

RXLR and CRN Effectors from the Sunflower Downy Mildew Pathogen Plasmopara halstedii Induce Hypersensitive-Like Responses in Resistant Sunflower Lines

Science.gov (United States)

Gascuel, Quentin; Buendia, Luis; Pecrix, Yann; Blanchet, Nicolas; Muños, Stéphane; Vear, Felicity; Godiard, Laurence

2016-01-01

Plasmopara halstedii is an obligate biotrophic oomycete causing downy mildew disease on sunflower, Helianthus annuus, an economically important oil crop. Severe symptoms of the disease (e.g., plant dwarfism, leaf bleaching, sporulation and production of infertile flower) strongly impair seed yield. Pl resistance genes conferring resistance to specific P. halstedii pathotypes were located on sunflower genetic map but yet not cloned. They are present in cultivated lines to protect them against downy mildew disease. Among the 16 different P. halstedii pathotypes recorded in France, pathotype 710 is frequently found, and therefore continuously controlled in sunflower by different Pl genes. High-throughput sequencing of cDNA from P. halstedii led us to identify potential effectors with the characteristic RXLR or CRN motifs described in other oomycetes. Expression of six P. halstedii putative effectors, five RXLR and one CRN, was analyzed by qRT-PCR in pathogen spores and in the pathogen infecting sunflower leaves and selected for functional analyses. We developed a new method for transient expression in sunflower plant leaves and showed for the first time subcellular localization of P. halstedii effectors fused to a fluorescent protein in sunflower leaf cells. Overexpression of the CRN and of 3 RXLR effectors induced hypersensitive-like cell death reactions in some sunflower near-isogenic lines resistant to pathotype 710 and not in susceptible corresponding lines, suggesting they could be involved in Pl loci-mediated resistances. PMID:28066456

RXLR and CRN effectors from the sunflower downy mildew pathogen Plasmopara halstedii induce hypersensitive-like responses in resistant sunflower lines

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Quentin Gascuel

2016-12-01

Full Text Available Plasmopara halstedii is an obligate biotrophic oomycete causing downy mildew disease on sunflower, Helianthus annuus, an economically important oil crop. Severe symptoms of the disease (e.g. plant dwarfism, leaf bleaching, sporulation and production of infertile flower strongly impair seed yield. Pl resistance genes conferring resistance to specific P. halstedii pathotypes were located on sunflower genetic map but yet not cloned. They are present in cultivated lines to protect them against downy mildew disease. Among the 16 different P. halstedii pathotypes recorded in France, pathotype 710 is frequently found, and therefore continuously controlled in sunflower by different Pl genes. High-throughput sequencing of cDNA from P. halstedii led us to identify potential effectors with the characteristic RXLR or CRN motifs described in other oomycetes. Expression of six P. halstedii putative effectors, five RXLR and one CRN, was analysed by qRT-PCR in pathogen spores and in the pathogen infecting sunflower leaves and these six effectors were selected for functional analyses. We developed a new method for transient expression in sunflower plant leaves and showed for the first time subcellular localization of P. halstedii effectors fused to a fluorescent protein in sunflower leaf cells. Overexpression of the CRN and of 3 RXLR effectors induced hypersensitive-like cell death reactions in some sunflower near-isogenic lines resistant to pathotype 710 and not in susceptible corresponding lines, suggesting they could be involved in Pl loci-mediated resistances.

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

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

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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.

Overexpression of VpPR10.1 by an efficient transformation method enhances downy mildew resistance in V. vinifera.

Science.gov (United States)

Su, Hang; Jiao, Yun-Tong; Wang, Fang-Fang; Liu, Yue-E; Niu, Wei-Li; Liu, Guo-Tian; Xu, Yan

2018-05-01

Putrescine and spermidine increase the transformation efficiency of Vitis vinifera L. cv. Thompson seedless. Accumulation of VpPR10.1 in transgenic V. vinifera Thompson seedless, likely increases its resistance to downy mildew. A more efficient method is described for facilitating Agrobacterium-mediated transformation of Vitis vinifera L. cv. Thompson Seedless somatic embryogenesis using polyamines (PAs). The efficacies of putrescine, spermidine and spermine are identified at a range of concentrations (10 µM, 100 µM and 1 mM) added to the culture medium during somatic embryo growth. Putrescine (PUT) and spermidine (SPD) promote the recovery of proembryonic masses (PEM) and the development of somatic embryos (SE) after co-cultivation. Judging from the importance of the time-frame in genetic transformation, PAs added at the co-cultivation stage have a stronger effect than delayed selection treatments, which are superior to antibiotic treatments in the selection stage. Best embryogenic responses are with 1 mM PUT and 100 µM SPD added to the co-culture medium. Using the above method, a pathogenesis-related gene (VpPR10.1) from Chinese wild Vitis pseudoreticulata was transferred into Thompson Seedless for functional evaluation. The transgenic line, confirmed by western blot analysis, was inoculated with Plasmopara viticola to test for downy mildew resistance. Based on observed restrictions of hyphal growth and increases in H 2 O 2 accumulation in the transgenic plants, the accumulation of VpPR10.1 likely enhanced the transgenic plants resistance to downy mildew.

Influência das condições climáticas sobre a eficácia de fungicidas empregados para o controle do míldio em Vitis vinifera

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Czermainski Ana Beatriz Costa

2004-01-01

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.

A Framework for Optimizing Phytosanitary Thresholds in Seed Systems.

Science.gov (United States)

Choudhury, Robin Alan; Garrett, Karen A; Klosterman, Steven J; Subbarao, Krishna V; McRoberts, Neil

2017-10-01

Seedborne pathogens and pests limit production in many agricultural systems. Quarantine programs help prevent the introduction of exotic pathogens into a country, but few regulations directly apply to reducing the reintroduction and spread of endemic pathogens. Use of phytosanitary thresholds helps limit the movement of pathogen inoculum through seed, but the costs associated with rejected seed lots can be prohibitive for voluntary implementation of phytosanitary thresholds. In this paper, we outline a framework to optimize thresholds for seedborne pathogens, balancing the cost of rejected seed lots and benefit of reduced inoculum levels. The method requires relatively small amounts of data, and the accuracy and robustness of the analysis improves over time as data accumulate from seed testing. We demonstrate the method first and illustrate it with a case study of seedborne oospores of Peronospora effusa, the causal agent of spinach downy mildew. A seed lot threshold of 0.23 oospores per seed could reduce the overall number of oospores entering the production system by 90% while removing 8% of seed lots destined for distribution. Alternative mitigation strategies may result in lower economic losses to seed producers, but have uncertain efficacy. We discuss future challenges and prospects for implementing this approach.

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.

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

Energy Technology Data Exchange (ETDEWEB)

Mohammadi, H.; Sarcheshmehpour, M.; Mafi, E.

2015-07-01

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, Phaeoacremonium cinereum, 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. (Author)

A Gene Expressed during Sexual and Asexual Sporulation in Phytophthora infestans is a Member of the Puf Family of Translational Regulators

DEFF Research Database (Denmark)

Cvitanich, Cristina; Judelson, Howard S.

2003-01-01

and persisted in mature oospores. Expression was also observed in hyphal tips just prior to asexual sporulation, in sporangiophores, in mature sporangia, and in zoospores. The signal quickly disappeared once spores made the transition to hyphae after germination. Nutrient limitation did not induce the gene...

Concentration- and Time-Dependent Effects of Isothiocyanates Produced from Brassicaceae Shoot Tissues on the Pea Root Rot Pathogen Aphanomyces euteiches

NARCIS (Netherlands)

Hossain, S.; Bergkvist, G.; Berglund, K.; Glinwood, R.; Kabouw, P.; Martensson, A.; Persson, P.

2014-01-01

Isothiocyanates (ITCs) hydrolyzed from glucosinolates (GSLs) in Brassicaceae tissue are toxic to soil organisms. In this study, the effect of aliphatic and aromatic ITCs from hydrated dry Brassicaceae shoot tissues on the mycelium and oospores of the pea root rot pathogen Aphanomyces euteiches was

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 (pHeffects 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 work best in acidic soils low in organic carbon and with low or no compost addition although this might

The Use of Copper Pesticides in Germany and the Search for Minimization and Replacement Strategies

Directory of Open Access Journals (Sweden)

Stefan Kuehne

2017-02-01

Full Text Available Copper pesticides used to control fungal and bacterial diseases such as grapes downy mildew (Plasmopara viticola, downy mildew of hops (Pseudoperonospora humili, apple scab (Venturia spp., fireblight (Erwinia amylovora and potato late blight (Phytophthora infestans, play an important role in plant protection. In a 2013 survey of copper application in Germany we found, that while the amounts of copper used per hectare in conventional grape (0.8 kg ha−1, hop (1.7 kg ha−1 and potato-farming (0.8 kg ha−1 were well below those used in organic farming (2.3, 2.6 and 1.4 kg ha−1, respectively, they were nearly identical to those used in apple growing (1.4 kg ha−1. Due to the smaller farming area, only 24% (26.5 tonnes of the total amount of copper was applied in organic farming compared to 76% (84.8 tonnes in conventional farming. Since 2001, the Federal Agency for Agriculture and Food (BLE promoted a copper research and minimization strategy which was funded with a total of C10.2 million. Our status quo analysis of research in this field shows that some progress is being made concerning alternative compounds, resistant varieties and decision support systems. However, it also shows that new approaches are not yet able to replace copper pesticides completely, especially in organic farming. In integrated pest management, copper preparations are important for the necessary active substance rotation and successful resistance management. The availability of such products is often essential for organic grapes, hops and fruit production and for extending the organic farming of these crops. We conclude that the complete elimination of copper pesticides is not yet practicable in organic farming as the production of several organic crops would become unprofitable and may lead to organic farmers reverting to conventional production. Several existing copper reduction strategies were, however, identified, and some, like modified forecast models adapted to

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African Journals Online (AJOL)

a heterothalic fungus that reproduces sexually in the presence of two compatible mating types. ... reproduce only asexuallyin other countries (Tooley et .... Sexual reproduction in P inſeftans leads to Oospore production wherever the two mating types A1 and A2 are presentin a popu- lation and come into contact inside a ...

A Sensitive Assay for Rapid Detection and Quantification of Aphanomyces euteiches in Soil.

Science.gov (United States)

Gangneux, Christophe; Cannesan, Marc-Antoine; Bressan, Mélanie; Castel, Lisa; Moussart, Anne; Vicré-Gibouin, Maïté; Driouich, Azeddine; Trinsoutrot-Gattin, Isabelle; Laval, Karine

2014-10-01

Aphanomyces euteiches is a widespread oomycete pathogen causing root rot in a wide range of leguminous crops. Losses can reach up to 100% for pea culture and there is currently no registered pesticide for its control. Crop management remains the most efficient tool to control root rot, and avoidance of infested soil seems to be the optimal solution. A test was developed to identify fields suitable for pea crops, consisting of the determination of the inoculum potential of soil using baiting plants. A new rapid, specific, and sensitive molecular method is described allowing the quantification of less than 10 oospores per gram of soil. This challenge is achieved by a real-time polymerase chain reaction procedure targeting internal transcribed spacer 1 from the ribosomal DNA operons. A preliminary study based on typical soils from northwestern France demonstrated that the A. euteiches oospore density in soil is related to the inoculum potential. Furthermore, this method has proved sensitive enough to accurately study the influence of biotic factors that may govern the actual emergence of root rot.

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

BACKGROUND: There has been an increasing interest in the use of selected non-Saccharomyces yeasts in co-culture with Saccharomyces cerevisiae. In this work, three non-Saccharomyces yeast strains (Metschnikowia viticola, Metschnikowia fructicola and Hanseniaspora uvarum) indigenously isolated...... in Denmark were used in sequential fermentations with S. cerevisiae on three cool-climate grape cultivars, Bolero, Rondo and Regent. During the fermentations, the yeast growth was determined as well as key oenological parameters, volatile compounds and sensory properties of finished rosé wines. RESULTS......: 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...

Control of grapevine wood fungi in commercial nurseries

Directory of Open Access Journals (Sweden)

C. Rego

2009-05-01

Full Text Available Previous surveys conducted in commercial nurseries found that different wood fungi, namely Cylindrocarpon spp., Botryosphaeriaceae, Phomopsis viticola and Phaeomoniella chlamydospora infect grapevine cuttings. Two field trials were carried out to evaluate the effectiveness of cyprodinil + fludioxonil, pyraclostrobin + metiram, fludioxonil and cyprodinil to prevent or reduce natural infections caused by such fungi. Rootstock and scion cuttings were soaked in fungicidal suspensions for 50 min prior to grafting. After callusing, the grafted cuttings were planted in two commercial field nurseries with and without a previous history of grapevine cultivation. After nine months in the nursery, the plants were uprooted and analysed for the incidence and severity of the wood fungi. Plants uprooted from the field without a previous history of grapevine cultivation were generally less strongly infected by wood fungi. Under this condition, only the mixture cyprodinil + fludioxonil simultaneously reduced the incidence of Cylindrocarpon and Botryosphaeriaceae fungi, as well as the severity of Cylindrocarpon infections. Treatments did not produce significant differences in the incidence and severity of P. viticola, and Pa. chlamydospora. For plants grown in the field with a grapevine history, all fungicides except cyprodinil significantly reduced the incidence and severity of Cylindrocarpon fungi. Also, the incidence and severity of Botryosphaeriaceae pathogens were significantly decreased both by cyprodinil + fludioxonil and by cyprodinil. No significant differences were noticed for P. viticola incidence and severity, and Pa. chlamydospora was not detected again. These results suggest that the practice of soaking grapevine cuttings in selected fungicides prior to grafting significantly reduces Cylindrocarpon spp. and Botryosphaeriaceae infections, thus improving the quality of planting material.

Monitoring the infective process of the downy mildew causal agent within micropropagated rose plants

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Sonia Yamile Gómez

2012-08-01

Full Text Available Downy mildew in the rose caused by a species of the Peronospora genus is a very restrictive disease for the Colombian greenhouse rose production. The damage observed in the susceptible varieties of commercial rose include symptoms affect young steams and tiny leaves causing reddish and brown spots and defoliation; leading to 10% production losses. The infective behavior of this pathogen was studied with the aim of increasing the knowledge about the biology of the rose downy mildew. The study of the infective process was performed on the Charlotte variety using micropropagated roses inoculated with suspensions of sporangia. A germinal tube was observed during the germination process, it came from a lateral papilla and reached up to 300 microns in length. During this study, the ability of the pathogen to use vascular sieves as communication systems within the plant was determined. Oogonia and antheridia were also observed inside the epidermal cells, and oospores inside the parenchymal tissue close to xylem vessels. To the best of our knowledge, these sexual structures have not been reported on in Colombia before. This study verifies the ability of the downy mildew causal agent to move through the xylem vessels and produce sexual structures, such as oogonia, antheridia and oospores within those tissues.

Effect of phosphate 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...... to measure the activity of the pathogen in roots. The enzyme activity increased with disease severity and disease incidence, except in plants supplemented with P at the highest level, where a peak in activity was seen 12 days after inoculation with the pathogen, followed by a decrease in activity...

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. © 2014 BSPP AND JOHN WILEY & SONS LTD.

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

Science.gov (United States)

Liu, Jing; Arneborg, Nils; Toldam-Andersen, Torben B; Petersen, Mikael A; Bredie, Wender Lp

2017-08-01

There has been an increasing interest in the use of selected non-Saccharomyces yeasts in co-culture with Saccharomyces cerevisiae. In this work, three non-Saccharomyces yeast strains (Metschnikowia viticola, Metschnikowia fructicola and Hanseniaspora uvarum) indigenously isolated in Denmark were used in sequential fermentations with S. cerevisiae on three cool-climate grape cultivars, Bolero, Rondo and Regent. During the fermentations, the yeast growth was determined as well as key oenological parameters, volatile compounds and sensory properties of finished rosé wines. 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 in sequential fermentation with S. cerevisiae resulted in richer berry and fruity flavours in wines. The sensory plot showed a more clear separation among wine samples by grape cultivars compared with yeast strains. Knowledge on the influence of indigenous non-Saccharomyces strains and grape cultivars on the flavour generation contributed to producing diverse wines in cool-climate wine regions. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Antifungal Activity Against Plant Pathogens of Metabolites from the Endophytic Fungus Cladosporium cladosporioides

OpenAIRE

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

2013-01-01

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-diacetyl cladosporin, 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, c...

Two-dimensional profiling of Xanthomonas campestris pv. viticola ...

African Journals Online (AJOL)

However, the analysis of the 2D-PAGE gel images revealed a larger number of spots in the lysis method when compared to the others. Taking ... Keywords: Bacterial canker, Vitis vinifera, proteomics, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), two-dimensional gel electrophoresis (2D-PAGE).

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

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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

Incidência de doenças e necessidade de controle em cultivo protegido de videira Incidence of diseases and needs of control in overhead covered grapes

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Geraldo Chavarria

2007-01-01

Full Text Available O cultivo protegido de videira no Brasil tem-se expandido, em área, visando principalmente à diminuição de danos por adversidades climáticas sobre a produção e a maturação das uvas. Entretanto, não se dispõe de informações sobre o microclima e as necessidades de controle fitossanitário que são impostas por essa tecnologia, as quais constituem os objetivos deste trabalho. O experimento foi instalado no ciclo 2005-2006, em Flores da Cunha-RS, em um vinhedo de 'Moscato Giallo', conduzido em "Y", com cobertura plástica impermeável (160µm, em 12 fileiras com 35m, deixando-se 5 fileiras sem cobertura (controle. Em ambas áreas, avaliou-se o microclima quanto à presença de água livre (registro visual, temperatura (T, umidade relativa (UR do ar, radiação fotossinteticamente ativa (RFA e velocidade do vento (VV próximos ao dossel vegetativo e aos cachos. Na área coberta, foram aplicados fungicidas quando necessário, enquanto na área descoberta foram realizadas aplicações por calendário. Durante a floração e a maturação, avaliaram-se a incidência e a severidade de míldio (Plasmopara viticola, oídio (Uncinula necator, podridão-cinzenta-da-uva (Botrytis cinerea, podridão-da-uva-madura (Glomerella cingulata e podridão ácida (leveduras imperfeitas e leveduras esporógenas. A cobertura plástica aumentou a temperatura diurna próxima ao dossel vegetativo, não influenciou na umidade relativa do ar, diminuiu a radiação fotossinteticamente ativa e a velocidade do vento e restringiu drasticamente a água livre sobre as folhas e cachos. Nessas condições, na área coberta, realizaram-se apenas duas aplicações para o controle do oídio, enquanto na área descoberta foram realizadas 17 aplicações para o controle de doenças fúngicas. Não houve incidência de doenças na avaliação realizada na floração, nos dois sistemas de cultivo; contudo, no período de maturação, houve decréscimos significativos de incid

Evaluation of biocontrol agents for grapevine pruning wound protection against trunk pathogen infection.

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Charl KOTZE

2011-12-01

Full Text Available Trunk diseases of grapevine are caused by numerous pathogens, including Eutypa lata, Phaeomoniella chlamydospora, and species of Botryosphaeriaceae (incl. Botryosphaeria and aggregate genera, Phomopsis and Phaeoacremonium. Since infections occur mainly through pruning wounds, that have been shown by previous research to stay susceptible for up to 16 weeks after pruning, long-term pruning wound protection is required for prevention of infection. This study evaluated several biocontrol agents against a range of trunk disease pathogens in dual plate laboratory trials to determine macroscopic and microscopic interactions. The biocontrol agents had a substantial effect on all the pathogens, with a wide range of macroscopic and microscopic interactions observed. The best performing biocontrol agents were tested in two field trials. Fresh pruning wounds were treated with benomyl, Trichoderma products (Biotricho®, Vinevax® and ECO 77® and isolates (USPP-T1 and -T2, identified as T. atroviride and Bacillus subtilis. Seven days after treatment the pruning wounds were inoculated by spraying with spore suspensions of Neofusicoccum australe, N. parvum, Diplodia seriata, Lasiodiplodia theobromae, Eutypa lata, Phaeomoniella chlamydospora or Phomopsis viticola. Eight months after inoculation, the treatments were evaluated by isolation onto potato dextrose agar. The efficacy of the biocontrol agents was in most cases similar or superior to that observed for benomyl. Isolate USPP-T1, in particular, was very effective, reducing incidence of Ph. viticola, E. lata, Pa. chlamydospora, N. australe, N. parvum, D. seriata and L. theobromae by 69, 76, 77, 78, 80, 85 and 92%, respectively. This is the first report of biological protection of grapevine pruning wounds against this group of grapevine trunk disease pathogens.

Evaluarea rezistenţei unor genotipuri de floarea-soarelui la mană şi rugină în condiţii naturale de infectare

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Aliona CUCEREAVÎI

2017-12-01

Full Text Available In field experiments conducted in 2013-2014 sunflower breeding material (maternal and paternal lines of different origin and hybrids from the collection of the company “AMG-Agroselect Comert” (Republic of Moldova were evaluated for their resistance to natural infection by downy mildew (Plasmopara halstedii and rust (Puccinia helianthi. During the experiments the number of attacked plants and the percentage of attack per plant were recorded as well as disease frequency and intensity and the degree of disease attack were calculated. The data of the field observations were correlated with the results of the molecular studies on the presence or absence of downy mildew and rust resistance genes (Pl6, Pl5/8 and R1, which were performed previously. Thus, the genotypes which contain Pl6, Pl5/8 genes were not infected at all or exhibited an insignificant level of infection by Plasmopara halstedii (maximum 2.4%. The infection by Puccinia helianthi was detected in all maternal lines with cytoplasmic male sterility which lack rust resistance gene (R1. The degree of attack ranged from 0.1 to 93.5%. The paternal lines and the hybrids that possess R1 gene were not attacked by the pathogen. The established correlations denote the effectiveness of molecular screening in rapid and accurate evaluation of the material used in plant breeding and of the obtained hybrids. The native MS-2570C line (adapted to the pedoclimate and cultivation conditions specific to the region and characterized by a low degree of attack or the complete lack thereof is of a particular interest to plant breeding programs as a source of downy mildew and rust resistance. Rezumat.În experienţe de câmp realizate în a. 2013-2014 s-a testat materialul de ameliorare (linii materne şi paterne de provenienţă diferită şi unii hibrizi de floarea-soarelui din colecţia companiei „AMG-Agroselect Comerţ” în scopul evaluării rezistenţei la infecţia naturală cu mană (Plasmopara

Microfungi of the Tatra Mts. 6. Fungus-like organisms: Albuginales, Peronosporales and Pythiales

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Wiesław Mułenko

2014-06-01

Full Text Available A list and the distribution of Oomycota species in the Tatra Mts (Western Carpathian Mts are presented. Revised herbarium vouchers and literature data were used for analysis. Thirty two species of oomycetes on fifty seven plant species were noted in the area, including two species of the order Albuginales (genera: Albugo and Pustula, on nine plant species, 29 species of the order Peronosporales (genera: Bremia, Hyaloperonospora, Peronospora and Plasmopara, on 49 plant species, and one species of the order Pythiales (genus: Myzocytium, on one species of algae. Twenty nine species were collected on the Polish side of the Tatra Mts and ten species were collected on the Slovak side. The oomycetes were collected at 185 localities.

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.

Diaporthe species associated with Vaccinium, with specific reference to Europe

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Lorenzo LOMBARD

2014-09-01

Full Text Available Species of the genus Vaccinium are commercially cultivated in Europe for their berries, which are highly valued for dietary and pharmaceutical properties. Cultivation is severely limited due to a range of fungal diseases, especially those caused by species of Diaporthe. A number of Diaporthe isolates have been collected from Vaccinium growing regions in Europe, and initially identified as D. vaccinii based on host association. Using DNA sequence inference of the combined β-tubulin, calmodulin, translation elongation factor 1-alpha and the internal transcribed spacer region of the nuclear rDNA, along with morphological characteristics, six species were characterised. Diaporthe eres, D. vaccinii and D. viticola are known species and three novel taxa are described here as D. asheicola, D. baccae and D. sterilis. This study is the first confirmed report of D. vaccinii in Latvia and the Netherlands.

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.

Fine mapping of the sunflower resistance locus Pl(ARG) introduced from the wild species Helianthus argophyllus.

Science.gov (United States)

Wieckhorst, S; Bachlava, E; Dussle, C M; Tang, S; Gao, W; Saski, C; Knapp, S J; Schön, C-C; Hahn, V; Bauer, E

2010-11-01

Downy mildew, caused by Plasmopara halstedii, is one of the most destructive diseases in cultivated sunflower (Helianthus annuus L.). The dominant resistance locus Pl(ARG) originates from silverleaf sunflower (H. argophyllus Torrey and Gray) and confers resistance to all known races of P. halstedii. We mapped Pl(ARG) on linkage group (LG) 1 of (cms)HA342 × ARG1575-2, a population consisting of 2,145 F(2) individuals. Further, we identified resistance gene candidates (RGCs) that cosegregated with Pl(ARG) as well as closely linked flanking markers. Markers from the target region were mapped with higher resolution in NDBLOS(sel) × KWS04, a population consisting of 2,780 F(2) individuals that does not segregate for Pl(ARG). A large-insert sunflower bacterial artificial chromosome (BAC) library was screened with overgo probes designed for markers RGC52 and RGC151, which cosegregated with Pl(ARG). Two RGC-containing BAC contigs were anchored to the Pl(ARG) region on LG 1.

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. Copyright © 2015 Elsevier Inc. All rights reserved.

Morphological and Genetic Analyses of the Invasive Forest Pathogen Phytophthora austrocedri Reveal that Two Clonal Lineages Colonized Britain and Argentina from a Common Ancestral Population.

Science.gov (United States)

Henricot, Béatrice; Pérez-Sierra, Ana; Armstrong, April C; Sharp, Paul M; Green, Sarah

2017-12-01

Phytophthora austrocedri is causing widespread mortality of Austrocedrus chilensis in Argentina and Juniperus communis in Britain. The pathogen has also been isolated from J. horizontalis in Germany. Isolates from Britain, Argentina, and Germany are homothallic, with no clear differences in the dimensions of sporangia, oogonia, or oospores. Argentinian and German isolates grew faster than British isolates across a range of media and had a higher temperature tolerance, although most isolates, regardless of origin, grew best at 15°C and all isolates were killed at 25°C. Argentinian and British isolates caused lesions when inoculated onto both A. chilensis and J. communis; however, the Argentinian isolate caused longer lesions on A. chilensis than on J. communis and vice versa for the British isolate. Genetic analyses of nuclear and mitochondrial loci showed that all British isolates are identical. Argentinian isolates and the German isolate are also identical but differ from the British isolates. Single-nucleotide polymorphisms are shared between the British and Argentinian isolates. We concluded that British isolates and Argentinian isolates conform to two distinct clonal lineages of P. austrocedri founded from the same as-yet-unidentified source population. These lineages should be recognized and treated as separate risks by international plant health legislation.

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

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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.

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.

Physiological and molecular characterization of Phytophthora infestans isolates from the Central Colombian Andean Region.

Science.gov (United States)

Céspedes, María C; Cárdenas, Martha E; Vargas, Angela M; Rojas, Alejandro; Morales, Juan G; Jiménez, Pedro; Bernal, Adriana J; Restrepo, Silvia

2013-01-01

Late blight, caused by Phytophthora infestans, is one of the most devastating diseases found in potato and tomato crops worldwide. In Colombia it also attacks other important crops: cape gooseberry and tree tomato. The knowledge of the pathogen population is determinant to effectively design control strategies. To determine the physiological and molecular characteristics of a set of Colombian P. infestans isolates. Strains isolated from Cundinamarca and Boyacá were examined for the level of resistance to mefenoxam and cymoxanil. Virulence was tested for all strains and crosses between A1 mating type, from different hosts, and the Colombian A2 mating type were tested for the production and viability of oospores in different substrates. Additionally, the molecular diversity of the avirulence gene Avr3a, the β-tubulin gene, and two single copy genes showing RxLR motif, was assessed. We found all levels of mefenoxam sensitivity, with 48% of the strains resistant. A high diversity of races was detected and the population was genetically clonal. Colombian strains had the possibility of sexual reproduction. These results will help in optimizing the use of fungicides and deployment of resistance as control strategies and will contribute to broader studies on diversity of this pathogen. Copyright © 2012 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

AQUATIC PLANT SPECIATION AFFECTED BY DIVERSIFYING SELECTION OF ORGANELLE DNA REGIONS(1).

Science.gov (United States)

Kato, Syou; Misawa, Kazuharu; Takahashi, Fumio; Sakayama, Hidetoshi; Sano, Satomi; Kosuge, Keiko; Kasai, Fumie; Watanabe, Makoto M; Tanaka, Jiro; Nozaki, Hisayoshi

2011-10-01

Many of the genes that control photosynthesis are carried in the chloroplast. These genes differ among species. However, evidence has yet to be reported revealing the involvement of organelle genes in the initial stages of plant speciation. To elucidate the molecular basis of aquatic plant speciation, we focused on the unique plant species Chara braunii C. C. Gmel. that inhabits both shallow and deep freshwater habitats and exhibits habitat-based dimorphism of chloroplast DNA (cpDNA). Here, we examined the "shallow" and "deep" subpopulations of C. braunii using two nuclear DNA (nDNA) markers and cpDNA. Genetic differentiation between the two subpopulations was measured in both nDNA and cpDNA regions, although phylogenetic analyses suggested nuclear gene flow between subpopulations. Neutrality tests based on Tajima's D demonstrated diversifying selection acting on organelle DNA regions. Furthermore, both "shallow" and "deep" haplotypes of cpDNA detected in cultures originating from bottom soils of three deep environments suggested that migration of oospores (dormant zygotes) between the two habitats occurs irrespective of the complete habitat-based dimorphism of cpDNA from field-collected vegetative thalli. Therefore, the two subpopulations are highly selected by their different aquatic habitats and show prezygotic isolation, which represents an initial process of speciation affected by ecologically based divergent selection of organelle genes. © 2011 Phycological Society of America.

Downy mildew intensity in tolerant grapes varieties in highlands of southern Brazil

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de Bem Betina

2016-01-01

Full Text Available The aim of this study was to evaluate the different degrees of tolerance to infection by P. viticolaamong three genotypes with constitutive resistance in comparison to susceptible varieties Vitis vinifera. For this purpose two experiments was conducted at EPAGRI Experimental Station, located in the city of São Joaquim, Santa Catarina State, at 2015/16 cycle. In the first experiment on the field, were quantified the incidence and severity and downy mildew intensity was compared by epidemiological variables, on the tolerant varieties Bronner, Regent, Cabernet Cortis and the susceptible Sangiovese. On the second experiment forty leaf discs for the same tolerant genotypes and the susceptible variety Chardonnay were artificially infected with P. viticola sporangia suspension and after seven days of incubation the discs were examined and the degree of infection was estimated based on the intensity of sporangiophore formation. Sangiovese showed highest downy mildew intensity in comparison to the tolerant varieties Cabernet Carbon, Regent and Bronner. Under controlled conditions, the susceptible variety Chardonnay showed higher sporangiophore formation on discs leafs in comparison to the tolerant varieties. All the downy mildew tolerant varieties evaluated showed lower disease development in comparison with V. vinifera varieties.

Grapevine bunch rots: impacts on wine composition, quality, and potential procedures for the removal of wine faults.

Science.gov (United States)

Steel, Christopher C; Blackman, John W; Schmidtke, Leigh M

2013-06-05

Bunch rot of grape berries causes economic loss to grape and wine production worldwide. The organisms responsible are largely filamentous fungi, the most common of these being Botrytis cinerea (gray mold); however, there are a range of other fungi responsible for the rotting of grapes such as Aspergillus spp., Penicillium spp., and fungi found in subtropical climates (e.g., Colletotrichum spp. (ripe rot) and Greeneria uvicola (bitter rot)). A further group more commonly associated with diseases of the vegetative tissues of the vine can also infect grape berries (e.g., Botryosphaeriaceae, Phomopsis viticola ). The impact these fungi have on wine quality is poorly understood as are remedial practices in the winery to minimize wine faults. Compounds found in bunch rot affected grapes and wine are typically described as having mushroom, earthy odors and include geosmin, 2-methylisoborneol, 1-octen-3-ol, 2-octen-1-ol, fenchol, and fenchone. This review examines the current state of knowledge about bunch rot of grapes and how this plant disease complex affects wine chemistry. Current wine industry practices to minimize wine faults and gaps in our understanding of how grape bunch rot diseases affect wine production and quality are also identified.

Fine mapping of the sunflower resistance locus PlARG introduced from the wild species Helianthus argophyllus

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Wieckhorst, S.; Bachlava, E.; Dußle, C. M.; Tang, S.; Gao, W.; Saski, C.; Knapp, S. J.; Schön, C.-C.; Hahn, V.

2010-01-01

Downy mildew, caused by Plasmopara halstedii, is one of the most destructive diseases in cultivated sunflower (Helianthus annuus L.). The dominant resistance locus PlARG originates from silverleaf sunflower (H. argophyllus Torrey and Gray) and confers resistance to all known races of P. halstedii. We mapped PlARG on linkage group (LG) 1 of (cms)HA342 × ARG1575-2, a population consisting of 2,145 F2 individuals. Further, we identified resistance gene candidates (RGCs) that cosegregated with PlARG as well as closely linked flanking markers. Markers from the target region were mapped with higher resolution in NDBLOSsel × KWS04, a population consisting of 2,780 F2 individuals that does not segregate for PlARG. A large-insert sunflower bacterial artificial chromosome (BAC) library was screened with overgo probes designed for markers RGC52 and RGC151, which cosegregated with PlARG. Two RGC-containing BAC contigs were anchored to the PlARG region on LG 1. Electronic supplementary material The online version of this article (doi:10.1007/s00122-010-1416-4) contains supplementary material, which is available to authorized users. PMID:20700574

Species from within the Phytophthora cryptogea complex and related species, P. erythroseptica and P. sansomeana, readily hybridize.

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Safaiefarahani, Banafsheh; Mostowfizadeh-Ghalamfarsa, Reza; Hardy, Giles E St J; Burgess, Treena I

2016-08-01

During a study on the phylogenetic relationships between species in the Phytophthora cryptogea complex and related species, Phytophthora erythroseptica and Phytophthora sansomeana, 19 hybrid isolates with multiple polymorphisms in the nuclear sequences were observed. Molecular characterization of hybrids was achieved by sequencing three nuclear (internal transcribed spacers, β-tubulin (TUB), heat shock protein 90) and two mitochondrial (cytochrome c oxidase subunit I (coxI), NADH dehydrogenase subunit I (NADH)) gene regions and cloning of the single-copy nuclear gene, TUB. Based on the molecular studies the hybrid isolates belonged to six distinct groups between P. cryptogea, P. erythroseptica, Phytophthora pseudocryptogea, P. sansomeana, and Phytophthora sp. kelmania. In all cases, only a single coxI and NADH allele was detected and nuclear genes were biparentally inherited, suggesting that the hybrids arose from sexual recombination events. Colony morphology, growth rate, cardinal temperatures, breeding system, and morphology of sporangia, oogonia, oospores, and antheridia were also determined. Some morphological differences between the hybrids and the parental species were noted; however, they were not sufficient to reliably distinguish the taxa and DNA markers from nuclear and mitochondrial genes will to be necessary for their identification. The parental species are all important pathogens of agricultural fields that have been transported globally. With the apparent ease of hybridization within this group there is ample opportunity for virulent hybrids to form, perhaps with extended host ranges. Copyright © 2016 British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Disentangling Peronospora on Papaver: Phylogenetics, Taxonomy, Nomenclature and Host Range of Downy Mildew of Opium Poppy (Papaver somniferum) and Related Species

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Voglmayr, Hermann; Montes-Borrego, Miguel; Landa, Blanca B.

2014-01-01

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. PMID:24806292

A novel, multiplexed, probe-based quantitative PCR assay for the soybean root- and stem-rot pathogen, Phytophthora sojae, utilizes its transposable element.

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Haudenshield, James S; Song, Jeong Y; Hartman, Glen L

2017-01-01

Phytophthora root rot of soybean [Glycine max (L.) Merr.] is caused by the oomycete Phytophthora sojae (Kaufm. & Gerd.). P. sojae has a narrow host range, consisting primarily of soybean, and it is a serious pathogen worldwide. It exists in root and stem tissues as mycelium, wherein it can form oospores which subsequently germinate to release motile, infectious zoospores. Molecular assays detecting DNA of P. sojae are useful in disease diagnostics, and for determining the presence of the organism in host tissues, soils, and runoff or ponded water from potentially infested fields. Such assays as published have utilized ITS sequences from the nuclear ribosomal RNA genes in conventional PCR or dye-binding quantitative PCR (Q-PCR) but are not amenable to multiplexing, and some of these assays did not utilize control strategies for type I or type II errors. In this study, we describe primers and a bifunctional probe with specificity to a gypsy-like retroelement in the P. sojae genome to create a fluorogenic 5'-exonuclease linear hydrolysis assay, with a multiplexed internal control reaction detecting an exogenous target to validate negative calls, and with uracil-deglycosylase-mediated protection against carryover contamination. The assay specifically detected 13 different P. sojae isolates, and excluded 17 other Phytophthora species along with 20 non-Phytophthora fungal and oomycete species pathogenic on soybean. A diagnostic limit of detection of 34 fg total P. sojae DNA was observed in serial dilutions, equivalent to 0.3 genome, and a practical detection sensitivity of four zoospores per sample was achieved, despite losses during DNA extraction.

A new oomycete species parasitic in nematodes, Chlamydomyzium dictyuchoides sp. nov.: developmental biology and phylogenetic studies.

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Beakes, Gordon W; Glockling, Sally L; James, Timothy Y

2014-07-01

The genus Chlamydomyzium is a little studied holocarpic oomycete parasite of nematodes of uncertain phylogenetic and taxonomic position. A new holocarpic species, Chlamydomyzium dictyuchoides, is described which has usually refractile cytoplasm and a dictyuchoid pattern of spore release. This new species infects bacteriotrophic rhabditid nematodes and was isolated from diverse geographical locations. Infection was initiated by zoospore encystment on the host surface and direct penetration of the cuticle. A sparsely branched, constricted, refractile thallus was formed which eventually occupied almost the entire host body cavity, often accompanied by complete dissolution of the host cuticle. Walled primary cysts formed throughout the thallus and each cyst released a single zoospore via an individual exit papillum, leaving a characteristic dictyuchoid wall net behind. At later stages of infection some thalli formed thick-walled stellate resting spores in uniseriate rows. Resting spore formation appeared to be parthenogenetic and was not accompanied by the formation of antheridial compartments. These spores had ooplast-like vacuoles and thick multi-layered walls, both of which suggest they were oospores. The maximum likelihood tree of sequences of the small ribosomal subunit (SSU) gene placed this new isolate in a clade before the main saprolegnialean and peronosporalean lines diverge. A second undescribed Chlamydomyzium sp., which has direct spore release forms a paraphyletic clade, close to C. dictyuchoides and Sapromyces. The fine structure of other documented Chlamydomyzium species was compared, including an undescribed (but sequenced) isolate, SL02, from Japan, Chlamydomyzium anomalum and Chlamydomyzium oviparasiticum. Chlamydomyzium as currently constituted is a paraphyletic genus that is part of a group of phylogenetically problematic early diverging clades that lie close to both the Leptomitales and Rhipidiales. Crown Copyright © 2014. Published by Elsevier

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

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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.

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

2017-06-01

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 F 2 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.

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

Genotyping-by-sequencing targeting of a novel downy mildew resistance gene Pl 20 from wild Helianthus argophyllus for sunflower (Helianthus annuus L.).

Science.gov (United States)

Ma, G J; Markell, S G; Song, Q J; Qi, L L

2017-07-01

Genotyping-by-sequencing revealed a new downy mildew resistance gene, Pl 20 , from wild Helianthus argophyllus located on linkage group 8 of the sunflower genome and closely linked to SNP markers that facilitate the marker-assisted selection of resistance genes. Downy mildew (DM), caused by Plasmopara halstedii, is one of the most devastating and yield-limiting diseases of sunflower. Downy mildew resistance identified in wild Helianthus argophyllus accession PI 494578 was determined to be effective against the predominant and virulent races of P. halstedii occurring in the United States. The evaluation of 114 BC 1 F 2:3 families derived from the cross between HA 89 and PI 494578 against P. halstedii race 734 revealed that single dominant gene controls downy mildew resistance in the population. Genotyping-by-sequencing analysis conducted in the BC 1 F 2 population indicated that the DM resistance gene derived from wild H. argophyllus PI 494578 is located on the upper end of the linkage group (LG) 8 of the sunflower genome, as was determined single nucleotide polymorphism (SNP) markers associated with DM resistance. Analysis of 11 additional SNP markers previously mapped to this region revealed that the resistance gene, named Pl 20 , co-segregated with four markers, SFW02745, SFW09076, S8_11272025, and S8_11272046, and is flanked by SFW04358 and S8_100385559 at an interval of 1.8 cM. The newly discovered P. halstedii resistance gene has been introgressed from wild species into cultivated sunflower to provide a novel gene with DM resistance. The homozygous resistant individuals were selected from BC 2 F 2 progenies with the use of markers linked to the Pl 20 gene, and these lines should benefit the sunflower community for Helianthus improvement.

Pl(17) is a novel gene independent of known downy mildew resistance genes in the cultivated sunflower (Helianthus annuus L.).

Science.gov (United States)

Qi, L L; Long, Y M; Jan, C C; Ma, G J; Gulya, T J

2015-04-01

Pl 17, a novel downy mildew resistance gene independent of known downy mildew resistance genes in sunflowers, was genetically mapped to linkage group 4 of the sunflower genome. 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 P. halstedii currently identified in the USA. To determine the chromosomal location of this resistance, 186 F 2:3 families derived from a cross of HA 458 with HA 234 were phenotyped for their resistance to race 734 of P. halstedii. The segregation ratio of the population supported that the resistance was controlled by a single dominant gene, Pl 17. Simple sequence repeat (SSR) and single nucleotide polymorphism (SNP) primers were used to identify molecular markers linked to Pl 17. Bulked segregant analysis using 849 SSR markers located Pl 17 to linkage group (LG) 4, which is the first DM gene discovered in this linkage group. An F2 population of 186 individuals was screened with polymorphic SSR and SNP primers from LG4. Two flanking markers, SNP SFW04052 and SSR ORS963, delineated Pl 17 in an interval of 3.0 cM. The markers linked to Pl 17 were validated in a BC3 population. A search for the physical location of flanking markers in sunflower genome sequences revealed that the Pl 17 region had a recombination frequency of 0.59 Mb/cM, which was a fourfold higher recombination rate relative to the genomic average. This region can be considered amenable to molecular manipulation for further map-based cloning of Pl 17.

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

Sunflower Hybrid Breeding: From Markers to Genomic Selection.

Science.gov (United States)

Dimitrijevic, Aleksandra; Horn, Renate

2017-01-01

In sunflower, molecular markers for simple traits as, e.g., fertility restoration, high oleic acid content, herbicide tolerance or resistances to Plasmopara halstedii, Puccinia helianthi , or Orobanche cumana have been successfully used in marker-assisted breeding programs for years. However, agronomically important complex quantitative traits like yield, heterosis, drought tolerance, oil content or selection for disease resistance, e.g., against Sclerotinia sclerotiorum have been challenging and will require genome-wide approaches. Plant genetic resources for sunflower are being collected and conserved worldwide that represent valuable resources to study complex traits. Sunflower association panels provide the basis for genome-wide association studies, overcoming disadvantages of biparental populations. Advances in technologies and the availability of the sunflower genome sequence made novel approaches on the whole genome level possible. Genotype-by-sequencing, and whole genome sequencing based on next generation sequencing technologies facilitated the production of large amounts of SNP markers for high density maps as well as SNP arrays and allowed genome-wide association studies and genomic selection in sunflower. Genome wide or candidate gene based association studies have been performed for traits like branching, flowering time, resistance to Sclerotinia head and stalk rot. First steps in genomic selection with regard to hybrid performance and hybrid oil content have shown that genomic selection can successfully address complex quantitative traits in sunflower and will help to speed up sunflower breeding programs in the future. To make sunflower more competitive toward other oil crops higher levels of resistance against pathogens and better yield performance are required. In addition, optimizing plant architecture toward a more complex growth type for higher plant densities has the potential to considerably increase yields per hectare. Integrative approaches

Sunflower Hybrid Breeding: From Markers to Genomic Selection

Directory of Open Access Journals (Sweden)

Aleksandra Dimitrijevic

2018-01-01

Full Text Available In sunflower, molecular markers for simple traits as, e.g., fertility restoration, high oleic acid content, herbicide tolerance or resistances to Plasmopara halstedii, Puccinia helianthi, or Orobanche cumana have been successfully used in marker-assisted breeding programs for years. However, agronomically important complex quantitative traits like yield, heterosis, drought tolerance, oil content or selection for disease resistance, e.g., against Sclerotinia sclerotiorum have been challenging and will require genome-wide approaches. Plant genetic resources for sunflower are being collected and conserved worldwide that represent valuable resources to study complex traits. Sunflower association panels provide the basis for genome-wide association studies, overcoming disadvantages of biparental populations. Advances in technologies and the availability of the sunflower genome sequence made novel approaches on the whole genome level possible. Genotype-by-sequencing, and whole genome sequencing based on next generation sequencing technologies facilitated the production of large amounts of SNP markers for high density maps as well as SNP arrays and allowed genome-wide association studies and genomic selection in sunflower. Genome wide or candidate gene based association studies have been performed for traits like branching, flowering time, resistance to Sclerotinia head and stalk rot. First steps in genomic selection with regard to hybrid performance and hybrid oil content have shown that genomic selection can successfully address complex quantitative traits in sunflower and will help to speed up sunflower breeding programs in the future. To make sunflower more competitive toward other oil crops higher levels of resistance against pathogens and better yield performance are required. In addition, optimizing plant architecture toward a more complex growth type for higher plant densities has the potential to considerably increase yields per hectare

Sunflower Hybrid Breeding: From Markers to Genomic Selection

Science.gov (United States)

Dimitrijevic, Aleksandra; Horn, Renate

2018-01-01

In sunflower, molecular markers for simple traits as, e.g., fertility restoration, high oleic acid content, herbicide tolerance or resistances to Plasmopara halstedii, Puccinia helianthi, or Orobanche cumana have been successfully used in marker-assisted breeding programs for years. However, agronomically important complex quantitative traits like yield, heterosis, drought tolerance, oil content or selection for disease resistance, e.g., against Sclerotinia sclerotiorum have been challenging and will require genome-wide approaches. Plant genetic resources for sunflower are being collected and conserved worldwide that represent valuable resources to study complex traits. Sunflower association panels provide the basis for genome-wide association studies, overcoming disadvantages of biparental populations. Advances in technologies and the availability of the sunflower genome sequence made novel approaches on the whole genome level possible. Genotype-by-sequencing, and whole genome sequencing based on next generation sequencing technologies facilitated the production of large amounts of SNP markers for high density maps as well as SNP arrays and allowed genome-wide association studies and genomic selection in sunflower. Genome wide or candidate gene based association studies have been performed for traits like branching, flowering time, resistance to Sclerotinia head and stalk rot. First steps in genomic selection with regard to hybrid performance and hybrid oil content have shown that genomic selection can successfully address complex quantitative traits in sunflower and will help to speed up sunflower breeding programs in the future. To make sunflower more competitive toward other oil crops higher levels of resistance against pathogens and better yield performance are required. In addition, optimizing plant architecture toward a more complex growth type for higher plant densities has the potential to considerably increase yields per hectare. Integrative approaches

Genetics and mapping of a novel downy mildew resistance gene, Pl(18), introgressed from wild Helianthus argophyllus into cultivated sunflower (Helianthus annuus L.).

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Qi, L L; Foley, M E; Cai, X W; Gulya, T J

2016-04-01

A novel downy mildew resistance gene, Pl(18), was introgressed from wild Helianthus argophyllus into cultivated sunflower and genetically mapped to linkage group 2 of the sunflower genome. The new germplasm, HA-DM1, carrying Pl(18) has been released to the public. Sunflower downy mildew (DM) is considered to be the most destructive foliar disease that has spread to every major sunflower-growing country of the world, except Australia. A new dominant downy mildew resistance gene (Pl 18) transferred from wild Helianthus argophyllus (PI 494573) into cultivated sunflower was mapped to linkage group (LG) 2 of the sunflower genome using bulked segregant analysis with 869 simple sequence repeat (SSR) markers. Phenotyping 142 BC1F2:3 families derived from the cross of HA 89 and H. argophyllus confirmed the single gene inheritance of resistance. Since no other Pl gene has been mapped to LG2, this gene was novel and designated as Pl (18). SSR markers CRT214 and ORS203 flanked Pl(18) at a genetic distance of 1.1 and 0.4 cM, respectively. Forty-six single nucleotide polymorphism (SNP) markers that cover the Pl(18) region were surveyed for saturation mapping of the region. Six co-segregating SNP markers were 1.2 cM distal to Pl(18), and another four co-segregating SNP markers were 0.9 cM proximal to Pl(18). The new BC2F4-derived germplasm, HA-DM1, carrying Pl(18) has been released to the public. This new line is highly resistant to all Plasmopara halstedii races identified in the USA providing breeders with an effective new source of resistance against downy mildew in sunflower. The molecular markers that were developed will be especially useful in marker-assisted selection and pyramiding of Pl resistance genes because of their close proximity to the gene and the availability of high-throughput SNP detection assays.

Discovery and introgression of the wild sunflower-derived novel downy mildew resistance gene Pl 19 in confection sunflower (Helianthus annuus L.).

Science.gov (United States)

Zhang, Z W; Ma, G J; Zhao, J; Markell, S G; Qi, L L

2017-01-01

A new downy mildew resistance gene, Pl 19 , was identified from wild Helianthus annuus accession PI 435414, introduced to confection sunflower, and genetically mapped to linkage group 4 of the sunflower genome. Wild Helianthus annuus accession PI 435414 exhibited resistance to downy mildew, which is one of the most destructive diseases to sunflower production globally. Evaluation of the 140 BC 1 F 2:3 families derived from the cross of CMS CONFSCLB1 and PI 435414 against Plasmopara halstedii race 734 revealed that a single dominant gene controls downy mildew resistance in the population. Bulked segregant analysis conducted in the BC 1 F 2 population with 860 simple sequence repeat (SSR) markers indicated that the resistance derived from wild H. annuus was associated with SSR markers located on linkage group (LG) 4 of the sunflower genome. To map and tag this resistance locus, designated Pl 19 , 140 BC 1 F 2 individuals were used to construct a linkage map of the gene region. Two SSR markers, ORS963 and HT298, were linked to Pl 19 within a distance of 4.7 cM. After screening 27 additional single nucleotide polymorphism (SNP) markers previously mapped to this region, two flanking SNP markers, NSA_003564 and NSA_006089, were identified as surrounding the Pl 19 gene at a distance of 0.6 cM from each side. Genetic analysis indicated that Pl 19 is different from Pl 17 , which had previously been mapped to LG4, but is closely linked to Pl 17 . This new gene is highly effective against the most predominant and virulent races of P. halstedii currently identified in North America and is the first downy mildew resistance gene that has been transferred to confection sunflower. The selected resistant germplasm derived from homozygous BC 2 F 3 progeny provides a novel gene for use in confection sunflower breeding programs.

Carbon stable isotope composition of charophyte organic matter in a small and shallow Spanish water body as a baseline for future trophic studies

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María Antonia Rodrigo

2015-12-01

Full Text Available Quantitative descriptions of foodweb structure based on isotope niche space require knowledge of producers’ isotopic signatures. In freshwater ecosystems charophytes are one of the main components of submerged vegetation and the feeding base for many herbivorous consumers, but knowledge about their organic carbon isotopic signatures is sparse. In this study, the δ13C organic values (and organic %C and %N of the four species of submerged macrophytes (three charophytes - Chara hispida, Nitella hyalina and Tolypella glomerata - and one angiosperm, Myriophyllum spicatum growing in a newly created shallow pond were measured monthly over a period of one year, to discern if i all charophyte species susceptible to being food for consumers and growing in the same waterbody have the same C isotopic composition; ii the δ13C values of a charophyte species change on a seasonal and spatial scale; iii the different parts (apical nodes, internodes, rhizoids, reproductive organs, oospores of a charophyte species have the same isotopic composition. The δ13C, %C and %N values of organic matter in the sediments where the plants were rooted were also measured as well as several limnological variables. The δ13C values for the angiosperm (-13.7±0.7‰ indicated 13C-enrichment, whereas the N. hyalina δ13C values were the most negative (-22.4±0.7‰. The mean δ13C value for C. hispida was -19.0±1.0‰ and -20.7±0.8‰ for T. glomerata. C. hispida δ13C values had a significant seasonal variation with 13C-poor values in the cold season, and slight spatial differences. Statistically significant differences were found between charophyte rhizoids (13C-enriched and the other parts of the thalli. The δ13C values in the sediments varied throughout time (-13‰ to -26‰. The C content was lower in the charophytes than in the angiosperm and there were no large differences among the charophytes. Charophyte fructifications were enriched in organic C compared to the

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

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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