Investigation of Sclerotinia sclerotiorum strains variability in Brazil.

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Abreu, M J; Souza, E A

2015-06-18

White mold is a common bean disease caused by the fungus Sclerotinia sclerotiorum, resulting in economic losses in Brazil and worldwide. Lack of knowledge about the population structure of the pathogen makes it difficult to control the disease. The aim of this study was to characterize strains of S. sclerotiorum obtained from ex-perimental and commercial common bean fields in Brazil. We analyzed 50 strains of S. sclerotiorum collected at several locations in the state of Minas Gerais. The strains were characterized according to their ability and time for developing apothecia. Morphological and physiological analyses such as the mycelial growth index, colony color, the time re-quired to form the first sclerotia on media, the number of sclerotia per plate, average sclerotium size, and sclerotium shape were performed. We determined the mycelial compatibility, conducted molecular analy-sis of microsatellites, and evaluated the aggressiveness of 28 strains. Most strains had the ability to form apothecia. A small group of strains showed mycelial compatibility, and the strains showed different aggres-siveness levels. Overall, the population studied here demonstrated wide variability based on the morphological, physiological, and molecular traits analyzed. The average size and shape of sclerotia presented a cor-relation of 0.617, whereas the times required to form sclerotia and the number of sclerotia per plate showed a correlation of -0.455. The char-acterization of the pathogen population described herein will provide an important tool for promoting the development of bean cultivars re-sistant to white mold.

Formulations of Bacillus subtilis BY-2 suppress Sclerotinia sclerotiorum on oilseed rape in the field

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We are developing a collection of Bacillus strains, isolated from different environments, for use in controlling Sclerotinia sclerotiorum on oilseed rape in China and elsewhere. Strain BY-2, isolated from internal tissues of an oilseed rape root, was demonstrated to be Bacillus subtilis based on bi...

Soybean production and carpogenic germination of Sclerotinia sclerotiorum under different cover cropsProdução de soja e germinação carpogênica de Sclerotinia sclerotiorum sob diferentes coberturas de solo

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Luciano Reis Venturoso

2013-05-01

Full Text Available The aim of this work was to evaluate the influence of different soil cover crops on the carpogenic germination of Sclerotinia sclerotiorum and the development and yield of soybean. The treatments consisted of mulches of brachiaria, canola, safflower, crambe, sunflower and forage radish mulch, and a control with no cover mulch. The crops were sown in pots containing 4.4 dm³ of soil type Rhodic Acrustox. After 45 days the plant material was cut into pieces in order to standardize the amount of straw to 2800 kg ha-1. Soybean seeds were sown and seven days after seedling emergence two sclerotia were allocated in each pot. With regard to carpogenic germination, we analyzed the time to germination of sclerotia and formation of apothecia, number of stipes and apothecia per sclerotia and the percentage apothecia formed. In the soybean crop was determined plant height at flowering and harvest, relative chlorophyll index, dry matter mass and root, number of pods per plant, number of seeds per pod, grain yield, number of nodules per plant and dry mass of nodules. With the exception of safflower mulch, the use of cover crops reduced the formation of stipes and apothecia of S. sclerotiorum. The covers with brachiaria, sunflower and forage radish mulch increased by 16, 10 and 6 days respectively the overall period of apothecium formation, but only brachiaria reduced the percentage of apothecia formed. The sunflower mulch hindered soybean development and yield. O trabalho teve por objetivo avaliar a influência de diferentes coberturas vegetais de solo sobre a germinação carpogênica de Sclerotinia sclerotiorum e sobre o desenvolvimento e rendimento da cultura da soja. Os tratamentos constaram da palhada de braquiária, canola, cártamo, crambe, girassol e nabo forrageiro, mais um controle sem cobertura. As culturas de cobertura foram semeadas em vasos contendo 4,4 dm³ de solo do tipo Latossolo Vermelho Distroférrico. Após 45 dias o material vegetal foi

Random T-DNA mutagenesis identifies a Cu-Zn-superoxide dismutase gene as a virulence factor of Sclerotinia sclerotiorum

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Agrobacterium-mediated transformation (AMT) was used to identify potential virulence factors in Sclerotinia sclerotiorum. Screening AMT transformants identified two mutants showing significantly reduced virulence. The mutants showed similar growth rate, colony morphology, and sclerotial and oxalate ...

A multiplex PCR assay for the detection and quantification of Sclerotinia sclerotiorum and Botrytis cinerea.

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Reich, J D; Alexander, T W; Chatterton, S

2016-05-01

Traditional culture methods for identifying the plant fungal pathogens Sclerotinia sclerotiorum (Lib.) de Bary and Botrytis cinerea Pers.:Fr. are slow and laborious. The goal of this study was to develop a multiplex real-time PCR (qPCR) assay to detect and quantify DNA from S. sclerotiorum and B. cinerea. A primer set (SsIGS_5) for S. sclerotiorum was designed that targeted the intergenic spacer (IGS) regions of the ribosomal DNA. Addition of a probe to the assay increased its specificity: when the primer/probe set was tested against 21 fungal species (35 strains), amplification was detected from all S. sclerotiorum strains and no other species. For qPCR, the SsIGS_5 primer and probe set exhibited a linear range from 7·0 ng to 0·07 pg target DNA (R(2)  = 0·99). SsIGS_5 was then multiplexed with a previously published primer/probe set for B. cinerea to develop a high-throughput method for the detection and quantification of DNA from both pathogens. When multiplexed, the sensitivity and specificity of both assays were not different from individual qPCR reactions. The multiplex assay is currently being used to detect and quantify S. sclerotiorum and B. cinerea DNA from aerosol samples collected in commercial seed alfalfa fields. A primer and probe set for the quantification of Sclerotinia sclerotiorum DNA in a PCR assay was developed. The probe-based nature of this assay signifies an improvement over previous assays for this species by allowing multiplex reactions while maintaining high sensitivity. The primer/probe set was used in a multiplex real-time PCR assay for the quantification of S. sclerotiorum and Botrytis cinerea DNA, enabling rapid analysis of environmental samples. In crops susceptible to both pathogens, this multiplex assay can be used to quickly quantify the presence of each pathogen. © 2016 Her Majesty the Queen in Right of Canada © 2016 The Society for Applied Microbiology. Reproduced with the permission of the Office of the

IDENTIFICAÇÃO E UTILIZAÇÃO DE Trichoderma spp. ARMAZENADOS E NATIVOS NO BIOCONTROLE DE Sclerotinia sclerotiorum

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GERARDA BEATRIZ PINTO DA SILVA; LEISE INÊS HECKLER; RICARDO FELICIANO DOS SANTOS; MIRIA ROSA DURIGO; ELENA BLUME

2015-01-01

The fungus Sclerotinia sclerotiorum , handles significant losses in lettuce production. Being a soil borne fungus, its management is difficult, and an alternative is the use of biological control using species of the Trichoderma genus. Thus, the objectives of this study were to identify native species of Trichoderma spp. presents in the soil with (CP) and without white mold (SP), evaluate the growth rate and in vitro antagonism of Trichoderma spp. against S. sclerotiorum and to ver...

Calcium oxalate crystals: an integral component of the Sclerotinia sclerotiorum/Brassica carinata pathosystem.

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Margaret B Uloth

Full Text Available Oxalic acid is an important virulence factor for disease caused by the fungal necrotrophic pathogen Sclerotinia sclerotiorum, yet calcium oxalate (CaOx crystals have not been widely reported. B. carinata stems were infected with S. sclerotiorum and observed using light microscopy. Six hours post inoculation (hpi, CaOx crystals were evident on 46% of stem sections and by 72 hpi on 100%, demonstrating that the secretion of oxalic acid by S. sclerotiorum commences before hyphal penetration. This is the first time CaOx crystals have been reported on B. carinata infected with S. sclerotiorum. The shape of crystals varied as infection progressed. Long tetragonal rods were dominant 12 hpi (68% of crystal-containing samples, but by 72 hpi, 50% of stems displayed bipyramidal crystals, and only 23% had long rods. Scanning electron microscopy from 24 hpi revealed CaOx crystals in all samples, ranging from tiny irregular crystals (< 0.5 μm to large (up to 40 μm highly organized arrangements. Crystal morphology encompassed various forms, including tetragonal prisms, oval plates, crystal sand, and druses. Large conglomerates of CaOx crystals were observed in the hyphal mass 72 hpi and these are proposed as a strategy of the fungus to hold and detoxify Ca2+ions. The range of crystal morphologies suggests that S. sclerotiorum growth and infection controls the form taken by CaOx crystals.

Attack modes and defence reactions in pathosystems involving Sclerotinia sclerotiorum, Brassica carinata, B. juncea and B. napus.

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Uloth, Margaret B; Clode, Peta L; You, Ming Pei; Barbetti, Martin J

2016-01-01

Sclerotinia stem rot (SSR, Sclerotinia sclerotiorum) is a damaging disease of oilseed brassicas world-wide. Host resistance is urgently needed to achieve control, yet the factors that contribute to stem resistance are not well understood. This study investigated the mechanisms of resistance to SSR. Stems of 5-week-old Brassica carinata, B. juncea and B. napus of known resistance were infected via filter paper discs impregnated with S. sclerotiorum mycelium under controlled conditions. Transverse sections of the stem and portions of the stem surface were examined using optical and scanning electron microscopy. The association of anatomical features with the severity of disease (measured by mean lesion length) was determined. Several distinct resistance mechanisms were recorded for the first time in these Brassica-pathogen interactions, including hypersensitive reactions and lignification within the stem cortex, endodermis and in tissues surrounding the lesions. Genotypes showing a strong lignification response 72 h post-infection (hpi) tended to have smaller lesions. Extensive vascular invasion by S. sclerotiorum was observed only in susceptible genotypes, especially in the vascular fibres and xylem. Mean lesion length was negatively correlated with the number of cell layers in the cortex, suggesting progress of S. sclerotiorum is impeded by more cell layers. Hyphae in the centre of lesions became highly vacuolate 72 hpi, reflecting an ageing process in S. sclerotiorum hyphal networks that was independent of host resistance. The infection process of S. sclerotiorum was analogous in B. carinata and B. napus. Infection cushions of the highly virulent isolate of S. sclerotiorum MBRS-1 were grouped together in dense parallel bundles, while hyphae in the infection cushions of a less aggressive isolate WW-3 were more diffuse, and this was unaffected by host genotype. A variety of mechanisms contribute to host resistance against S. sclerotiorum across the three

Genomic analysis of the necrotrophic fungal pathogens Sclerotinia sclerotiorum and Botrytis cinerea.

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Joelle Amselem

2011-08-01

Full Text Available Sclerotinia sclerotiorum and Botrytis cinerea are closely related necrotrophic plant pathogenic fungi notable for their wide host ranges and environmental persistence. These attributes have made these species models for understanding the complexity of necrotrophic, broad host-range pathogenicity. Despite their similarities, the two species differ in mating behaviour and the ability to produce asexual spores. We have sequenced the genomes of one strain of S. sclerotiorum and two strains of B. cinerea. The comparative analysis of these genomes relative to one another and to other sequenced fungal genomes is provided here. Their 38-39 Mb genomes include 11,860-14,270 predicted genes, which share 83% amino acid identity on average between the two species. We have mapped the S. sclerotiorum assembly to 16 chromosomes and found large-scale co-linearity with the B. cinerea genomes. Seven percent of the S. sclerotiorum genome comprises transposable elements compared to <1% of B. cinerea. The arsenal of genes associated with necrotrophic processes is similar between the species, including genes involved in plant cell wall degradation and oxalic acid production. Analysis of secondary metabolism gene clusters revealed an expansion in number and diversity of B. cinerea-specific secondary metabolites relative to S. sclerotiorum. The potential diversity in secondary metabolism might be involved in adaptation to specific ecological niches. Comparative genome analysis revealed the basis of differing sexual mating compatibility systems between S. sclerotiorum and B. cinerea. The organization of the mating-type loci differs, and their structures provide evidence for the evolution of heterothallism from homothallism. These data shed light on the evolutionary and mechanistic bases of the genetically complex traits of necrotrophic pathogenicity and sexual mating. This resource should facilitate the functional studies designed to better understand what makes these

Isolation of oxalic acid tolerating fungi and decipherization of its potential to control Sclerotinia sclerotiorum through oxalate oxidase like protein.

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Yadav, Shivani; Srivastava, Alok K; Singh, Dhanajay P; Arora, Dilip K

2012-11-01

Oxalic acid plays major role in the pathogenesis by Sclerotinia sclerotiorum; it lowers the pH of nearby environment and creates the favorable condition for the infection. In this study we examined the degradation of oxalic acid through oxalate oxidase and biocontrol of Sclerotinia sclerotiorum. A survey was conducted to collect the rhizospheric soil samples from Indo-Gangetic Plains of India to isolate the efficient fungal strains able to tolerate oxalic acid. A total of 120 fungal strains were isolated from root adhering soils of different vegetable crops. Out of 120 strains a total of 80 isolates were able to grow at 10 mM of oxalic acid whereas only 15 isolates were grow at 50 mM of oxalic acid concentration. Then we examined the antagonistic activity of the 15 isolates against Sclerotinia sclerotiorum. These strains potentially inhibit the growth of the test pathogen. A total of three potential strains and two standard cultures of fungi were tested for the oxalate oxidase activity. Strains S7 showed the maximum degradation of oxalic acid (23 %) after 60 min of incubation with fungal extract having oxalate oxidase activity. Microscopic observation and ITS (internally transcribed spacers) sequencing categorized the potential fungal strains into the Aspergillus, Fusarium and Trichoderma. Trichoderma sp. are well studied biocontrol agent and interestingly we also found the oxalate oxidase type activity in these strains which further strengthens the potentiality of these biocontrol agents.

pH Dependency of sclerotial development and pathogenicity revealed by using genetically defined oxalate-minus mutants of Sclerotinia sclerotiorum

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The devastating plant pathogen Sclerotinia sclerotiorum produces copious (up to 50mM) amounts of oxalic acid, which, for over a quarter century, has been claimed as the pathogenicity determinant based on UV-induced mutants that concomitantly lost oxalate production and pathogenicity. Such a claim wa...

Transformation of Sclerotinia Sclerotiorum with the Green Fluorescent Protein Gene and Fluorescence of Hyphae in Four Inoculated Hosts

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Sclerotinia sclerotiorum is an important pathogen of a wide variety of crops. To obtain a genetic marker to observe and study the interaction of the pathogen with its hosts, isolates ND30 and ND21 were transformed using pCT74 and gGFP constructs both containing genes for the green fluorescent protei...

Components of a rice-oilseed rape production system augmented with trichoderma sp. Tri-1 control sclerotinia sclerotiorum on oilseed rape

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Sclerotinia sclerotiorum causes serious yield losses on many crops throughout the world. In two field trials conducted at the same location in consecutive years, a treatment containing formulated Trichoderma harzianum-1 (Tri-1) resulted in oilseed rape seed yield that was significantly greater than...

Rhizosphere competent Mesorhizobiumloti MP6 induces root hair curling, inhibits Sclerotinia sclerotiorum and enhances growth of Indian mustard (Brassica campestris Mesorhizobium loti MP6 rizosférico competente induz encurvamento do pelo daraiz, inibe Sclerotinia sclerotiorum e estimula o crescimento de mostarda indiana (Brassica campestris

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Shikha Chandra

2007-03-01

Full Text Available The bacterial strain Mesorhizobium loti MP6, isolated from root nodules of Mimosa pudica induced growth and yield of Brassica campestris. The isolate MP6 secreted hydroxamate type siderophore in Chrom-Azurol Siderophore (CAS agar medium. Production of hydrocyanic acid (HCN, indole acetic acid (IAA and phosphate solubilizing ability was also recorded under normal growth conditions. Root hair curling was observed through simple glass-slide technique. In vitro study showed a significant increase in population of M. loti MP6 in rhizosphere due to root exudates of B. campestris. In dual culture technique the strain showed a strong antagonistic effect against Sclerotinia sclerotiorum, a white rot pathogen of Brassica campestris. The growth of S. sclerotiorum was inhibited by 75% after prolonged incubation. Efficient root colonization of mustard seedlings was confirmed by using a streptomycin-resistant marker M. loti MP6strep+. The M. loti MP6 coated seeds proved enhanced seed germination, early vegetative growth and grain yield as compared to control. Also, a drastic decline (99% in the incidence of white rot was observed due to application of M. loti MP6.A cepa bacteriana Mesorhizobium loti MP6 isolada de nódulos de raiz de Mimosa pudica induziu o crescimento e o rendimento de Brassica campestris. A cepa MP6 secretou sideróforo do tipo hidroxamato em meio sólido Chrom-Azurol Siderophore (CAS. Em condições normais de crescimento, a cepa foi também capaz de produzir de ácido cianídrico (HCN e acido indolacético (AIA e solubilizar fosfato. O encurvamento do pelo da raiz foi observado usando a simples técnica de lâmina e lamínula. Estudos in vitro mostraram um aumento significativo na população de M. loti MP6 na rizosfera devido aos exsudatos de B. campestris. Empregando-se técnica de co-cultura, a cepa mostrou um grande efeito antagônico contra o fungo Sclerotinia sclerotiorum, o patógeno da podridão branca de Brassica campestris. Ap

Ca-Lignosulphonate and sclerotial viability of Sclerotinia sclerotiorum

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MATTEO MONTANARI

2012-01-01

Full Text Available Lignosulphonates, low cost by-products of the pulping process, have shown suppressive effects against some diseases caused by soil-borne pathogens. In this study, the effect of 1.5% v/v calcium lignosulphonate (Ca-Ls amendment to two commercial potting mixes (peat + coconut fibres; PC; and municipal compost + peat + pumice; MCPP on the viability of Sclerotinia sclerotiorum sclerotia was investigated. Sclerotia were buried in the Ca-Ls amended substrates for 30 days. Non-amended PC and MCPP, sterile sand and sterile PC with and without Ca-Ls were used as controls. The viability of sclerotia recovered from PC and MCPP amended with Ca-Ls was reduced by 50 and 42% respectively compared to control treatments. Ca-Ls amendment decreased sclerotial viability by enhancing the activity of the indigenous mycoparasitic fungi, Fusarium oxysporum, Mucor spp. and Trichoderma spp. The biocontrol ability of Ca-Ls against sclerotia was due to the stimulation of microbial activity and is, therefore, strictly dependent on the microbial composition of the substrate.

1,8-Dihydroxynaphthalene monoglucoside, a new metabolite of Sclerotinia sclerotiorum, and the effect of tricyclazole on its production.

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Starratt, A N; Ross, L M; Lazarovits, G

2002-04-01

Isolate SS7 of Sclerotinia sclerotiorum was previously shown to produce and excrete into agar medium copious amounts of the melanin precursor 1,8-dihydroxynaphthalene. Much reduced quantities of this product were produced in the presence of tricyclazole, an inhibitor of pentaketide melanin biosynthesis. In this study, we demonstrate that young cultures of isolate SS7 produce 1,8-dihydroxynaphthalene monoglucoside, a new natural product not previously reported from fungi. When cultured in the presence of tricyclazole, such young cultures also accumulated two new monoglucosides of 1,3,8-trihydroxynaphthalene, which, as well as 1,8-dihydroxynaphthalene monoglucoside, were also obtained from cultures of two other isolates of S. sclerotiorum. It is proposed that rapid glucosylation of 1,3,8-trihydroxynaphthalene in young tricyclazole-inhibited S. sclerotiorum cultures accounts for the failure to observe 2-hydroxyjuglone or other metabolites usually associated with blockage of the pentaketide pathway to melanin in fungi.

Efeito da freqüência de rega e da umidade do solo sobre a germinação carpogênica de sclerotinia sclerotiorum Effect of watering frequency and soil moisture status on carpogenic germination of Sclerotina sclerotiorum

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Reginaldo Napoleão

2007-03-01

Full Text Available Os efeitos da freqüência de rega e da umidade do solo na germinação carpogênica de Sclerotinia sclerotiorum foram estudados em condições ambientais controladas. Solo e escleródios foram acondicionados em caixas tipo gerbox e umedecidos uma, duas, três e cinco vezes por semana até os níveis de 75 e 100% da saturação. O solo regado uma vez por semana até 75% da saturação não permitiu a germinação dos escleródios, enquanto o solo molhado até 100% da saturação permitiu a germinação de até 70% dos escleródios, assim como um grande número de apotécios. Regas mais freqüentes, nos dois níveis de umidade do solo, aumentaram a germinação de escleródios e a produção de apotécios. Tão importante quanto a umidade do solo foi o intervalo entre regas, pois regas mais freqüentes, mesmo com volumes menores de água, favoreceram a maior germinação carpogênica do patógeno.The effects of watering frequency and soil moisture status on the carpogenic germination of Sclerotinia sclerotiorum were studied under controlled environmental conditions. Soil and sclerotia were placed in gerbox and periodically watered up to 75 and 100% of saturation, one, two, three and five times a week. There was no sclerotium germination when the soil was watered up to 75% of saturation once a week. On the other hand, soils saturated frequently allowed higher percentage of sclerotia germination, until 70%, as well as higher numbers of apothecia. Independently of the soil water content, germination of sclerotia and production of apothecia increased with watering frequency. Important factors on the carpogenic germination of S. sclerotiorum were the soil moisture and the watering frequency. Frequent watering, even with low water volumes applied, also increased the carpogenic germination.

A secretory protein of necrotrophic fungus Sclerotinia sclerotiorum that suppresses host resistance.

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Wenjun Zhu

Full Text Available SSITL (SS1G_14133 of Sclerotinia sclerotiorum encodes a protein with 302 amino acid residues including a signal peptide, its secretion property was confirmed with immunolocalization and immunofluorescence techniques. SSITL was classified in the integrin alpha N-terminal domain superfamily, and its 3D structure is similar to those of human integrin α4-subunit and a fungal integrin-like protein. When S. sclerotiorum was inoculated to its host, high expression of SSITL was detected during the initial stages of infection (1.5-3.0 hpi. Targeted silencing of SSITL resulted in a significant reduction in virulence; on the other hand, inoculation of SSITL silenced transformant A10 initiated strong and rapid defense response in Arabidopsis, the highest expressions of defense genes PDF1.2 and PR-1 appeared at 3 hpi which was 9 hr earlier than that time when plants were inoculated with the wild-type strain of S. sclerotiorum. Systemic resistance induced by A10 was detected by analysis of the expression of PDF1.2 and PR-1, and confirmed following inoculation with Botrytis cinerea. A10 induced much larger lesions on Arabidopsis mutant ein2 and jar1, and slightly larger lesions on mutant pad4 and NahG in comparison with the wild-type plants. Furthermore, both transient and constitutive expression of SSITL in Arabidopsis suppressed the expression of PDF1.2 and led to be more susceptible to A10 and the wild-type strain of S. sclerotiorum and B. cinerea. Our results suggested that SSITL is an effector possibly and plays significant role in the suppression of jasmonic/ethylene (JA/ET signal pathway mediated resistance at the early stage of infection.

Mycoparasitism studies of Trichoderma harzianum against Sclerotinia sclerotiorum: evaluation of antagonism and expression of cell wall-degrading enzymes genes.

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Troian, Rogério Fraga; Steindorff, Andrei Stecca; Ramada, Marcelo Henrique Soller; Arruda, Walquiria; Ulhoa, Cirano José

2014-10-01

Trichoderma spp. are known for their biocontrol activity against several plant pathogens. A specific isolate of Trichoderma harzianum, 303/02, has the potential to inhibit the growth of Sclerotinia sclerotiorum, an important agent involved in several crop diseases. In this study, the interaction between T. harzianum 303/02 and mycelia, sclerotia and apothecia of S. sclerotiorum was studied by scanning electron microscopy. RT-qPCR was used to examine the expression of 11 genes potentially involved in biocontrol. T. harzianum 303/02 parasitizes S. sclerotiorum by forming branches that coil around the hyphae. The fungus multiplied abundantly at the sclerotia and apothecia surface, forming a dense mycelium that penetrated the inner surface of these structures. The levels of gene expression varied according to the type of structure with which T. harzianum was interacting. The data also showed the presence of synergistic action between the cell-wall degrading enzymes.

IDENTIFICAÇÃO E UTILIZAÇÃO DE Trichoderma spp. ARMAZENADOS E NATIVOS NO BIOCONTROLE DE Sclerotinia sclerotiorum

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GERARDA BEATRIZ PINTO DA SILVA

2015-01-01

Full Text Available The fungus Sclerotinia sclerotiorum , handles significant losses in lettuce production. Being a soil borne fungus, its management is difficult, and an alternative is the use of biological control using species of the Trichoderma genus. Thus, the objectives of this study were to identify native species of Trichoderma spp. presents in the soil with (CP and without white mold (SP, evaluate the growth rate and in vitro antagonism of Trichoderma spp. against S. sclerotiorum and to verify the biocontrol potential of Trichoderma spp. microbi- olized lettuce seeds, growing in substrate infested with S. sclerotiorum . Trichoderma spp. isolates were obtained from areas with and without history of white mold or stored in water. Mycelial growth rate and sporu- lation of the Trichoderma spp. isolates and control of Trichoderma spp. versus S. sclerotiorum in the in vitro essays. For the in vivo essay, lettuce seeds were microbiolized with Trichoderma spp. and the substrate was infested with S. sclerotiorum . The native isolates of Trichoderma identified belong to T. koningiopsis and T. asperellum species. The CP isolates had higher mycelial growth rates when compared to the SP isolates and stored while the stored isolates showed better responses in confrontation. The application of Trichoderma spp. promoted higher seedlings quality compared to control, as well as good seedlings development in the presence of the pathogen.

Alternative products in the "in vitro" inhibition of Sclerotinia sclerotiorum

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Mello Alexandre Furtado Silveira

2005-01-01

Full Text Available The white mold, caused by Sclerotinia sclerotiorum, is a very important disease in tomato crops. The objective of this work was to study the effect of plant extracts, animal residues and industrial by-products extracts on the fungus in vitro growth. Treatments consisted of different concentrations of pyrolignous oil, neem oil, monosodium glutamate, sewage sludge and organic compost [coffee residue (50% coal residue (10%, maize residue (25%, poultry waste (12.5%, poultry meal (2.5%]. Positive control consisted of Petri dishes with PDA medium and negative control treatment consisted of PDA medium with procymidone. Fungus colonies were incubated at 22ºC and light intensity of 260 lux. Variables such as mycelium growth rate, sclerotia production, and viability 7 and 17 days after the transfer of mycelium disc to neon media were assessed. The extract of organic compost at 30% was effective in controlling mycelial growth and sclerotia production. This treatment, as well as neem oil at 0.5% increased soil respiration.

Molecular characterization of a bipartite double-stranded RNA virus and its satellite-like RNA co-infecting the phytopathogenic fungus Sclerotinia sclerotiorum

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Lijiang eLiu

2015-05-01

Full Text Available A variety of mycoviruses have been found in Sclerotinia sclerotiorum. In this study, we report a novel mycovirus Sclerotinia sclerotiorum botybirnavirus 1 (SsBRV1 that was originally isolated from the hypovirulent strain SCH941 of S. sclerotiorum. SsBRV1 has rigid spherical virions that are ~38 nm in diameter, and three dsRNA segments (dsRNA1, 2 and 3 with lengths of 6.4, 6.0 and 1.7 kbp, respectively were packaged in the virions. dsRNA1 encodes a cap-pol fusion protein, and dsRNA2 encodes a polyprotein with unknown functions but contributes to the formation of virus particles. The dsRNA3 is dispensable and may be a satellite-like RNA (SatlRNA of SsBRV1. Although phylogenetic analysis of the RdRp domain demonstrated that SsBRV1 is related to Botrytis porri RNA virus 1 (BpRV1 and Ustilago maydis dsRNA virus-H1 (UmV-H1, the structure proteins of SsBRV1 do not have any significant sequence similarities with other known viral proteins with the exception of those of BpRV1. SsBRV1 carrying dsRNA3 seems to have no obvious effects on the colony morphology, but can significantly reduce the growth rate and virulence of S. sclerotiorum. Notably, a growth hormone receptor binding domain (GHBP, Pfam12772 is detected in ORF2-encoded protein of SsBRV1, which have not been reported in any other viruses. These findings provide new insights into the virus taxonomy, virus evolution and the interactions between SsBRV1 and the fungal hosts.

Discovery of Sclerotinia sclerotiorum Hypovirulence-Associated Virus-1 in Urban River Sediments of Heathcote and Styx Rivers in Christchurch City, New Zealand.

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Kraberger, Simona; Stainton, Daisy; Dayaram, Anisha; Zawar-Reza, Peyman; Gomez, Christopher; Harding, Jon S; Varsani, Arvind

2013-08-08

In samples of benthic and bank river sediments of two urban rivers in Christchurch city (New Zealand), we identified and recovered isolates of Sclerotinia sclerotiorum hypovirulence-associated virus-1 (SsHADV-1), a fungus-infecting circular single-stranded DNA virus. This is the first report of SsHADV-1 outside of China and in environmental samples.

Genetic Diversity Studies Based on Morphological Variability, Pathogenicity and Molecular Phylogeny of the Sclerotinia sclerotiorum Population From Indian Mustard (Brassica juncea

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Pankaj Sharma

2018-06-01

Full Text Available White mold or stem rot disease are ubiquitously distributed throughout the world and the causal organism of this disease Sclerotinia sclerotiorum (Lib. de Bary, is known to infect over 400 plant species. Sclerotinia stem rot is one of the most devastating fungal diseases and poses a serious threat to the worldwide cultivation of oilseed Brassica including India. S. sclerotiorum pathogen usually infects the stem but in severe cases leaves and pods also affected at different developmental stages that deteriorate not only the oil quality but also causing the seed and oil yield losses up to 90% depending on the severity of the disease infestation. This study investigated the morphological and molecular characterization of pathogenic S. sclerotiorum (Lib de Bary geographical isolates from oilseed Brassica including Brassica juncea (Indian mustard. The aim of this study was to compare isolates of S. sclerotiorum originated from different agro-climatic conditions and to analyse similarity or differences between them as well as to examine the virulence of this pathogen specifically in Brassica for the first time. The collection of S. sclerotiorum isolates from symptomatic Brassica plants was done and analyzed for morphological features, and molecular characterization. The virulence evaluation test of 65 isolates on four Brassica cultivars has shown 5 of them were highly virulent, 46 were virulent and 14 were moderately virulent. Phylogenetic analysis encompassing all the morphological features, SSR polymorphism, and ITS sequencing has shown the existence of high genetic diversity among the isolates that categorized all the isolates in three evolutionary lineages in the derived dendrogram. Further, genetic variability analysis based on sequences variation in ITS region of all the isolates has shown the existence of either insertions or deletions of the nucleotides in the ITS region has led to the interspecies variability and observed the variation were

Formulations of the endophytic bacterium Bacillus subtilis Tu-100 suppress Sclerotinia sclerotiorum on oilseed rape and improve plant vigor in field trials conducted at separate locations

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Sclerotinia sclerotiorum causes serious yield losses in crops in The People’s Republic of China. Two formulations of oilseed rape seed containing the endophytic bacterium Bacillus subtilis Tu-100 were evaluated for suppression of this pathogen in field trials conducted at two independent locations....

Selection of endophytic fungi from comfrey (Symphytum officinale L.) for in vitro biological control of the phytopathogen Sclerotinia sclerotiorum (Lib.).

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Rocha, Rafaeli; da Luz, Daniela Eleutério; Engels, Cibelle; Pileggi, Sônia Alvim Veiga; de Souza Jaccoud Filho, David; Matiello, Rodrigo Rodrigues; Pileggi, Marcos

2009-01-01

Biological control consists of using one organism to attack another that may cause economic damage to crops. Integrated Pest Management (IPM) is a very common strategy. The white mold produced by Sclerotinia sclerotiorum (Lib.) causes considerable damage to bean crops. This fungus is a soil inhabitant, the symptoms of which are characterized by water-soaked lesions covered by a white cottony fungal growth on the soil surface and/or the host plant. Possible biological control agents taken from plants are being investigated as phytopathogen inhibitors. These are endophytic microorganisms that inhabit the intercellular spaces of vegetal tissues and are often responsible for antimicrobial production. The objective of the present study was to select endophytic fungi isolated from comfrey (Symphytum officinale L.) leaves with in vitro antagonist potential against the phytopathogenic fungus S. sclerotiorum. Twelve isolates of endophytic fungi and a pathogenic strain of S. sclerotiorum were used in the challenge method. With the aid of this method, four endophytes with the best antagonistic activity against S. sclerotiorum were selected. Pathogen growth inhibition zones were considered indicative of antibiosis. The percentages of pathogenic mycelia growth were measured both with and without the antagonist, resulting in growth reductions of 46.7% to 50.0% for S. sclerotiorum. These analyses were performed by evaluating the endophytic/pathogenic mycelia growth in mm/day over an eight-day period of antagonistic tests.

Analysis of genetic diversity of Sclerotinia sclerotiorum from eggplant by mycelial compatibility, random amplification of polymorphic DNA (RAPD and simple sequence repeat (SSR analyses

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Fatih Mehmet Tok

2016-09-01

Full Text Available The genetic diversity and pathogenicity/virulence among 60 eggplant Sclerotinia sclerotiorum isolates collected from six different geographic regions of Turkey were analysed using mycelial compatibility groupings (MCGs, random amplified polymorphic DNA (RAPD and simple sequence repeat (SSR polymorphism. By MCG tests, the isolates were classified into 22 groups. Out of 22 MCGs, 36% were represented each by a single isolate. The isolates showed great variability for virulence regardless of MCG and geographic origin. Based on the results of RAPD and SSR analyses, 60 S. sclerotiorum isolates representing 22 MCGs were grouped in 2 and 3 distinct clusters, respectively. Analyses using RAPD and SSR markers illustrated that cluster groupings or genetic distance of S. sclerotiorum populations from eggplant were not distinctly relative to the MCG, geographical origin and virulence diversity. The patterns obtained revealed a high heterogeneity of genetic composition and suggested the occurrence of clonal and sexual reproduction of S. sclerotiorum on eggplant in the areas surveyed.

The Sclerotinia sclerotiorum Mating Type Locus (MAT) Contains a 3.6-kb Region That Is Inverted in Every Meiotic Generation

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Maruthachalam, Karunakaran; Wu, Bo-Ming; Subbarao, Krishna V.

2013-01-01

Sclerotinia sclerotiorum is a fungal plant pathogen and the causal agent of lettuce drop, an economically important disease of California lettuce. The structure of the S. sclerotiorum mating type locus MAT has previously been reported and consists of two idiomorphs that are fused end-to-end as in other homothallics. We investigated the diversity of S. sclerotiorum MAT using a total of 283 isolates from multiple hosts and locations, and identified a novel MAT allele that differed by a 3.6-kb inversion and was designated Inv+, as opposed to the previously known S. sclerotiorum MAT that lacked the inversion and was Inv-. The inversion affected three of the four MAT genes: MAT1-2-1 and MAT1-2-4 were inverted and MAT1-1-1 was truncated at the 3’-end. Expression of MAT genes differed between Inv+ and Inv- isolates. In Inv+ isolates, only one of the three MAT1-2-1 transcript variants of Inv- isolates was detected, and the alpha1 domain of Inv+ MAT1-1-1 transcripts was truncated. Both Inv- and Inv+ isolates were self-fertile, and the inversion segregated in a 1∶1 ratio regardless of whether the parent was Inv- or Inv+. This suggested the involvement of a highly regulated process in maintaining equal proportions of Inv- and Inv+, likely associated with the sexual state. The MAT inversion region, defined as the 3.6-kb MAT inversion in Inv+ isolates and the homologous region of Inv- isolates, was flanked by a 250-bp inverted repeat on either side. The 250-bp inverted repeat was a partial MAT1-1-1 that through mediation of loop formation and crossing over, may be involved in the inversion process. Inv+ isolates were widespread, and in California and Nebraska constituted half of the isolates examined. We speculate that a similar inversion region may be involved in mating type switching in the filamentous ascomycetes Chromocrea spinulosa, Sclerotinia trifoliorum and in certain Ceratocystis species. PMID:23457637

Defense to Sclerotinia sclerotiorum in oilseed rape is associated with the sequential activations of salicylic acid signaling and jasmonic acid signaling.

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Wang, Zheng; Tan, Xiaoli; Zhang, Zhiyan; Gu, Shoulai; Li, Guanying; Shi, Haifeng

2012-03-01

Signaling pathways mediated by salicylic acid (SA) and jasmonic acid (JA) are widely studied in various host-pathogen interactions. For oilseed rape (Brassica napus)-Sclerotinia sclerotiorum interaction, little information of the two signaling molecules has been described in detail. In this study, we showed that the level of SA and JA in B. napus leaves was increased with a distinct temporal profile, respectively, after S. sclerotiorum infection. The application of SA or methyl jasmonate enhanced the resistance to the pathogen. Furthermore, a set of SA and JA signaling marker genes were identified from B. napus and were used to monitor the signaling responses to S. sclerotiorum infection by examining the temporal expression profiles of these marker genes. The SA signaling was activated within 12h post inoculation (hpi) followed by the JA signaling which was activated around 24 hpi. In addition, SA-JA crosstalk genes were activated during this process. These results suggested that defense against S. sclerotiorum in oilseed rape is associated with a sequential activation of SA signaling and JA signaling, which provide important clues for designing strategies to curb diseases caused by S. sclerotioru. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

EFEITO DA APLICAÇÃO DE FUNGICIDAS NO SOLO SOBRE A GERMINAÇÃO CARPOGÊNICA E MICELIOGÊNICA DE ESCLERÓDIOS DE Sclerotinia sclerotiorum EFFECT OF FUNGICIDE APPLICATION IN THE SOIL ON THE CARPOGENIC AND MYCELIOGENIC GERMINATION OF Sclerotinia sclerotiorum

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Gesimária Ribeiro Costa

2007-09-01

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Os fungicidas benomyl, procymidone, iprodione, vinclozolin, fluazinan e tiofanato metílico foram utilizados para verificar seus efeitos na germinação carpogênica e miceliogênica de escleródios de <em>Sclerotinia sclerotiorum>, no solo. O solo coletado foi distribuído em caixas gerbox. Em cada caixa foram colocados 300 g de solo e, posteriormente, foi efetuado o enterrio de 25 escleródios à profundidade de 2,0 cm. Os fungicidas foram aplicados no solo, na dose de 0,5 kg.ha-1 de i.a., simulando uma lâmina de água de 6,0 mm. Em geral, o vinclozolin foi o melhor tratamento, apresentando uma eficiência de 100% na inibição de estipes e apotécios. O fluazinan permitiu apenas a formação de estipes inviáveis, resultando na ausência de apotécios. Na germinação miceliogênica dos escleródios, observou-se que após quinze dias de incubação, o tiofanato metílico reduziu em 75% a germinação miceliogênica, seguido dos fungicidas procymidone e vinclozolin, que apresentaram, em média, 60% de inibição. Os fungicidas benomyl, fluazinan e iprodione foram os menos eficientes na inibição da germinação miceliogênica dos escleródios, não diferindo estatisticamente do controle. Após trinta dias de incubação dos escleródios no solo, os fungicidas procymidone, tiofanato metílico, vinclozolin, iprodione e fluazinam não diferiram significativamente entre si quanto à inibição da germinação miceliogênica, mas diferiram do fungicida benomyl e do tratamento testemunha.

PALAVRAS-CHAVE: Controle químico; mofo branco; Phaseolus vulgaris.

The effects of benomyl, procymidone, iprodione, vinclozolin, fluazinan and methyl thiophanate on the carpogenic and myceliogenic germination of <em>Sclerotinia> <em>sclerotiorum sclerotiaem>, in soil, were

UTILIZAÇÃO DE Trichoderma spp. NO CONTROLE DE Sclerotinia sclerotiorum (Lib.) de Bary E NO CRESCIMENTO DE ALFACE

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Gerarda Beatriz Pinto da Silva

2013-01-01

O fungo Sclerotinia sclerotiorum é responsável por perdas significativas na produção de alface. Por se tratar de um fungo de solo, seu manejo é dificultado, sendo uma alternativa, o uso do controle biológico utilizando espécies do gênero Trichoderma. Dentre os muitos fatores que podem interferir no desempenho desse antagonista, a origem dos isolados e o armazenamento são ainda pouco estudados. Pouco se sabe também a respeito de como isolados de Trichoderma spp. obtidos de áreas com e sem hist...

Effects of ozone treatment on Botrytis cinerea and Sclerotinia sclerotiorum in relation to horticultural product quality.

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Sharpe, Deana; Fan, Lihua; McRae, Ken; Walker, Brad; MacKay, Ron; Doucette, Craig

2009-08-01

Botrytis cinerea and Sclerotinia sclerotiorum are fungal pathogens that cause the decay of many fruits and vegetables. Ozone may be used as an antimicrobial agent to control the decay. The effect of gaseous ozone on spore viability of B. cinerea and mycelial growth of B. cinerea and S. sclerotiorum were investigated. Spore viability of B. cinerea was reduced by over 99.5% (P < 0.01) and height of the aerial mycelium was reduced from 4.7 mm in the control to less than 1 mm after exposure to 450 or 600 ppb ozone for 48 h at 20 degrees C. Sporulation of B. cinerea was also substantially inhibited by ozone treatments. However, ozone had no significant effect on mycelial growth of S. sclerotiorum in vitro. Decay and quality parameters including color, chlorophyll fluorescence (CF), and ozone injury were further assessed for various horticultural commodities (apple, grape, highbush blueberry, and carrot) treated with 450 ppb of ozone for 48 h at 20 degrees C over a period of 12 d. Lesion size and height of the aerial mycelium were significantly reduced by the ozone treatment on carrots inoculated with mycelial agar plugs of B. cinerea or S. sclerotiorum. Lesion size was also reduced on treated apples inoculated with 5 x 10(6) spores/mL of B. cinerea, and decay incidence of treated grapes was reduced. The 450 ppb ozone for 48 h treatment had no significant effect on color of carrots and apples or on CF of apples and grapes. Ozone, an environmentally sound antimicrobial agent, inactivates microorganisms through oxidization and residual ozone spontaneously decomposes to nontoxic products. It may be applied to fruits and vegetables to reduce decay and extend shelf life.

Genetic characterization of resistance to Sclerotinia in lettuce cultivar Eruption

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Lettuce drop caused by the fungal pathogens Sclerotinia minor and S. sclerotiorum is a serious disease of lettuce. The use of genetic resistance as part of an integrated lettuce drop management strategy should have a significant economic advantage in mitigating yield loss. Sclerotinia resistance is ...

A new point mutation in the iron-sulfur subunit of succinate dehydrogenase confers resistance to boscalid in Sclerotinia sclerotiorum.

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Wang, Yong; Duan, Yabing; Wang, Jianxin; Zhou, Mingguo

2015-09-01

Research has established that mutations in highly conserved amino acids of the succinate dehydrogenase (SDH) complex in various fungi confer SDH inhibitor (SDHI) resistance. For Sclerotinia sclerotiorum (Lib.) de Bary, a necrotrophic fungus with a broad host range and a worldwide distribution, boscalid resistance has been attributed to the mutation H132R in the highly conserved SdhD subunit protein of the SDH complex. In our previous study, however, only one point mutation, A11V in SdhB (GCA to GTA change in SdhB), was detected in S. sclerotiorum boscalid-resistant (BR) mutants. In the current study, replacement of the SdhB gene in a boscalid-sensitive (BS) S. sclerotiorum strain with the mutant SdhB gene conferred resistance. Compared with wild-type strains, BR and GSM (SdhB gene in the wild-type strain replaced by the mutant SdhB gene) mutants were more sensitive to osmotic stress, lacked the ability to produce sclerotia and exhibited lower expression of the pac1 gene. Importantly, the point mutation was not located in the highly conserved sequence of the iron-sulfur subunit of SDH. These results suggest that resistance based on non-conserved vs. conserved protein domains differs in mechanism. In addition to increasing our understanding of boscalid resistance in S. sclerotiorum, the new information will be useful for the development of alternative antifungal drugs. © 2014 BSPP AND JOHN WILEY & SONS LTD.

Germinação carpogênica de Sclerotinia sclerotiorum sob diferentes resíduos e extratos de plantas cultivadas

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Francimar Perez Matheus da Silva

2011-09-01

Full Text Available Os efeitos de resíduos de plantas cultivadas e seus extratos sobre Sclerotinia sclerotiorum são poucos conhecidos. Três experimentos foram conduzidos, com resíduos de plantas cultivadas e seus extratos etanólicos e suas partições. Resíduos e extratos de culturas de aveia, ervilhaca, feijão, milheto, milho e trigo foram avaliados em condições controladas. Escleródios cobertos com resíduos de aveia, ervilhaca, feijão e milheto não germinaram carpogenicamente. Extratos etanólicos de resíduos de aveia e ervilhaca mostraram-se eficientes na inibição da germinação carpogênica, enquanto que do milheto e do trigo não diferiram da testemunha. Todas as partições de extratos etanólicos avaliadas reduziram a germinação carpogênica. Resíduos vegetais afetaram negativamente o número de apotécios emitidos por escleródio.

Oxalic acid has an additional, detoxifying function in Sclerotinia sclerotiorum pathogenesis.

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Annerose Heller

Full Text Available The mechanism of the diseases caused by the necrotroph plant pathogen Sclerotinia sclerotiorum is not well understood. To investigate the role of oxalic acid during infection high resolution, light-, scanning-, transmission electron microscopy and various histochemical staining methods were used. Our inoculation method allowed us to follow degradation of host plant tissue around single hyphae and to observe the reaction of host cells in direct contact with single invading hyphae. After penetration the outer epidermal cell wall matrix appeared degraded around subcuticular hyphae (12-24 hpi. Calcium oxalate crystals were detected in advanced (36-48 hpi and late (72 hpi infection stages, but not in early stages. In early infection stages, surprisingly, no toxic effect of oxalic acid eventually secreted by S. sclerotiorum was observed. As oxalic acid is a common metabolite in plants, we propose that attacked host cells are able to metabolize oxalic acid in the early infection stage and translocate it to their vacuoles where it is stored as calcium oxalate. The effects, observed on healthy tissue upon external application of oxalic acid to non-infected, living tissue and cell wall degradation of dead host cells starting at the inner side of the walls support this idea. The results indicate that oxalic acid concentrations in the early stage of infection stay below the toxic level. In plant and fungi oxalic acid/calcium oxalate plays an important role in calcium regulation. Oxalic acid likely could quench calcium ions released during cell wall breakdown to protect growing hyphae from toxic calcium concentrations in the infection area. As calcium antimonate-precipitates were found in vesicles of young hyphae, we propose that calcium is translocated to the older parts of hyphae and detoxified by building non-toxic, stable oxalate crystals. We propose an infection model where oxalic acid plays a detoxifying role in late infection stages.

Application of Hyperspectral Imaging to Detect Sclerotinia sclerotiorum on Oilseed Rape Stems

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Wenwen Kong

2018-01-01

Full Text Available Hyperspectral imaging covering the spectral range of 384–1034 nm combined with chemometric methods was used to detect Sclerotinia sclerotiorum (SS on oilseed rape stems by two sample sets (60 healthy and 60 infected stems for each set. Second derivative spectra and PCA loadings were used to select the optimal wavelengths. Discriminant models were built and compared to detect SS on oilseed rape stems, including partial least squares-discriminant analysis, radial basis function neural network, support vector machine and extreme learning machine. The discriminant models using full spectra and optimal wavelengths showed good performance with classification accuracies of over 80% for the calibration and prediction set. Comparing all developed models, the optimal classification accuracies of the calibration and prediction set were over 90%. The similarity of selected optimal wavelengths also indicated the feasibility of using hyperspectral imaging to detect SS on oilseed rape stems. The results indicated that hyperspectral imaging could be used as a fast, non-destructive and reliable technique to detect plant diseases on stems.

Emerging trends in molecular interactions between plants and the broad host range fungal pathogens Botrytis cinerea and Sclerotinia sclerotiorum

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Malick eMbengue

2016-03-01

Full Text Available Fungal plant pathogens are major threats to food security worldwide. Sclerotinia sclerotiorum and Botrytis cinerea are closely related Ascomycete plant pathogens causing mold diseases on hundreds of plant species. There is no genetic source of complete plant resistance to these broad host range pathogens known to date. Instead, natural plant populations show a continuum of resistance levels controlled by multiple genes, a phenotype designated as quantitative disease resistance. Little is known about the molecular mechanisms controlling the interaction between plants and S. sclerotiorum and B. cinerea but significant advances were made on this topic in the last years. This minireview highlights a selection of nine themes that emerged in recent research reports on the molecular bases of plant-S. sclerotiorum and plant-B. cinerea interactions. On the fungal side, this includes progress on understanding the role of oxalic acid, on the study of fungal small secreted proteins. Next, we discuss the exchanges of small RNA between organisms and the control of cell death in plant and fungi during pathogenic interactions. Finally on the plant side, we highlight defense priming by mechanical signals, the characterization of plant Receptor-like proteins and the hormone abscisic acid in the response to B. cinerea and S. sclerotiorum , the role of plant general transcription machinery and plant small bioactive peptides. These represent nine trends we selected as remarkable in our understanding of fungal molecules causing disease and plant mechanisms associated with disease resistance to two devastating broad host range fungi.

Genetic analysis of partial resistance to basal stem rot (Sclerotinia sclerotiorum in sunflower

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Amouzadeh Masoumeh

2013-01-01

Full Text Available Basal stem rot, caused by Sclerotinia sclerotiorum (Lib. de Bary, is one of the major diseases of sunflower (Helianthus annuus L. in the world. Quantitative trait loci (QTLs implicated in partial resistance to basal stem rot disease were identified using 99 recombinant inbred lines (RILs from the cross between sunflower parental lines PAC2 and RHA266. The study was undertaken in a completely randomized design with three replications under controlled conditions. The RILs and their parental lines were inoculated with a moderately aggressive isolate of S. sclerotiorum (SSKH41. Resistance to disease was evaluated by measuring the percentage of necrosis area three days after inoculation. QTLs were mapped using an updated high-density SSR and SNP linkage map. ANOVA showed significant differences among sunflower lines for resistance to basal stem rot (P≤0.05. The frequency distribution of lines for susceptibility to disease showed a continuous pattern. Composite interval mapping analysis revealed 5 QTLs for percentage of necrotic area, localized on linkage groups 1, 3, 8, 10 and 17. The sign of additive effect was positive in 5 QTLs, suggesting that the additive allele for partial resistance to basal stem rot came from the paternal line (RHA266. The phenotypic variance explained by QTLs (R2 ranged from 0.5 to 3.16%. Identified genes (HUCL02246_1, GST and POD, and SSR markers (ORS338, and SSL3 encompassing the QTLs for partial resistance to basal stem rot could be good candidates for marker assisted selection.

Control biológico de la pudrición basal del tallo en Crisantemo (Dendranthema grandiflorum ocasionada por Sclerotinia sclerotiorum con algunos aislamientos de Trichoderma sp. y Gliocladium sp.

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Valencia Jenny

1999-12-01

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EI hongo <em>Sclerotinia sclerotiorum em>ocasiona perdidas apreciables en diversos cultivos de flores y hortalizas. La investigación consistió en evaluar la capacidad antagónica de algunos aislamientos de <em>Trichoderma em>sp. Y <em>Gliocladium em>sp. Sobre <em>Sclerotinia sclerotiorum, em>tanto in vitro, como en plantas de crisantemo. Los mejores antagonistas del patógeno in vitro fueron los aislamientos G-8B y G-98 de <em>Gliocladium em>sp. y los aislamientos T-2IB y T-34B de <em>Trichoderma em>sp. Los dos aislamientos de <em>Gliocladium em>sp. Formaron un halo de inhibición alrededor de la colonia del patógeno, mientras que los aislamientos de <em>Trichoderma em>sp. Produjeron esporulación abundante sobre las colonias del patógeno. Los mejores tratamientos en el control de la enfermedad en plantas de crisantemo fueron los aislamientos T-2IB y T-34B de <em>Trichoderma em>sp. y el aislamiento G-8B de <em>Gliocladium em>sp., con un menor índice y una menor severidad de la enfermedad. La metodología de evaluación in vitro utilizada en la investigación fue confiable, ya que los resultados encontrados en el laboratorio fueron bastante similares a los obtenidos en el ensayo de invernadero. Además, los aislamientos de <em>Trichoderma em>y de <em>Gliocladium em>utilizados ocasionaron un estimulo apreciable en el crecimiento de las plantas y un adelanto en la floración.

Pre-treatment of soybean plants with calcium stimulates ROS responses and mitigates infection by Sclerotinia sclerotiorum.

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Arfaoui, Arbia; El Hadrami, Abdelbasset; Daayf, Fouad

2018-01-01

Considering the high incidence of white mold caused by Sclerotinia sclerotiorum in a variety of field crops and vegetables, different control strategies are needed to keep the disease under economical threshold. This study assessed the effect of foliar application of a calcium formulation on disease symptoms, oxalic acid production, and on the oxidative stress metabolism in soybean plants inoculated with each of two isolates of the pathogen that have contrasting aggressiveness (HA, highly-aggressive versus WA, weakly-aggressive). Changes in reactive oxygen species (ROS) levels in soybean plants inoculated with S. sclerotiorum isolates were assessed at 6, 24, 48 and 72 h post inoculation (hpi). Generation of ROS including hydrogen peroxide (H 2 O 2 ), anion superoxide (O 2 - ) and hydroxyl radical (OH) was evaluated. Inoculation with the WA isolate resulted in more ROS accumulation compared to the HA isolate. Pre-treatment with the calcium formulation restored ROS production in plants inoculated with the HA isolate. We also noted a marked decrease in oxalic acid content in the leaves inoculated with the HA isolate in presence of calcium, which coincided with an increase in plant ROS production. The expression patterns of genes involved in ROS detoxification in response to the calcium treatments and/or inoculation with S. Sclerotiorum isolates were monitored by RT-qPCR. All of the tested genes showed a higher expression in response to inoculation with the WA isolate. The expression of most genes tested peaked at 6 hpi, which preceded ROS accumulation in the soybean leaves. Overall, these data suggest that foliar application of calcium contributes to a decrease in oxalic acid production and disease, arguably via modulation of the ROS metabolism. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

Overexpression of Three Glucosinolate Biosynthesis Genes in Brassica napus Identifies Enhanced Resistance to Sclerotinia sclerotiorum and Botrytis cinerea.

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Yuanyuan Zhang

Full Text Available Sclerotinia sclerotiorum and Botrytis cinerea are notorious plant pathogenic fungi with an extensive host range including Brassica crops. Glucosinolates (GSLs are an important group of secondary metabolites characteristic of the Brassicales order, whose degradation products are proving to be increasingly important in plant protection. Enhancing the defense effect of GSL and their associated degradation products is an attractive strategy to strengthen the resistance of plants by transgenic approaches. We generated the lines of Brassica napus with three biosynthesis genes involved in GSL metabolic pathway (BnMAM1, BnCYP83A1 and BnUGT74B1, respectively. We then measured the foliar GSLs of each transgenic lines and inoculated them with S. sclerotiorum and B. cinerea. Compared with the wild type control, over-expressing BnUGT74B1 in B. napus increased the aliphatic and indolic GSL levels by 1.7 and 1.5 folds in leaves respectively; while over-expressing BnMAM1 or BnCYP83A1 resulted in an approximate 1.5-fold higher only in the aliphatic GSL level in leaves. The results of plant inoculation demonstrated that BnUGT74B1-overexpressing lines showed less severe disease symptoms and tissue damage compared with the wild type control, but BnMAM1 or BnCYP83A1-overexpressing lines showed no significant difference in comparison to the controls. These results suggest that the resistance to S. sclerotiorum and B. cinerea in B. napus could be enhanced through tailoring the GSL profiles by transgenic approaches or molecular breeding, which provides useful information to assist plant breeders to design improved breeding strategies.

Tipping the balance: Sclerotinia sclerotiorum secreted oxalic acid suppresses host defenses by manipulating the host redox environment.

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Brett Williams

2011-06-01

Full Text Available Sclerotinia sclerotiorum is a necrotrophic ascomycete fungus with an extremely broad host range. This pathogen produces the non-specific phytotoxin and key pathogenicity factor, oxalic acid (OA. Our recent work indicated that this fungus and more specifically OA, can induce apoptotic-like programmed cell death (PCD in plant hosts, this induction of PCD and disease requires generation of reactive oxygen species (ROS in the host, a process triggered by fungal secreted OA. Conversely, during the initial stages of infection, OA also dampens the plant oxidative burst, an early host response generally associated with plant defense. This scenario presents a challenge regarding the mechanistic details of OA function; as OA both suppresses and induces host ROS during the compatible interaction. In the present study we generated transgenic plants expressing a redox-regulated GFP reporter. Results show that initially, Sclerotinia (via OA generates a reducing environment in host cells that suppress host defense responses including the oxidative burst and callose deposition, akin to compatible biotrophic pathogens. Once infection is established however, this necrotroph induces the generation of plant ROS leading to PCD of host tissue, the result of which is of direct benefit to the pathogen. In contrast, a non-pathogenic OA-deficient mutant failed to alter host redox status. The mutant produced hypersensitive response-like features following host inoculation, including ROS induction, callose formation, restricted growth and cell death. These results indicate active recognition of the mutant and further point to suppression of defenses by the wild type necrotrophic fungus. Chemical reduction of host cells with dithiothreitol (DTT or potassium oxalate (KOA restored the ability of this mutant to cause disease. Thus, Sclerotinia uses a novel strategy involving regulation of host redox status to establish infection. These results address a long-standing issue

Comparative proteomic analysis in pea treated with microbial consortia of beneficial microbes reveals changes in the protein network to enhance resistance against Sclerotinia sclerotiorum.

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Jain, Akansha; Singh, Akanksha; Singh, Surendra; Singh, Vinay; Singh, Harikesh Bahadur

2015-06-15

Microbial consortia may provide protection against pathogenic ingress via enhancing plant defense responses. Pseudomonas aeruginosa PJHU15, Trichoderma harzianum TNHU27 and Bacillus subtilis BHHU100 were used either singly or in consortia in the pea rhizosphere to observe proteome level changes upon Sclerotinia sclerotiorum challenge. Thirty proteins were found to increase or decrease differentially in 2-DE gels of pea leaves, out of which 25 were identified by MALDI-TOF MS or MS/MS. These proteins were classified into several functional categories including photosynthesis, respiration, phenylpropanoid metabolism, protein synthesis, stress regulation, carbohydrate and nitrogen metabolism and disease/defense-related processes. The respective homologue of each protein identified was trapped in Pisum sativum and a phylogenetic tree was constructed to check the ancestry. The proteomic view of the defense response to S. sclerotiorum in pea, in the presence of beneficial microbes, highlights the enhanced protection that can be provided by these microbes in challenged plants. Copyright © 2015 Elsevier GmbH. All rights reserved.

Components of a Rice-Oilseed Rape Production System Augmented with Trichoderma sp. Tri-1 Control Sclerotinia sclerotiorum on Oilseed Rape.

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Hu, Xiaojia; Roberts, Daniel P; Xie, Lihua; Maul, Jude E; Yu, Changbing; Li, Yinshui; Zhang, Yinbo; Qin, Lu; Liao, Xing

2015-10-01

Sclerotinia sclerotiorum causes serious yield losses on many crops throughout the world. A multicomponent treatment that consisted of the residual rice straw remaining after rice harvest and Trichoderma sp. Tri-1 (Tri-1) formulated with the oilseed rape seedcake fertilizer was used in field soil infested with S. sclerotiorum. This treatment resulted in oilseed rape seed yield that was significantly greater than the nontreated control or when the fungicide carbendizem was used in the presence of this pathogen in field trials. Yield data suggested that the rice straw, oilseed rape seedcake, and Tri-1 components of this treatment all contributed incrementally. Similar treatment results were obtained regarding reduction in disease incidence. Slight improvements in yield and disease incidence were detected when this multicomponent treatment was combined with a fungicide spray. Inhibition of sclerotial germination by this multicomponent treatment trended greater than the nontreated control at 90, 120, and 150 days in field studies but was not significantly different from this control. This multicomponent treatment resulted in increased yield relative to the nontreated control in the absence of pathogen in a greenhouse pot study, while the straw alone and the straw plus oilseed rape seedcake treatments did not; suggesting that Tri-1 was capable of promoting growth. Experiments reported here indicate that a treatment containing components of a rice-oilseed rape production system augmented with Tri-1 can control S. sclerotiorum on oilseed rape, be used in integrated strategies containing fungicide sprays for control of this pathogen, and promote plant growth.

Plant hormones in defense response of Brassica napus to Sclerotinia sclerotiorum - reassessing the role of salicylic acid in the interaction with a necrotroph.

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Nováková, Miroslava; Sašek, Vladimír; Dobrev, Petre I; Valentová, Olga; Burketová, Lenka

2014-07-01

According to general model, jasmonic acid (JA) and ethylene (ET) signaling pathways are induced in Arabidopsis after an attack of necrotroph, Sclerotinia sclerotiorum (Lib.) de Bary. However, abscisic acid (ABA) and salicylic acid (SA) also seem to play a role. While signaling events in Arabidopsis have been intensively studied recently, information for the natural host Brassica napus is limited. In this study, multiple plant hormone quantification and expression analysis of marker genes of the signaling pathways was used to gain a complete view of the interaction of B. napus with S. sclerotiorum. Strong response of ET biosynthetic gene ACS2 was observed, accompanied by increases of SA and JA levels that correspond to the elevated expression of marker genes PR1 and LOX3. Interestingly, the level of ABA and the expression of its marker gene RD26 were also elevated. Furthermore, induction of the SA-dependent defense decreased disease symptoms. In addition, SA signaling is suggested as a possible target for manipulation by S. sclerotiorum. A gene for putative chorismate mutase SS1G_14320 was identified that is highly expressed during infection but not in vitro. Our results bring the evidence of SA involvement in the interaction of plant with the necrotroph that conflict with the current model. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

A Model for Sclerotinia sclerotiorum Infection and Disease Development in Lettuce, Based on the Effects of Temperature, Relative Humidity and Ascospore Density

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Clarkson, John P.; Fawcett, Laura; Anthony, Steven G.; Young, Caroline

2014-01-01

The plant pathogen Sclerotinia sclerotiorum can cause serious losses on lettuce crops worldwide and as for most other susceptible crops, control relies on the application of fungicides, which target airborne ascospores. However, the efficacy of this approach depends on accurate timing of these sprays, which could be improved by an understanding of the environmental conditions that are conducive to infection. A mathematical model for S. sclerotiorum infection and disease development on lettuce is presented here for the first time, based on quantifying the effects of temperature, relative humidity (RH) and ascospore density in multiple controlled environment experiments. It was observed that disease can develop on lettuce plants inoculated with dry ascospores in the absence of apparent leaf wetness (required for spore germination). To explain this, the model conceptualises an infection court area containing microsites (in leaf axils and close to the stem base) where conditions are conducive to infection, the size of which is modified by ambient RH. The model indicated that minimum, maximum and optimum temperatures for ascospore germination were 0.0, 29.9 and 21.7°C respectively and that maximum rates of disease development occurred at spore densities >87 spores cm−2. Disease development was much more rapid at 80–100% RH at 20°C, compared to 50–70% RH and resulted in a greater proportion of lettuce plants infected. Disease development was also more rapid at 15–27°C compared to 5–10°C (85% RH). The model was validated by a further series of independent controlled environment experiments where both RH and temperature were varied and generally simulated the pattern of disease development well. The implications of the results in terms of Sclerotinia disease forecasting are discussed. PMID:24736409

Caracterización de los mecanismos de defensa a sclerotinia sclerotiorum, agente causal de la podredumbre del estudio de perfiles metabólicos y transcripcionales

OpenAIRE

Peluffo, Lucila

2010-01-01

En Argentina, la podredumbre húmeda del capítulo, causada por el patógeno necrotrófico Sclerotinia sclerotiorum, es una enfermedad que provoca serias mermas en la producción y tiene una incidencia anual promedio sobre la producción de la pampa húmeda del 10-20%. Los síntomas de la enfermedad se manifiestan al final de la etapa de floración o durante el llenado de granos por lesiones en el receptáculo que pueden extenderse y afectar todo el capítulo, produciendo la caída del mismo. En estados ...

Reaction of common bean lines and aggressiveness of Sclerotinia sclerotiorum isolates.

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Silva, P H; Santos, J B; Lima, I A; Lara, L A C; Alves, F C

2014-11-07

The aims of this study were to evaluate the reaction of common bean lines to white mold, the aggressiveness of different Sclerotinia sclerotiorum isolates from various common bean production areas in Brazil, and comparison of the diallel and GGE (genotype main effect plus genotype-by-environment interaction) biplot analysis procedures via study of the line-by-isolate interaction. Eleven common bean (Phaseolus vulgaris) lines derived from 3 backcross populations were used. Field experiments were performed in the experimental area of the Departamento de Biologia of the Universidade Federal de Lavras, Lavras, MG, Brazil, in the 2011 and 2012 dry crop season and 2011 winter crop season through a randomized block design with 3 replications. This study was also set up in a greenhouse. Inoculations were performed 28 days after sowing by means of the straw test method. The reaction of the bean lines to white mold was assessed according to a diagrammatic scale from 1 (plant without symptoms) to 9 (dead plant). Estimations of general reaction capacity (lines) and general aggressiveness capacity (isolates) indicated different horizontal levels of resistance in the lines and levels of aggressiveness in the isolates. Therefore, it was possible to select more resistant lines and foresee those crosses that are the most promising for increasing the level of resistance. It was also possible to identify the most aggressive isolates that were more efficient in distinguishing the lines. Both diallel and GGE biplot analyses were useful in identifying the genotypic values of lines and isolates.

Interspecific Transmission of Double-Stranded RNA and Hypovirulence from Sclerotinia sclerotiorum to S. minor.

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Melzer, M S; Ikeda, S S; Boland, G J

2002-07-01

ABSTRACT Interspecific transmission of a hypovirulence-associated double-stranded RNA (dsRNA) and hypovirulent phenotype was attempted from hypovirulent isolate Ss275 of Sclerotinia sclerotiorum to five virulent isolates of S. minor. dsRNA and the hypovirulent phenotype were successfully transmitted to one of the five isolates, Sm10. Three putative converted isolates of Sm10 were slow growing and developed atypical colony morphologies characteristic of the hypovirulent phenotype. These isolates were assayed for virulence and produced significantly smaller lesions than isolate Sm10 on detached leaves of Romaine lettuce. One of these putative converted isolates, designated Sm10T, was tested to confirm interspecific transmission of dsRNA. In northern hybridizations, dsRNA isolated from Sm10T hybridized with a digoxigenin-labeled cDNA probe prepared from dsRNA isolated from Ss275. Random amplified polymorphic DNA analysis confirmed that isolate Sm10T was derived from Sm10 and not from Ss275 or a hybrid of the two species. The dsRNA and hypovirulent phenotype were subsequently transmitted intraspecifically from Sm10T to Sm8. To our knowledge, this is the first report of interspecific transmission of dsRNA and an associated hypovirulent phenotype between fungal plant pathogens by hyphal anastomosis.

Genotyping-by-sequencing uncovers the introgression alien segments associated with Sclerotinia basal stalk rot resistance from wild species—I. Helianthus argophyllus and H. petiolaris

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Basal stalk rot (BSR), caused by Sclerotinia Sclerotiorum, is a devastating disease in sunflower worldwide. The progress of breeding for Sclerotinia BSR resistance has been hampered due to the lack of effective sources of resistance for cultivated sunflower. Our objective was to transfer BSR resista...

Mycelial compatibility and aggressiveness of Sclerotinia sclerotiorum isolates from Brazil and the United States

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Lucimara Junko Koga

2014-04-01

Full Text Available The objective of this work was to evaluate the genetic diversity among Sclerotinia sclerotiorum isolates from Brazil and the USA, assess their aggressiveness variability, and verify the existence of an isolate-cultivar interaction. Isolate variability was determined by mycelial compatibility grouping (MCG, and isolate aggressiveness by cut-stem inoculations of soybean cultivars. Two experiments for MCGs and two for aggressiveness were conducted with two sets of isolates. The first set included nine isolates from the same soybean field in Brazil and nine from the Midwest region of the USA. The second set included 16 isolates from several regions of Brazil and one from the USA. In the first set, 18 isolates formed 12 different MCGs. In the second set, 81% of the isolates from Brazil grouped into a single MCG. No common MCGs were observed among isolates from Brazil and the USA. The isolates showed aggressiveness differences in the first set, but not in the second. Although aggressiveness differed in the first set, soybean cultivars and isolates did not interact significantly. Cultivar rank remained the same, regardless of the genetic diversity, aggressiveness difference, and region or country of origin of the isolate. Results from screening of soybean cultivars, performed by the cut-stem method in the USA, can be used as reference for researchers in Brazil.

Análise dos níveis de poligalacturonases e glucanases expressas durante os processos de interação patogênica e saprofítica de Sclerotinia sclerotiorum

OpenAIRE

BARBOSA, Silvio Romero Costa

2008-01-01

O fungo Sclerotinia sclerotiorum, pode interagir com uma grande gama de espécies vegetais bem como obedecer à alta especificidade e especialização patogênica. Ele é capaz de digerir a parede celular de plantas hospedeiras utilizando para tal uma série de mecanismos bioquímicos e morfogenéticos que otimizam a invasão. Várias enzimas são produzidas durante a interação planta-hospedeiro e dentre elas se destacam a família das poligalacturonases (PGs) e as beta glucanases. As PGs catalisam a hidr...

Selection of endophytic fungi from comfrey (Symphytum officinale L. for in vitro biological control of the phytopathogen Sclerotinia sclerotiorum (Lib. Seleção de fungos endofíticos de confrei (Symphytum officinale L., buscando controle biológico in vitro do fitopatógeno Sclerotinia sclerotiorum (Lib.

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Rafaeli Rocha

2009-03-01

Full Text Available Biological control consists of using one organism to attack another that may cause economic damage to crops. Integrated Pest Management (IPM is a very common strategy. The white mold produced by Sclerotiniasclerotiorum (Lib. causes considerable damage to bean crops. This fungus is a soil inhabitant, the symptoms of which are characterized by water-soaked lesions covered by a white cottony fungal growth on the soil surface and/or the host plant. Possible biological control agents taken from plants are being investigated as phytopathogen inhibitors. These are endophytic microorganisms that inhabit the intercellular spaces of vegetal tissues and are often responsible for antimicrobial production. The objective of the present study was to select endophytic fungi isolated from comfrey (Symphytumofficinale L. leaves with in vitro antagonist potential against the phytopathogenic fungus S. sclerotiorum. Twelve isolates of endophytic fungi and a pathogenic strain of S. sclerotiorum were used in the challenge method. With the aid of this method, four endophytes with the best antagonistic activity against S. sclerotiorum were selected. Pathogen growth inhibition zones were considered indicative of antibiosis. The percentages of pathogenic mycelia growth were measured both with and without the antagonist, resulting in growth reductions of 46.7% to 50.0% for S. sclerotiorum. These analyses were performed by evaluating the endophytic/pathogenic mycelia growth in mm/day over an eight-day period of antagonistic tests.O controle biológico consiste no uso de organismos que atacam outros que causam danos a culturas de plantas. Esta é uma estratégia muito utilizada no Controle Integrado de Pragas (CIP. O mofo branco, causado por Sclerotiniasclerotiorum (Lib., causa danos em culturas de feijão. Este fungo é encontrado no solo e seus sintomas são caracterizados por lesões úmidas cobertas por micélios algodonosos, crescidos a partir do solo e/ou da planta

Characterization of mechanisms underlying degradation of sclerotia of Sclerotinia sclerotiorum by Aspergillus aculeatus Asp-4 using a combined qRT-PCR and proteomic approach.

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Hu, Xiaojia; Qin, Lu; Roberts, Daniel P; Lakshman, Dilip K; Gong, Yangmin; Maul, Jude E; Xie, Lihua; Yu, Changbing; Li, Yinshui; Hu, Lei; Liao, Xiangsheng; Liao, Xing

2017-08-31

The biological control agent Aspergillus aculeatus Asp-4 colonizes and degrades sclerotia of Sclerotinia sclerotiorum resulting in reduced germination and disease caused by this important plant pathogen. Molecular mechanisms of mycoparasites underlying colonization, degradation, and reduction of germination of sclerotia of this and other important plant pathogens remain poorly understood. An RNA-Seq screen of Asp-4 growing on autoclaved, ground sclerotia of S. sclerotiorum for 48 h identified 997 up-regulated and 777 down-regulated genes relative to this mycoparasite growing on potato dextrose agar (PDA) for 48 h. qRT-PCR time course experiments characterized expression dynamics of select genes encoding enzymes functioning in degradation of sclerotial components and management of environmental conditions, including environmental stress. This analysis suggested co-temporal up-regulation of genes functioning in these two processes. Proteomic analysis of Asp-4 growing on this sclerotial material for 48 h identified 26 up-regulated and 6 down-regulated proteins relative to the PDA control. Certain proteins with increased abundance had putative functions in degradation of polymeric components of sclerotia and the mitigation of environmental stress. Our results suggest co-temporal up-regulation of genes involved in degradation of sclerotial compounds and mitigation of environmental stress. This study furthers the analysis of mycoparasitism of sclerotial pathogens by providing the basis for molecular characterization of a previously uncharacterized mycoparasite-sclerotial interaction.

Detecting the Hormonal Pathways in Oilseed Rape behind Induced Systemic Resistance by Trichoderma harzianum TH12 to Sclerotinia sclerotiorum.

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Alkooranee, Jawadayn Talib; Aledan, Tamarah Raad; Ali, Ali Kadhim; Lu, Guangyuan; Zhang, Xuekun; Wu, Jiangsheng; Fu, Chunhua; Li, Maoteng

2017-01-01

Plants have the ability to resist pathogen attack after infection or treatment with biotic and abiotic elicitors. In oilseed rape plant Brassica napus AACC and in the artificially synthesized Raphanus alboglabra RRCC, the root-colonizing Trichoderma harzianum TH12 fungus triggers induced systemic resistance (ISR), and its culture filtrate (CF) triggers a systemic acquired resistance (SAR) response against infection by the Sclerotinia sclerotiorum. Salicylic acid (SA) and jasmonic acid/ethylene (JA/ET) are plant hormone signals that play important roles in the regulation of ISR and SAR. In this study, at six different time points (1, 2, 4, 6, 8 and 10 days post-infection [dpi]), six resistance genes were used as markers of signaling pathways: JA/ET signaling used AOC3, PDF1.2 and ERF2 genes, while PR-1, TGA5 and TGA6 genes were used as markers of SA signaling. The results of quantitative real-time polymerase chain reaction (qRT-PCR) showed that AOC3, PDF1.2 and ERF2 expression levels in infected leaves of AACC and RRCC increase at 1 and 2 dpi with S. sclerotiorum or inoculation with TH12. PR-1, TGA5 and TGA6 expression levels increased at 8 and 10 dpi in infected leaves. PR-1, TGA5 and TGA6 expression levels increased early in plants treated with CF in both of the healthy genotypes. Furthermore, induction of SA- and JA/ET-dependent defense decreased disease symptoms in infected leaves at different times. The results suggest that the RRCC genotype exhibits resistance to disease and that the ability of TH12 and its CF to induce systemic resistance in susceptible and resistant oilseed rape genotypes exists. In addition, the results indicate for the first time that in RRCC the SA signaling pathway is involved in resistance to necrotrophic pathogens.

Candidate gene association mapping of Sclerotinia stalk rot resistance in sunflower (Helianthus annuus L.) uncovers the importance of COI1 homologs.

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Talukder, Zahirul I; Hulke, Brent S; Qi, Lili; Scheffler, Brian E; Pegadaraju, Venkatramana; McPhee, Kevin; Gulya, Thomas J

2014-01-01

Functional markers for Sclerotinia basal stalk rot resistance in sunflower were obtained using gene-level information from the model species Arabidopsis thaliana. Sclerotinia stalk rot, caused by Sclerotinia sclerotiorum, is one of the most destructive diseases of sunflower (Helianthus annuus L.) worldwide. Markers for genes controlling resistance to S. sclerotiorum will enable efficient marker-assisted selection (MAS). We sequenced eight candidate genes homologous to Arabidopsis thaliana defense genes known to be associated with Sclerotinia disease resistance in a sunflower association mapping population evaluated for Sclerotinia stalk rot resistance. The total candidate gene sequence regions covered a concatenated length of 3,791 bp per individual. A total of 187 polymorphic sites were detected for all candidate gene sequences, 149 of which were single nucleotide polymorphisms (SNPs) and 38 were insertions/deletions. Eight SNPs in the coding regions led to changes in amino acid codons. Linkage disequilibrium decay throughout the candidate gene regions declined on average to an r (2) = 0.2 for genetic intervals of 120 bp, but extended up to 350 bp with r (2) = 0.1. A general linear model with modification to account for population structure was found the best fitting model for this population and was used for association mapping. Both HaCOI1-1 and HaCOI1-2 were found to be strongly associated with Sclerotinia stalk rot resistance and explained 7.4 % of phenotypic variation in this population. These SNP markers associated with Sclerotinia stalk rot resistance can potentially be applied to the selection of favorable genotypes, which will significantly improve the efficiency of MAS during the development of stalk rot resistant cultivars.

Registration of an oilseed sunflower germplasm line HA-BSR1 highly tolerant to Sclerotinia basal stalk rot

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Basal stalk rot (BSR) caused by Sclerotinia sclerotiorum (Lib.) de Bary is a devastating disease that causes a significant damage to worldwide sunflower (Helianthus annuus L.) production by reducing seed yield and quality. The objective of this research was to develop highly BSR tolerant sunflower g...

Association mapping in sunflower for sclerotinia head rot resistance

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Fusari Corina M

2012-06-01

Full Text Available Abstract Background Sclerotinia Head Rot (SHR is one of the most damaging diseases of sunflower in Europe, Argentina, and USA, causing average yield reductions of 10 to 20 %, but leading to total production loss under favorable environmental conditions for the pathogen. Association Mapping (AM is a promising choice for Quantitative Trait Locus (QTL mapping, as it detects relationships between phenotypic variation and gene polymorphisms in existing germplasm without development of mapping populations. This article reports the identification of QTL for resistance to SHR based on candidate gene AM. Results A collection of 94 sunflower inbred lines were tested for SHR under field conditions using assisted inoculation with the fungal pathogen Sclerotinia sclerotiorum. Given that no biological mechanisms or biochemical pathways have been clearly identified for SHR, 43 candidate genes were selected based on previous transcript profiling studies in sunflower and Brassica napus infected with S. sclerotiorum. Associations among SHR incidence and haplotype polymorphisms in 16 candidate genes were tested using Mixed Linear Models (MLM that account for population structure and kinship relationships. This approach allowed detection of a significant association between the candidate gene HaRIC_B and SHR incidence (P  Conclusions These results suggest that AM will be useful in dissecting other complex traits in sunflower, thus providing a valuable tool to assist in crop breeding.

Chemical and biological control of Sclerotinia stem rot in the soybean crop

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Ciro Hideki Sumida

2015-05-01

Full Text Available It was evaluated the effect of fungicides and the microbial control agent Trichoderma harzianum on the inhibition of the carpogenic and ascospore germination of Sclerotinia sclerotiorum. This study also evaluated the chemical, fungicidal and microbial control of white mold or Sclerotinia stem rot of soybean in the field. Three experiments were conducted, as follows: 1 inhibition of carpogenic germination of sclerotia, 2 inhibition of ascospore germination, and 3 control of Sclerotinia stem rot in a soybean crop under field conditions. The treatments evaluated were fluazinam, procymidone, iprodione, thiophanate-methyl, carbendazim, benzalkonium chloride + fluazinam, and T. harzianum. Procymidone resulted in an inhibition of 13.5% and benzalkonium chloride in an inhibition of 13.9% in an ascospore germination test. Fluazinam and procymidone were the most effective in reducing the production of ascospores/apothecium, representing 65.6% and 82.4% of inhibition. Procymidone and fluazinam if combined or not with benzalkonium chloride were the most effective in controlling sclerotinia stem rot under field conditions when applied at the onset of flowering and 15 days later. In the 2009-10 harvest, these two fungicides reduced the incidence of Sclerotinia stem rot by 73.1 and 71.6% and in the 2010-11 harvest by 75.7 and 77.6%, respectively.

Use of formulated Trichoderma sp. Tri-1 in combination with reduced rates of chemical pesticide for control of Sclerotinia sclerotiorium on oilseed rape

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Sustainable strategies for control of Sclerotinia sclerotiorum on oilseed rape are needed. Here we tested combinations of Trichoderma sp. Tri-1, formulated with oilseed rape seedcake and straw, with reduced application rates of the chemical pesticide Carbendazim for control of this pathogen on oils...

Development of a novel Sinapis arvensis disomic addition line in Brassica napus containing the restorer gene for Nsa CMS and improved resistance to Sclerotinia sclerotiorum and pod shattering.

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Wei, Wenhui; Li, Yunchang; Wang, Lijun; Liu, Shengyi; Yan, Xiaohong; Mei, Desheng; Li, Yinde; Xu, Yusong; Peng, Pengfei; Hu, Qiong

2010-04-01

An allo-cytoplasmic male sterile line, which was developed through somatic hybridization between Brassica napus and Sinapis arvensis (thus designated as Nsa CMS line), possesses high potential for hybrid production of rapeseed. In order to select for restorer lines, fertile plants derived from the same somatic hybridization combination were self-pollinated and testcrossed with the parental Nsa CMS line for six generations. A novel disomic alien addition line, B. napus-S. arvensis, has been successfully developed. GISH analysis showed that it contains one pair of chromosomes from S. arvensis and 19 pairs from B. napus, and retains stable and regular mitotic and meiotic processes. The addition line displays very strong restoration ability to Nsa CMS line, high resistance to Sclerotinia sclerotiorum and a low incidence of pod shattering. Because the addition line shares these very important agricultural characters, it is a valuable restorer to Nsa CMS line, and is named NR1 here (Nsa restorer no. 1).

Interference and Mechanism of Dill Seed Essential Oil and Contribution of Carvone and Limonene in Preventing Sclerotinia Rot of Rapeseed

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Huang, Bo; He, Jingsheng; Tian, Jun; Zeng, Hong; Chen, Yuxin; Wang, Youwei

2015-01-01

This study aimed to evaluate the inhibitory effects of dill (Anethum graveolens L.) seed essential oil against Sclerotinia sclerotiorum and its mechanism of action. The antifungal activities of the two main constituents, namely carvone and limonene, were also measured. Mycelial growth and sclerotial germination were thoroughly inhibited by dill seed essential oil at the 1.00 μL/mL under contact condition and 0.125μL/mL air under vapor condition. Carvone also contributed more than limonene in inhibiting the growth of S. sclerotiorum. Carvone and limonene synergistically inhibited the growth of the fungus. In vivo experiments, the essential oil remarkably suppressed S. sclerotiorum, and considerable morphological alterations were observed in the hyphae and sclerotia. Inhibition of ergosterol synthesis, malate dehydrogenase, succinate dehydrogenase activities, and external medium acidification were investigated to elucidate the antifungal mechanism of the essential oil. The seed essential oil of A. graveolens can be extensively used in agriculture for preventing the oilseed crops fungal disease. PMID:26133771

Interference and Mechanism of Dill Seed Essential Oil and Contribution of Carvone and Limonene in Preventing Sclerotinia Rot of Rapeseed.

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Bingxin Ma

Full Text Available This study aimed to evaluate the inhibitory effects of dill (Anethum graveolens L. seed essential oil against Sclerotinia sclerotiorum and its mechanism of action. The antifungal activities of the two main constituents, namely carvone and limonene, were also measured. Mycelial growth and sclerotial germination were thoroughly inhibited by dill seed essential oil at the 1.00 μL/mL under contact condition and 0.125μL/mL air under vapor condition. Carvone also contributed more than limonene in inhibiting the growth of S. sclerotiorum. Carvone and limonene synergistically inhibited the growth of the fungus. In vivo experiments, the essential oil remarkably suppressed S. sclerotiorum, and considerable morphological alterations were observed in the hyphae and sclerotia. Inhibition of ergosterol synthesis, malate dehydrogenase, succinate dehydrogenase activities, and external medium acidification were investigated to elucidate the antifungal mechanism of the essential oil. The seed essential oil of A. graveolens can be extensively used in agriculture for preventing the oilseed crops fungal disease.

A cerato-platanin protein SsCP1 targets plant PR1 and contributes to virulence of Sclerotinia sclerotiorum

DEFF Research Database (Denmark)

Yang, Guogen; Tang, Liguang; Gong, Yingdi

2018-01-01

in the apoplast to facilitate infection by S. sclerotiorum. Overexpressing PR1 enhanced resistance to the wild-type strain, but not to the Sscp1 knockout strain of S. sclerotiorum. Sscp1-expressing transgenic plants showed increased concentrations of salicylic acid (SA) and higher levels of resistance to several...

Effects of Sublethal Fungicides on Mutation Rates and Genomic Variation in Fungal Plant Pathogen, Sclerotinia sclerotiorum.

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Amaradasa, B Sajeewa; Everhart, Sydney E

2016-01-01

Pathogen exposure to sublethal doses of fungicides may result in mutations that may represent an important and largely overlooked mechanism of introducing new genetic variation into strictly clonal populations, including acquisition of fungicide resistance. We tested this hypothesis using the clonal plant pathogen, Sclerotinia sclerotiorum. Nine susceptible isolates were exposed independently to five commercial fungicides with different modes of action: boscalid (respiration inhibitor), iprodione (unclear mode of action), thiophanate methyl (inhibition of microtubulin synthesis) and azoxystrobin and pyraclostrobin (quinone outside inhibitors). Mycelium of each isolate was inoculated onto a fungicide gradient and sub-cultured from the 50-100% inhibition zone for 12 generations and experiment repeated. Mutational changes were assessed for all isolates at six neutral microsatellite (SSR) loci and for a subset of isolates using amplified fragment length polymorphisms (AFLPs). SSR analysis showed 12 of 85 fungicide-exposed isolates had a total of 127 stepwise mutations with 42 insertions and 85 deletions. Most stepwise deletions were in iprodione- and azoxystrobin-exposed isolates (n = 40/85 each). Estimated mutation rates were 1.7 to 60-fold higher for mutated loci compared to that expected under neutral conditions. AFLP genotyping of 33 isolates (16 non-exposed control and 17 fungicide exposed) generated 602 polymorphic alleles. Cluster analysis with principal coordinate analysis (PCoA) and discriminant analysis of principal components (DAPC) identified fungicide-exposed isolates as a distinct group from non-exposed control isolates (PhiPT = 0.15, P = 0.001). Dendrograms based on neighbor-joining also supported allelic variation associated with fungicide-exposure. Fungicide sensitivity of isolates measured throughout both experiments did not show consistent trends. For example, eight isolates exposed to boscalid had higher EC50 values at the end of the experiment, and

Effects of Sublethal Fungicides on Mutation Rates and Genomic Variation in Fungal Plant Pathogen, Sclerotinia sclerotiorum.

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B Sajeewa Amaradasa

Full Text Available Pathogen exposure to sublethal doses of fungicides may result in mutations that may represent an important and largely overlooked mechanism of introducing new genetic variation into strictly clonal populations, including acquisition of fungicide resistance. We tested this hypothesis using the clonal plant pathogen, Sclerotinia sclerotiorum. Nine susceptible isolates were exposed independently to five commercial fungicides with different modes of action: boscalid (respiration inhibitor, iprodione (unclear mode of action, thiophanate methyl (inhibition of microtubulin synthesis and azoxystrobin and pyraclostrobin (quinone outside inhibitors. Mycelium of each isolate was inoculated onto a fungicide gradient and sub-cultured from the 50-100% inhibition zone for 12 generations and experiment repeated. Mutational changes were assessed for all isolates at six neutral microsatellite (SSR loci and for a subset of isolates using amplified fragment length polymorphisms (AFLPs. SSR analysis showed 12 of 85 fungicide-exposed isolates had a total of 127 stepwise mutations with 42 insertions and 85 deletions. Most stepwise deletions were in iprodione- and azoxystrobin-exposed isolates (n = 40/85 each. Estimated mutation rates were 1.7 to 60-fold higher for mutated loci compared to that expected under neutral conditions. AFLP genotyping of 33 isolates (16 non-exposed control and 17 fungicide exposed generated 602 polymorphic alleles. Cluster analysis with principal coordinate analysis (PCoA and discriminant analysis of principal components (DAPC identified fungicide-exposed isolates as a distinct group from non-exposed control isolates (PhiPT = 0.15, P = 0.001. Dendrograms based on neighbor-joining also supported allelic variation associated with fungicide-exposure. Fungicide sensitivity of isolates measured throughout both experiments did not show consistent trends. For example, eight isolates exposed to boscalid had higher EC50 values at the end of the

Effects of Sublethal Fungicides on Mutation Rates and Genomic Variation in Fungal Plant Pathogen, Sclerotinia sclerotiorum

Science.gov (United States)

Amaradasa, B. Sajeewa

2016-01-01

Pathogen exposure to sublethal doses of fungicides may result in mutations that may represent an important and largely overlooked mechanism of introducing new genetic variation into strictly clonal populations, including acquisition of fungicide resistance. We tested this hypothesis using the clonal plant pathogen, Sclerotinia sclerotiorum. Nine susceptible isolates were exposed independently to five commercial fungicides with different modes of action: boscalid (respiration inhibitor), iprodione (unclear mode of action), thiophanate methyl (inhibition of microtubulin synthesis) and azoxystrobin and pyraclostrobin (quinone outside inhibitors). Mycelium of each isolate was inoculated onto a fungicide gradient and sub-cultured from the 50–100% inhibition zone for 12 generations and experiment repeated. Mutational changes were assessed for all isolates at six neutral microsatellite (SSR) loci and for a subset of isolates using amplified fragment length polymorphisms (AFLPs). SSR analysis showed 12 of 85 fungicide-exposed isolates had a total of 127 stepwise mutations with 42 insertions and 85 deletions. Most stepwise deletions were in iprodione- and azoxystrobin-exposed isolates (n = 40/85 each). Estimated mutation rates were 1.7 to 60-fold higher for mutated loci compared to that expected under neutral conditions. AFLP genotyping of 33 isolates (16 non-exposed control and 17 fungicide exposed) generated 602 polymorphic alleles. Cluster analysis with principal coordinate analysis (PCoA) and discriminant analysis of principal components (DAPC) identified fungicide-exposed isolates as a distinct group from non-exposed control isolates (PhiPT = 0.15, P = 0.001). Dendrograms based on neighbor-joining also supported allelic variation associated with fungicide-exposure. Fungicide sensitivity of isolates measured throughout both experiments did not show consistent trends. For example, eight isolates exposed to boscalid had higher EC50 values at the end of the experiment

Evaluación de tres métodos de control del Moho blanco (Sclerotinia sclerotiorum (Lib. de Bary en lechuga (Lactuca sativa L.

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Arias Luis Alejandro

2007-06-01

Full Text Available

El principal limitante fitosanitario del cultivo de lechuga (Lactuca sativa L. es la enfermedad conocida como moho blanco causada por el hongo Sclerotinia sclerotiorum (Lib. de Bary. Con el fin de realizar un aporte al conocimiento de esta enfermedad y establecer protocolos para su control en campo se realizó un experimento entre los meses de febrero y mayo de 2006 en el cual se compararon tres métodos de control: aplicación de 0,5 kg· ha^-1 de procimidona, dosis de conidiosporas (Trichoderma harzianum DSM 14944 de 0,25 g· m^-2 de suelo, solarización, integración de solarización con procimidona y solarización con T. harzianum. Se evaluó la incidencia de la enfermedad, densidad de inóculo en el suelo antes y al finalizar el ensayo, temperatura del suelo durante la solarización y productividad. La densidad promedio del inóculo fue de 1,67 esclerocios de S. sclerotiorum por100 g de suelo al inicio y 2,07 esclerocios por 100 g de suelo al finalizar el ensayo. La temperatura del suelo solarizado se incrementó hasta 50°C con un promedio de 34,69°C hacia el medio día. Los porcentajes de incidencia de la enfermedad fueron: con procimidona, 3,3%; con T. harzianum, 10,0%; con solarización, 15,8%; con solarización y procimidona, 8,8%; con solarización y T. harzianum, 9,2% y testigo 18,3%. No se presentaron diferencias significativas en la productividad. En conclusión el control de la enfermedad fue mejor con la aplicación de procimidona y deficiente con T. harzianum y la solarización.

Identification of mycoparasitism-related genes against the phytopathogen Sclerotinia sclerotiorum through transcriptome and expression profile analysis in Trichoderma harzianum.

Science.gov (United States)

Steindorff, Andrei Stecca; Ramada, Marcelo Henrique Soller; Coelho, Alexandre Siqueira Guedes; Miller, Robert Neil Gerard; Pappas, Georgios Joannis; Ulhoa, Cirano José; Noronha, Eliane Ferreira

2014-03-18

The species of T. harzianum are well known for their biocontrol activity against plant pathogens. However, few studies have been conducted to further our understanding of its role as a biological control agent against S. sclerotiorum, a pathogen involved in several crop diseases around the world. In this study, we have used RNA-seq and quantitative real-time PCR (RT-qPCR) techniques in order to explore changes in T. harzianum gene expression during growth on cell wall of S. sclerotiorum (SSCW) or glucose. RT-qPCR was also used to examine genes potentially involved in biocontrol, during confrontation between T. harzianum and S. sclerotiorum. Data obtained from six RNA-seq libraries were aligned onto the T. harzianum CBS 226.95 reference genome and compared after annotation using the Blast2GO suite. A total of 297 differentially expressed genes were found in mycelia grown for 12, 24 and 36 h under the two different conditions: supplemented with glucose or SSCW. Functional annotation of these genes identified diverse biological processes and molecular functions required during T. harzianum growth on SSCW or glucose. We identified various genes of biotechnological value encoding proteins with functions such as transporters, hydrolytic activity, adherence, appressorium development and pathogenesis. To validate the expression profile, RT-qPCR was performed using 20 randomly chosen genes. RT-qPCR expression profiles were in complete agreement with the RNA-Seq data for 17 of the genes evaluated. The other three showed differences at one or two growth times. During the confrontation assay, some genes were up-regulated during and after contact, as shown in the presence of SSCW which is commonly used as a model to mimic this interaction. The present study is the first initiative to use RNA-seq for identification of differentially expressed genes in T. harzianum strain TR274, in response to the phytopathogenic fungus S. sclerotiorum. It provides insights into the mechanisms of

Sensibilidad in vitro de dos especies de Sclerotinia spp. y Sclerotium cepivorum a agentes de control biológico y fungicidas

OpenAIRE

Pérez-Moreno, Luis; Belmonte-Vargas, José Roberto; Núñez-Palenius, Héctor Gordon; Guzmán-Mendoza, Rafael; Mendoza-Celedón, Briseida

2015-01-01

Se evaluó la respuesta in vitro de dos aislados de Sclerotinia minor,cinco de S. sclerotiorum,y dos de Sclerotium cepivoruma 16 agentes biológicos y ocho fungicidas en un diseño experimental de parcelas divididas con arreglo factorial por cada patógeno. El factor A correspondió a los aislados del hongo y el factor B a los productos de control. La comparación de medias se realizó con la prueba de Tukey (P < 0.05). Se hicieron 11 evaluaciones cada 24 horas del crecimiento promedio radial miceli...

Isolation of Stem rot Disease Causing Organism of Brinjal and their in-vitro Inhibition with Fungicides and Bio-control Agents

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Shaily Javeria

2014-09-01

Full Text Available Different strains of Sclerotinia sclerotiorum were isolated from the diseased samples collected from different hosts and locations. Among the 14 isolates, 12 isolates colonies covered the entire Petri plates within 96 hours but, two isolates from fababean and yellow mustard showed slow colony growth within 96 hours. All isolates produced sclerotia which were varied in number, but the fenugreek isolate produced maximum (43 number of sclerotia and lambs quarter isolate produced minimum number of sclerotia (12 on PDA medium. To examine inhibitory effect of fungicide on the mycelial growth of the pathogen, 9 fungicides were tested in vitro against Sclerotinia sclerotiorum, of those carbendazim, carboxin, topsin-M and carbendazim+ mancozeb (SAAF were found most effective and inhibited the mycelial growth of pathogen up to 100 per cent at 0.05%, 0.1%, and 0.2% concentration. The effect of different bioagents viz., Trichoderma harzianum, T. viride, T. koningii, T. atroviride, T. longibraciatum, Aspergillus niger, Chaetomium globosome and Penicillium notatum in inhibiting the growth of Sclerotinia sclerotiorum was studied through “Dual Culture Technique”. The data showed that among the eight biocontrol agent six were fond effective. The maximum inhibition was found by T. harzianum causing 70.82% inhibition of mycelial growth of the pathogen S. sclerotiorum.

Identification of QTLs for resistance to sclerotinia stem rot and BnaC.IGMT5.a as a candidate gene of the major resistant QTL SRC6 in Brassica napus.

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Jian Wu

Full Text Available Stem rot caused by Sclerotinia sclerotiorum in many important dicotyledonous crops, including oilseed rape (Brassica napus, is one of the most devastating fungal diseases and imposes huge yield loss each year worldwide. Currently, breeding for Sclerotinia resistance in B. napus, as in other crops, can only rely on germplasms with quantitative resistance genes. Thus, the identification of quantitative trait locus (QTL for S. sclerotiorum resistance/tolerance in this crop holds immediate promise for the genetic improvement of the disease resistance. In this study, ten QTLs for stem resistance (SR at the mature plant stage and three QTLs for leaf resistance (LR at the seedling stage in multiple environments were mapped on nine linkage groups (LGs of a whole genome map for B. napus constructed with SSR markers. Two major QTLs, LRA9 on LG A9 and SRC6 on LG C6, were repeatedly detected across all environments and explained 8.54-15.86% and 29.01%-32.61% of the phenotypic variations, respectively. Genotypes containing resistant SRC6 or LRA9 allele showed a significant reduction in disease lesion after pathogen infection. Comparative mapping with Arabidopsis and data mining from previous gene profiling experiments identified that the Arabidopsis homologous gene of IGMT5 (At1g76790 was related to the SRC6 locus. Four copies of the IGMT5 gene in B. napus were isolated through homologous cloning, among which, only BnaC.IGMT5.a showed a polymorphism between parental lines and can be associated with the SRC6. Furthermore, two parental lines exhibited a differential expression pattern of the BnaC.IGMT5.a gene in responding to pathogen inoculation. Thus, our data suggested that BnaC.IGMT5.a was very likely a candidate gene of this major resistance QTL.

Moléstias em tomateiro cultivado em estufas plásticas em quatro municípios da região central do Rio Grande do Sul, Brasil

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Blume Elena

2004-01-01

Full Text Available Moléstias podem atacar severamente hortaliças cultivadas em cultivos protegidos e, para o manejo integrado dessas moléstias, o conhecimento das condições climáticas que favorecem sua incidência nesses cultivos é fundamental. O trabalho teve o objetivo de identificar as moléstias em tomateiro cultivado em estufas plásticas em quatro municípios na região central do Rio Grande do Sul e as condições de temperatura e umidade relativa (UR do ar nas quais elas ocorrem. O estudo foi realizado nos municípios de São Pedro do Sul, São Sepé, Ivorá e Santa Maria (RS, no período de março a outubro de 1998. A temperatura e umidade relativa do ar foram medidas diariamente com um psicrômetro. As moléstias observadas e sua incidência máxima foram: requeima (Phytophthora infestans: 100,0%, pinta-preta (Alternaria solani: 98,1%, mofo cinzento (Botrytis cinerea: 55,4%, cladosporiose (Cladosporuim fulvum: 48,9%, septoriose (Septoria lycopersici: 37,5%, talo-oco (Erwinia spp.: 33,0%, murcha de fusarium (Fusarium oxysporum f. sp. lycopersici: 8,4% e podridão de esclerotínia (Sclerotinia sclerotiorum: 1,3%. Valores de UR superiores a 80%, de maneira geral, aumentaram a incidência das moléstias, em sua maioria de origem fúngica. A incidência de moléstias varia entre municípios de uma mesma região climática de acordo com os fatores meteorológicos e práticas de manejo da cultura.

Isolation and characterization of rhizosphere bacteria for the biocontrol of the damping-off disease of tomatoes in Tunisia.

Science.gov (United States)

Hammami, Inés; Ben Hsouna, Anis; Hamdi, Naceur; Gdoura, Radhouane; Triki, Mohamed Ali

2013-01-01

Fluorescent Pseudomonas spp., isolated from tomato and pepper plants rhizosphere soil, was evaluated in vitro as a potential antagonist of fungal pathogens. Pseudomonas strains were tested against the causal agents of tomatoes damping-off (Sclerotinia sclerotiorum), root rot (Fusarium solani), and causal agents of stem canker and leaf blight (Alternaria alternata). For this purpose, dual culture antagonism assays were carried out on 25% tryptic soy agar, King B medium and potato dextrose agar to determine the effect of the strains on mycelial growth of the pathogens. In addition, strains were screened for their ability to produce exoenzymes and siderophores. All the strains significantly inhibited Alternaria alternata, particularly in 25% TSA medium. Antagonistic effect on Sclerotinia sclerotiorum and Fusarium solani was greater on King B medium. Protease was produced by 30% of the strains, but no strain produced cellulase or chitinase. Finally, the selected Pseudomonas strain, Psf5, was evaluated on tomato seedling development and as a potential candidate for controlling tomato damping-off caused by Sclerotinia sclerotiorum, under growth chamber conditions. In vivo studies resulted in significant increases in plant stand as well as in root dry weight. Psf5 was able to establish and survive in tomato plants rhizosphere after 40days following the planting of bacterized seeds. © 2013 Académie des sciences. Published by Elsevier SAS. All rights reserved.

Produção in vitro de enzimas extracelulares por fungos e sua relação com os sintomas descritos em planta hospedeira

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Alice Elias

2015-12-01

Full Text Available RESUMOOs fungos fitopatogênicos habitantes de solo causam perdas econômicas em muitas culturas e são difíceis de serem controlados. Esses fungos podem ser agrupados pelos sintomas comuns que causam nas plantas, bem como pelas enzimas extracelulares que podem produzir. O objetivo do trabalho foi verificar a produção in vitro de enzimas extracelulares por fungos de solo e tentar relacionar essas enzimas com os sintomas que cada fungo causa em planta hospedeira. O ensaio foi delineado em esquema inteiramente casualizado, com dois fatores, sete fungos (Cylindrocladium spathiphylli, Rhizoctonia solani, Sclerotium rolfsii, Ceratocystis fimbriata, Sclerotinia sclerotiorum, Fusarium oxysporum f. sp. dianthi e Verticillium dahliae mais testemunha e seis enzimas (amilase, carboximetilcelulase, lipase, lacase, catalase e gelatinase com 10 repetições. Catalase e gelatinase foram mensuradas por escala de notas, enquanto que as demais pelo cálculo da área da coroa circular. O ensaio foi repetido e a análise foi realizada com os dados de dois ensaios. Os fungos que causam podridão na raiz ou no colo da planta apresentaram maior produção de lacase, enquanto os que causam obstrução, fendas ou até a destruição do sistema vascular demonstraram a prevalência da lipase.

A plant defensin gene from Orychophragmus violaceus can improve ...

African Journals Online (AJOL)

Quantitative RT-PCR showed that the expression of Ovd in the sense plant line was stronger than non-transformed plant and antisense plant. The lesion size on detached leaves of the transgenic plants and nontransformed control caused by Sclerotinia sclerotiorum mycelia was examined. Lesion size was reduced in sense ...

Genome size variation in the pine fusiform rust pathogen Cronartium quercuum f.sp. fusiforme as determined by flow cytometry

Science.gov (United States)

Claire L Anderson; Thomas L Kubisiak; C Dana Nelson; Jason A Smith; John M Davis

2010-01-01

The genome size of the pine fusiform rust pathogen Cronartium quercuum f.sp. fusiforme (Cqf) was determined by flow cytometric analysis of propidium iodide-stained, intact haploid pycniospores with haploid spores of two genetically well characterized fungal species, Sclerotinia sclerotiorum and Puccinia graminis f.sp. tritici, as size standards. The Cqf haploid genome...

White mold intensity on common bean in response to plant density, irrigation frequency, grass mulching, Trichoderma spp., and fungicide Intensidade do mofobranco em feijão em função de densidade de plantas, freqüência de irrigação, cobertura vegetal do solo, Trichoderma spp. e fungicida

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Trazilbo José de Paula Júnior

2009-02-01

Full Text Available The purpose of this study was to evaluate the efficiency of integrated managements on white mold control on common bean. Initially, in vitro testing was made to assess the antagonism of 11 Trichoderma isolates against Sclerotinia sclerotiorum and to investigate fungicides (fluazinam and procymidone inhibitory effects on those fungi. In two field experiments the following combinations were tested: irrigation frequencies (seven or 14 days, plant densities (six or 12 plants per meter, and three disease controls (untreated control, fungicide or Trichoderma spp.. In a third experiment plant densities were replaced by grass mulching treatments (with or without mulching. Fluazinam was applied at 45 and 55 days after emergence (DAE. The antagonists T. harzianum (experiments 1 and 3 and T. stromatica (experiment 2 were applied through sprinkler irrigation at 10 and 25 DAE, respectively. Most of the Trichoderma spp. were effective against the pathogen in vitro. Fluazinam was more toxic than procymidone to both the pathogen and the antagonist. Fungicide applications increased yield between 32 % and 41 %. In field one application of Trichoderma spp. did not reduce disease intensity and did not increase yield. The reduction from 12 to six plants per meter did not decrease yield, and disease severity diminished in one of the two experiments. It is concluded that of the strategies for white mold control just reduction of plant density and applications of fungicide were efficient.Objetivou-se estudar a eficácia de técnicas de manejo integrado no controle do mofo-branco em feijão. Inicialmente, foram feitos testes in vitro para avaliar o antagonismo de 11 isolados de Trichoderma contra Sclerotinia sclerotiorum e investigar os efeitos de fungicidas (fluazinam e procymidone sobre esses fungos. Em dois ensaios de campo, foram testadas estas combinações: freqüências de irrigação (sete ou 14 dias, densidades de plantas (seis ou 12 por metro e três controles

Mycelial compatibility groups and pathogenicity of Sclerotinia ...

African Journals Online (AJOL)

... was determined by mycelial compatibility grouping (MCG) and isolate aggressiveness comparisons. MCG, host specificity and aggressiveness of S. sclerotiorum isolates were assessed. Isolate pairs were designated compatible when no barrage zone formed at sites of contact. They were designated incompatible when a ...

Study on the Occurrence and Epidemic Model of Rape Sclerotinia Stem Rot of ‘Zheyou 50’

OpenAIRE

Xu Sen-fu; Wang Hui-fu; Yu Shanhong; Wang En-guo

2013-01-01

In order to investigate invading and epidemic rules of rape sclerotinia stem rot of ‘Zheyou 50’ and promote the development of brassica campestris industry, this paper studied the outbreak regularity and epidemic model of rape sclerotinia stem rot according to field investigation and infection. The result showed that machinery direct seeding rape was good for the occurrence of sclerotinia stem rot for the reason of late seeding and high density. The period from water damage appeared to wiltin...

Identification Of Protein Vaccine Candidates Using Comprehensive Proteomic Analysis Strategies

Science.gov (United States)

2007-12-01

that fascinating fungus known as Coccidioides. I also want to thank the UA Mass Spectrometry Facility and the UA Proteomics Consortium, especially...W. & N. N. Kav. 2006. The proteome of the phytopathogenic fungus Sclerotinia sclerotiorum. Proteomics 6: 5995-6007. 127. de Godoy, L. M., J. V...IDENTIFICATION OF PROTEIN VACCINE CANDIDATES USING COMPREHENSIVE PROTEOMIC ANALYSIS STRATEGIES by James G. Rohrbough

In vitro selection of rape variants resistant to oxalic acid using haploid stem apexes

International Nuclear Information System (INIS)

Wang Yifei; Huang Jianhua; Lu Ruiju; Sun Yuefang; Zhou Runmei; Zhou Zhijiang; Xie Zhujie; Liu Chenghong

2002-01-01

Mutagenic treatment was made of the haploid stem apexes rape strain '9841' and '9885' with Pingyangmycin. As a result of positive selection with oxalic acid providing selection pressure, variants with significantly higher tolerance to oxalic acid than the original ones were obtained. 3 germplasm with significantly higher resistance to Sclerotinia sclerotiorum than cultivar Hu You 12 were selected from field test

Biological Control of Lettuce Drop and Host Plant Colonization by Rhizospheric and Endophytic Streptomycetes

Science.gov (United States)

Chen, Xiaoyulong; Pizzatti, Cristina; Bonaldi, Maria; Saracchi, Marco; Erlacher, Armin; Kunova, Andrea; Berg, Gabriele; Cortesi, Paolo

2016-01-01

Lettuce drop, caused by the soil borne pathogen Sclerotinia sclerotiorum, is one of the most common and serious diseases of lettuce worldwide. Increased concerns about the side effects of chemical pesticides have resulted in greater interest in developing biocontrol strategies against S. sclerotiorum. However, relatively little is known about the mechanisms of Streptomyces spp. as biological control agents against S. sclerotiorum on lettuce. Two Streptomyces isolates, S. exfoliatus FT05W and S. cyaneus ZEA17I, inhibit mycelial growth of Sclerotinia sclerotiorum by more than 75% in vitro. We evaluated their biocontrol activity against S. sclerotiorum in vivo, and compared them to Streptomyces lydicus WYEC 108, isolated from Actinovate®. When Streptomyces spp. (106 CFU/mL) were applied to S. sclerotiorum inoculated substrate in a growth chamber 1 week prior lettuce sowing, they significantly reduced the risk of lettuce drop disease, compared to the inoculated control. Interestingly, under field conditions, S. exfoliatus FT05W and S. cyaneus ZEA17I protected lettuce from drop by 40 and 10% respectively, whereas S. lydicus WYEC 108 did not show any protection. We further labeled S. exfoliatus FT05W and S. cyaneus ZEA17I with the enhanced GFP (EGFP) marker to investigate their rhizosphere competence and ability to colonize lettuce roots using confocal laser scanning microscopy (CLSM). The abundant colonization of young lettuce seedlings by both strains demonstrated Streptomyces' capability to interact with the host from early stages of seed germination and root development. Moreover, the two strains were detected also on 2-week-old roots, indicating their potential of long-term interactions with lettuce. Additionally, scanning electron microscopy (SEM) observations showed EGFP-S. exfoliatus FT05W endophytic colonization of lettuce root cortex tissues. Finally, we determined its viability and persistence in the rhizosphere and endorhiza up to 3 weeks by quantifying its

Biological control of lettuce drop and host plant colonization by rhizospheric and endophytic streptomycetes

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Xiaoyulong eChen

2016-05-01

Full Text Available Lettuce drop, caused by the soil borne pathogen Sclerotinia sclerotiorum, is one of the most common and serious diseases of lettuce worldwide. Increased concerns about the side effects of chemical pesticides have resulted in greater interest in developing biocontrol strategies against S. sclerotiorum. However, relatively little is known about the mechanisms of Streptomyces spp. as biological control agents against S. sclerotiorum on lettuce. Two Streptomyces isolates, S. exfoliatus FT05W and S. cyaneus ZEA17I, inhibit mycelial growth of Sclerotinia sclerotiorum by more than 75% in vitro. We evaluated their biocontrol activity against S. sclerotiorum in vivo, and compared them to Streptomyces lydicus WYEC 108, isolated from Actinovate®. When Streptomyces spp. (106 CFU/mL were applied to S. sclerotiorum inoculated substrate in a growth chamber one week prior lettuce sowing, they significantly reduced the risk of lettuce drop disease, compared to the inoculated control. Interestingly, under field conditions, S. exfoliatus FT05W and S. cyaneus ZEA17I protected lettuce from drop by 40% and 10% respectively, whereas S. lydicus WYEC 108 did not show any protection. We further labeled S. exfoliatus FT05W and S. cyaneus ZEA17I with the enhanced GFP (EGFP marker to investigate their rhizosphere competence and ability to colonize lettuce roots using confocal laser scanning microscopy (CLSM. The abundant colonization of young lettuce seedlings by both strains demonstrated Streptomyces’ capability to interact with the host from early stages of seed germination and root development. Moreover, the two strains were detected also on two-week-old roots, indicating their potential of long-term interactions with lettuce. Additionally, scanning electron microscopy (SEM observations showed EGFP-S. exfoliatus FT05W endophytic colonization of lettuce root cortex tissues. Finally, we determined its viability and persistence in the rhizosphere and endorhiza up to

Colonization of lettuce rhizosphere and roots by tagged Streptomyces

OpenAIRE

Maria eBonaldi; Xiaoyulong eChen; Andrea eKunova; Cristina ePizzatti; Marco eSaracchi; Paolo eCortesi

2015-01-01

Beneficial microorganisms are increasingly used in agriculture, but their efficacy often fails due to limited knowledge of their interactions with plants and other microorganisms present in rhizosphere. We studied spatio-temporal colonization dynamics of lettuce roots and rhizosphere by genetically modified Streptomyces spp. Five Streptomyces strains, strongly inhibiting in vitro the major soil-borne pathogen of horticultural crops, Sclerotinia sclerotiorum, were transformed with pIJ8641 plas...

Biogenic silver nanoparticles based on trichoderma harzianum: synthesis, characterization, toxicity evaluation and biological activity

Science.gov (United States)

Guilger, Mariana; Pasquoto-Stigliani, Tatiane; Bilesky-Jose, Natália; Grillo, Renato; Abhilash, P. C.; Fraceto, Leonardo Fernandes; Lima, Renata De

2017-03-01

White mold is an agricultural disease caused by the fungus Sclerotinia sclerotiorum, which affects important crops. There are different ways of controlling this organism, but none provides inhibition of its resistance structures (sclerotia). Nanotechnology offers promising applications in agricultural area. Here, silver nanoparticles were biogenically synthesized using the fungus Trichoderma harzianum and characterized. Cytotoxicity and genotoxicity were evaluated, and the nanoparticles were initially tested against white mold sclerotia. Their effects on soybean were also investigated with no effects observed. The nanoparticles showed potential against S. sclerotiorum, inhibiting sclerotia germination and mycelial growth. Nanoparticle characterization data indicated spherical morphology, satisfactory polydispersity and size distribution. Cytotoxicity and genotoxicity assays showed that the nanoparticles caused both the effects, although, the most toxic concentrations were above those applied for white mold control. Given the potential of the nanoparticles against S. sclerotiorum, we conclude that this study presents a first step for a new alternative in white mold control.

Design, synthesis and fungicidal evaluation of novel pyraclostrobin analogues.

Science.gov (United States)

Wang, Lili; Zhao, Shuangshuang; Kong, Xiaotian; Cao, Lingling; Tian, Sheng; Ye, Yonghao; Qiao, Chunhua

2018-02-15

A series of novel pyraclostrobin derivatives were designed and prepared as antifungal agents. Their antifungal activities were tested in vitro with five important phytopathogenic fungi, namely, Batrylis cinerea, Phytophthora capsici, Fusarium sulphureum, Gloeosporium pestis and Sclerotinia sclerotiorum using the mycelium growth inhibition method. Among these compounds, 5s displayed IC 50 value of 0.57 μg/mL against Batrylis cinerea and 5k-II displayed IC 50 value of 0.43 μg/mL against Sclerotinia sclerotiorum, which were close to that of the positive control pyraclostrobin (0.18 μg/mL and 0.15 μg/mL). Other compounds 5f, 5k-II, 5j, 5m and 5s also exhibited strong antifungal activity. Further enzymatic assay demonstrated compound 5s inhibited porcine bc 1 complex with IC 50 value of 0.95 μM. The statistical results from an integrated computational pipeline demonstrated the predicted total binding free energy for compound 5s is the highest. Consequently, compound 5s with the biphenyl-4-methoxyl side chain could serve as a new motif as inhibitors of bc 1 complex and deserve to be further investigated. Copyright © 2018 Elsevier Ltd. All rights reserved.

Soybean diseases in Poland

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J. Marcinkowska

2013-12-01

Full Text Available Field observations on the occurrence of soybean diseases were undertaken in the southern and central regions of Poland in the period 1976-1980. Most prevalent were foliage diseases caused by Peronospora manshurica, Pseudomonas syrinqae pv. glycinea and soybean mosaic virus (SMV. Sclerotinia sclerotiorum and Ascochyta sojaecola were reported as pathogens of local importance. The following pathogenic fungi: Botrytis cinerea, Fusarium culmorum, F. oxysporum and Rhizoctonia solani were also isolated from soybean.

The fungi causin damping-off of carrot seedlings

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Bogdan Nowicki

2013-12-01

Full Text Available When 136 samples of dying carrot seedlings from several fields were analyzed Alternaria rudicina proved to be the most common seedling pathogen (41%, followed by some Fusarium species (27%, mostly F. avenaceum.The less common seedling pathogens were Pythium spp. (13%, Phoma spp.(2,5% and Botrytis cinerea (1,4%. Some other fungi (Bipolaris sorokiniana, Sclerotinia sclerotiorum, Stemphylium botryosym and Ulocladium consortiale were found in less than 1% of seedlings examined.

Synthesis, crystal structure and biological activity of n-(5-(o-tolyl)-1, 3, 4-thiadiazol-2-yl)cyclopropanecarboxamide

International Nuclear Information System (INIS)

Tong, J.Y.; Sun, N.B.; Wu, H.K.

2013-01-01

A new 1, 3, 4-thiadiazole compound, N-(5-(o-tolyl)-1,3,4-thiadiazol-2-yl) cyclopropanecarboxamide, was synthesized and its structure was confirmed by 1H NMR, MS and HRMS. The single crystal structure of the title compound was determined by X-ray diffraction. The preliminary biological test showed that the synthesized compound has moderate herbicidal activity against Brassica campestris and fungicidal activities against Sclerotinia sclerotiorum(Lib.) de Bary, Rhizoctonia solanii, Fusarium oxysporum, Corynespora cassiicola, and Botrytis cinerea. (author)

Physiological and sanity seed quality of common beans (Phaseolus vulgaris L. from Goias state / Qualidade fisiológica e sanitária de sementes de feijoeiro (Phaseolus vulgaris L. provenientes do estado de Goiás

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Myrna Hilal Moraes

2008-08-01

Full Text Available The common beans (Phaseolus vulgaris L. is a fabacea sufficiently spread out in all domestic territory. However, the quality of its seeds represents one of the main causes of low productivity in the beans farmings in Brazil. The objective of this work was to evaluate physiological and sanitary seed qualities of eleven bean cultivars. The physiological seed quality was evaluated trough standard germination and vigor tests. The sanitary seed quality was evaluated through two tests: blotter test was employed to evaluate fungi incidence and “Koch & Menten” method was employed to observe Sclerotinia sclerotiorum (Lib de Bary occurrence. Xamego, BRS Valente, Bambu and Pérola had the best results of physiological tests. Jalo Precoce, Roxo 90, Corrente and Aporé had no good results of vigor and germination, besides presenting the lowest indices of died seeds. Fusarium sp., Aspergillus spp., Penicillium sp., Phoma sp., Rhizopus sp. and Botrytis sp. were the fungi detected in the sanity tests.O feijoeiro comum (Phaseolus vulgaris L. é uma fabacea bastante difundida em todo território nacional. A baixa qualidade de suas sementes representa uma das principais causas de baixa produtividade nas lavouras de feijão no Brasil. O objetivo deste trabalho foi avaliar a qualidade fisiológica e sanitária de sementes de nove cultivares de feijão provenientes do Estado de Goiás. A qualidade fisiológica das sementes foi avaliada através dos testes de germinação e vigor, e a análise sanitária, através dos métodos de papel de filtro, para verificar a ocorrência de fungos em geral, e do método de Koch e Menten, para a avaliação de Sclerotinia sclerotiorum (Lib de Bary. As cultivares que tiveram os melhores desempenhos nos testes fisiológicos foram Xamego, BRS Radiante, Bambu e Pérola. As cultivares Jalo Precoce, Roxo 90, Corrente e Aporé apresentaram baixos índices de vigor e germinação de plântulas normais, além de apresentarem os maiores

Synthesis and antifungal activity of nicotinamide derivatives as succinate dehydrogenase inhibitors.

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Ye, Yong-Hao; Ma, Liang; Dai, Zhi-Cheng; Xiao, Yu; Zhang, Ying-Ying; Li, Dong-Dong; Wang, Jian-Xin; Zhu, Hai-Liang

2014-05-07

Thirty-eight nicotinamide derivatives were designed and synthesized as potential succinate dehydrogenase inhibitors (SDHI) and precisely characterized by (1)H NMR, ESI-MS, and elemental analysis. The compounds were evaluated against two phytopathogenic fungi, Rhizoctonia solani and Sclerotinia sclerotiorum, by mycelia growth inhibition assay in vitro. Most of the compounds displayed moderate activity, in which, 3a-17 exhibited the most potent antifungal activity against R. solani and S. sclerotiorum with IC50 values of 15.8 and 20.3 μM, respectively, comparable to those of the commonly used fungicides boscalid and carbendazim. The structure-activity relationship (SAR) of nicotinamide derivatives demonstrated that the meta-position of aniline was a key position contributing to the antifungal activity. Inhibition activities against two fungal SDHs were tested and achieved the same tendency with the data acquired from in vitro antifungal assay. Significantly, 3a-17 was demonstrated to successfully suppress disease development in S. sclerotiorum infected cole in vivo. In the molecular docking simulation, sulfur and chlorine of 3a-17 were bound with PHE291 and PRO150 of the SDH homology model, respectively, which could explain the probable mechanism of action between the inhibitory and target protein.

New record of Phytophthora root and stem rot of Lavandula angustifolia

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Leszek B. Orlikowski

2013-12-01

Full Text Available Phytophthora cinnamomi was isolated from rotted root and stem parts of lavender as well as from soil taken from containers with diseased plants. Additionally Botrytis cinerea, Fusarium spp. and Sclerotinia sclerotiorum were often isolated from diseased tissues. P. cinnamomi colonised leaves and stem parts of 4 lavender species in laboratory trials and caused stem rot of plants in greenhouse experiments. Cardinal temperature for in vitro growth were about 7,5 and 32°C with optimum 25-27,5°C. The species colonised stem tissues at temperature ranged from 10° to 32°C.

Diseases of herbs from Apiaceae family

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Ewa Dorota Zalewska

2013-04-01

Full Text Available The largest participation in causing the disease of herbal plants have fungi. Studies on their occurrence on plants of the family Apiaceae are conducted in the Lublin region since 2001. The observations of plant healthiness are carried out directly on the plantations. Plants with symptoms of disease are studied in the laboratory. Identification of the fungi is performed based on etiological symptoms and on the base of fungal cultures isolated from plants. Among the many species of fungi obtained from diseased plants to the particularly harmful belong: Septoria carvi, Colletotrichum gloeosporioidesand C. dematium, Sclerotinia sclerotiorum, Passalora puncta(Cercosporidium punctum and Erysiphe umbelliferarum.

Advance in the biological handling of pathogen forming sclerocios

International Nuclear Information System (INIS)

Avila de Moreno, L.C.E.; Velandia Mosalve, J.

1995-01-01

Field rehearsals in the Center of Investigations Tibaitata allowed to determine that the mushrooms Trichoderma sp., Trichogramma search. Konigui No.1, Trichoderma search Konigui No.2 and Gliocladium sp. They have antagonistic effects against Rhizoctonia solani, that which showed in low percentages of tubers affected by R. solani, when the soil was treated with the mentioned mushrooms. In the control of Sclerotinia sclerotiorum, it was determined that Trichoderma barzianum should be applied in the moment in that it is presented more than 3 days of continuous humidity in the soil; lower these conditions 14.3 percent of plants it was only observed affected by the pathogen

Patogenic fungi associated with blue lupine seeds

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Bogdan Nowicki

2013-12-01

Full Text Available Over 10% ofseeds harvested in 1991 and 1992 (50 samples, 400 seeds in each sample proved to be infested with various fungi. Fusarium spp. and Botrytis cinerea were the most common pathogens isolated. Fusarium avenaceum was the most common and highIy pathogenic species. Fusarium semitectum and F. tricinctum were highly pathogenic to lupin seedlings but they were the least common Fusarium isolated from seeds. Similarily, Sclerotinia sclerotiorum was isolated only from 0,2% seeds tested but this fungus was highly pathogenic to lupin seedlings. Some other fungi know as lupin pathogens (F. oxysporum, Stemphylium botryosum, Pleiochaeta setosa and Phomopsis leptostromiformis were also noted in tested seeds.

Fungicides and Application Timing for Control of Early Leafspot, Southern Blight, and Sclerotinia Blight of Peanut

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W. James Grichar

2016-01-01

Full Text Available Field studies were conducted in 2013 and 2014 in south Texas near Yoakum and from 2008 to 2011 in central Texas near Stephenville to evaluate various fungicides for foliar and soilborne disease control as well as peanut yield response under irrigation. Control of Sclerotinia blight caused by Sclerotinia minor Jagger with penthiopyrad at 1.78 L/ha was comparable to fluazinam or boscalid; however, the 1.2 L/ha dose of penthiopyrad did not provide consistent control. Peanut yield was reduced with the lower penthiopyrad dose when compared with boscalid, fluazinam, or the high dose of penthiopyrad. Control of early leaf spot, caused by Cercospora arachidicola S. Hori or southern blight, caused by Sclerotium rolfsii Sacc., with penthiopyrad in a systems approach was comparable with propiconazole, prothioconazole, or pyraclostrobin systems and resulted in disease control that was higher than the nontreated control. Peanut yield was also comparable with the penthiopyrad, propiconazole, prothioconazole, or pyraclostrobin systems and reflects the ability of the newer fungicides to control multiple diseases found in Texas peanut production.

Virus-mediated suppression of host non-self recognition facilitates horizontal transmission of heterologous viruses.

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Songsong Wu

2017-03-01

Full Text Available Non-self recognition is a common phenomenon among organisms; it often leads to innate immunity to prevent the invasion of parasites and maintain the genetic polymorphism of organisms. Fungal vegetative incompatibility is a type of non-self recognition which often induces programmed cell death (PCD and restricts the spread of molecular parasites. It is not clearly known whether virus infection could attenuate non-self recognition among host individuals to facilitate its spread. Here, we report that a hypovirulence-associated mycoreovirus, named Sclerotinia sclerotiorum mycoreovirus 4 (SsMYRV4, could suppress host non-self recognition and facilitate horizontal transmission of heterologous viruses. We found that cell death in intermingled colony regions between SsMYRV4-infected Sclerotinia sclerotiorum strain and other tested vegetatively incompatible strains was markedly reduced and inhibition barrage lines were not clearly observed. Vegetative incompatibility, which involves Heterotrimeric guanine nucleotide-binding proteins (G proteins signaling pathway, is controlled by specific loci termed het (heterokaryon incompatibility loci. Reactive oxygen species (ROS plays a key role in vegetative incompatibility-mediated PCD. The expression of G protein subunit genes, het genes, and ROS-related genes were significantly down-regulated, and cellular production of ROS was suppressed in the presence of SsMYRV4. Furthermore, SsMYRV4-infected strain could easily accept other viruses through hyphal contact and these viruses could be efficiently transmitted from SsMYRV4-infected strain to other vegetatively incompatible individuals. Thus, we concluded that SsMYRV4 is capable of suppressing host non-self recognition and facilitating heterologous viruses transmission among host individuals. These findings may enhance our understanding of virus ecology, and provide a potential strategy to utilize hypovirulence-associated mycoviruses to control fungal diseases.

Mycoparasitism studies of Trichoderma species against three phytopathogenic fungi: evaluation of antagonism and hydrolytic enzyme production.

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Qualhato, Thiago Fernandes; Lopes, Fabyano Alvares Cardoso; Steindorff, Andrei Stecca; Brandão, Renata Silva; Jesuino, Rosália Santos Amorim; Ulhoa, Cirano José

2013-09-01

Trichoderma spp. are used for biocontrol of several plant pathogens. However, their efficient interaction with the host needs to be accompanied by production of secondary metabolites and cell wall-degrading enzymes. Three parameters were evaluated after interaction between four Trichoderma species and plant-pathogenic fungi: Fusarium solani, Rhizoctonia solani and Sclerotinia sclerotiorum. Trichoderma harzianum and T. asperellum were the most effective antagonists against the pathogens. Most of the Trichoderma species produced toxic volatile metabolites, having significant effects on growth and development of the plant pathogens. When these species were grown in liquid cultures with cell walls from these plant pathogens, they produced and secreted β-1,3-glucanase, NAGAse, chitinase, acid phosphatase, acid proteases and alginate lyase.

Antifungal efficacy of hydrogen peroxide in dental unit waterline disinfection.

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Szymańska, Jolanta

2006-01-01

The concentration and composition of fungal flora in dental unit waterlines (DUWL) were evaluated. For this purpose, water samples from unit reservoirs and high-speed handpieces, and biofilm samples from the waterline walls from units were collected. Subsequently, analogous samples from DUWL were taken before and after disinfection using agent containing hydrogen peroxide. In the examined samples, the yeast-like fungi Candida albicans and Candida curvata were found. The following species of mould were also identified: Aspergillus amstelodami, Aspergillus fumigatus, Aspergillus glaucus group, Aspergillus (=Eurotium herbariorum) repens, Citromyces spp., Geotrichum candidum, Penicillium (glabrum) frequentans, Penicillium pusillum, Penicillium turolense and Sclerotium sclerotiorum (Sclerotinia sclerotiorum). Before disinfection, Candida curvata and Candida albicans constituted the greatest proportion of the total fungi in the reservoirs water; in the water of handpieces--Candida albicans and Aspergillus glaucus group; and in the biofilm samples--Aspergillus glaucus group and Candida albicans. After disinfection, in all 3 kinds of samples, Candida albicans prevailed, constituting from 31.2-85.7 % of the total fungi. The application of agent containing hydrogen peroxide caused a significant decrease both in the number of total fungi and individual fungal species, which confirms the product effectiveness in fungal decontamination of DUWL.

RNA interference of endochitinases in the sugarcane endophyte Trichoderma virens 223 reduces its fitness as a biocontrol agent of pineapple disease.

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Aline S Romão-Dumaresq

Full Text Available The sugarcane root endophyte Trichoderma virens 223 holds enormous potential as a sustainable alternative to chemical pesticides in the control of sugarcane diseases. Its efficacy as a biocontrol agent is thought to be associated with its production of chitinase enzymes, including N-acetyl-ß-D-glucosaminidases, chitobiosidases and endochitinases. We used targeted gene deletion and RNA-dependent gene silencing strategies to disrupt N-acetyl-ß-D-glucosaminidase and endochitinase activities of the fungus, and to determine their roles in the biocontrol of soil-borne plant pathogens. The loss of N-acetyl-ß-D-glucosaminidase activities was dispensable for biocontrol of the plurivorous damping-off pathogens Rhizoctonia solani and Sclerotinia sclerotiorum, and of the sugarcane pathogen Ceratocystis paradoxa, the causal agent of pineapple disease. Similarly, suppression of endochitinase activities had no effect on R. solani and S. sclerotiorum disease control, but had a pronounced effect on the ability of T. virens 223 to control pineapple disease. Our work demonstrates a critical requirement for T. virens 223 endochitinase activity in the biocontrol of C. paradoxa sugarcane disease, but not for general antagonism of other soil pathogens. This may reflect its lifestyle as a sugarcane root endophyte.

Effects of Fungicides, Time of Application, and Application Method on Control of Sclerotinia Blight in Peanut

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Jason E. Woodward

2015-01-01

Full Text Available Field studies were conducted from 2007 to 2010 to evaluate the response of peanut cultivars to different fungicides, application timings, and methods. Overall, fungicides reduced Sclerotinia blight incidence and increased pod yields when applied to susceptible and partially resistant cultivars. Disease suppression was greater when full fungicide rates were applied preventatively; however, yields between fungicide treated plots were similar. Lower levels of disease and higher yields were achieved with the partially resistant cultivar Tamrun OL07 compared to the susceptible cultivars Flavor Runner 458 and Tamrun OL 02. Despite possessing improved resistance Tamrun OL07 responded to all fungicide applications. While similar levels of disease control were achieved with broadcast or banded applications made during the day or at night, the yield response for the different application methods was inconsistent among years. A negative relationship (slope = −73.8; R2=0.73; P<0.01 was observed between final disease incidence ratings and yield data from studies where a fungicide response was observed. These studies suggest that both boscalid and fluazinam are effective at controlling Sclerotinia blight in peanuts. Alternative management strategies such as nighttime and banded applications could allow for lower fungicide rates to be used; however, additional studies are warranted.

Isolation of fungal homokaryotic lines from heterokaryotic transformants by sonic disruption of mycelia.

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Bashi, Zafer Dallal; Khachatourians, George; Hegedus, Dwayne Daniel

2010-01-01

Fungal hyphae--and in some cases, spores--are multi-nucleate. During genetic transformation of these spores or mycelia, only one nucleus generally receives the transferred T-DNA generating heterokaryotic colonies. Characterization of genetic changes, such as the effects of gene disruption in the transformants, requires purified homokaryotic lines. Hyphal tip transfer has conventionally been used to isolate homokaryons. We developed an alternative method for purification of fungal homokaryons from transformed heterokaryotic lines of Sclerotinia sclerotiorum. Ultrasound pulses were used to generate bi-septate, unicellular hyphal fragments that regenerate under selection to produce homokaryotic lines that can be easily identified using colony PCR. This technique facilitates the purification of transformed lines, which allows for routine genome manipulation, and should be adaptable for other filamentous fungi.

Efeito da temperatura e reação de genótipos de quiabeiro ao mofo branco

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Ivan Herman Fischer

2014-03-01

Full Text Available O quiabeiro é suscetível a várias doenças que podem causar prejuízos ao agricultor, caso não sejam manejadas. Objetivaram-se avaliar o efeito da temperatura e a reação de genótipos de quiabeiro ao mofo branco. Plantas de 30 dias inoculadas na região do colo, com e sem ferimento, com dois isolados de Sclerotinia sclerotiorum, foram mantidas por duas semanas nas temperaturas de 15; 20; 25; 30 ou 35ºC. Quatorze genótipos de quiabeiro com um e dois meses de emergência, assim como frutos no ponto de colheita comercial tiveram seus comportamentos avaliados quanto à doença. Avaliaram-se a incidência de plantas com tombamento ou mortas e o comprimento da lesão nos frutos inoculados. A doença foi favorecida por temperaturas entre 15 e 20ºC e, de maneira geral, não foram observadas diferenças entre os isolados e influência do ferimento na ocorrência da doença. Não foram encontrados genótipos de quiabeiro resistentes ao mofo branco, pois todos apresentaram incidência superior a 30% de plantas mortas, sendo as plantas mais novas mais suscetíveis, e comprimento de lesão nos frutos >6,0 cm. Entretanto, diferenças de suscetibilidade entre os genótipos foram observadas em plantas de 30 e de 60 dias e podem ser informações úteis aos produtores para escolha de cultivares em clima ameno, assim como ser exploradas em programas de melhoramento.

A "footprint" of plant carbon fixation cycle functions during the development of a heterotrophic fungus.

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Lyu, Xueliang; Shen, Cuicui; Xie, Jiatao; Fu, Yanping; Jiang, Daohong; Hu, Zijin; Tang, Lihua; Tang, Liguang; Ding, Feng; Li, Kunfei; Wu, Song; Hu, Yanping; Luo, Lilian; Li, Yuanhao; Wang, Qihua; Li, Guoqing; Cheng, Jiasen

2015-08-11

Carbon fixation pathway of plants (CFPP) in photosynthesis converts solar energy to biomass, bio-products and biofuel. Intriguingly, a large number of heterotrophic fungi also possess enzymes functionally associated with CFPP, raising the questions about their roles in fungal development and in evolution. Here, we report on the presence of 17 CFPP associated enzymes (ten in Calvin-Benson-Basham reductive pentose phosphate pathway and seven in C4-dicarboxylic acid cycle) in the genome of Sclerotinia sclerotiorum, a heterotrophic phytopathogenic fungus, and only two unique enzymes: ribulose-1, 5-bisphosphate carboxylase-oxygenase (Rubisco) and phosphoribulokinase (PRK) were absent. This data suggested an incomplete CFPP-like pathway (CLP) in fungi. Functional profile analysis demonstrated that the activity of the incomplete CLP was dramatically regulated during different developmental stages of S. sclerotiorum. Subsequent experiments confirmed that many of them were essential to the virulence and/or sclerotial formation. Most of the CLP associated genes are conserved in fungi. Phylogenetic analysis showed that many of them have undergone gene duplication, gene acquisition or loss and functional diversification in evolutionary history. These findings showed an evolutionary links in the carbon fixation processes of autotrophs and heterotrophs and implicated the functions of related genes were in course of continuous change in different organisms in evolution.

Colonization of lettuce rhizosphere and roots by tagged Streptomyces.

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Bonaldi, Maria; Chen, Xiaoyulong; Kunova, Andrea; Pizzatti, Cristina; Saracchi, Marco; Cortesi, Paolo

2015-01-01

Beneficial microorganisms are increasingly used in agriculture, but their efficacy often fails due to limited knowledge of their interactions with plants and other microorganisms present in rhizosphere. We studied spatio-temporal colonization dynamics of lettuce roots and rhizosphere by genetically modified Streptomyces spp. Five Streptomyces strains, strongly inhibiting in vitro the major soil-borne pathogen of horticultural crops, Sclerotinia sclerotiorum, were transformed with pIJ8641 plasmid harboring an enhanced green fluorescent protein marker and resistance to apramycin. The fitness of transformants was compared to the wild-type strains and all of them grew and sporulated at similar rates and retained the production of enzymes and selected secondary metabolites as well as in vitro inhibition of S. sclerotiorum. The tagged ZEA17I strain was selected to study the dynamics of lettuce roots and rhizosphere colonization in non-sterile growth substrate. The transformed strain was able to colonize soil, developing roots, and rhizosphere. When the strain was inoculated directly on the growth substrate, significantly more t-ZEA17I was re-isolated both from the rhizosphere and the roots when compared to the amount obtained after seed coating. The re-isolation from the rhizosphere and the inner tissues of surface-sterilized lettuce roots demonstrated that t-ZEA17I is both rhizospheric and endophytic.

Transgenic Brassica juncea plants expressing MsrA1, a synthetic cationic antimicrobial peptide, exhibit resistance to fungal phytopathogens.

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Rustagi, Anjana; Kumar, Deepak; Shekhar, Shashi; Yusuf, Mohd Aslam; Misra, Santosh; Sarin, Neera Bhalla

2014-06-01

Cationic antimicrobial peptides (CAPs) have shown potential against broad spectrum of phytopathogens. Synthetic versions with desirable properties have been modeled on these natural peptides. MsrA1 is a synthetic chimera of cecropin A and melittin CAPs with antimicrobial properties. We generated transgenic Brassica juncea plants expressing the msrA1 gene aimed at conferring fungal resistance. Five independent transgenic lines were evaluated for resistance to Alternaria brassicae and Sclerotinia sclerotiorum, two of the most devastating pathogens of B. juncea crops. In vitro assays showed inhibition by MsrA1 of Alternaria hyphae growth by 44-62 %. As assessed by the number and size of lesions and time taken for complete leaf necrosis, the Alternaria infection was delayed and restricted in the transgenic plants with the protection varying from 69 to 85 % in different transgenic lines. In case of S. sclerotiorum infection, the lesions were more severe and spread profusely in untransformed control compared with transgenic plants. The sclerotia formed in the stem of untransformed control plants were significantly more in number and larger in size than those present in the transgenic plants where disease protection of 56-71.5 % was obtained. We discuss the potential of engineering broad spectrum biotic stress tolerance by transgenic expression of CAPs in crop plants.

Transferring of Sclerotinia resistance from wild into cultivated sunflower: Combining of conventional and laboratory techniques

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Vasić Dragana M.

2002-01-01

Full Text Available Five populations of each H. molis, H. maximiliani, H. rigidus and H. tuberosus were screened for resistance to stem form of Sclerotinia. On the basis of the results obtained by screening, nine crosses of resistant populations with either other wild species populations or with cultivated sunflower were made. As in some crosses a small quantity of seed was produced and the seeds germinated poorly, modified tissue culture methods were used to enhance germination and produce clones of interesting plants. These methods were found to be efficient both for seed germination and plant production and multiplication.

SANIDADE DE SEMENTES DE GIRASSOL PROVENIENTES DE TRÊS MUNICÍPIOS DO ESTADO DO MARANHÃO

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Delineide Pereira Gomes

2008-01-01

Full Text Available The increasing importance of sunflower leads to studies on seed pathogen, to guarantee crop sanity and to provide identification of pathogens in new areas. Genotypes seeds lots produced in Embrapa Soja assays carried out in tree cities of the State of Maranhão, Brazil (Balsas, São Luís and Timon were analyzed, with the objective of evaluating sanitary quality of sunflower seeds. Sanitary analysis was performed by blotter test method and identification of fungi genera was based on morphological features. The occurrence of Fusarium sp., Alternaria spp., Curvularia sp., Dreschelera sp., Aspergillus sp., Penicillium sp., Phoma sp., Trichoderma sp., Botrytis sp., Rhizoctonia sp., Rhizopus sp., Colletotrichum sp., Chaetomium sp., Cladosporium sp. and Sclerotinia sclerotiorum was observed in seeds of sunflower, with variable incidences.

Biological control of white mold by Trichoderma harzianum in common bean under field conditions

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Daniel Diego Costa Carvalho

2015-12-01

Full Text Available Abstract: The objective of this work was to evaluate Trichoderma harzianum isolates for biological control of white mold in common bean (Phaseolus vulgaris. Five isolates were evaluated for biocontrol of white mold in 'Perola' common bean under field conditions, in the 2009 and 2010 crop seasons. A commercial isolate (1306 and a control treatment were included. Foliar applications at 2x109 conidia mL-1 were performed at 42 and 52 days after sowing (DAS, in 2009, and at 52 DAS in 2010. The CEN287, CEN316, and 1306 isolates decreased the number of Sclerotinia sclerotiorum apothecia per square meter in comparison to the control, in both crop seasons. CEN287, CEN316, and 1306 decreased white mold severity during the experimental period, when compared to the control.

Desinfestação de substratos para produção de mudas, utilizando vapor de água Disinfesting substrate for transplants production employing hot water steam

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João Bosco C. Silva

2001-07-01

Full Text Available O tratamento sanitário de substratos é uma operação importante no processo de produção de mudas e no cultivo de plantas em vasos ou outros contentores. Tradicionalmente tem-se utilizado o gás brometo de metila como agente desinfetante. Entretanto, a produção deste gás deverá ser abolida até o ano 2010, forçando-se a busca de novas opções. Desenvolveu-se na Embrapa Hortaliças um equipamento que utiliza o vapor de água à baixa pressão, produzido por uma caldeira industrial, com capacidade para evaporar 30 L/h de água, para aquecer o substrato contido em uma caixa metálica cilíndrica com capacidade de 2000 L. O vapor é aplicado no fundo da caixa que contém uma camada de brita coberta com uma tela metálica de malha de 2 mm, que favorece a distribuição uniforme do vapor por toda a massa de substrato. O tempo de aquecimento é de aproximadamente 3 horas e o calor armazenado durante este período mantém a massa de substrato aquecida a temperaturas pasteurizantes, por até 4 horas após a aplicação do vapor. Para testar a eficácia do sistema avaliou-se a sobrevivência dos patógenos Ralstonia solanacearum, Fusarium oxysporum, Sclerotinia sclerotiorum e Rhizoctonia solani. Aplicou-se vapor por uma hora, não considerando o período de aquecimento, e coletaram-se as amostras após uma, duas, três ou quatro horas o início da aplicação de vapor. O tratamento por uma hora, em adição ao período de aquecimento, resultou na eliminação dos patógenos.The disinfestation of substrate is an important process for transplanting production and for plant cultivation in pots or boxes. Traditionally, metyl bromide gas has been employed to eliminate microorganisms. However the production of bromide gas in Brazil will be interrupted by the year 2010 and it is necessary to search for new options. We have devised an equipment that utilizes hot steam water at low pressure produced into a boiler machine with the capacity of evaporating

Colonization of lettuce rhizosphere and roots by tagged Streptomyces

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Maria eBonaldi

2015-02-01

Full Text Available Beneficial microorganisms are increasingly used in agriculture, but their efficacy often fails due to limited knowledge of their interactions with plants and other microorganisms present in rhizosphere. We studied spatio-temporal colonization dynamics of lettuce roots and rhizosphere by genetically modified Streptomyces spp. Five Streptomyces strains, strongly inhibiting in vitro the major soil-borne pathogen of horticultural crops, Sclerotinia sclerotiorum, were transformed with pIJ8641 plasmid harboring an enhanced green fluorescent protein marker and resistance to apramycin. The fitness of transformants was compared to the wild-type strains and all of them grew and sporulated at similar rates and retained the production of enzymes and selected secondary metabolites as well as in vitro inhibition of S. sclerotiorum. The tagged ZEA17I strain was selected to study the dynamics of lettuce roots and rhizosphere colonization in non-sterile growth substrate. The transformed strain was able to colonize soil, developing roots and rhizosphere. When the strain was inoculated directly on the growth substrate, significantly more t-ZEA17I was re-isolated both from the rhizosphere and the roots when compared to the amount obtained after seed coating. The re-isolation from the rhizosphere and the inner tissues of surface-sterilized lettuce roots demonstrated that t-ZEA17I is both rhizospheric and endophytic.

The effect of irrigation and foliar fertilization on the colonization of american ginseng (Panax quinquefolium l. diseased parts by different micro-organisms

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Alina Pastucha

2012-12-01

Full Text Available Field studies on the health of American ginseng cultivated in the Lublin district on poor sandy soil were conducted in the years 2004-2006. The studies involved treatment combinations with irrigation and without irrigation as well as foliar fertilization with Alkalin PK and Resistim of American ginseng plants. Mycological analysis was made of diseased ginseng parts with the aim of determining the quantitative and qualitative composition of fungi-like organisms and fungi threatening the cultivation of this plant. Fungi from the genera of Cylindrocarpon, Fusarium and the following species Alternaria alternata, Rhizoctonia solani, Sclerotinia sclerotiorum, as well as fungi-like organisms: Pythium irregulare and Phytophthora sp., were isolated from the infected parts of ginseng. The smallest number of fungi was isolated from the plants growing on the plots without irrigation and those where foliar application with Alkalin PK was applied.

The Cerato-Platanin protein Epl-1 from Trichoderma harzianum is involved in mycoparasitism, plant resistance induction and self cell wall protection.

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Gomes, Eriston Vieira; Costa, Mariana do Nascimento; de Paula, Renato Graciano; de Azevedo, Rafael Ricci; da Silva, Francilene Lopes; Noronha, Eliane F; Ulhoa, Cirano José; Monteiro, Valdirene Neves; Cardoza, Rosa Elena; Gutiérrez, Santiago; Silva, Roberto Nascimento

2015-12-09

Trichoderma harzianum species are well known as biocontrol agents against important fungal phytopathogens. Mycoparasitism is one of the strategies used by this fungus in the biocontrol process. In this work, we analyzed the effect of Epl-1 protein, previously described as plant resistance elicitor, in expression modulation of T. harzianum genes involved in mycoparasitism process against phytopathogenic fungi; self cell wall protection and recognition; host hyphae coiling and triggering expression of defense-related genes in beans plants. The results indicated that the absence of Epl-1 protein affects the expression of all mycoparasitism genes analyzed in direct confrontation assays against phytopathogen Sclerotinia sclerotiorum as well as T. harzianum itself; the host mycoparasitic coiling process and expression modulation of plant defense genes showing different pattern compared with wild type strain. These data indicated the involvement T. harzianum Epl-1 in self and host interaction and also recognition of T. harzianum as a symbiotic fungus by the bean plants.

Biodegradable and bioactive CGP/PVA film for fungal growth inhibition.

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Silva, Bárbara Dumas S; Ulhoa, Cirano J; Batista, Karla A; Di Medeiros, Maria Carolina; Da Silva Filho, Rômulo Roosevelt; Yamashita, Fabio; Fernandes, Kátia F

2012-07-01

In this study, chitinolytic enzymes produced by Trichoderma asperellum were immobilized on a biodegradable film manufactured with a blend of cashew gum polysaccharide (CGP) and polyvinyl alcohol (PVA), and tested as a fungal growth inhibitor. The film was produced by casting a blend of CGP and PVA solution on glass molds. The CGP/PVA film showed 68% water solubility, tensile strength of 23.7 MPa, 187.2% elongation and 52% of mass loss after 90 days in soil. The presence of T-CWD enzymes immobilized by adsorption or covalent attachment resulted in effective inhibition of fungal growth. Sclerotinia sclerotiorum was the most sensitive organism, followed by Aspergillus niger and Penicillium sp. SEM micrograph showed that the presence of immobilized T-CWD enzymes on CGP/PVA film produced morphological modifications on vegetative and germinative structures of the microorganisms, particularly hyphae disruption and changes of spores shape. Copyright © 2012 Elsevier Ltd. All rights reserved.

Disease control effect of strevertenes produced by Streptomyces psammoticus against tomato fusarium wilt.

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Kim, Jeong Do; Han, Jae Woo; Lee, Sung Chul; Lee, Dongho; Hwang, In Cheon; Kim, Beom Seok

2011-03-09

During screening of microorganisms producing antifungal metabolites, Streptomyces psammoticus strain KP1404 was isolated. The culture extract of this strain showed potent disease control efficacy against Fusarium wilt on tomato plants. The antifungal metabolites ST-1 and ST-2 were isolated from the culture extract using a variety of chromatographic procedures. On the basis of MS and NMR spectrometric analysis, the structures of the antifungal active compounds ST-1 and ST-2 were determined to be the polyene antibiotics strevertene A and strevertene B, respectively. In vitro, strevertenes A and B showed inhibitory effects against the mycelial growth of Alternaria mali , Aspergillus oryzae , Cylindrocarpon destructans , Colletotrichum orbiculare , Fusarium oxysporum f.sp. lycopersici, and Sclerotinia sclerotiorum , even at concentrations of 4-16 μg/mL. Fusarium wilt development on tomato plants was strongly retarded by treatment with 1 μg/mL of these strevertenes. The disease control efficacies of strevertenes on Fusarium wilt were as remarkable as that of benomyl.

Fungal Endophyte Diversity and Bioactivity in the Indian Medicinal Plant Ocimum sanctum Linn.

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Kanika Chowdhary

Full Text Available Endophytic mycopopulation isolated from India's Queen of herbs Tulsi (Ocimum sanctum were explored and investigated for their diversity and antiphytopathogenic activity against widespread plant pathogens Botrytis cinerea, Sclerotinia sclerotiorum, Rhizoctonia solani and Fusarium oxysporum. 90 fungal isolates, representing 17 genera were recovered from 313 disease-free and surface sterilised plant segments (leaf and stem tissues from three different geographic locations (Delhi, Hyderabad and Mukteshwar during distinct sampling times in consequent years 2010 and 2011 in India. Fungal endophytes were subjected to molecular identification based on rDNA ITS sequence analysis. Plant pathogens such as F. verticillioides, B. maydis, C. coarctatum, R. bataticola, Hypoxylon sp., Diaporthe phaseolorum, Alternaria tenuissima and A. alternata have occurred as endophyte only during second sampling (second sampling in 2011 in the present study. Bi-plot generated by principal component analysis suggested tissue specificity of certain fungal endophytes. Dendrogram revealed species abundance as a function of mean temperature of the location at the time of sampling. Shannon diversity in the first collection is highest in Hyderabad leaf tissues (H' = 1.907 whereas in second collection it was highest from leaf tissues of Delhi (H' = 1.846. Mukteshwar (altitude: 7500 feet reported least isolation rate in second collection. Nearly 23% of the total fungal isolates were considered as potent biocontrol agent. Hexane extract of M. phaseolina recovered from Hyderabad in first collection demonstrated highest activity against S. sclerotiorum with IC50 value of 0.38 mg/ml. Additionally, its components 2H-pyran-2-one, 5,6-dihydro-6-pentyl and palmitic acid, methyl ester as reported by GC-MS Chromatogram upon evaluation for their antiphytopathogenic activity exhibited IC50 value of 1.002 and 0.662 against respectively S. sclerotiorum indicating their significant role in

Current Status and Challenges in Identifying Disease Resistance Genes in Brassica napus

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Ting Xiang Neik

2017-11-01

Full Text Available Brassica napus is an economically important crop across different continents including temperate and subtropical regions in Europe, Canada, South Asia, China and Australia. Its widespread cultivation also brings setbacks as it plays host to fungal, oomycete and chytrid pathogens that can lead to serious yield loss. For sustainable crop production, identification of resistance (R genes in B. napus has become of critical importance. In this review, we discuss four key pathogens affecting Brassica crops: Clubroot (Plasmodiophora brassicae, Blackleg (Leptosphaeria maculans and L. biglobosa, Sclerotinia Stem Rot (Sclerotinia sclerotiorum, and Downy Mildew (Hyaloperonospora parasitica. We first review current studies covering prevalence of these pathogens on Brassica crops and highlight the R genes and QTL that have been identified from Brassica species against these pathogens. Insights into the relationships between the pathogen and its Brassica host, the unique host resistance mechanisms and how these affect resistance outcomes is also presented. We discuss challenges in identification and deployment of R genes in B. napus in relation to highly specific genetic interactions between host subpopulations and pathogen pathotypes and emphasize the need for common or shared techniques and research materials or tighter collaboration between researchers to reconcile the inconsistencies in the research outcomes. Using current genomics tools, we provide examples of how characterization and cloning of R genes in B. napus can be carried out more effectively. Lastly, we put forward strategies to breed resistant cultivars through introgressions supported by genomic approaches and suggest prospects that can be implemented in the future for a better, pathogen-resistant B. napus.

Current Status and Challenges in Identifying Disease Resistance Genes in Brassica napus

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Neik, Ting Xiang; Barbetti, Martin J.; Batley, Jacqueline

2017-01-01

Brassica napus is an economically important crop across different continents including temperate and subtropical regions in Europe, Canada, South Asia, China and Australia. Its widespread cultivation also brings setbacks as it plays host to fungal, oomycete and chytrid pathogens that can lead to serious yield loss. For sustainable crop production, identification of resistance (R) genes in B. napus has become of critical importance. In this review, we discuss four key pathogens affecting Brassica crops: Clubroot (Plasmodiophora brassicae), Blackleg (Leptosphaeria maculans and L. biglobosa), Sclerotinia Stem Rot (Sclerotinia sclerotiorum), and Downy Mildew (Hyaloperonospora parasitica). We first review current studies covering prevalence of these pathogens on Brassica crops and highlight the R genes and QTL that have been identified from Brassica species against these pathogens. Insights into the relationships between the pathogen and its Brassica host, the unique host resistance mechanisms and how these affect resistance outcomes is also presented. We discuss challenges in identification and deployment of R genes in B. napus in relation to highly specific genetic interactions between host subpopulations and pathogen pathotypes and emphasize the need for common or shared techniques and research materials or tighter collaboration between researchers to reconcile the inconsistencies in the research outcomes. Using current genomics tools, we provide examples of how characterization and cloning of R genes in B. napus can be carried out more effectively. Lastly, we put forward strategies to breed resistant cultivars through introgressions supported by genomic approaches and suggest prospects that can be implemented in the future for a better, pathogen-resistant B. napus. PMID:29163558

Nucleic adaptability of heterokaryons to fungicides in a multinucleate fungus, Sclerotinia homoeocarpa.

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Kessler, Dylan; Sang, Hyunkyu; Bousquet, Amanda; Hulvey, Jonathan P; Garcia, Dawlyn; Rhee, Siyeon; Hoshino, Yoichiro; Yamada, Toshihiko; Jung, Geunhwa

2018-06-01

Sclerotinia homoeocarpa is the causal organism of dollar spot in turfgrasses and is a multinucleate fungus with a history of resistance to multiple fungicide classes. Heterokaryosis gives rise to the coexistence of genetically distinct nuclei within a cell, which contributes to genotypic and phenotypic plasticity in multinucleate fungi. We demonstrate that field isolates, resistant to either a demethylation inhibitor or methyl benzimidazole carbamate fungicide, can form heterokaryons with resistance to each fungicide and adaptability to serial combinations of different fungicide concentrations. Field isolates and putative heterokaryons were assayed on fungicide-amended media for in vitro sensitivity. Shifts in fungicide sensitivity and microsatellite genotypes indicated that heterokaryons could adapt to changes in fungicide pressure. Presence of both nuclei in heterokaryons was confirmed by detection of a single nucleotide polymorphism in the β-tubulin gene, the presence of microsatellite alleles of both field isolates, and the live-cell imaging of two different fluorescently tagged nuclei using laser scanning confocal microscopy. Nucleic adaptability of heterokaryons to fungicides was strongly supported by the visualization of changes in fluorescently labeled nuclei to fungicide pressure. Results from this study suggest that heterokaryosis is a mechanism by which the pathogen adapts to multiple fungicide pressures in the field. Copyright © 2018 Elsevier Inc. All rights reserved.

RNA-Seq analysis of the Sclerotinia homoeocarpa--creeping bentgrass pathosystem.

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Angela M Orshinsky

Full Text Available Sclerotinia homoeocarpa causes dollar spot disease, the predominate disease on highly-maintained turfgrass. Currently, there are major gaps in our understanding of the molecular interactions between S. homoeocarpa and creeping bentgrass. In this study, 454 sequencing technology was used in the de novo assembly of S. homoeocarpa and creeping bentgrass transcriptomes. Transcript sequence data obtained using Illumina's first generation sequencing-by-synthesis (SBS were mapped to the transcriptome assemblies to estimate transcript representation in different SBS libraries. SBS libraries included a S. homoeocarpa culture control, a creeping bentgrass uninoculated control, and a library for creeping bentgrass inoculated with S. homoeocarpa and incubated for 96 h. A Fisher's exact test was performed to determine transcripts that were significantly different during creeping bentgrass infection with S. homoeocarpa. Fungal transcripts of interest included glycosyl hydrolases, proteases, and ABC transporters. Of particular interest were the large number of glycosyl hydrolase transcripts that target a wide range of plant cell wall compounds, corroborating the suggested wide host range and saprophytic abilities of S. homoeocarpa. Several of the multidrug resistance ABC transporters may be important for resistance to both fungicides and plant defense compounds. Creeping bentgrass transcripts of interest included germins, ubiquitin transcripts involved in proteasome degradation, and cinnamoyl reductase, which is involved in lignin production. This analysis provides an extensive overview of the S. homoeocarpa-turfgrass pathosystem and provides a starting point for the characterization of potential virulence factors and host defense responses. In particular, determination of important host defense responses may assist in the development of highly resistant creeping bentgrass varieties.

Incidência de ocratoxina A em diferentes frações do café (Coffea arabica L.: bóia, mistura e varrição após secagem em terreiros de terra, asfalto e cimento Incidence of ochratoxin A in fraction diferents coffee beans (Coffea Arabica L: "boia", mixes and "varrição"

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Luís Roberto Batista

2007-06-01

Full Text Available A incidência de ocratoxina A foi estudada em café mistura, bóia e varrição secas em três tipos de terreiro: terra, cimento e asfalto. Foram analisadas 238 amostras coletadas em 11 municípios da região sul do Estado de Minas Gerais, sendo 35 bóia, 97 - mistura e 106 varrição. Das amostras analisadas, em 40% não foi detectada a presença de ocratoxina A, em 31%, foram detectadas a presença de ocratoxina A em níveis que variaram de 0,1 a 5,0 µg/Kg de café. Estes resultados demonstram que 169 amostras (71% analisadas estariam dentro dos limites em estudo da Legislação Européia que regulamenta a concentração máxima de ocratoxina A em grãos de café torrado. As espécies de Aspergillus identificadas como produtoras de ocratoxina A foram Aspergillus ochraceus, A. sclerotiorum e A. sulphureus. Os níveis de contaminação de ocratoxina A em grãos de café foram maiores na fração varrição e nas frações bóia e mistura, secas em terreiro de terra. Os resultados deste estudo concluem que o terreiro de terra aumenta o risco de contaminação com ocratoxina A em grãos de café. A fração varrição devido aos riscos de exposição a ocratoxina A, deve ser reduzida através da adoção de boas práticas agrícolas e não ser utilizada para fins de consumo humano e animal.The ochratoxin incidence was studied in coffee it mixes, it "bóia" and "varrição" dry in three yard types: earth, cement and asphalt. 238 samples were analyzed collected in 11 municipal districts of the south of Minas Gerais state, being 35 "bóia", 97 - it mixes and 106 varrição. Of the analyzed samples, in 40% the ochratoxin A presence it was not detected, in 74 samples, 31%, ochratoxin A presence were detected the in levels that varied from 0,1 to 5,0 µg/Kg of coffee beans. These results demonstrate that in 169 samples (71% analyzed they would be inside of the limits in study of the European Legislation that regulates the maximum concentration of

Sitios de infección por hongos más frecuentes en la zanahoria (Daucus carota L. y patogenicidad en sus diferentes tejidos.

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German Rivera

2016-03-01

Full Text Available Fungi from carrot roots collected in the field local market were isolated and identified. Sites of natural infection on the root were recorded. Sixteen genera were identified and their pathogenicity tested by  placing them on: not wounded periderm, pericyclic parenchyma, phloem parenchyma and xylem parenchyma. Half of the total fungi was isolated in both field and market samples. The most frequent natural infection sites were the crown, end of the tap root, and lateral roots. In the pathogenicity tests a gradient of resistance was shown varying from a very high level at periderm to a low level at the xylem parenchyma.  Sclerotinia sclerotiorum,  Sclerotium rolfsii, Fusarium tricinctum, F. nivale, F. solani, F. oxysporum, Fusarium sp., Trichoderma sp. y Gliocladium sp., all penetrated directly throught the periderm and also infected the other tissues. Geotrivhum candidum Rhizopus stolonifer . Verticillium sp., Penicillium so., and Candida sp. did no intect the periderm, but were pathogenic  to the pericyclic, phloem, and xylem parenchyma. Phoma sp. and Mucor sp. only infected phloem and xylem parenchyma.

Etiology of root parsley damping-off

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Bogdan Nowicki

2013-12-01

Full Text Available The investigations were done between 1990-1994. Seedlings collected from 120 plantations were evaluated. The fungi responsible for seedling damping-off occurrening most often were Alternariu spp., Fusarium spp. and Pythium spp. isolated from 46,3, 32,2 and 16,6% of infected plants, respectively. The most important pathogens were A.petroselini which infected 33% of seedlings and A.radicina - 11%„ Among Fusarium species the most common was F.avenaceum, comprising 61% of total Fusarium isolates. The next were following: F.culmorum - 21%, F.solani - 12,6% and 3% for both F.equiseti and F.oxysporum. Damping-off of se,edlings was also caused by the other fungi but they were noted in low intensity. Among them were following: Phoma spp., A.alternata and Rhizoctonia solani on 2,8; 2,3 and 1,2% of tested seedlings. respectively. The species: Sclerotinia sclerotiorum, Botrytis cinerea, Bipolaris sorokinianu and Septoria petroselini were isolated in total from 0,9% of seedlings. Drechslera biseptata and Stemphylium botryosum caused seedling damping-off sporadically.

Mycopopulation of marshmallow (Althaea officinalis L

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Pavlović Snežana Đ.

2007-01-01

Full Text Available Marshmallow is an important medicinal plant in Serbia. Because of increasing demands on the market, cultivation has been started. Through regular quality control of commercial seeds and plantations, mycopopulation of marshmallow was recorded in the period 2000-2006. Seeds of marshmallow were dominated by Alternaria alternata and species from the genus Fusarium (Fusarium verticillioides, F. proliferatum, F. semitectum, F. oxysporum and F. solani. Species belonging to Fusarium genus are the cause of rot of seeds and roots of marshmallow, causing chlorosis and fading, and therefore deterioration and necrosis of plants, as well as decrease of seed germination of seeds. Leaves and stalks of marshmallow were from time to time under massive attack of Puccinia malvacearum, and that was the reason why leaves were unuseful as a herbal drug. On roots and lower part of the stalks, massive appearance of Sclerotinia sclerotiorum, a causal agent of the white rot, in cases when marshmallow was cultivated after sunflower, was recorded, too. From other fungi in roots, species belonging to the genus Fusarium (F. oxysporum, F. solani and F. verticillioides were dominant.

Toprak Solucanlarından Elde Edilen Vermikompostun Bazı Bitki Patojenleri Üzerindeki Antimikrobiyal Aktivitelerinin Araştırılması

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Uğur Tutar

2012-12-01

Full Text Available Toprak solucanlarının, organik atıkları biyolojik olarak parçalayarak ayrıştırmaları ile oluşturdukları “vemikompost” un, bazı patojen bakteri ve funguslara karşı etkili oldukları yapılan çeşitli araştırmalarla saptanmıştır. Bu çalışmada, Eisenia fetida türü toprak solucanlarından elde edilen vermikompostun; etanol ve kloroform solventleri kullanılarak elde edilen ekstrelerinin, bitkilerde hastalıklara neden olan toprak kaynaklı patojen 9 adet bakteri ve 9 adet fungusa karşı etkinliklerinin belirlenmesi amacıyla “disk difüzyon” ve “MIC” testleri uygulanmıştır. Çalışma sonuçlarına göre, toprak solucanlarından elde edilen vermikompostun kloroform ile elde edilen ekstrelerinin Pseudomonas syringae, Xhantomonas carotae, Sclerotinia sclerotiorum, Fusarim oxysporum, Aspergillus humicola ve Aspergillus fumigatus’ a karşı etkileri güçlü olurken Erwinia chrysanthemi, Pseudomonas fluorescens, ve Penicillium brevicompactum’ a karşı etkilerinin daha zayıf olduğu görülmüştür. Vermikompostun etanol ile elde edilen ekstrelerinin ise Pseudomonas syringae, Xhantomonas campestris ve Aspergillus fumigatus’ a karşı etkilerinin güçlü olduğu, Erwinia herbicola, Erwinia chrysanthemi ve Sclerotinia sclerotiorum’ a  karşı ise daha zayıf bir etki gösterdiği saptanmıştır.

Triazole Fungicides Sensitivity of Sclerotinia homoeocarpa in Korean Golf Courses

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Ji Won Lee

2017-12-01

Full Text Available Chemical management of dollar spot in turf may lead to the development of Sclerotinia homoeocarpa populations with reduced fungicide sensitivity. The objective of this study was to investigate resistance of S. homoeocarpa isolates to triazole fungicides and to test cross-resistance among three triazole fungicides. A total of 66 isolates of S. homoeocarpa were collected from 15 golf courses across Korea, and tested via in vitro sensitivity assay against hexaconazole, propiconazole and tebuconazole. EC₅₀ values of the isolates to these fungicides were distributed in the range of 0.001–1.1 a. i. μg ml−1. Based on the EC₅₀ values, twelve representative strains were selected as sensitive isolates including control and insensitive isolates with respect to each fungicide. At a concentration of 0.1 a. i. μg ml−1 for all fungicides, the selected strains were distinguished as sensitive or resistant isolates with the mycelial growth inhibition rate of 50% as the criterion. The EC₅₀ values of resistant strains exposed to hexaconazole, propiconazole and tebuconazole were 20–50 times, 50–70 times, and 77 times greater, respectively, than that of the control strains. Two isolates of S. homoeocarpa S0–41 and Sh14-2-1 showed sensitivity toward all the fungicides used, while two other isolates Sh7-5-1 and Sh2-1-1 showed resistance to all fungicides. Each isolate showed similar resistance to the three types of triazole fungicides, whereby cross-resistance of isolates was confirmed in the present study; all three triazole fungicide combinations displayed significant correlation coefficients equivalent to or greater than 0.8.

Evidence for Genetic Similarity of Vegetative Compatibility Groupings in Sclerotinia homoeocarpa

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Seog Won Chang

2014-12-01

Full Text Available Vegetative compatibility groups (VCGs are determined for many fungi to test for the ability of fungal isolates to undergo heterokaryon formation. In several fungal plant pathogens, isolates belonging to a VCG have been shown to share significantly higher genetic similarity than those of different VCGs. In this study we sought to examine the relationship between VCG and genetic similarity of an important cool season turfgrass pathogen, Sclerotinia homoeocarpa. Twenty-two S. homoeocarpa isolates from the Midwest and Eastern US, which were previously characterized in several studies, were all evaluated for VCG using an improved nit mutant assay. These isolates were also genotyped using 19 microsatellites developed from partial genome sequence of S. homoeocarpa. Additionally, partial sequences of mitochondrial genes cytochrome oxidase II and mitochondrial small subunit (mtSSU rRNA, and the atp6-rns intergenic spacer, were generated for isolates from each nit mutant VCG to determine if mitochondrial haplotypes differed among VCGs. Of the 22 isolates screened, 15 were amenable to the nit mutant VCG assay and were grouped into six VCGs. The 19 microsatellites gave 57 alleles for this set. Unweighted pair group methods with arithmetic mean (UPGMA tree of binary microsatellite data were used to produce a dendrogram of the isolate genotypes based on microsatellite alleles, which showed high genetic similarity of nit mutant VCGs. Analysis of molecular variance of microsatellite data demonstrates that the current nit mutant VCGs explain the microsatellite genotypic variation among isolates better than the previous nit mutant VCGs or the conventionally determined VCGs. Mitochondrial sequences were identical among all isolates, suggesting that this marker type may not be informative for US populations of S. homoeocarpa.

Antagonistic studies and hyphal interactions of the new antagonist Aspergillus piperis against some phytopathogenic fungi in vitro in comparison with Trichoderma harzianum.

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El-Debaiky, Samah A

2017-12-01

The present study represents, for the first time, the detailed studies about the hyphal interactions of Aspergillus piperis, as a new antagonist, against some isolated plant pathogenic fungi (Alternaria alternata, Alternaria solani, Botrytis cinerea, Sclerotium cepivorum and Sclerotinia sclerotiorum) in vitro. The bio-controlling capability of A. piperis against the tested phytopathogens was tested using the dual culture method. This experiment revealed that A. piperis had antagonistic activity and reduced the growth of the tested phytopathogens and grew over their mycelia in the paired plates. Also, several antagonistic mechanisms were recorded, in this study, between A. piperis and the tested phytopathogens using the microscopic examination. The bio-controlling activity and the antagonistic mechanisms exhibited by the new antagonist, A. piperis were compared with those obtained by the common antagonist, Trichoderma harzianum against the same phytopathogens. The obtained results showed that, A. piperis was more effective than T. harzianum in inhibiting all the tested species in the dual culture plates. The best result was 81.85% inhibition percentage against S. sclerotiorum by A. piperis while, T. harzianum exhibits only 45.18%. Moreover, several antagonistic mechanisms and hyphal interactions were investigated among the hyphae of both A.piperis and T. harzianum and the hyphae of the tested phytopathogens. These mechanisms were summarized as; mycoparasitism (coiling and penetration of the hyphae) and antibiosis in the form of lysis of the hyphal cells and spores, denaturation and breaking of the hyphae. The indirect interaction (antibiosis) and the direct mycoparasitism were observed by A. piperis against all the tested phytopathogens, but it attacked the hyphae and conidiophores of A. alternata by only the antibiosis interaction. The microscopic examination revealed also that T. harzianum attacked the tested phytopathogens by both antibiosis and mycoparasitism

Reactive Oxygen Species Play a Role in the Infection of the Necrotrophic Fungi, Rhizoctonia solani in Wheat.

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Foley, Rhonda C; Kidd, Brendan N; Hane, James K; Anderson, Jonathan P; Singh, Karam B

2016-01-01

Rhizoctonia solani is a nectrotrophic fungal pathogen that causes billions of dollars of damage to agriculture worldwide and infects a broad host range including wheat, rice, potato and legumes. In this study we identify wheat genes that are differentially expressed in response to the R. solani isolate, AG8, using microarray technology. A significant number of wheat genes identified in this screen were involved in reactive oxygen species (ROS) production and redox regulation. Levels of ROS species were increased in wheat root tissue following R. solani infection as determined by Nitro Blue Tetrazolium (NBT), 3,3'-diaminobenzidine (DAB) and titanium sulphate measurements. Pathogen/ROS related genes from R. solani were also tested for expression patterns upon wheat infection. TmpL, a R. solani gene homologous to a gene associated with ROS regulation in Alternaria brassicicola, and OAH, a R. solani gene homologous to oxaloacetate acetylhydrolase which has been shown to produce oxalic acid in Sclerotinia sclerotiorum, were highly induced in R. solani when infecting wheat. We speculate that the interplay between the wheat and R. solani ROS generating proteins may be important for determining the outcome of the wheat/R. solani interaction.

Reactive Oxygen Species Play a Role in the Infection of the Necrotrophic Fungi, Rhizoctonia solani in Wheat.

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Rhonda C Foley

Full Text Available Rhizoctonia solani is a nectrotrophic fungal pathogen that causes billions of dollars of damage to agriculture worldwide and infects a broad host range including wheat, rice, potato and legumes. In this study we identify wheat genes that are differentially expressed in response to the R. solani isolate, AG8, using microarray technology. A significant number of wheat genes identified in this screen were involved in reactive oxygen species (ROS production and redox regulation. Levels of ROS species were increased in wheat root tissue following R. solani infection as determined by Nitro Blue Tetrazolium (NBT, 3,3'-diaminobenzidine (DAB and titanium sulphate measurements. Pathogen/ROS related genes from R. solani were also tested for expression patterns upon wheat infection. TmpL, a R. solani gene homologous to a gene associated with ROS regulation in Alternaria brassicicola, and OAH, a R. solani gene homologous to oxaloacetate acetylhydrolase which has been shown to produce oxalic acid in Sclerotinia sclerotiorum, were highly induced in R. solani when infecting wheat. We speculate that the interplay between the wheat and R. solani ROS generating proteins may be important for determining the outcome of the wheat/R. solani interaction.

Antifungal activity of indigenous bacillus sp. isolate Q3 against marshmallow mycobiota

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Jošić Dragana Lj.

2011-01-01

Full Text Available Marshmallow is a host of a number of saprophytic and parasitic fungi in Serbia. The seeds of marshmallow are contaminated with fungi from different genera, especially Alternaria and Fusarium, which significantly reduced seed germination and caused seedling decay. In this study we investigate antagnonism of indigenous Bacillus sp. isolate Q3 against marshmallow mycopopulation. Bacillus sp. Q3 was isolated from maize rhizosphere, characterized by polyphasic approch and tested for plant growth promoting treats. Bacillus sp. Q3 produced antifungal metabolites with growth inhibition activity against numerous fungi in dual culture: 61.8% of Alternaria alternata, 74.8% of Myrothecium verrucaria and 33.6% of Sclerotinia sclerotiorum. That effect could be caused by different antifungal metabolites including siderophores, hydrolytic enzymes, organic acids and indole acetic acid (IAA. Suppression of natural marshmallow seed infection by Q3 isolate was observed. The seeds were immersed in different concentrations of bacterial suspension during 2h and their infections by phytopathogenic fungi were estimated. The results showed significant reduction of seed infection by Alternaria spp. The presented results indicate possible application of this isolate as promising biological agent for control of marshmallow seed pathogenic fungi.

Contribution of proteomics to the study of plant pathogenic fungi.

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Gonzalez-Fernandez, Raquel; Jorrin-Novo, Jesus V

2012-01-01

Phytopathogenic fungi are one of the most damaging plant parasitic organisms, and can cause serious diseases and important yield losses in crops. The study of the biology of these microorganisms and the interaction with their hosts has experienced great advances in recent years due to the development of moderm, holistic and high-throughput -omic techniques, together with the increasing number of genome sequencing projects and the development of mutants and reverse genetics tools. We highlight among these -omic techniques the importance of proteomics, which has become a relevant tool in plant-fungus pathosystem research. Proteomics intends to identify gene products with a key role in pathogenicity and virulence. These studies would help in the search of key protein targets and in the development of agrochemicals, which may open new ways for crop disease diagnosis and protection. In this review, we made an overview on the contribution of proteomics to the knowledge of life cycle, infection mechanisms, and virulence of the plant pathogenic fungi. Data from current, innovative literature, according to both methodological and experimental systems, were summarized and discussed. Specific sections were devoted to the most studied fungal phytopathogens: Botrytis cinerea, Sclerotinia sclerotiorum, and Fusarium graminearum.

A Novel Fungal Metabolite with Beneficial Properties for Agricultural Applications

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Francesco Vinale

2014-07-01

Full Text Available Trichoderma are ubiquitous soil fungi that include species widely used as biocontrol agents in agriculture. Many isolates are known to secrete several secondary metabolites with different biological activities towards plants and other microbes. Harzianic acid (HA is a T. harzianum metabolite able to promote plant growth and strongly bind iron. In this work, we isolated from the culture filtrate of a T. harzianum strain a new metabolite, named isoharzianic acid (iso-HA, a stereoisomer of HA. The structure and absolute configuration of this compound has been determined by spectroscopic methods, including UV-Vis, MS, 1D and 2D NMR analyses. In vitro applications of iso-HA inhibited the mycelium radial growth of Sclerotinia sclerotiorum and Rhizoctonia solani. Moreover, iso HA improved the germination of tomato seeds and induced disease resistance. HPLC-DAD experiments showed that the production of HA and iso HA was affected by the presence of plant tissue in the liquid medium. In particular, tomato tissue elicited the production of HA but negatively modulated the biosynthesis of its analogue iso-HA, suggesting that different forms of the same Trichoderma secondary metabolite have specific roles in the molecular mechanism regulating the Trichoderma plant interaction.

A novel fungal metabolite with beneficial properties for agricultural applications.

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Vinale, Francesco; Manganiello, Gelsomina; Nigro, Marco; Mazzei, Pierluigi; Piccolo, Alessandro; Pascale, Alberto; Ruocco, Michelina; Marra, Roberta; Lombardi, Nadia; Lanzuise, Stefania; Varlese, Rosaria; Cavallo, Pierpaolo; Lorito, Matteo; Woo, Sheridan L

2014-07-08

Trichoderma are ubiquitous soil fungi that include species widely used as biocontrol agents in agriculture. Many isolates are known to secrete several secondary metabolites with different biological activities towards plants and other microbes. Harzianic acid (HA) is a T. harzianum metabolite able to promote plant growth and strongly bind iron. In this work, we isolated from the culture filtrate of a T. harzianum strain a new metabolite, named isoharzianic acid (iso-HA), a stereoisomer of HA. The structure and absolute configuration of this compound has been determined by spectroscopic methods, including UV-Vis, MS, 1D and 2D NMR analyses. In vitro applications of iso-HA inhibited the mycelium radial growth of Sclerotinia sclerotiorum and Rhizoctonia solani. Moreover, iso HA improved the germination of tomato seeds and induced disease resistance. HPLC-DAD experiments showed that the production of HA and iso HA was affected by the presence of plant tissue in the liquid medium. In particular, tomato tissue elicited the production of HA but negatively modulated the biosynthesis of its analogue iso-HA, suggesting that different forms of the same Trichoderma secondary metabolite have specific roles in the molecular mechanism regulating the Trichoderma plant interaction.

Efficient Synthesis and Bioactivity of Novel Triazole Derivatives.

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Hu, Boyang; Zhao, Hanqing; Chen, Zili; Xu, Chen; Zhao, Jianzhuang; Zhao, Wenting

2018-03-21

Triazole pesticides are organic nitrogen-containing heterocyclic compounds, which contain 1,2,3-triazole ring. In order to develop potential glucosamine-6-phosphate synthase (GlmS) inhibitor fungicides, forty compounds of triazole derivatives were synthesized in an efficient way, thirty nine of them were new compounds. The structures of all the compounds were confirmed by high resolution mass spectrometer (HRMS), ¹H-NMR and 13 C-NMR. The fungicidal activities results based on means of mycelium growth rate method indicated that some of the compounds exhibited good fungicidal activities against P. CapasiciLeonian , Sclerotinia sclerotiorum (Lib.) de Bary, Pyricularia oryzae Cav. and Fusarium oxysporum Schl. F.sp. vasinfectum (Atk.) Snyd. & Hans. at the concentration of 50 µg/mL, especially the inhibitory rates of compounds 1-d and 1-f were over 80%. At the same time, the preliminary studies based on the Elson-Morgan method indicated that the compounds exhibited some inhibitory activity toward glucosamine-6-phosphate synthase (GlmS). These compounds will be further studied as potential antifungal lead compounds. The structure-activity relationships (SAR) were discussed in terms of the effects of the substituents on both the benzene and the sugar ring.

Efficient Synthesis and Bioactivity of Novel Triazole Derivatives

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Boyang Hu

2018-03-01

Full Text Available Triazole pesticides are organic nitrogen-containing heterocyclic compounds, which contain 1,2,3-triazole ring. In order to develop potential glucosamine-6-phosphate synthase (GlmS inhibitor fungicides, forty compounds of triazole derivatives were synthesized in an efficient way, thirty nine of them were new compounds. The structures of all the compounds were confirmed by high resolution mass spectrometer (HRMS, 1H-NMR and 13C-NMR. The fungicidal activities results based on means of mycelium growth rate method indicated that some of the compounds exhibited good fungicidal activities against P. CapasiciLeonian, Sclerotinia sclerotiorum (Lib. de Bary, Pyricularia oryzae Cav. and Fusarium oxysporum Schl. F.sp. vasinfectum (Atk. Snyd. & Hans. at the concentration of 50 µg/mL, especially the inhibitory rates of compounds 1-d and 1-f were over 80%. At the same time, the preliminary studies based on the Elson-Morgan method indicated that the compounds exhibited some inhibitory activity toward glucosamine-6-phosphate synthase (GlmS. These compounds will be further studied as potential antifungal lead compounds. The structure-activity relationships (SAR were discussed in terms of the effects of the substituents on both the benzene and the sugar ring.

Host Resistance and Chemical Control for Management of Sclerotinia Stem Rot of Soybean in Ohio.

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Huzar-Novakowiski, Jaqueline; Paul, Pierce A; Dorrance, Anne E

2017-08-01

Recent outbreaks of Sclerotinia stem rot (SSR) of soybean in Ohio, along with new fungicides and cultivars with resistance to this disease, have led to a renewed interest in studies to update disease management guidelines. The effect of host resistance (in moderately resistant [MR] and moderately susceptible [MS] cultivars) and chemical control on SSR and yield was evaluated in 12 environments from 2014 to 2016. The chemical treatments evaluated were an untreated check, four fungicides (boscalid, picoxystrobin, pyraclostrobin, and thiophanate-methyl), and one herbicide (lactofen) applied at soybean growth stage R1 (early flowering) alone or at R1 followed by a second application at R2 (full flowering). SSR developed in 6 of 12 environments, with mean disease incidence in the untreated check of 2.5 to 41%. The three environments with high levels of SSR (disease incidence in the untreated check >20%) were used for further statistical analysis. There were significant effects (P Pyraclostrobin increased SSR compared with the untreated check in the three environments with high levels of disease. In the six fields where SSR did not develop, chemical treatment did not increase yield, nor was the yield from the MR cultivar significantly different from the MS cultivar. For Ohio, MR cultivars alone were effective for management of SSR in soybean fields where this disease has historically occurred.

The Shewanella algae strain YM8 produces volatiles with strong inhibition activity against Aspergillus pathogens and aflatoxins

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Andong eGong

2015-10-01

Full Text Available Aflatoxigenic Aspergillus fungi and associated aflatoxins are ubiquitous in the production and storage of food/feed commodities. Controlling these pests is a challenge. In this study, the Shewanella algae strain YM8 was found to produce volatiles that have strong antifungal activity against Aspergillus pathogens. Gas chromatography-mass spectrometry profiling revealed 15 volatile organic compounds (VOCs emitted from YM8, of which dimethyl trisulfide was the most abundant. We obtained authentic reference standards for six of the VOCs; these all significantly reduced mycelial growth and conidial germination in Aspergillus; dimethyl trisulfide and 2,4-bis(1,1-dimethylethyl-phenol showed the strongest inhibitory activity. YM8 completely inhibited Aspergillus growth and aflatoxin biosynthesis in maize and peanut samples stored at different water activity levels, and scanning electron microscopy revealed severely damaged conidia and a complete lack of mycelium development and conidiogenesis. YM8 also completely inhibited the growth of eight other agronomically important species of phytopathogenic fungi: A. parasiticus, A. niger, Alternaria alternate, Botrytis cinerea, Fusarium graminearum, Fusarium oxysporum, Monilinia fructicola, and Sclerotinia sclerotiorum. This study demonstrates the susceptibility of Aspergillus and other fungi to VOCs from marine bacteria and indicates a new strategy for effectively controlling these pathogens and the associated mycotoxin production in the field and during storage.

Effect of polyacetylenic acids from Prunella vulgaris on various plant pathogens.

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Yoon, M-Y; Choi, G J; Choi, Y H; Jang, K S; Park, M S; Cha, B; Kim, J-C

2010-11-01

This study is aiming at characterizing antifungal substances from the methanol extract of Prunella vulgaris and at investigating those substances' antifungal and antioomycete activities against various plant pathogens. Two polyacetylenic acids were isolated from P. vulgaris as active principles and identified as octadeca-9,11,13-triynoic acid and trans-octadec-13-ene-9,11-diynoic acid. These two compounds inhibited the growth of Magnaporthe oryzae, Rhizoctonia solani, Phytophthora infestans, Sclerotinia sclerotiorum, Fusarium oxysporum f. sp. raphani, and Phytophthora capsici. In addition, these two compounds and the wettable powder-type formulation of an n-hexane fraction of P. vulgaris significantly suppressed the development of rice blast, tomato late blight, wheat leaf rust, and red pepper anthracnose. These data show that the extract of P. vulgaris and two polyacetylenic acids possess antifungal and antioomycete activities against a broad spectrum of tested plant pathogens. This is the first report on the occurrence of octadeca-9,11,13-triynoic acid and trans-octadec-13-ene-9,11-diynoic acid in P. vulgaris and their efficacy against plant diseases. The crude extract containing the two polyacetylenic acids can be used as a natural fungicide for the control of various plant diseases. © 2010 The Authors. © 2010 The Society for Applied Microbiology.

Present status of the development of mycoherbicides against water hyacinth: successes and challenges. A review

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Jijakli, MH.

2012-01-01

Full Text Available Recent trends in the implementation of bioherbicide use in the control of water hyacinth (Eichhornia crassipes [Martius] Solms Laubach have depended primarily on several strategies. The use of bioherbicides has been stimulated as part of the search for alternatives to chemical control, as the use of these more environmentally-friendly formulations minimizes hazards resulting from herbicide residue to both human and animal health, and to the ecology. In addition, one of the major strategies in the concept of biological control is the attempt to incorporate biological weed control methods as a component of integrated weed management, in order to achieve satisfactory results while reducing herbicide application to a minimum. Several fungal pathogens with mycoherbicide potential (Sclerotinia sclerotiorum in Hyakillä and Cercospora rodmanii, named ABG-5003 have been discovered on diseased water hyacinth plants, but none has become commercially available in the market. Biological, technological, and commercial constraints have hindered progress in this area. Many of these constraints are being addressed, but there is a critical need to better understand the biochemical and physiological data regarding the pathogenesis of these new bioherbicides. Oil emulsions are recognized as a way to increase both efficiency of application and efficacy of biocontrol agents.

Antifungal activity, main active components and mechanism of Curcuma longa extract against Fusarium graminearum

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Zhang, Fusheng; Chen, Qin; Chen, Cheng; Yu, Xiaorui; Liu, Qingya; Bao, Jinku

2018-01-01

Curcuma longa possesses powerful antifungal activity, as demonstrated in many studies. In this study, the antifungal spectrum of Curcuma longa alcohol extract was determined, and the resulting EC50 values (mg/mL) of its extract on eleven fungi, including Fusarium graminearum, Fusarium chlamydosporum, Alternaria alternate, Fusarium tricinctum, Sclerotinia sclerotiorum, Botrytis cinerea, Fusarium culmorum, Rhizopus oryzae, Cladosporium cladosporioides, Fusarium oxysporum and Colletotrichum higginsianum, were 0.1088, 0.1742, 0.1888, 0.2547, 0.3135, 0.3825, 0.4229, 1.2086, 4.5176, 3.8833 and 5.0183, respectively. Among them, F. graminearum was selected to determine the inhibitory effects of the compounds (including curdione, isocurcumenol, curcumenol, curzerene, β-elemene, curcumin, germacrone and curcumol) derived from Curcuma longa. In addition, the antifungal activities of curdione, curcumenol, curzerene, curcumol and isocurcumenol and the synergies of the complexes of curdione and seven other chemicals were investigated. Differential proteomics of F. graminearum was also compared, and at least 2021 reproducible protein spots were identified. Among these spots, 46 were classified as differentially expressed proteins, and these proteins are involved in energy metabolism, tRNA synthesis and glucose metabolism. Furthermore, several fungal physiological differences were also analysed. The antifungal effect included fungal cell membrane disruption and inhibition of ergosterol synthesis, respiration, succinate dehydrogenase (SDH) and NADH oxidase. PMID:29543859

Sunflower Hybrid Breeding: From Markers to Genomic Selection.

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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

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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

Antifungal mechanisms of ZnO and Ag nanoparticles to Sclerotinia homoeocarpa

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Li, Junli; Sang, Hyunkyu; Guo, Huiyuan; Popko, James T.; He, Lili; White, Jason C.; Parkash Dhankher, Om; Jung, Geunhwa; Xing, Baoshan

2017-04-01

Fungicides have extensively been used to effectively combat fungal diseases on a range of plant species, but resistance to multiple active ingredients has developed in pathogens such as Sclerotinia homoeocarpa, the causal agent of dollar spot on cool-season turfgrasses. Recently, ZnO and Ag nanoparticles (NPs) have received increased attention due to their antimicrobial activities. In this study, the NPs’ toxicity and mechanisms of action were investigated as alternative antifungal agents against S. homoeocarpa isolates that varied in their resistance to demethylation inhibitor (DMI) fungicides. S. homoeocarpa isolates were treated with ZnO NPs and ZnCl2 (25-400 μg ml-1) and Ag NPs and AgNO3 (5-100 μg ml-1) to test antifungal activity of the NPs and ions. The mycelial growth of S. homoeocarpa isolates regardless of their DMI sensitivity was significantly inhibited on ZnO NPs (≥200 μg ml-1), Ag NPs (≥25 μg ml-1), Zn2+ ions (≥200 μg ml-1), and Ag+ ions (≥10 μg ml-1) amended media. Expression of stress response genes, glutathione S-transferase (Shgst1) and superoxide dismutase 2 (ShSOD2), was significantly induced in the isolates by exposure to the NPs and ions. In addition, a significant increase in the nucleic acid contents of fungal hyphae, which may be due to stress response, was observed upon treatment with Ag NPs using Raman spectroscopy. We further observed that a zinc transporter (Shzrt1) might play an important role in accumulating ZnO and Ag NPs into the cells of S. homoeocarpa due to overexpression of Shzrt1 significantly induced by ZnO or Ag NPs within 3 h of exposure. Yeast mutants complemented with Shzrt1 became more sensitive to ZnO and Ag NPs as well as Zn2+ and Ag+ ions than the control strain and resulted in increased Zn or Ag content after exposure. This is the first report of involvement of the zinc transporter in the accumulation of Zn and Ag from NP exposure in filamentous plant pathogenic fungi. Understanding the molecular

AISLAMIENTO DE CEPAS NATIVAS DE TRICHODERMA SPP DE SUELOS HORTICOLAS DEL VALLE DE TOLUCA, COMO BIOCONTROL POTENCIAL DE SCLEROTINIA

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Hilda G. García-Nuñez

2012-08-01

Full Text Available Se evaluó la presencia de Trichoderma en siete localidades en la zona sur del Valle de Toluca, Estado de México. Esta es un área con un alto potencial en la producción de hortalizas. El estudio se dirigió al aislamiento de cepas nativas de Trichoderma a partir de muestras de suelo, identificación de factores fisiográficos, así como las propiedades físicas y químicas del suelo que determinan la ocurrencia de Trichoderma. Se evaluó el potencial de las cepas de Trichoderma para el control de Sclerotinia spp., hongo patógeno causante de la pudrición blanda en lechuga. Se aislaron once cepas, el mayor número de ellas asociadas al tipo de suelo se encontró en las localidades de San Francisco Putla y San Francisco Tetetla. El análisis de regresión logística mostró que no hay una relación entre las propiedades del suelo (materia orgánica y pH y la presencia de Trichoderma. La prueba de Tukey (p

In vitro attachment of phylloplane yeasts to Botrytis cinerea, Rhizoctonia solani, and Sclerotinia homoeocarpa.

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Allen, Tom W; Burpee, Leon L; Buck, James W

2004-12-01

The ability of yeasts to attach to hyphae or conidia of phytopathogenic fungi has been speculated to contribute to biocontrol activity on plant surfaces. Attachment of phylloplane yeasts to Botrytis cinerea, Rhizoctonia solani, and Sclerotinia homoeocarpa was determined using in vitro attachment assays. Yeasts were incubated for 2 d on potato dextrose agar (PDA) prior to experimentation. A total of 292 yeasts cultured on PDA were screened for their ability to attach to conidia of B. cinerea; 260 isolates (89.1%) attached to conidia forming large aggregates of cells, and 22 isolates (7.5%) weakly attached to conidia with 1 or 2 yeast cells attached to a few conidia. Ten yeasts (3.4%), including 8 isolates of Cryptococcus laurentii, 1 isolate of Cryptococcus flavescens, and an unidentified species of Cryptococcus, failed to attach to conidia. All non-attaching yeasts produced copious extracellular polysaccharide (EPS) on PDA. Seventeen yeast isolates did not attach to hyphal fragments of B. cinerea, R. solani, and S. homoeocarpa after a 1 h incubation, but attachment was observed after 24 h. Culture medium, but not culture age, significantly affected the attachment of yeast cells to conidia of B. cinerea. The 10 yeast isolates that did not attach to conidia when grown on agar did attach to conidia (20%-57% of conidia with attached yeast cells) when cultured in liquid medium. Attachment of the biocontrol yeast Rhodotorula glutinis PM4 to conidia of B. cinerea was significantly greater at 1 x 10(7) yeast cells x mL(-1) than at lower concentrations of yeast cells. The ability of yeast cells to attach to fungal conidia or hyphae appears to be a common phenotype among phylloplane yeasts.

Doenças da mandioquinha-salsa e sua situação atual no Brasil Present situation of arracacha (Arracacia xanthorrhiza diseases in Brazil

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Gilmar P. Henz

2002-06-01

Full Text Available Este artigo tem como objetivo principal relatar as doenças registradas para a mandioquinha-salsa até a data presente e descrever sua situação atual no País. Na literatura, foram registrados para a cultura 27 gêneros de fungos, três de bactérias, doze de nematóides e cinco espécies de vírus. Destes, já foram relatados no Brasil treze fungos, e todos nematóides e bactérias, enquanto nenhum vírus foi oficialmente registrado, embora já tenham sido observadas plantas com sintomas típicos de viroses a campo. A maior parte dos registros estão na forma de resumos, capítulos de livros ou publicações genéricas, desprovidos de uma série de dados relevantes, como provas de patogenicidade, importância da doença, perdas, identificação mais acurada, entre outras. A cultura da mandioquinha-salsa ainda é tida como rústica, sendo poucas as doenças consideradas limitantes. O nematóide das galhas (Meloidogyne spp. e a podridão-mole pós-colheita causada por Erwinia spp. são os principais problemas atualmente, e causam perdas significativas. Também ocorrem com muita frequência manchas foliares causadas por Septoria, Cercospora e Xanthomonas campestris pv. arracaciae, e em algumas regiões podridão de plantas a campo causadas por Sclerotinia sclerotiorum e Sclerotium rolfsii. Como não existe nenhum produto químico oficialmente registrado para a mandioquinha-salsa no Ministério da Agricultura, Pecuária e Abastecimento, as medidas de controle passíveis de recomendação incluem ações preventivas, como o uso de material propagativo sadio (preferencialmente mudas pré-enraizadas de origem conhecida, a adoção de rotação de culturas, a eliminação de plantas ou partes doentes, e a adubação e a irrigação adequadas. O estudo sistematizado das doenças e o registro cuidadoso de surtos ou novas enfermidades para a cultura são importantes para subsidiar futuras medidas de controle.The diseases of arracacha that have already

Evaluation of <em>HER2em> Gene Amplification in Breast Cancer Using Nuclei Microarray <em>in em>S>itu em>Hybridization

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Xuefeng Zhang

2012-05-01

Full Text Available Fluorescence<em> em>>in situ em>hybridization (FISH assay is considered the “gold standard” in evaluating <em>HER2/neu (HER2em> gene status. However, FISH detection is costly and time consuming. Thus, we established nuclei microarray with extracted intact nuclei from paraffin embedded breast cancer tissues for FISH detection. The nuclei microarray FISH (NMFISH technology serves as a useful platform for analyzing <em>HER2em> gene/chromosome 17 centromere ratio. We examined <em>HER2em> gene status in 152 cases of invasive ductal carcinomas of the breast that were resected surgically with FISH and NMFISH. <em>HER2em> gene amplification status was classified according to the guidelines of the American Society of Clinical Oncology and College of American Pathologists (ASCO/CAP. Comparison of the cut-off values for <em>HER2em>/chromosome 17 centromere copy number ratio obtained by NMFISH and FISH showed that there was almost perfect agreement between the two methods (κ coefficient 0.920. The results of the two methods were almost consistent for the evaluation of <em>HER2em> gene counts. The present study proved that NMFISH is comparable with FISH for evaluating <em>HER2em> gene status. The use of nuclei microarray technology is highly efficient, time and reagent conserving and inexpensive.

Neonatal Phosphate Nutrition Alters <em>in em>Vivo> and <em>in em>Vitro> Satellite Cell Activity in Pigs