scholarly journals A fluorescence-based viability assay for Phytophthora agathidicida oospores

2021 ◽  
Author(s):  
Michael J. Fairhurst ◽  
Julie R. Deslippe ◽  
Monica L. Gerth
Keyword(s):  
Plant Pathogens ◽  
Fluorescent Dyes ◽  
Viable Cell ◽  
Automated Image Analysis ◽  
Fluorescein Diacetate ◽  
Term Survival ◽  
Viability Staining ◽  
Viability Assay ◽  
Wide Range ◽  
Syto 9

Viability staining is an essential tool in many fields of microbiology. In this study, we aimed to establish a dual fluorescence method for detecting the viability of Phytophthora oospores. Phytophthora is a genus of plant pathogens in the class Oomycete that cause disease in a wide range of agriculturally and ecologically important plants. During the Phytophthora disease cycle, thick-walled oospores can be produced via sexual reproduction. These oospores are essential for long-range dispersal and long-term survival of the pathogen, and therefore methods for the study of oospores are of great interest. In this study, we tested five fluorescent dyes for their ability to stain Phytophthora agathidicida oospores: SYTO 9, FUN-1, fluorescein diacetate, propidium iodide, and TOTO-3 iodide. These dyes represented a selection of total, viable and non-viable cell stains Each dye was assessed individually, and then the best dyes were combined for dual viability staining. In this study, we have identified three dyes — SYTO 9, fluorescein diacetate and TOTO-3 — that can be used to report total, viable and non-viable oospores respectively. We further demonstrate that fluorescein diacetate and TOTO-3 can be used together for dual viability staining. This new method is quantitative and compatible with automated image analysis, allowing oospores to be rapidly and accurately assessed for viability.

Studies on the EC50 of Natural Monoterpenes as Fungal Inhibitors with Quantitative Structure-Activity Relationships (QSARs)

2020 ◽  
Vol 10 (1) ◽  
pp. 44-60
Author(s):  
Mohamed E.I. Badawy ◽  
Entsar I. Rabea ◽  
Samir A.M. Abdelgaleil
Keyword(s):  
Biological Activity ◽  
Plant Pathogens ◽  
Molecular Descriptors ◽  
Microbial Pathogens ◽  
Quantitative Structure ◽  
Qsar Study ◽  
Qsar Analysis ◽  
Pharmacophore Modelling ◽  
Structure Activity ◽  
Wide Range

Background:Monoterpenes are the main constituents of the essential oils obtained from plants. These natural products offered wide spectra of biological activity and extensively tested against microbial pathogens and other agricultural pests.Methods:Antifungal activity of 10 monoterpenes, including two hydrocarbons (camphene and (S)- limonene) and eight oxygenated hydrocarbons ((R)-camphor, (R)-carvone, (S)-fenchone, geraniol, (R)-linalool, (+)-menthol, menthone, and thymol), was determined against fungi of Alternaria alternata, Botrytis cinerea, Botryodiplodia theobromae, Fusarium graminearum, Phoma exigua, Phytophthora infestans, and Sclerotinia sclerotiorum by the mycelia radial growth technique. Subsequently, Quantitative Structure-Activity Relationship (QSAR) analysis using different molecular descriptors with multiple regression analysis based on systematic search and LOOCV technique was performed. Moreover, pharmacophore modelling was carried out using LigandScout software to evaluate the common features essential for the activity and the hypothetical geometries adopted by these ligands in their most active forms.Results:The results showed that the antifungal activities were high, but depended on the chemical structure and the type of microorganism. Thymol showed the highest effect against all fungi tested with respective EC50 in the range of 10-86 mg/L. The QSAR study proved that the molecular descriptors HBA, MR, Pz, tPSA, and Vp were correlated positively with the biological activity in all of the best models with a correlation coefficient (r) ≥ 0.98 and cross-validated values (Q2) ≥ 0.77.Conclusion:The results of this work offer the opportunity to choose monoterpenes with preferential antimicrobial activity against a wide range of plant pathogens.

scholarly journals Advancements in Assessments of Bio-Tissue Engineering and Viable Cell Delivery Matrices Using Bile Acid-Based Pharmacological Biotechnologies

Nanomaterials ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1861
Author(s):  
Armin Mooranian ◽  
Melissa Jones ◽  
Corina Mihaela Ionescu ◽  
Daniel Walker ◽  
Susbin Raj Wagle ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Viable Cell ◽  
Cell Delivery ◽  
Artificial Organ ◽  
Bioartificial Pancreas ◽  
Advantages And Disadvantages ◽  
Wide Range ◽  
Bioartificial Organs ◽  
Significant Interest ◽  
Modern Technologies

The utilisation of bioartificial organs is of significant interest to many due to their versatility in treating a wide range of disorders. Microencapsulation has a potentially significant role in such organs. In order to utilise microcapsules, accurate characterisation and analysis is required to assess their properties and suitability. Bioartificial organs or transplantable microdevices must also account for immunogenic considerations, which will be discussed in detail. One of the most characterized cases is the investigation into a bioartificial pancreas, including using microencapsulation of islets or other cells, and will be the focus subject of this review. Overall, this review will discuss the traditional and modern technologies which are necessary for the characterisation of properties for transplantable microdevices or organs, summarizing analysis of the microcapsule itself, cells and finally a working organ. Furthermore, immunogenic considerations of such organs are another important aspect which is addressed within this review. The various techniques, methodologies, advantages, and disadvantages will all be discussed. Hence, the purpose of this review is providing an updated examination of all processes for the analysis of a working, biocompatible artificial organ.

scholarly journals Microbial antagonists against plant pathogens in Iran: A review

2020 ◽  
Vol 5 (1) ◽  
pp. 404-440 ◽  
Author(s):  
Mehrdad Alizadeh ◽  
Yalda Vasebi ◽  
Naser Safaie
Keyword(s):  
Biological Control ◽  
Chemical Control ◽  
Plant Disease ◽  
Plant Pathogens ◽  
Plant Diseases ◽  
Agricultural Products ◽  
Control Measures ◽  
Wide Range ◽  
Plant Disease Management ◽  
Published Research

AbstractThe purpose of this article was to give a comprehensive review of the published research works on biological control of different fungal, bacterial, and nematode plant diseases in Iran from 1992 to 2018. Plant pathogens cause economical loss in many agricultural products in Iran. In an attempt to prevent these serious losses, chemical control measures have usually been applied to reduce diseases in farms, gardens, and greenhouses. In recent decades, using the biological control against plant diseases has been considered as a beneficial and alternative method to chemical control due to its potential in integrated plant disease management as well as the increasing yield in an eco-friendly manner. Based on the reported studies, various species of Trichoderma, Pseudomonas, and Bacillus were the most common biocontrol agents with the ability to control the wide range of plant pathogens in Iran from lab to the greenhouse and field conditions.

scholarly journals From young massive star clusters to old globulars: long-term survival chances

2006 ◽  
Vol 2 (S237) ◽  
pp. 408-408
Author(s):  
Richard de Grijs
Keyword(s):  
Galaxy Formation ◽  
Massive Star ◽  
Cluster Formation ◽  
Star Clusters ◽  
Term Survival ◽  
Star Forming ◽  
Distant Galaxies ◽  
Wide Range ◽  
Galaxy Collisions

Young, massive star clusters (YMCs) are the most notable and significant end products of violent star-forming episodes triggered by galaxy collisions and close encounters. The question remains, however, whether or not at least a fraction of the compact YMCs seen in abundance in extragalactic starbursts, are potentially the progenitors of (≳10 Gyr) old globular cluster (GC)-type objects. If we could settle this issue convincingly, one way or the other, the implications of such a result would have far-reaching implications for a wide range of astrophysical questions, including our understanding of the process of galaxy formation and assembly, and the process and conditions required for star (cluster) formation. Because of the lack of a statistically significant sample of YMCs in the Local Group, however, we need to resort to either statistical arguments or to the painstaking approach of case-by-case studies of individual objects in more distant galaxies.

Optimization of Growth Conditions of Citrus Anthracnose Agent Colletotrichum gloeosporioides Isolates in Tunisia

2021 ◽  
Vol 3 (1) ◽  
Author(s):  
Ben Hadj-Daoud H ◽  
◽  
Ben Salem I ◽  
Boughalleb-M’Hamdi N ◽  
◽  
...  
Keyword(s):  
Morphological Traits ◽  
Laboratory Experiments ◽  
Plant Pathogens ◽  
Colletotrichum Gloeosporioides ◽  
Optimal Growth ◽  
Growth Conditions ◽  
Fruit Drop ◽  
Nutritional Conditions ◽  
Solid Media ◽  
Wide Range

Background: Colletotrichum gloeosporioides is important plant pathogens on a wide range of plant hosts such as citrus causing pre- or post-harvest infections as anthracnose, post-bloom fruit drop, tearstain and stem-end rot on fruit, or wither-tip of twigs. Method: The optimization of growth conditions of this pathogen was performed (solid media, temperature, pH and water potential under laboratory experiments). Results: Our results revealed that the maximum radial growth of C. gloeosporioides was recorded on SDA medium. All isolates were able to grow on PDA at temperatures of 15 and 30°C (over 0.7cm/day). Optimal growth radial was recorded at pH 5, 6, 7 and 8. Similar responses were obtained with both salt types, but, in general, C. gloeosporioides was more tolerant to KCl than NaCl. Conclusion: Studies of cultural, morphological traits of the pathogen are prominent to understand the response of the pathogen in different environmental and nutritional conditions.

Simultaneous Measurement of Temperature and Velocity in Turbulent Buoyant Plume by Combined LIF and PIV Technique

2006 ◽  
Author(s):  
Yoshie Watanabe ◽  
Yuji Hashizume ◽  
Nobuyuki Fujisawa
Keyword(s):  
Simultaneous Measurement ◽  
Particle Image ◽  
Fluorescent Dyes ◽  
Buoyant Plume ◽  
Thermal Flow ◽  
Piv Technique ◽  
Image Velocimetry ◽  
Wide Range ◽  
High Temperature Sensitivity ◽  
Surrounding Fluid

An experimental technique for simultaneous measurement of temperature and velocity in a thermal flow is described. This technique is based on the two-color laser-induced fluorescence technique combined with the particle image velocimetry. Illumination is provided from Nd:YAG laser and the fluorescent dyes are chosen as Rhodamine B and Fluorescent Sodium, which combination allows the accurate velocity measurement in a wide range of flow velocity and high temperature sensitivity in temperature measurement. The measurement of temperature and velocity in turbulent buoyant plume is carried out by this method, and the structure of the plume is studied in connection with the entrainment of surrounding fluid at the interface.

scholarly journals Identification and Characterization of Diaporthe ambigua, D. australafricana, D. novem, and D. rudis Causing a Postharvest Fruit Rot in Kiwifruit

Plant Disease ◽  
2017 ◽  
Vol 101 (8) ◽  
pp. 1402-1410 ◽  
Author(s):  
Gonzalo A. Díaz ◽  
Bernardo A. Latorre ◽  
Mauricio Lolas ◽  
Enrique Ferrada ◽  
Paulina Naranjo ◽  
...  
Keyword(s):  
San Francisco ◽  
Plant Pathogens ◽  
Elongation Factor ◽  
Its Region ◽  
Fruit Rot ◽  
Lesion Length ◽  
Translation Elongation ◽  
Wide Range ◽  
Postharvest Rot ◽  
Diaporthe Ambigua

Diaporthe spp. are important plant pathogens causing wood cankers, blight, dieback, and fruit rot in a wide range of hosts. During surveys conducted during the 2013 and 2014 seasons, a postharvest rot in Hayward kiwifruit (Actinidia deliciosa) was observed in Chile. In order to identify the species of Diaporthe associated with this fruit rot, symptomatic fruit were collected from seven kiwifruit packinghouses located between San Francisco de Mostazal and Curicó (central Chile). Twenty-four isolates of Diaporthe spp. were identified from infected fruit based on morphological and cultural characters and analyses of nucleotides sequences of three loci, including the internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2), a partial sequences of the β-tubulin, and translation elongation factor 1-α genes. The Diaporthe spp. identified were Diaporthe ambigua, D. australafricana, D. novem, and D. rudis. Multilocus phylogenetic analysis revealed that Chilean isolates were grouped in separate clades with their correspondent ex-types species. All species of Diaporthe were pathogenic on wounded kiwifruit after 30 days at 0°C under normal and controlled-atmosphere (2% O2 and 5% CO2) storage and they were sensitive to benomyl, pyraclostrobin, and tebuconazole fungicides. D. ambigua isolates were the most virulent based on the lesion length measured in inoculated Hayward and Jintao kiwifruit. These findings confirm D. ambigua, D. australafricana, D. novem, and D. rudis as the causal agents of kiwifruit rot during cold storage in Chile. The specie D. actinidiae, a common of Diaporthe sp. found associated with kiwifruit rot, was not identified in the present study.

scholarly journals Origins and Evolution of the Global RNA Virome

mBio ◽  
2018 ◽  
Vol 9 (6) ◽  
Author(s):  
Yuri I. Wolf ◽  
Darius Kazlauskas ◽  
Jaime Iranzo ◽  
Adriana Lucía-Sanz ◽  
Jens H. Kuhn ◽  
...  
Keyword(s):  
Plant Pathogens ◽  
Rna Viruses ◽  
Rna Virus ◽  
Phylogenomic Analysis ◽  
Rna Helicases ◽  
Sequence Alignments ◽  
Recent Advances ◽  
Wide Range ◽  
Positive Sense ◽  
Dsrna Viruses

ABSTRACTViruses with RNA genomes dominate the eukaryotic virome, reaching enormous diversity in animals and plants. The recent advances of metaviromics prompted us to perform a detailed phylogenomic reconstruction of the evolution of the dramatically expanded global RNA virome. The only universal gene among RNA viruses is the gene encoding the RNA-dependent RNA polymerase (RdRp). We developed an iterative computational procedure that alternates the RdRp phylogenetic tree construction with refinement of the underlying multiple-sequence alignments. The resulting tree encompasses 4,617 RNA virus RdRps and consists of 5 major branches; 2 of the branches include positive-sense RNA viruses, 1 is a mix of positive-sense (+) RNA and double-stranded RNA (dsRNA) viruses, and 2 consist of dsRNA and negative-sense (−) RNA viruses, respectively. This tree topology implies that dsRNA viruses evolved from +RNA viruses on at least two independent occasions, whereas −RNA viruses evolved from dsRNA viruses. Reconstruction of RNA virus evolution using the RdRp tree as the scaffold suggests that the last common ancestors of the major branches of +RNA viruses encoded only the RdRp and a single jelly-roll capsid protein. Subsequent evolution involved independent capture of additional genes, in particular, those encoding distinct RNA helicases, enabling replication of larger RNA genomes and facilitating virus genome expression and virus-host interactions. Phylogenomic analysis reveals extensive gene module exchange among diverse viruses and horizontal virus transfer between distantly related hosts. Although the network of evolutionary relationships within the RNA virome is bound to further expand, the present results call for a thorough reevaluation of the RNA virus taxonomy.IMPORTANCEThe majority of the diverse viruses infecting eukaryotes have RNA genomes, including numerous human, animal, and plant pathogens. Recent advances of metagenomics have led to the discovery of many new groups of RNA viruses in a wide range of hosts. These findings enable a far more complete reconstruction of the evolution of RNA viruses than was attainable previously. This reconstruction reveals the relationships between different Baltimore classes of viruses and indicates extensive transfer of viruses between distantly related hosts, such as plants and animals. These results call for a major revision of the existing taxonomy of RNA viruses.

scholarly journals Constitutive expression of bcl-2 in B cells causes a lethal form of lupuslike autoimmune disease after induction of neonatal tolerance to H-2b alloantigens.

1996 ◽  
Vol 183 (6) ◽  
pp. 2523-2531 ◽  
Author(s):  
M López-Hoyos ◽  
R Carrió ◽  
R Merino ◽  
L Buelta ◽  
S Izui ◽  
...  
Keyword(s):  
B Cells ◽  
Autoimmune Disease ◽  
Constitutive Expression ◽  
Systemic Autoimmune Disease ◽  
Spleen Cells ◽  
Term Survival ◽  
F1 Hybrid ◽  
Wide Range

The bcl-2 protooncogene has been shown to provide a survival signal to self-reactive B cells, but it fails to override their developmental arrest after encounter with antigen. Furthermore, constitutive expression of bcl-2 in B cells does not promote the development of autoimmune disease in most strains of mice, indicating that signals other than those conferred by bcl-2 are required for long-term survival and differentiation of self-reactive B cells in vivo. To further examine the factors that are required for the pathogenesis of autoimmune disease, we have assessed the effect of bcl-2 overexpression on the development of host-versus-graft disease, a self-limited model of systemic autoimmune disease. In this model, injection of spleen cells from (C57BL/6 x BALB/c)F1 hybrid mice into BALB/c newborn parental mice induces immunological tolerance to donor tissues and activation of autoreactive F1 donor B cells through interactions provided by allogeneic host CD4+ T cells. BALB/c newborns injected with spleen cells from (C57BL/6 x BALB/c)F1 mice expressing a bcl-2 transgene in B cells developed high levels of anti-single-stranded DNA and a wide range of pathogenic autoantibodies that were not or barely detectable in mice injected with nontransgenic spleen cells. In mice injected with transgenic B cells, the levels of pathogenic autoantibodies remained high during the course of the study and were associated with long-term persistence of donor B cells, development of a severe autoimmune disease, and accelerated mortality. These results demonstrate that bcl-2 can provide survival signals for the maintenance and differentiation of autoreactive B cells, and suggest that both increased B cell survival and T cell help play critical roles in the development of certain forms of systemic autoimmune disease.

scholarly journals Droplet-based microfluidics platform for antifungal analysis against filamentous fungi

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Sehrish Iftikhar ◽  
Aurélie Vigne ◽  
Julia Elisa Sepulveda-Diaz
Keyword(s):  
Fungal Pathogens ◽  
Antimicrobial Agents ◽  
Pearson Correlation ◽  
Hyphal Growth ◽  
Bulk Analysis ◽  
Single Spore ◽  
Microfluidic Platform ◽  
Viability Assay ◽  
Wide Range ◽  
Potential Applications

AbstractFungicides are extensively used in agriculture to control fungal pathogens which are responsible for significant economic impact on plant yield and quality. The conventional antifungal screening techniques, such as water agar and 96-well plates, are based on laborious protocols and bulk analysis, restricting the analysis at the single spore level and are time consuming. In this study, we present a droplet-based microfluidic platform that enables antifungal analysis of single spores of filamentous fungus Alternaria alternata. A droplet-based viability assay was developed, allowing the germination and hyphal growth of single A. alternata spores within droplets. The viability was demonstrated over a period of 24 h and the antifungal screening was achieved using Kunshi/Tezuma as antifungal agent. The efficacy results of the droplet-based antifungal analysis were compared and validated with the results obtained from conventional protocols. The percentage inhibitions assessed by the droplet-based platform were equivalent with those obtained by the other two methods, and the Pearson correlation analysis showed high correlation between the three assays. Taken together, this droplet-based microfluidic platform provides a wide range of potential applications for the analysis of fungicide resistance development as well as combinatorial screening of other antimicrobial agents and even antagonistic fungi.

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