Fusarium sp., causal agent of vascular wilt in citrus and its sensitivity to fungicides

2021 ◽  
Vol 40 (1) ◽  
Author(s):  
Luis Enrique Ortiz-Martínez ◽  
Leticia Robles-Yerena ◽  
Santos Gerardo Leyva-Mir ◽  
Moisés Camacho-Tapia ◽  
Lucia Juárez-Rodríguez
Keyword(s):  
Bacillus Subtilis ◽  
Vascular System ◽  
Culture Media ◽  
Negative Control ◽  
Causal Agent ◽  
Biological Agent ◽  
Economic Losses ◽  
Vascular Wilt ◽  
Apical Bud

<p>Citrus wilt is a disease of recent appearance in the northern area of Veracruz that causes economic losses to producers in the region. The present work aimed to identify the causative agent of this disease and evaluate different fungicides to determine its<em> in vitro</em> sensitivity. A fungus was consistently isolated in plants with wilt symptoms; it was morphologically identified indifferent culture media and molecularly identified by PCR using the EF1-728F/EF1-986R primers. The fungus was inoculated in three varieties of citrus under greenhouse conditions. The sensitivity test was carried out with the fungicides chlorothalonil, benomyl thiabendazole, prochloraz, and a biological agent (<em>Bacillus subtilis</em>) at different concentrations, plus a negative control. <em>Fusarium</em> sp. (Accession No. MW438335) was morphologically and molecularly identified as the causal agent of vascular wilt in citrus fruits, causing growth retardation, decreased number of roots, wilting of the apical bud, and necrosis in the vascular system of the three varieties inoculated. The most effective fungicides in inhibiting mycelial growth were thiabendazole, prochloraz, and the biological agent <em>Bacillus subtilis</em>.</p>

scholarly journals Dual Negative Control of spx Transcription Initiation from the P3 Promoter by Repressors PerR and YodB in Bacillus subtilis

2006 ◽  
Vol 189 (5) ◽  
pp. 1736-1744 ◽  
Author(s):  
Montira Leelakriangsak ◽  
Kazuo Kobayashi ◽  
Peter Zuber
Keyword(s):  
Oxidative Stress ◽  
Bacillus Subtilis ◽  
Transcription Initiation ◽  
Transcriptional Start Site ◽  
Regulatory Region ◽  
Point Mutations ◽  
Negative Control ◽  
Response To Treatment ◽  
Additive Increase

ABSTRACT The spx gene encodes an RNA polymerase-binding protein that exerts negative and positive transcriptional control in response to oxidative stress in Bacillus subtilis. It resides in the yjbC-spx operon and is transcribed from at least five promoters located in the yjbC regulatory region or in the yjbC-spx intergenic region. Induction of spx transcription in response to treatment with the thiol-specific oxidant diamide is the result of transcription initiation at the P3 promoter located upstream of the spx coding sequence. Previous studies conducted elsewhere and analyses of transcription factor mutants using transformation array technology have uncovered two transcriptional repressors, PerR and YodB, that target the cis-acting negative control elements of the P3 promoter. Expression of an spx-bgaB fusion carrying the P3 promoter is elevated in a yodB or perR mutant, and an additive increase in expression was observed in a yodB perR double mutant. Primer extension analysis of spx RNA shows the same additive increase in P3 transcript levels in yodB perR mutant cells. Purified YodB and PerR repress spx transcription in vitro when wild-type spx P3 promoter DNA was used as a template. Point mutations at positions within the P3 promoter relieved YodB-dependent repression, while a point mutation at position +24 reduced PerR repression. DNase I footprinting analysis showed that YodB protects a region that includes the P3 −10 and −35 regions, while PerR binds to a region downstream of the P3 transcriptional start site. The binding of both repressors is impaired by the treatment of footprinting reactions with diamide or hydrogen peroxide. The study has uncovered a mechanism of dual negative control that relates to the oxidative stress response of gram-positive bacteria.

scholarly journals Integrated management of the Fusarium vascular wilt disease of Cucurbita pepo in Iraq

2019 ◽  
Vol 23 (1) ◽  
pp. 40
Author(s):  
Safaa N. Hussein
Keyword(s):  
Cucurbita Pepo ◽  
Disease Incidence ◽  
Dry Weight ◽  
Negative Control ◽  
Wilt Disease ◽  
Pathogenicity Test ◽  
Vascular Wilt ◽  
Universal Primer ◽  
Vascular Wilt Disease

Fusarium vascular wilt disease is one of the most harmful disease that affected broad range of plant species including zucchini (Cucurbita pepo). The objective of the research was to investigate the presence of the phytopathogenic gungus Fusarium oxysporum the causal agent of the disease in zucchini fields. Forty five isolates of F. oxysporum were isolated from four locations in province of Dyala during 2014-2015. Isolate Foq9 was most virulent in the pathogenicity test in vitro. Twenty nine isolates of them amplified their DNA positively with the universal primer of F. oxysporum in the polymerase chain reaction technique (PCR). In vitro significant suppression efficiency were observed of the fungicides Topsin M 70 WP (Tm) and Tecto 500 SC (Tc) and two botanical extract solution extracted from Garlic (Gr) and Ginger (Gn) against the pathogen on the potato sucrose agar (PSA). In greenhouse experiments all of the agents decreased the percentage of disease incidence and severity significantly, while the tetra-inoculum (Tm+Tc+Gr+Gn) was superior which exhibited 0% disease incidence and severity compared to the negative control which was 95%, 79% respectively, also the combined application of the botanical extracts (Gr+Gn) reduced the disease incidence and severity significantly. All of the treatment increased plant growth criteria represented by dry weight of the plant compared to the control.

scholarly journals Daya Anthelmintik Infusa Biji Buah Pinang (Areca catechu) Terhadap Cacing Ascaridia galli Secara in Vitro

2019 ◽  
Vol 36 (2) ◽  
pp. 254
Author(s):  
Wida Wahidah Mubarokah ◽  
Wisnu Nurcahyo ◽  
Kurniasih Kurniasih
Keyword(s):  
Egg Production ◽  
Anthelmintic Resistance ◽  
Negative Control ◽  
Economic Losses ◽  
In Vitro Test ◽  
Ascaridia Galli ◽  
Group Vi ◽  
Group I ◽  
Areca Catechu

A. galli infection caused high economic losses and decreased the amount and quality of egg production. The existence of anthelmintic resistance lead to the development of research on alternative treatments for A. galli. This aim of the research was to know LC50 of Areca catechu on infestation of A. galli. In this research A. galli was taken from a chicken slaughterhouse in Terban, Yogyakarta, and was divided into 8 groups for in vitro research. Group I was treated with an 10% infusion of A. catechu; group II was treated with 12.5% infusion of A. catechu; group III was treated with a 15% infusion of A. catechu; group IV was treated with a 17.5% infusion of A. catechu; group V was treated with 20% infusion of A. catechu; group VI was treated with a 22.5% infusion of A. catechu; group VII was treated with 25% A. catechu infusion and group VIII as negative control (0.9% NaCl). A. galli mortality is recorded every one hour until worm mortality is 100%. The results were then analyzed using the Reed and Muench method. Based on the in vitro test the infusion of Areca catechu against Ascaridia galli can cause death in worms with an LC 50 calculation of 21. 18%.

scholarly journals PENGARUH EKSTRAK Sargassum cristaefolium PADA MULTIPLIKASI Dendrobium antennatum Rchb.f SECARA IN VITRO

2020 ◽  
Vol 8 (1) ◽  
pp. 18
Author(s):  
Faisal Ansyarif ◽  
Mursal Ghazali ◽  
Aida Muspiah ◽  
Rina Kurnianingsih
Keyword(s):  
Culture Media ◽  
Positive Control ◽  
Negative Control ◽  
Growth Media ◽  
Extract Concentration ◽  
Tissue Culture Media ◽  
Randomized Design ◽  
Completely Randomized Design ◽  
Natural Cytokinin

The purpose of this study was to determine the potential and concentration of Sargassum cristaefolium extract as a natural cytokinin in tissue culture media of Dendrobium antennatum Rchb.f. This study is experimental with a completely randomized design, using several extract concentrations compared with the positive control (BAP 1.5 ppm) and negative control (MS0 media). Extract concentrations used 5 ppm, 10 ppm, 15 ppm, 20 ppm, and 25 ppm. Based on analysis of variance (ANOVA), the effect of Sargassum cristaefolium extract on the growth media significant on all parameters. Sargassum cristaefolium extracts caused different responses at certain levels of concentration. Extract concentration of 10 ppm was able to initiate the highest number of shoots and leaves compared to other extract concentrations, where as the concentration 20 ppm was able to accelerate and increase root growth.

scholarly journals Analysis of the Antimicrobial Efficacy, In Vitro, of Oral Antiseptics on Fungi and Bacteria

2021 ◽  
Vol 2 (2) ◽  
pp. 1-6
Author(s):  
Mateus José Dutra
Keyword(s):  
Hydrogen Peroxide ◽  
Active Ingredient ◽  
Benzalkonium Chloride ◽  
Culture Media ◽  
Cetylpyridinium Chloride ◽  
Positive Control ◽  
Negative Control ◽  
Antimicrobial Action ◽  
Control Methodology

Objectives: The study aimed to evaluate the action, in vitro, of hydrogen peroxide in the concentrations of 1%, 1.5% and 3%, of cetylpyridinium chloride, of benzalkonium chloride of 1.3% and of chlorhexidine 0.12% associated with 1.5% hydrogen peroxide over standardized strains of microorganisms Enterococcus faecalis; Staphylococcus aureus; Candida albicans and Escherichia coli, aspiring to determine which is the best product. Chlorhexidine 0.12% was used as a positive control and sterile distilled water as a negative control. Methodology: Standardized suspensions of the microorganisms were prepared, poured and spread over the culture media. Sterile 4 mm paper discs were placed on the seeded agar and soaked with 15 microliters (uL) of each of the antiseptics. The plates were incubated in a bacteriological oven at 37º Celsius for 48 hours. The results were obtained through the growth inhibition halo. Results: The antiseptic that obtained the greatest antimicrobial action was the association of hydrogen peroxide at 1.5% with chlorhexidine at 0.12%, followed by benzalkonium chloride 1.3%. Hydrogen peroxide at 1% did not show any antimicrobial activity. Conclusions: The 0.12% chlorhexidine associated with 1.5% hydrogen peroxide demonstrated the best antimicrobial action, indicating a possible synergistic action between the products, increasing the efficiency when compared to the isolated use of each active ingredient. Benzalkonium chloride 1.3%, also showed an effective antimicrobial action, indicating that these products or combinations have a great potential to be used as an active ingredient in mouthwashes.

scholarly journals Morphometry of bovine blastocysts produced in vitro in culture media with antioxidants cysteamine or oily extract of Lippia origanoides

2021 ◽  
Vol 73 (4) ◽  
pp. 799-811
Author(s):  
N.V. Sollecito ◽  
R.N. Alves ◽  
M.E. Beletti ◽  
E.C.M. Pereira ◽  
M.S. Miranda ◽  
...  
Keyword(s):  
Inner Cell Mass ◽  
Culture Media ◽  
Cell Mass ◽  
Morphological Characteristics ◽  
Poor Quality ◽  
Blastocyst Stage ◽  
Negative Control ◽  
Transmission Microscopy ◽  
Electron Transmission Microscopy

ABSTRACT This study aimed to evaluate the ultrastructural morphometry of bovine embryos produced in vitro grown at different concentrations of antioxidants. After in vitro maturation and fertilization, the presumptive zygotes were assigned into five treatments. T1) without the addition of any antioxidants (negative control); T2) addition of 50μM/mL cysteamine; and T3, T4 and T5) adding 2.5μg/mL, 5.0μg/mL or 10.0μg/mL of the antioxidants derived from the oily extract from Lippia origanoides, respectively. On D7 of culture, the embryos in the blastocyst stage were fixed and prepared for electron transmission microscopy. These were evaluated for the proportion of cytoplasm-to-nucleus, cytoplasm-to-mitochondria, cytoplasm-to-vacuoles, cytoplasm-to-autophagic vacuoles and cytoplasm-to-lipid droplets. Blastocysts cultured in media containing oily extract of Lippia origanoides presented morphological characteristics such as high cell:mitochondria ratio and low cell:vacuoles and cell:autophagic vacuole ratio, possibly been morphological indicators of embryonic quality. Inner cell mass (ICM) from blastocysts cultured in media without any antioxidants had the highest cell:vacuole ratio. Similar results were found in the trophectoderm (TE) cells of blastocysts from treatment 2. Embryo culture media supplemented with antioxidants derived from Lippia origanoides oil produced embryos with a higher cytoplasmic proportion of organelles, such as mitochondria. Also, treatments without any antioxidants or with the addition of cysteamine presented cytoplasmic vacuolization, a characteristic related to production of poor-quality embryos.

scholarly journals Potency of Endophyte Microbes as an Antagonistic Agent for Colletotrichum capsici Causes Anthracnose Disease in Cayenne pepper (Capsicum frustescens)

el–Hayah ◽  
2018 ◽  
Vol 6 (3) ◽  
pp. 104-111
Author(s):  
Zahroul Afifah ◽  
Ulfah Utami
Keyword(s):  
Bacillus Cereus ◽  
Culture Media ◽  
Disease Incidence ◽  
Negative Control ◽  
Dual Culture ◽  
Anthracnose Disease ◽  
Colletotrichum Capsici ◽  
Trichoderma Sp

Background: An anthracnose disease caused by pathogenic fungal Colletotrichum capsici has been attacking the cayenne plants either harvested or has not been harvested. This disease must be handled appropriately and quickly because it can reduce the production of chili up to 90%. Recently, anthracnose disease prevention still use chemical fungicide that if applied for long time will cause new impact for environment. Objective: Trichoderma and Bacillus cereus endophytes may be used as antagonistic agents for C. capsici pathogens because they have various antibiotic compounds. Methods: This research uses experimental method. The stages of this study include sterilization of tools and materials, preparation of culture media of fungal and bacteria, rejuvenation of endophytic microbe culture Trichoderma sp. and Bacillus cereus, rejuvenation of  C. capsici pathogen, antagonistic test in vitro using dual culture method. Result and conclusion: The results of in vitro antagonistic tests showed that inhibition percentage of Trichoderma treatment (96%) and combination treatment Trichoderma and B. cereus (97%) is not significantly different. While in B. cereus treatment (11, 88%) significantly different with all of treatments. Endophytes are shown by its dominating growth in petri dishes than C. capsici pathogen or B. cereus endophytes. Furthermore,for endophytes Trichoderma sp continued on in vivo test because it was most effective.The result for incubation period is 3 days after inoculation compared with negative control 2 days. For disease incidence 100%, and for disease intensity that is 61,25% compared with negative control equal to 88,75%.

scholarly journals Development, Biological Characterization, and Immunological Evaluation of Virosome Vaccine against Newcastle Disease in Pakistan

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Muhammad Hidayat Rasool ◽  
Asif Mehmood ◽  
Muhammad Saqalein ◽  
Muhammad Atif Nisar ◽  
Ahmad Almatroudi ◽  
...  
Keyword(s):  
Newcastle Disease ◽  
Fluorescent Protein ◽  
Virulent Strain ◽  
Negative Control ◽  
Economic Losses ◽  
Experimental Conditions ◽  
In Vivo Evaluation ◽  
Inactivated Vaccines

Newcastle disease (ND) is a highly fatal, infectious, viral disease, and despite immunization with live and inactivated vaccines, the disease is still endemic, causing heavy morbidity and mortality leading to huge economic losses to the poultry industry in Pakistan. Therefore, the present study was aimed for the first time in the country at using novel virosomal technology to develop the ND vaccine using an indigenous highly virulent strain of the virus. ND virosome was prepared using Triton X-100, and SM2 Bio-Beads were used to remove the detergent and reconstitute the viral membrane into virosome. Confirmation was done by transmission electron microscopy and protein analysis by SDS-PAGE. In vitro cell adhesion property was observed by incorporating green fluorescent protein (GFP), producing plasmid into virosome and in vitro cell culture assay. Sterility, safety, and stability of the vaccine were tested before in vivo evaluation of immunogenicity and challenge protection study in commercial broiler. The virosome vaccine was administered (30 μg/bird) at days 7 and 14 through the intranasal route in comparison with commercially available live and inactivated ND vaccines. Results revealed significantly high ( p < 0.05 ) and clinically protective hemagglutination inhibition (HI) antibody titers at 7, 14, 21, and 28 days postimmunization with the virosome vaccine in comparison to the negative control. The GMTs were comparable to live and inactivated vaccines with nonsignificant ( p > 0.05 ) differences throughout the experiment. Antibody levels increased in all vaccinated groups gradually from the 7th day and were maximum at 28th-day postvaccination. In the virosome-administered group, GMT was 83.18 and 77.62 at 21st and 28th-days postvaccination, respectively. Challenge revealed 100%, 90%, and 80% protection in virosome, live, and inactivated vaccinated groups, respectively. Under given experimental conditions, we can conclude that ND virosome vaccine prepared from the indigenous virus was found to be safe and immunogenic.

scholarly journals Identification and control the causal agent of root rot disease of cucumber in Iraq: تشخيص ومكافحة مسبب مرض تعفن جذور محصول الخيار في العراق

2019 ◽  
Vol 3 (3) ◽  
Author(s):  
Safaa Neamat Hussein
Keyword(s):  
Root Rot ◽  
Disease Incidence ◽  
Morphological Characteristics ◽  
Negative Control ◽  
Significant Inhibition ◽  
Causal Agent ◽  
Root Rot Disease ◽  
Rot Disease ◽  
And Control

This study aimed to isolate the causal agent of the root rot disease of cucumber and control it biologically. Samples were collected in the cucumber fields in the Diyala and Saladin governorates of Iraq. Isolation test demonstrated associate fungi belong to the twelve geniuses. Fusarium solani exhibited highly percentage of appearance of 82.15% with frequency 54.00%. Seventy-seven isolates identified as F. solani according to their cultural and morphological characteristics while sixty-five isolates of them amplified successfully with specific primer of Fusarium spp using PCR technique. Isolate DF13 was most virulent isolated while exhibited 0% cucumber seed germination in vitro. The bio-agent Bacillus pumilus demonstrated significant inhibition ability against the fungal isolate DF13 in vitro of 100%. Under greenhouse condition B. pumilus decreased the disease incidence and severity to 30.55% and 20.75% respectively compared to the negative control which was 80.50%, 55.00% respectively.

scholarly journals Tunable Extracellular Self-Assembly of Multi-Protein Conjugates from Bacillus subtilis

2016 ◽  
Author(s):  
Charlie Gilbert ◽  
Mark Howarth ◽  
Colin R. Harwood ◽  
Tom Ellis
Keyword(s):  
Bacillus Subtilis ◽  
Self Assembly ◽  
Culture Media ◽  
Protein Complexes ◽  
Modular System ◽  
Protein Conjugation ◽  
Novel Approach ◽  
Multifunctional Enzymes

The ability to stably and specifically conjugate recombinant proteins to one another is a powerful in vitro technique for engineering multifunctional enzymes, protein therapeutics and novel biological materials. However, for many applications spontaneous in vivo protein conjugation would be preferable to in vitro methods. Exploiting the recently described SpyTag-SpyCatcher system, we describe here how enzymes and structural proteins can be genetically-encoded to covalently conjugate in culture media following programmable secretion by Bacillus subtilis. Using this novel approach, we demonstrate how self-conjugation of a secreted industrial enzyme, XynA, dramatically increases its resilience to boiling and we show that cellular consortia can be engineered to self-assemble functional multi-protein complexes with tunable composition. This genetically-encoded modular system provides a new, flexible strategy for protein conjugation harnessing the substantial advantages of extracellular self-assembly.

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