scholarly journals Colonization of Fusarium oxysporum transformed with the red fluorescence protein gene (tdTomato) mediated by Agrobacterium tumefaciens in roots of two avocado cultivars

2021 ◽  
Vol 10 (2) ◽  
pp. e22010212554
Author(s):  
Grace Estefanía Uribe Hurtado ◽  
William Fernando Viera Arroyo ◽  
Álvaro Xavier Sampedro Lema ◽  
Kang Jin Cho ◽  
Alicia Beatriz Villavicencio Pazos ◽  
...  
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Fusarium Oxysporum ◽  
Protein Gene ◽  
Phytophthora Cinnamomi ◽  
Pathogenic Fungus ◽  
Persea Americana ◽  
Soil Pathogens ◽  
Fluorescence Protein ◽  
Red Fluorescence Protein ◽  
Low Incidence

Avocado (Persea americana) is a fruit crop of economic importance in Ecuador. Currently, a low incidence of Phytophthora cinnamomi has been reported, however, there are other soil pathogens that can affect this crop, even at the initial stages of plant multiplication (nursery), for this reason the use of rootstocks that tolerate these biotic adversities is recommended. In this research, the fungus Fusarium oxysporum was isolated from roots of nursery seedlings with symptoms of necrosis. In addition, an isolate of this pathogenic fungus modified with a strain of Agrobacterium tumefaciens was used to determine the infection of F. oxysporum in the roots of the Fuerte (commercial) and Criollo (local) cultivars. The results allowed to infer that the cultivar Criollo presented a greater tolerance to F. oxysporum than the cultivar Fuerte, which corroborates its use as a rootstock for commercial avocado varieties. Furthermore, to our knowledge, this is the first report of F. oxysporum affecting avocado nursery seedlings in Ecuador.

Transformation of Metarhizium anisopliae mediated by Agrobacterium tumefaciens

2006 ◽  
Vol 52 (7) ◽  
pp. 623-626 ◽  
Author(s):  
Weiguo Fang ◽  
Yan Pei ◽  
Michael J Bidochka
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Metarhizium Anisopliae ◽  
Protein Gene ◽  
Fluorescent Protein ◽  
Pathogenic Fungus ◽  
Single Copy ◽  
Transformation Method ◽  
The Other ◽  
Green Fluorescent Protein Gene ◽  
Green Fluorescent

A simple, highly efficient, and reliable Agrobacterium tumefaciens-mediated transformation method was developed for the insect pathogenic fungus Metarhizium anisopliae. Expression of the green fluorescent protein gene, egfp, and the benomyl resistance gene, benA3, were used as markers in transformed M. anisopliae. Transformation efficiencies were dependent on the strain of A. tumefaciens used. With strain AGL-1, 17.0 ± 1.4 transformants per plate could be obtained using conidial concentrations of 106 conidia/mL and a 2 day co-cultivation in the presence of 200 µmol/L acetosyringone. On the other hand, transformations using strain LBA4404 were unsuccessful. Ten transformants were tested by Southern analysis and found to contain a single copy T-DNA. Twenty transformants were subcultured for five generations on nonselective media, and 95% of the transformants were mitotically stable. Agrobacterium tumefaciens-mediated transformation of M. anisopliae can serve as a useful tool to investigate genes involved in insect pathogenicity.Key words: entomopathogenic fungi, Metarhizium anisopliae, Agrobacterium tumefaciens, genetic transformation.

A Monitoring System for Green Fluorescence Protein Gene-transformed Fusarium oxysporum in Melon Seedlings

2001 ◽  
Vol 67 (4) ◽  
pp. 273-280 ◽  
Author(s):  
Teruo NONOMURA ◽  
Yoshinori MATSUDA ◽  
Mikako TAKASUGI ◽  
Takashi OOTANI ◽  
Tomoya HASEGAWA ◽  
...  
Keyword(s):  
Fusarium Oxysporum ◽  
Monitoring System ◽  
Protein Gene ◽  
Green Fluorescence Protein ◽  
Green Fluorescence ◽  
Fluorescence Protein ◽  
Green Fluorescence Protein Gene

scholarly journals Development of specific primers for Fusarium oxysporum causing damping off of Lilium formolongi

2020 ◽  
Vol 18 (1) ◽  
Author(s):  
Takeshi Toda ◽  
Shun Hanesaka ◽  
Kuniaki Shishido ◽  
Shin-ichi Fuji ◽  
Hiromitsu Furuya
Keyword(s):  
Fusarium Oxysporum ◽  
Pathogenic Fungus ◽  
Primer Design ◽  
Damping Off ◽  
Specific Primers ◽  
Forma Specialis

AbstractPrimers specific for the hypothetical forma specialis of Fusarium oxysporum were designed to amplify DNA from this pathogenic fungus that infects plants including lilies. The F. oxysporum sequence between the transposal elements han and hop was used for primer design. Three primer pairs designed from this region were confirmed as specific for 24 isolates of F. oxysporum pathogenic to lilies, except for one pathogenic isolates as extraordinary. No amplification was observed from F. oxysporum non-pathogenic to lily, from 12 forma specialis, and 14 fungi and oomycetes concerned with Liliaceae plants. We propose that specific primers designed from this region will be useful to detect isolates of F. oxysporum that are pathogenic to lilies.

Glycoconjugates on the surface of spores of the pathogenic fungus Phytophthora cinnamomi studied using fluorescence and electron microscopy and flow cytometry

1990 ◽  
Vol 36 (3) ◽  
pp. 183-192 ◽  
Author(s):  
A. R. Hardham ◽  
E. Suzaki
Keyword(s):  
Flow Cytometry ◽  
Plasma Membrane ◽  
Cell Surface ◽  
Phytophthora Cinnamomi ◽  
Pathogenic Fungus ◽  
Fluorescein Isothiocyanate ◽  
Pathogenic Fungi ◽  
Surface Flow ◽  
Soybean Agglutinin ◽  
Binding Material

Glycoconjugates on the surface of zoospores and cysts of the pathogenic fungus Phytophthora cinnamomi have been studied using fluorescein isothiocyanate labelled lectins for fluorescence microscopy and flow cytometry, and ferritin- and gold-labelled lectins for ultrastructural analysis. Of the five lectins used, only concanavalin A (ConA) binds to the surface of the zoospores, including the flagella and water expulsion vacuole. This suggests that of accessible saccharides, glucosyl or mannosyl residues predominate on the outer surface of the zoospore plasma membrane. Early in encystment, a system of flat disc-like cisternae, which underlie the zoospore plasma membrane, vesiculate. These and other small peripheral vesicles quickly disappear. After the induction of encystment, ConA is no longer localised close to the plasma membrane but binds to material loosely associated with the cell surface. Quantitative measurements by flow cytometry indicate that the ConA-binding material is gradually lost from the cell surface. The cyst wall is weakly labelled, but the site of germ tube emergence stains intensely. During the first 2 min after the induction of encystment, material that binds soybean agglutinin, Helix pommatia agglutinin, and peanut agglutinin appears on the surface of the fungal cells. The distribution of this material, rich in galactosyl or N-acetyl-D-galactosaminosyl residues, is initially patchy, but by 5 min the material evenly coats most of the cell surface. Labelling of zoospores in which intracellular sites are accessible indicates that the soybean agglutinin binding material is stored in vesicles that lie beneath the plasma membrane. Quantitation of soybean agglutinin labelling shows that maximum binding occurs 2–3 min after the induction of encystment. Key words: cell surface, flow cytometry, lectins, pathogenic fungi, Phytophthora cinnamomi.

USAGE OF TRANSCRIPTIONAL REGULATOR LRP BASED BIOSENSOR FOR RESEARCH OF VALINE BIOSYNTHESIS PATHWAYS IN CORYNEBACTERIUM GLUTAMICUM CELLS

2021 ◽  
Vol 1 (19) ◽  
pp. 318-320
Author(s):  
D.D. Derbikov ◽  
A.S. Yanenko
Keyword(s):  
Corynebacterium Glutamicum ◽  
Protein Gene ◽  
Transcriptional Regulator ◽  
Valine Biosynthesis ◽  
Fluorescence Protein ◽  
Gene Promotor

The biosensor construction based on Lrp gene was obtained, what contains a fluorescence protein gene under control of BrnFE gene promotor. It was displayed that the fluorescence of Corynebacterium glutamicum cells carrying the biosensor correlates with the production of valine, and with its help it is possible to study the biosynthesis of valine.

scholarly journals Transcriptome analysis of an incompatible Persea americana-Phytophthora cinnamomi interaction reveals the involvement of SA- and JA-pathways in a successful defense response

PLoS ONE ◽  
2018 ◽  
Vol 13 (10) ◽  
pp. e0205705 ◽  
Author(s):  
Noëlani van den Berg ◽  
Waheed Mahomed ◽  
Nicholas A. Olivier ◽  
Velushka Swart ◽  
Bridget G. Crampton
Keyword(s):  
Transcriptome Analysis ◽  
Defense Response ◽  
Phytophthora Cinnamomi ◽  
Persea Americana

scholarly journals Isolating promoters from Corynebacterium ammoniagenes ATCC 6871 and application in CoA synthesis

2019 ◽  
Author(s):  
Yingshuo Hou ◽  
Siyu Chen ◽  
Jianjun Wang ◽  
Guizhen Liu ◽  
Sheng Wu ◽  
...  
Keyword(s):  
Metabolic Engineering ◽  
Molecular Chaperone ◽  
Coenzyme A ◽  
Industrial Applications ◽  
Upstream Sequence ◽  
Activity Levels ◽  
Corynebacterium Ammoniagenes ◽  
Fluorescence Protein ◽  
Red Fluorescence Protein ◽  
Dna Promoters

Abstract Background：Corynebacterium ammoniagenes is an important industrial organism that is widely used to produce nucleotides and the potential for industrial production of coenzyme A by Corynebacterium ammoniagenes ATCC 6871 has been shown. However, the yield of coenzyme A needs to be improved and the available constitutive promoters are rather limited in this strain. Results：In this study, 20 putative DNA promoters derived from genes with high transcription levels and 6 promoters from molecular chaperone genes were identified. To evaluate the activity of each promoter, red fluorescence protein (RFP) was used as a reporter. We successfully isolated a range of promoters with different activity levels resulting in different fluorescent intensities. A fragment derived from the upstream sequence of the 50S ribosomal protein L21 (Prpl21) exhibited the strongest activity among the 26 identified promoters. Furthermore, type III pantothenate kinase from Pseudomonas putida (PpcoaA) was overexpressed in C. ammoniagenes under the control of Prpl21, CoA yield increased approximately 4.1 times. Conclusions：This study provides a paradigm for rational isolation of promoters with different activities and their application in metabolic engineering. These promoters will enrich the available promoter toolkit for C. ammoniagenes and should be valuable in current platforms for metabolic engineering and synthetic biology for the optimization of pathways to extend the product spectrum or improve the productivity in C. ammoniagenes ATCC 6871 for industrial applications.

scholarly journals UJI DAYAHAMBAT EUGENOL DARI DAUN CENGKEH HASIL FRAKSINASI TERHADAP PERTUMBUHAN JAMUR PATOGEN Fusarium oxysporum f.sp. Cubense Eugenol Inhibitory Test From Clove Leaf Fractionation Of Fungal Patogen Growth Fusarium oxysporum f.sp. cubense

AgriPeat ◽  
2019 ◽  
Vol 20 (02) ◽  
pp. 107-113
Author(s):  
Admin Journal
Keyword(s):  
Fusarium Oxysporum ◽  
Vacuum Distillation ◽  
Pathogenic Fungus ◽  
Lethal Concentration ◽  
In Vitro Tests ◽  
In Vitro Test ◽  
Microsoft Excel ◽  
Vitro Test

ABSTRACTThis study aims to determine the inhibition of eugenol derived from fractionation clove leaf essentialoils (CLEO) on the growth of pathogenic fungus Fusarium oxysporum f. sp. cubense (Foc) and LC50(Lethal Concentration 50). This research was in vitro, started with purification of clove leaf essentialoil, fractionation by vacuum distillation and bioassay. In vitro tests include exploration of minimuminhibition and preventability tests. Data were analyzed with Microsoft Excel 2010 program. Theresults of minimum inhibition showed at 218,75 ppm concentration of each level was able to inhibitthe growth of Foc fungi. The minimum inhibition exploration was carried out at 218,75 ppm, 109,38ppm, 54,69 ppm and 27,34 ppm. Exploration results showed that fractionated CLEO has been able toinhibit the growth of Foc fungi at 27,34 ppm in the amount of 15,60%. This concentration is used asthe lowest concentration in the inhibitory test. Furthermore, the inhibitory test was carried out startingat the highest concentration of 218,75 ppm, 109,38 ppm, 54,69 ppm and 27,34 ppm. Observationswere made for 7 days after inoculation (DAI). The results showed the best inhibition was at aconcentration of 218,75 ppm at 90,70% and LC50 at 11.17 µL.Keywords: CLEO, fractionation, Foc, in vitro test and LC50

scholarly journals The sss Colonization Gene of the Tomato-Fusarium oxysporum f. sp. radicis-lycopersici Biocontrol Strain Pseudomonas fluorescens WCS365 Can Improve Root Colonization of Other Wild-type Pseudomonas spp. Bacteria

2000 ◽  
Vol 13 (11) ◽  
pp. 1177-1183 ◽  
Author(s):  
Linda C. Dekkers ◽  
Ine H. M. Mulders ◽  
Claartje C. Phoelich ◽  
Thomas F. C. Chin-A-Woeng ◽  
André H. M. Wijfjes ◽  
...  
Keyword(s):  
Fusarium Oxysporum ◽  
Pseudomonas Fluorescens ◽  
Pathogenic Fungus ◽  
Cell Types ◽  
Disease Suppression ◽  
Systemic Resistance ◽  
Root Tip ◽  
Wild Type ◽  
Tomato Root ◽  
Colonization Ability

We show that the disease tomato foot and root rot caused by the pathogenic fungus Fusarium oxysporum f. sp. radicis-lycopersici can be controlled by inoculation of seeds with cells of the efficient root colonizer Pseudomonas fluorescens WCS365, indicating that strain WCS365 is a bio-control strain. The mechanism for disease suppression most likely is induced systemic resistance. P. fluorescens strain WCS365 and P. chlororaphis strain PCL1391, which acts through the production of the antibiotic phenazine-1-carboxamide, were differentially labeled using genes encoding autofluorescent proteins. Inoculation of seeds with a 1:1 mixture of these strains showed that, at the upper part of the root, the two cell types were present as microcolonies of either one or both cell types. Microcolonies at the lower root part were predominantly of one cell type. Mixed inoculation tended to improve biocontrol in comparison with single inoculations. In contrast to what was observed previously for strain PCL1391, mutations in various colonization genes, including sss, did not consistently decrease the biocontrol ability of strain WCS365. Multiple copies of the sss colonization gene in WCS365 improved neither colonization nor biocontrol by this strain. However, introduction of the sss-containing DNA fragment into the poor colonizer P. fluorescens WCS307 and into the good colonizer P. fluorescens F113 increased the competitive tomato root tip colonization ability of the latter strains 16- to 40-fold and 8- to 16-fold, respectively. These results show that improvement of the colonization ability of wild-type Pseudomonas strains by genetic engineering is a realistic goal.

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