scholarly journals The Incorporation of Amplified Metal-Enhanced Fluorescence in a CMOS-Based Biosensor Increased the Detection Sensitivity of a DNA Marker of the Pathogenic Fungus Colletotrichum gloeosporioides

Biosensors ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 204
Author(s):  
Dorin Harpaz ◽  
Noam Alkan ◽  
Evgeni Eltzov
Keyword(s):  
Glass Surface ◽  
Colletotrichum Gloeosporioides ◽  
Pathogenic Fungus ◽  
Light Signal ◽  
Detection Sensitivity ◽  
Silver Layer ◽  
Enhanced Fluorescence ◽  
Silver Deposition ◽  
Reaction Duration ◽  
Dna Strand

Half of the global agricultural fresh produce is lost, mainly because of rots that are caused by various pathogenic fungi. In this study, a complementary metal-oxide-semiconductor (CMOS)-based biosensor was developed, which integrates specific DNA strands that allow the detection of enoyl-CoA-hydratase/isomerase, which is a quiescent marker of Colletotrichum gloeosporioides fungi. The developed biosensor mechanism is based on the metal-enhanced fluorescence (MEF) phenomenon, which is amplified by depositing silver onto a glass surface. A surface DNA strand is then immobilized on the surface, and in the presence of the target mRNA within the sample, the reporter DNA strand that is linked to horseradish peroxidase (HRP) enzyme will also bind to it. The light signal that is later produced from the HRP enzyme and its substrate is enhanced and detected by the coupled CMOS sensor. Several parameters that affect the silver-deposition procedure were examined, including silver solution temperature and volume, heating mode, and the tank material. Moreover, the effect of blocking treatment (skim milk or bovine serum albumin (BSA)) on the silver-layer stability and nonspecific DNA absorption was tested. Most importantly, the effect of the deposition reaction duration on the silver-layer formation and the MEF amplification was also investigated. In the study findings a preferred silver-deposition reaction duration was identified as 5–8 min, which increased the deposition of silver on the glass surface up to 13-times, and also resulted in the amplification of the MEF phenomenon with a maximum light signal of 50 relative light units (RLU). It was found that MEF can be amplified by a customized silver-deposition procedure that results in increased detection sensitivity. The implementation of the improved conditions increased the biosensor sensitivity to 3.3 nM (4500 RLU) with a higher detected light signal as compared to the initial protocol (400 RLU). Moreover, the light signal was amplified 18.75-, 11.11-, 5.5-, 11.25-, and 3.75-times in the improved protocol for all the tested concentrations of the target DNA strand of 1000, 100, 10, 3.3, and 2 nM, respectively. The developed biosensor system may allow the detection of the pathogenic fungus in postharvest produce and determine its pathogenicity state.

Control of Winged Waterprimrose (Jussiaea decurrens) and Northern Jointvetch (Aeschynomene virginica) with Fungal Pathogens

Weed Science ◽  
1979 ◽  
Vol 27 (5) ◽  
pp. 497-501 ◽  
Author(s):  
C. D. Boyette ◽  
G. E. Templeton ◽  
R. J. Smith
Keyword(s):  
Oryza Sativa ◽  
Glycine Max ◽  
Fungal Pathogens ◽  
Liquid Culture ◽  
Field Experiments ◽  
Colletotrichum Gloeosporioides ◽  
Pathogenic Fungus ◽  
Field Tests ◽  
Wide Range ◽  
Growing Region

An indigenous, host-specific, pathogenic fungus that parasitizes winged waterprimrose [Jussiaea decurrens(Walt.) DC.] is endemic in the rice growing region of Arkansas. The fungus was isolated and identified asColletotrichum gloeosporioides(Penz.) Sacc. f.sp. jussiaeae(CGJ). It is highly specific for parasitism of winged waterprimrose and not parasitic on creeping waterprimrose (J. repensL. var.glabrescensKtze.), rice (Oryza sativaL.), soybeans [Glycine max(L.) Merr.], cotton (Gossypium hirsutumL.), or 4 other crops and 13 other weeds. The fungus was physiologically distinct from C.gloeosporioides(Penz.) Sacc. f. sp.aeschynomene(CGA), an endemic anthracnose pathogen of northern jointvetch[Aeschynomene virginica(L.) B.S.P.], as indicated by cross inoculations of both weeds. Culture in the laboratory and inoculation of winged waterprimrose in greenhouse, growth chamber and field experiments indicated that the pathogen was stable, specific, and virulent in a wide range of environments. The pathogen yielded large quantities of spores in liquid culture. It is suitable for control of winged waterprimrose. Winged waterprimrose and northern jointvetch were controlled in greenhouse and field tests by application of spore mixtures of CGJ and CGA at concentrations of 1 to 2 million spores/ml of each fungus in 94 L/ha of water; the fungi did not damage rice or nontarget crops.

Evaluation of Worm-Bed Leachate as an Antifungal Agent against Pathogenic Fungus,Colletotrichum gloeosporioides

2014 ◽  
Vol 22 (1) ◽  
pp. 23-32 ◽  
Author(s):  
Eduardo Contreras-Blancas ◽  
Victor M. Ruíz-Valdiviezo ◽  
Fortunata Santoyo-Tepole ◽  
Marco Luna-Guido ◽  
Rocío Meza-Gordillo ◽  
...  
Keyword(s):  
Antifungal Agent ◽  
Colletotrichum Gloeosporioides ◽  
Pathogenic Fungus

scholarly journals In Planta Production of Indole-3-Acetic Acid by Colletotrichum gloeosporioides f. sp. aeschynomene

2004 ◽  
Vol 70 (3) ◽  
pp. 1852-1854 ◽  
Author(s):  
Rudy Maor ◽  
Sefi Haskin ◽  
Hagit Levi-Kedmi ◽  
Amir Sharon
Keyword(s):  
Acetic Acid ◽  
Fungal Biomass ◽  
Colletotrichum Gloeosporioides ◽  
Pathogenic Fungus ◽  
Plant Colonization ◽  
In Planta ◽  
Iaa Production ◽  
Axenic Cultures ◽  
Indole 3 Acetic Acid ◽  
Iaa Biosynthesis

ABSTRACT The plant pathogenic fungus Colletotrichum gloeosporioides f. sp. aeschynomene utilizes external tryptophan to produce indole-3-acetic acid (IAA) through the intermediate indole-3-acetamide (IAM). We studied the effects of tryptophan, IAA, and IAM on IAA biosynthesis in fungal axenic cultures and on in planta IAA production by the fungus. IAA biosynthesis was strictly dependent on external tryptophan and was enhanced by tryptophan and IAM. The fungus produced IAM and IAA in planta during the biotrophic and necrotrophic phases of infection. The amounts of IAA produced per fungal biomass were highest during the biotrophic phase. IAA production by this plant pathogen might be important during early stages of plant colonization.

scholarly journals Validation of an eDNA-based method for the detection of wildlife pathogens in water

2020 ◽  
Vol 141 ◽  
pp. 171-184
Author(s):  
N Sieber ◽  
H Hartikainen ◽  
C Vorburger
Keyword(s):  
Water Samples ◽  
Pathogen Detection ◽  
Batrachochytrium Dendrobatidis ◽  
Tap Water ◽  
Pathogenic Fungus ◽  
Environmental Dna ◽  
Detection Sensitivity ◽  
Detection Rates ◽  
Filtration Step ◽  
Wildlife Pathogens

Monitoring the occurrence and density of parasites and pathogens can identify high infection-risk areas and facilitates disease control and eradication measures. Environmental DNA (eDNA) techniques are increasingly used for pathogen detection due to their relative ease of application. Since many factors affect the reliability and efficacy of eDNA-based detection, rigorous validation and assessment of method limitations is a crucial first step. We evaluated an eDNA detection method using in situ filtration of large-volume water samples, developed to detect and quantify aquatic wildlife parasites by quantitative PCR (qPCR). We assessed method reliability using Batrachochytrium dendrobatidis, a pathogenic fungus of amphibians and the myxozoan Tetracapsuloides bryosalmonae, causative agent of salmonid proliferative kidney disease, in a controlled experimental setup. Different amounts of parasite spores were added to tanks containing either clean tap water or water from a semi-natural mesocosm community. Overall detection rates were higher than 80%, but detection was not consistent among replicate samples. Within-tank variation in detection emphasises the need for increased site-level replication when dealing with parasites and pathogens. Estimated parasite DNA concentrations in water samples were highly variable, and a significant increase with higher spore concentrations was observed only for B. dendrobatidis. Despite evidence for PCR inhibition in DNA extractions from mesocosm water samples, the type of water did not affect detection rates significantly. Direct spiking controls revealed that the filtration step reduced detection sensitivity. Our study identifies sensitive quantification and sufficient replication as major remaining challenges for the eDNA-based methods for detection of parasites in water.

scholarly journals Molecular Characterization of a Trisegmented Mycovirus from the Plant Pathogenic Fungus Colletotrichum gloeosporioides

Viruses ◽  
2016 ◽  
Vol 8 (10) ◽  
pp. 268 ◽  
Author(s):  
Jie Zhong ◽  
Xi Pang ◽  
Hong Zhu ◽  
Bi Gao ◽  
Wen Huang ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Colletotrichum Gloeosporioides ◽  
Pathogenic Fungus ◽  
Plant Pathogenic Fungus

scholarly journals Dirhamnolipid Produced by the Pathogenic Fungus Colletotrichum gloeosporioides BWH-1 and Its Herbicidal Activity

Molecules ◽  
2019 ◽  
Vol 24 (16) ◽  
pp. 2969 ◽  
Author(s):  
Zhaolin Xu ◽  
Mengying Shi ◽  
Yongqing Tian ◽  
Pengfei Zhao ◽  
Yifang Niu ◽  
...  
Keyword(s):  
Weed Management ◽  
Colletotrichum Gloeosporioides ◽  
Pathogenic Fungus ◽  
Primary Root ◽  
Herbicidal Activity ◽  
Weed Species ◽  
Plant Weight ◽  
Phytotoxic Activity ◽  
Ic50 Values ◽  
Purity Check

Fungal phytotoxins used as ecofriendly bioherbicides are becoming efficient alternatives to chemical herbicides for sustainable weed management. Previous study found that cultures of the pathogenic fungus Colletotrichum gloeosporioides BWH-1 showed phytotoxic activity. This study further isolated the major phytotoxin from cultures of the strain BWH-1 using bioactivity-guided isolation, by puncturing its host plant for an activity test and analyzing on the HPLC-DAD-3D mode for a purity check. Then, the active and pure phytotoxin was characterized as a dirhamnolipid (Rha-Rha-C10-C10) using the NMR, ESIMS, IR and UV methods. The herbicidal activity of dirhamnolipid was evaluated by the inhibition rate on the primary root length and the fresh plant weight of nine test plants, and the synergistic effect when combining with commercial herbicides. Dirhamnolipid exhibited broad herbicidal activity against eight weed species with IC50 values ranging from 28.91 to 217.71 mg L−1 and no toxicity on Oryza sativa, and the herbicidal activity could be synergistically improved combining dirhamnolipid with commercial herbicides. Thus, dirhamnolipid that originated from C. gloeosporioides BWH-1 displayed the potential to be used as a bioherbicide alone, or as an adjuvant added into commercial herbicides, leading to a decrease in herbicides concentration and increased control efficiency.

Postharvest application of partitioned plant extracts from Sinaloa, Mexico for controlling papaya pathogenic fungus Colletotrichum gloeosporioides

2021 ◽  
Author(s):  
Jordi G. López-Velázquez ◽  
Francisco Delgado-Vargas ◽  
Lidia E. Ayón-Reyna ◽  
Gabriela López-Angulo ◽  
Silvia Bautista-Baños ◽  
...  
Keyword(s):  
Plant Extracts ◽  
Colletotrichum Gloeosporioides ◽  
Pathogenic Fungus ◽  
Postharvest Application

A novel ourmia-like mycovirus isolated from the plant pathogenic fungus Colletotrichum gloeosporioides

2019 ◽  
Vol 164 (10) ◽  
pp. 2631-2635 ◽  
Author(s):  
Jun Guo ◽  
Jun Zi Zhu ◽  
Xin Yu Zhou ◽  
Jie Zhong ◽  
Cai Hong Li ◽  
...  
Keyword(s):  
Colletotrichum Gloeosporioides ◽  
Pathogenic Fungus ◽  
Plant Pathogenic Fungus

Quantum-Dot/Dendrimer Based Functional Nanotubes for Sensitive Detection of DNA Hybridization

2008 ◽  
Vol 55 ◽  
pp. 84-90 ◽  
Author(s):  
Chuan Liang Feng ◽  
Ya Ming Yu ◽  
Xin Hua Zhong ◽  
Martin Steinhart ◽  
Jean P. Majoral ◽  
...  
Keyword(s):  
Quantum Dot ◽  
Resonance Energy Transfer ◽  
Fluorescence Emission ◽  
Resonance Energy ◽  
Detection Sensitivity ◽  
Layer By Layer ◽  
Enhanced Fluorescence ◽  
Highly Sensitive ◽  
Cascade Energy ◽  
Functionalized Nanotubes

A highly sensitive method for detecting oligonucleotide targets has been developed using quantum dot (QD) functionalized nanotubes (NTs) that contain a cascade energy band-gap architecture. Their assembly with different types of QDs by the layer-by-layer (LBL) allowed for the formation of fluorescence resonance energy transfer (FRET) structures in the NTs resulting in an enhanced fluorescence emission from dye labeled oligonucleotide targets captured by probe DNA through hybridization. This approach results in a significant enhancement of detection of dye labeled oligonucleotide targets. The promising increase in detection sensitivity allows for low detection limits by tuning the QD emission, adding other types of QDs, or adjusting the distance between all photoactive compounds in the NTs. Hence, the strategy used here to produce new hybrid materials may pave a way for broadening application of NTs in the field of biochemical materials, e.g. sensitive biosensors.

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