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Hyphae

Hyphae are the vegetative, tubular filaments that make up the body of a fungus.
They are responsible for absorbing nutrients, anchoring the fungus to its substrate, and facilitating reproduction.
Hyphae can form complex networks called mycelia, which allow fungi to explore and exploit their environment.
Understanding hyphae is crucial for research into fungal biology, ecology, and medical applications.
PubCompare.ai's AI-driven tools can help locate and optimize protocols related to hyphae, enhancing research reproducibilty and efficiency.
This concise overview provides an informative introduction to this important fungal structure.

Most cited protocols related to «Hyphae»

1
Quantifying Fungal Endocytosis and Adherence
The number of organisms or latex beads that were endocytosed or cell-associated with the various host cells was determined using our standard differential fluorescence assay [5 (link),7 (link),9 (link),51 (link)]. The host cells were grown to 95% confluency onto 12-mm diameter coverslips coated with fibronectin in 24-well tissue-culture plates. They were incubated with 105C. albicans hyphae in RPMI 1640 medium. After 45 min, the cells were rinsed twice with Hank's balanced salt solution (HBSS; Irvine Scientific) in a standardized manner and then fixed with 3% paraformaldehyde. In experiments performed with C. albicans, the adherent but nonendocytosed organisms were labeled with rabbit polyclonal anti–C. albicans antibodies (Biodesign International, http://www.biodesign.com) that had been conjugated with Alexa 568 (Invitrogen), which fluoresces red. Next, the cells were permeablized with 0.5% Triton X-100 (Sigma-Aldrich) in PBS, and then the cell-associated organisms (the endocytosed plus nonendocytosed organisms) were labeled with the anti–C. albicans antibodies conjugated with Alexa 488 (Invitrogen), which fluoresces green. The coverslips were viewed using an epifluorescent microscope, and the number of endocytosed organisms was determined by subtracting the number of nonendocytosed organisms (which fluoresced red) from the number of cell-associated organisms (which fluoresced green). At least 100 organisms were examined on each coverslip, and the results were expressed as the number of endocytosed or cell-associated organisms per high-powered field.
Experiments investigating the endocytosis and adherence of the yellow-green fluorescing latex beads were performed similarly, except that 3 × 105 beads were added to each well. The adherent, nonendocytosed control beads coated with biotinylated BSA were labeled with strepavidin Alexa 568. The adherent beads coated with either rAls1-N or rAls3-N were detected by indirect immunofluorescence using rabbit polyclonal anti-Als1 antibodies followed by Alexa 568–conjugated goat anti-rabbit antibodies.
Phan Q.T., Myers C.L., Fu Y., Sheppard D.C., Yeaman M.R., Welch W.H., Ibrahim A.S., Edwards JE J.r, & Filler S.G. (2007). Als3 Is a Candida albicans Invasin That Binds to Cadherins and Induces Endocytosis by Host Cells. PLoS Biology, 5(3), e64.
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Publication 2007
alexa 568 Amyotrophic lateral sclerosis 1 Anti-Antibodies anti-c antibody Biological Assay Cells Endocytosis Fibronectins Fluorescence Goat Hanks Balanced Salt Solution Hemoglobin, Sickle Hyphae Indirect Immunofluorescence Latex Microscopy paraform Rabbits Streptavidin Tissues Triton X-100
2
Fungal Colony Morphology and Growth Assay
Colony morphology and growth rate were assayed with potato dextrose agar (PDA) cultures grown at 25°C for three days. Conidiation was assayed with 5-day-old CMC cultures as described [20] (link), [70] (link). Conidium morphology was examined and photographed with an Olympus BX-51 microscope. For assaying conidium germination and germ tube growth, freshly harvested conidia were cultured in liquid YEPD medium for 12 h. Slab cultures grown on a thin layer of complete medium (CM) for 36 h were examined for defects in hyphal tip growth and branching [20] (link), [70] (link).
Wang C., Zhang S., Hou R., Zhao Z., Zheng Q., Xu Q., Zheng D., Wang G., Liu H., Gao X., Ma J.W., Kistler H.C., Kang Z, & Xu J.R. (2011). Functional Analysis of the Kinome of the Wheat Scab Fungus Fusarium graminearum. PLoS Pathogens, 7(12), e1002460.
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Publication 2011
Agar Conidia Germination Glucose Hyphae Microscopy Solanum tuberosum
3
Fungal Growth Characterization Protocol
Growth rates, the optimum temperature of growth, and colony characteristics
were determined on three different media at four different temperatures. The
strains were pregrown on CMD, or MEA (2 % malt extract, 2 % agar-agar, both
from Merck) where noted, until they reached a diameter of 55–65 mm. Agar
plugs 0.5 cm diam were then cut from the margin of the colonies and
transferred to fresh medium, 1.0–1.5 mm from the edge of the 9-cm-diam
Petri dish with the mycelium facing down on the surface of the agar. CMD, PDA
(potato dextrose agar, Merck, Darmstadt, Germany) and low nutrient agar (SNA,
Nirenberg 1976; pH adjusted to 5.5) were used. The tests were performed at 15
°C (with alternating 12 h weak UV light (Philips TL-D/08 blacklight blue)
and 12 h darkness), 25 °C (with alternating 12 h cool white fluorescent
light and 12 h darkness), and 30 °C and 35 °C (both in darkness). The
different lighting conditions were due to a simultaneous use of incubators to
culture for anamorph morphology. For growth at 25 °C, the Petri dishes
were sealed with Parafilm to avoid drying out of the agar caused by the
ventilator of the cooling incubator (MIR 153, Sanyo, Gunma, Japan). The
maximum colony radius was measured once daily for at least 7 d or until the
plates were entirely covered with mycelium. The growth rate was calculated by
linear regression of log t versus log r (t = time of incubation, r = radius
measured from the edge of the agar plug), using only measurements from the
phase where the logarithmic increase of the colony radius was linear over log
t. The data given are ranges obtained from 3–6 experiments for all media
and temperatures, except species with stipitate stromata, H. alutacea, H.
nybergiana and H. seppoi, where only single experiments were
carried out.
In addition, the plates were examined daily under the compound microscope
at low magnification (10× objective), and the time of first appearance
of conidia, autolytic behaviour of marginal hyphae, coilings in surface
hyphae, presence of chlamydospores, formation of pigments and odour, and the
colony appearance were noted.
, & Jaklitsch W.M. (2009). European species of Hypocrea Part I. The green-spored species. Studies in Mycology, 63, 1-91.
Publication 2009
Agar Autolysis Conidia Darkness Debility Glucose Hyperostosis, Diffuse Idiopathic Skeletal Hyphae Light Light Microscopy Mycelium Nutrients Odors Pigmentation Potato Radius Strains Ultraviolet Rays
4
High Molecular Weight Genomic DNA Extraction
QM6a was inoculated on petri-plates with malt extract agar (MEA) medium at 25 °C until full asexual sporulation was observed (~5 days). 2 × 108 conidial spores were collected, and then inoculated in 50 mL potato dextrose medium (PDB) at 30 °C for 6 h. The germinated hyphae were harvested by centrifugation at 3000g for 5 min at room temperature and incubated in 2 mL lysing enzyme buffer [0.1 M KH2PO4 (pH 5), 1.2 M Sorbitol, 5% lysing enzyme (Sigma, USA)] at 30 °C for 1.5 h. The protoplasts were harvested by centrifugation at 600g for 10 min at 4 °C, dissolved in 1.2 mL GuHCl solution (43% guanidine-HCl, 0.1 M EDTA pH 8.0, 0.15 M NaCl, 0.05% Sarkosyl) at 65 °C for 20 min and then mixed with 6.4 mL ice-cold ethanol to precipitate the genomic DNA. The pellet was dissolved in 10× TE with 0.6 mg/mL RNase H at 37 °C for 1 h and then in 0.4 mg/mL proteinase K at 65 °C for 1 h. The genomic DNA was purified with the phenol:chloroform:isoamyl alcohol (25:24:1) method and then recovered by standard precipitation with ethanol. Next, the quality of high molecular weight genomic DNA for Illumina MiSeq and PacBio sequencing was validated by PFGE. The genomic DNA was separated in a 1% agarose gel in 0.5× TBE buffer, using a CHEF DR II (Biorad) with 0.5× TBE running buffer, continuously refrigerated at 14 °C and 6 V/cm (current 110–125 mA) for 18 h. The Lambda DNA-Mono Cut Mix (New England Biolabs, N3019S) was used as size marker. Visualization was performed after staining with ethidium bromide after the electrophoresis.
Li W.C., Huang C.H., Chen C.L., Chuang Y.C., Tung S.Y, & Wang T.F. (2017). Trichoderma reesei complete genome sequence, repeat-induced point mutation, and partitioning of CAZyme gene clusters. Biotechnology for Biofuels, 10, 170.
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Publication 2017
Agar Buffers Centrifugation Chloroform Cold Temperature Conidia Edetic Acid Electrophoresis Electrophoresis, Gel, Pulsed-Field Endopeptidase K Enzymes Ethanol Ethidium Bromide Genome Glucose Guanidine Hyphae isopentyl alcohol Phenols Protoplasts Ribonuclease H Sepharose Sodium Chloride sodium lauroyl sarcosinate Solanum tuberosum Sorbitol Spores Tris-borate-EDTA buffer
5
Isolation and Cultivation of Sporothrix Strains
All isolates used throughout this study are listed in Table 1, including the geographical, genotype and clinical origin when pertinent. The yeast phase of each strain was obtained by cultivation in Brain Heart Infusion broth (Oxoid, Hampshire, UK) at 37 °C for 7 days, as previously described [30 (link)]. For in vivo experiments, the mycelia phase of each strain was grown in Sabouraud broth (Difco, Detroit, USA) for 5 days at room temperature, and the conidia were isolated from hyphae using a Buchner funnel and gauze (see Figure S2). For the scanning electron microscopy (SEM) experiments, the mycelial phase of S. schenckii isolates 1099-18 and IPEC 15383 and S. brasiliensis isolates IPEC 17943 and 5110 were cultivated in Sabouraud-agar discs placed between coverslips and incubated at room temperature for 15 days.
Castro R.A., Kubitschek-Barreira P.H., Teixeira P.A., Sanches G.F., Teixeira M.M., Quintella L.P., Almeida S.R., Costa R.O., Camargo Z.P., Felipe M.S., de Souza W, & Lopes-Bezerra L.M. (2013). Differences in Cell Morphometry, Cell Wall Topography and Gp70 Expression Correlate with the Virulence of Sporothrix brasiliensis Clinical Isolates. PLoS ONE, 8(10), e75656.
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Publication 2013
Agar Brain Conidia Genotype Heart Hyphae Mycelium Scanning Electron Microscopy Strains Yeasts

Most recents protocols related to «Hyphae»

1
Microscopic Examination of Fungal Infections
The wet mount preparation of urine and HVS sample were observed and the presence of pus cells, epithelial cells, red blood cells and Candida albicans positive were estimated under 10 × and 40 × objective lens.
The confirmation of candida infections was performed by observing the colonies from culture for their morphological characteristics such as size, cream-coloured pastry colonies, and morphology of the colonies and production of hyphae examined under a 40 × objective lens. Gram stain to identified Candida sp and staining with lactophenol cotton blue, Candida sp were finally examined under 40 × and 100 × under oil emersion [20 ].
Asare K.K., Bentil H.A., Gyesi E., Amoah S., Bentsi-Enchill F, & Opoku Y.K. (2023). Candidiasis profile at the outpatient department of the university of cape coast hospital in the central region of Ghana: a retrospective study. BMC Women's Health, 23, 101.
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Publication 2023
Candida Candida albicans Candidiasis Epithelial Cells Erythrocytes Gram's stain Hyphae lactophenol cotton blue Lens, Crystalline Urine
2
Visualizing Hyphal Development Using Trypan Blue
The hyphal development was visualized using the Trypan Blue staining technique (Koch and Slusarenko, 1990 (link)). In brief, the PM-infected melon leaves were soaked in Trypan Blue staining solution and immediately heated in 100 °C water for 2–5 min. After allowing the solution to cool down to room temperature, the staining solution was discarded and the leaves were decolored using a 2.5 mg/ml chloral hydrate solution, which was replaced every 24 hours until the leaves were completely decolored.
Zhang T., Xu N., Amanullah S, & Gao P. (2023). Genome-wide identification, evolution, and expression analysis of MLO gene family in melon (Cucumis melo L.). Frontiers in Plant Science, 14, 1144317.
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Publication 2023
Hydrate, Chloral Hyphae Melons Trypan Blue
3
Pot Experiment on Nitrogen Dynamics
Pot expt 1 was harvested 6 days after the addition of inorganic nitrogen. Pot expt 2 was harvested twice, on day 24 (i.e., 45 days after maize planting) and day 34 (i.e., 55 days after maize planting) after patch placement. The details of the harvest procedure and determination of soil water content, dissolved organic carbon (DOC), total dissolved nitrogen (TDN), mineral N concentrations, hyphal length density (HLD), TC, TN, ammonium (NH4+-N), and nitrate (NO3−-N) concentrations are shown in the Supplementary Information.
Li X., Zhao R., Li D., Wang G., Bei S., Ju X., An R., Li L., Kuyper T.W., Christie P., Bender F.S., Veen C., van der Heijden M.G., van der Putten W.H., Zhang F., Butterbach-Bahl K, & Zhang J. (2023). Mycorrhiza-mediated recruitment of complete denitrifying Pseudomonas reduces N2O emissions from soil. Microbiome, 11, 45.
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Publication 2023
Ammonium Dissolved Organic Carbon Hyphae Minerals Nitrates Nitrogen Zea mays
4
Microcosm Design for Hyphal Tracing
Microcosms with two chambers, one root chamber for plant growth (3 × 10 × 15 cm3) and one hyphal chamber for hyphal growth (7 × 10 × 15 cm3), were constructed (Fig. 1). The two chambers were separated by a 30-μm or a 0.45-μm mesh that allowed or prevented AMF hyphae access to the hyphal chamber. In all cases, the roots were not allowed to access the hyphal chamber.

Flow chart of the experiments. The study comprised several experiments: two pot experiments, in vitro experiments, and an inoculation experiment. The two pot experiments (pot expts 1 and 2) tested whether the proliferation of AMF into microsites of residues may reduce N2O emissions and disassemble the regulation pathway. We also tested the abundance of the microbiome in the hyphosphere; the in vitro experiment assessed the importance of P. fluorescens co-colonization with AMF for reduced N2O emissions. We isolated denitrifiers and identified the key components of hyphal exudates. We then examined the chemotaxis, growth, N2O emission, and denitrifying gene expression of P. fluorescens in response to AMF exudates and key compounds under in vitro culture conditions; finally, the inoculation experiment validated the effects of AMF or citrate exuded by AMF on N2O emissions and nosZ gene expression of P. fluorescens in pot culture. A conceptual model is used to illustrate the pathways by which AMF interact with P. fluorescens to mitigate N2O production

In each hyphal chamber, we introduced a patch with a 30-μm pore nylon mesh bag (4 × 7.5 cm2, 5 cm high) that could be filled with residues. A gas probe was inserted into the patch to collect gas samples to measure the N2O concentration as an indicator of N2O production in the patch (Fig. 1).
Li X., Zhao R., Li D., Wang G., Bei S., Ju X., An R., Li L., Kuyper T.W., Christie P., Bender F.S., Veen C., van der Heijden M.G., van der Putten W.H., Zhang F., Butterbach-Bahl K, & Zhang J. (2023). Mycorrhiza-mediated recruitment of complete denitrifying Pseudomonas reduces N2O emissions from soil. Microbiome, 11, 45.
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Publication 2023
Chemotaxis Citrates Exudate Gene Expression Hyphae Microbiome Nylons Plant Development Plant Roots Vaccination
5
Impacts of Hyphal Exudates on N2O Production
A sealed serum bottle assay was conducted to examine the effects of hyphal exudates and major compounds on net N2O production by P. fluorescens JL1. Hyphal exudate was applied as one treatment. Fructose, trehalose, citrate, malate, glutamine, or glutamic acid was selected as the carbon source treatment because these compounds were detected at high concentrations in hyphal exudates. Glucose was used as the control. There were 8 treatments in total. The same liquid MSR medium [17 (link)] as that used for the collection of hyphal exudates (see Supplementary Information) was used to dissolved specific carbon source. The carbon and nitrogen contents in the medium were adjusted to the same level as those in the hyphal exudate solutions (7.16 mM C and 2.35 mM N). The hyphal exudate medium and specific compound medium were supplemented with 10% FeNaEDTA (relative to MSR medium) to ensure denitrification. The medium pH was then adjusted to 7.2, and the medium was filtered through an Acrodisc syringe filter (0.22-μm Super Membrane, Pall Corporation, Port Washington, NY) to obtain carbon-based medium (CB medium). The CB medium was supplemented with 92.84 mM glucose to reach an initial C concentration of 100 mM. NO3−-N was supplemented to reach a level of 10 mM to ensure denitrification. The pellet obtained from the centrifugation of 1 mL P. fluorescens JL1 suspension was re-suspended in 10 mL modified CB medium and transferred to a 120-mL anaerobic serum bottle. All serum bottles were shaken at 180 rpm and maintained at 30 °C. The gas was measured after 0.5, 1, 2, 3, 6, 8, 10, and 12 h. Each treatment was set up in triplicate. Details are shown in the Supplementary Information.
Li X., Zhao R., Li D., Wang G., Bei S., Ju X., An R., Li L., Kuyper T.W., Christie P., Bender F.S., Veen C., van der Heijden M.G., van der Putten W.H., Zhang F., Butterbach-Bahl K, & Zhang J. (2023). Mycorrhiza-mediated recruitment of complete denitrifying Pseudomonas reduces N2O emissions from soil. Microbiome, 11, 45.
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Publication 2023
Biological Assay Carbon Centrifugation Citrates Denitrification Exudate Fructose Glucose Glutamic Acid Glutamine Hyphae malate Nitrogen Serum Syringes Tissue, Membrane Trehalose

Top products related to «Hyphae»

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Calcofluor white is a fluorescent brightening agent used in microscopy and staining applications. It binds to cellulose and chitin in cell walls, allowing for the visualization of fungal and other microbial structures.
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TRIzol reagent is a monophasic solution of phenol, guanidine isothiocyanate, and other proprietary components designed for the isolation of total RNA, DNA, and proteins from a variety of biological samples. The reagent maintains the integrity of the RNA while disrupting cells and dissolving cell components.
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DAPI is a fluorescent dye that binds strongly to adenine-thymine (A-T) rich regions in DNA. It is commonly used as a nuclear counterstain in fluorescence microscopy to visualize and locate cell nuclei.

More about "Hyphae"

Hyphae are the fundamental structural and functional units of fungi, the diverse group of eukaryotic organisms that play critical roles in ecosystems, agriculture, and human health.
These tubular, thread-like filaments make up the vegetative body of a fungus, known as the mycelium.
Hyphae are responsible for a variety of essential functions, including nutrient absorption, anchoring the fungus to its substrate, and facilitating reproduction through the formation of spores.
Understanding the biology and behavior of hyphae is crucial for research in fields such as fungal ecology, microbiology, and biotechnology.
Calcofluor white is a fluorescent dye commonly used to visualize and study the cell walls of hyphae and other fungal structures under a microscope.
The TRIzol reagent, on the other hand, is a powerful tool for extracting high-quality RNA from fungal samples, enabling researchers to investigate gene expression and other molecular aspects of hyphae.
FBS, or fetal bovine serum, is a commonly used growth supplement in fungal cell culture media, providing essential nutrients and growth factors.
Microscopy techniques, such as the BX51 and Eclipse 80i models, allow researchers to observe the intricate morphology and dynamic behavior of hyphae in detail.
The DNeasy Plant Mini Kit and RNeasy Plant Mini Kit are used to extract high-quality DNA and RNA, respectively, from fungal samples, facilitating genetic and transcriptomic analyses of hyphae.
Miracloth is a specialized filtration material often used in fungal research to separate hyphae and other fungal structures from liquid media or soil samples.
By leveraging these tools and techniques, researchers can gain deeper insights into the structure, function, and ecological significance of hyphae, ultimately expanding our understanding of the fascinating world of fungi and their impact on our environment, agriculture, and human health.
PubCompare.ai's AI-driven protocol optimization tools can further enhance research reproducibility and efficiency in this field, helping scientists locate and optimize the most effective protocols for studying hyphae and related fungal structures.