المؤلفان بالشكر كارين Hilburn حصول على الدعم الفني الممتاز. وقد أيد هذا المشروع من قبل مبادرة القومي للبحوث التعاونية للدولة وزارة الزراعة الأميركية للتعليم والبحوث والإرشاد (جائزة 2004-35604-14327 #). معتمد من قبل ماتياس باسكوالي برانكو فايس الزمالة. ومن المسلم به أيضا الولايات المتحدة والقمح والشعير والجلبة مبادرة للدعم المتواصل من البحوث.

<ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> آغار الجزرة (Klittich ويزلي ، 1988) وقد أعدت في 9 سم ، وقطرها أطباق بتري. </li><li style=";text-align:right;direction:rtl"> تم تطبيق مكعب من 2 ملم تحتوي على أجار أفطورة جديدة نمت على أجار دكستروز البطاطس الى وسط كل طبق بتري. </li><li style=";text-align:right;direction:rtl"> وقد نمت الثقافات عند 25 درجة مئوية والإضاءة 12 ساعة لمدة 96 ساعة. </li><li style=";text-align:right;direction:rtl"> طبق واحد مل من توين مائي 2.5 ٪ من 60 إلى السطح من الثقافة (تريل والمشتركة عام 2000 ،) والمنتشرة في الثقافة بقضيب من البلاستيك ، عن طريق الضغط بلطف. </li><li style=";text-align:right;direction:rtl"> Perithecia اطلاق النار ascospores المتقدمة 7-10 أيام بعد هذا العلاج التعريفي عندما أبقى في بيئة الرطوبة العالية في 25 درجة مئوية مع الإضاءة نفسها. </li><li style=";text-align:right;direction:rtl"> يتم تنفيذ جمع بوغ عن طريق تغيير غطاء صحن بيتري ، وترك perithecia لاطلاق النار لفترة محدودة (على سبيل المثال 12 ساعة) في الظلام. </li><li style=";text-align:right;direction:rtl"> يضاف الماء المعقم إلى غطاء ، وتعليق ascospores برفق عن طريق تحريك المياه على السطح. </li><li style=";text-align:right;direction:rtl"> يتم جمع Ascospores في أنابيب معقمة الصقر ، وغسلها واستخدامها لمزيد من التحليل. </li></ol>

<ol>
	<li>Beyer, M., Verreet, J. A., Francl, L., Shaner, G., Bergstrom, G., Gilbert, J., Pedersen, W., Dill-Macky, R., Sweets, L., Corwin, B., Jin, Y., Gallenberg, D., Wiersma, J., Goswami, R. S., Kistler, H. C., Klittich, C. J. R., Leslie, J. F., Trail, F., Common, R., Tschantz, A. T., Horst, R. L., Nelson, P. E., Tschanz, A. T., Horst, R. K., Nelson, P. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Germination+of+Gibberella+zeae+ascospores+as+affected+by+age+of+spores+after+discharge+and+environmental+factors.">Germination of Gibberella zeae ascospores as affected by age of spores after discharge and environmental factors.</a> Genetics. 118, 417-2013 (2005).</li></ol>

ويستند البروتوكول المقدمة هنا على الإجراءات السابقة المستخدمة لإنتاج perithecial لSPP الفيوزاريوم. (Klittich ويزلي ، 1988 ؛ تريل والمشتركة عام 2000 ،). وتوحيد الإجراءات التي تم إنشاؤها على كميات كبيرة من ascospores (تكفي لتحليل ميكروأري) ضروري لاستنساخ التجربة. وقد أفيد أن العوامل البيئية ، والعمر?...

<table><tbody><tr><td>Petri dish 9 cm diameter</td><td>Tool</td><td></td><td></td><td></td></tr><tr><td>Tween 60 (20%)</td><td>Reagent</td><td></td><td></td><td></td></tr><tr><td>Carrot agar (20% w/v organic carrots and 1.5% w/v agar) </td><td>Reagent</td><td></td><td></td><td></td></tr></tbody></table>

gibberella zeae ascospore production and collection for microarray experiments.

فيوزاريوم graminearum شواب (teleomorph Gibberella zeae) هو الممرض مصنع يسبب مرض الجرب على القمح والشعير الذي يقلل من المحصول وجودة الحبوب. F. graminearum يتسبب أيضا في ساق والأذن وتتعفن من الذرة هو منتج للسموم الفطرية مثل trichothecenes التي تلوث الحبوب ومضرة للانسان والماشية (جوسوامي وكيستلر ، 2004). الفطريات تنتج نوعين من الجراثيم. Ascospores ، وبطرق الزراعه الناتجة عن التكاثر الجنسي ، هي المصدر الرئيسي لنقل العدوى الأولية. يتم تفريغها قسرا من هذه الجراثيم perithecia ناضجة ومشتتة بفعل الرياح (Francl وآخرون 1999). هي سبب رئيسي العدوى الثانوية macroconidia التي تنتجها وسائل اجنسي على سطح المصنع. لدراسة العمليات التنموية للascospores في هذا الفطر ، وكان المطلوب إجراء لجمع كمية كبيرة في ظل ظروف معقمة. لدينا بروتوكول تم تصويره من أجل توليد أكبر قدر من المعلومات لفهم واستنساخ ، والجوانب الحاسمة عندما يتم إنشاؤها بالكامل ملامح التعبير الجيني الجينوم وتفسيرها. على وجه الخصوص ، تقلب بوغ زقي الإنبات والنشاط البيولوجي تعتمد على التلاعب مسبقة من المواد. ويقترح استخدام الفيديو لتوثيق كل خطوة من خطوات الإنتاج بوغ زقي من أجل زيادة التوحيد القياسي ، والامتثال للمتطلبات الصارمة المتزايدة للتحليل [ميكروأري. الإجراء يتطلب معدات المختبرات فقط القياسية. تظهر الخطوات اللازمة لمنع التلوث وصالح تزامن وقت ascospores.

Watch this Scientific Journal Video about Gibberella zeae Ascospore Production and Collection for Microarray Experiments. at JoVE.com

Gibberella zeae Ascospore Production and Collection for Microarray Experiments.

لدراسة العمليات التنموية في ascospores zeae Gibberella ، وهو إجراء لجمع تحت ظروف معقمة تم تصويره من أجل توليد أكبر قدر من المعلومات لوصف البروتوكول. هذا يجب تسهيل استنساخ التجربة ، جانبا حاسما عندما تنفذ بالكامل الاختبارات الشخصية الجينوم التعبير.

بوغ زقي zeae Gibberella الإنتاج والتحصيل للتجارب ميكروأري.

Fusarium graminearum Schwabe (teleomorph Gibberella zeae) is a plant pathogen causing scab disease on wheat and barley that reduces crop yield and grain ...

gibberella-zeae-ascospore-production-collection-for-microarray

Research

JoVE Journal

Biology

14.2K Views.  USDA. لدراسة العمليات التنموية في ascospores zeae Gibberella ، وهو إجراء لجمع تحت ظروف معقمة تم تصويره من أجل توليد أكبر قدر من المعلومات لوصف البروتوكول. هذا يجب تسهيل استنساخ التجربة ، جانبا حاسما عندما تنفذ بالكامل الاختبارات الشخصية الجينوم التعبير.

Video: Gibberella zeae Ascospore Production and Collection for Microarray Experiments.

Quantification of Fungal Colonization, Sporogenesis, and Production of Mycotoxins Using Kernel Bioassays

The rotting of grains by seed-infecting fungi poses one of the greatest economic challenges to cereal production worldwide, not to mention serious risks to human and animal health. Among cereal production, maize is arguably the most affected crop, due to pathogen-induced losses in grain integrity and mycotoxin seed contamination. The two most prevalent and problematic mycotoxins for maize growers and food and feed processors are aflatoxin and fumonisin, produced by Aspergillus flavus and Fusarium verticillioides, respectively.
Recent studies in molecular plant-pathogen interactions have demonstrated promise in understanding specific mechanisms associated with plant responses to fungal infection and mycotoxin contamination1,2,3,4,5,6. Because many labs are using kernel assays to study plant-pathogen interactions, there is a need for a standardized method for quantifying different biological parameters, so results from different laboratories can be cross-interpreted. For a robust and reproducible means for quantitative analyses on seeds, we have developed in-lab kernel assays and subsequent methods to quantify fungal growth, biomass, and mycotoxin contamination. Four sterilized maize kernels are inoculated in glass vials with a fungal suspension (106) and incubated for a predetermined period. Sample vials are then selected for enumeration of conidia by hemocytometer, ergosterol-based biomass analysis by high performance liquid chromatography (HPLC), aflatoxin quantification using an AflaTest fluorometer method, and fumonisin quantification by HPLC.

The devastation of cereal crops by seed-infecting fungi has prompted numerous research efforts to better understand plant-pathogen interactions. To study seed-fungal interactions in a laboratory setting, we developed a robust method for the quantification of fungal reproduction, biomass, and mycotoxin contamination using kernel bioassays.

الكمي للاستعمار الفطرية، تولد الأبواغ، وإنتاج السموم الفطرية عن طريق اختبارات بيولوجية على النواة

The rotting of grains by seed-infecting fungi poses one of the greatest economic challenges to cereal production worldwide, not to mention serious risks ...

Immunology and Infection

Inhibition of Aspergillus flavus Growth and Aflatoxin Production in Transgenic Maize Expressing the &#945;-amylase Inhibitor from Lablab purpureus L.

Aflatoxin contamination in food and feed crops is a major challenge worldwide. Aflatoxins, produced by the fungus Aspergillus flavus (A. flavus) are potent carcinogens that substantially reduce crop value in maize and other oil rich crops like peanut besides posing serious threat to human and animal health. Different approaches, including traditional breeding, transgenic expression of resistance associated proteins, and RNA interference (RNAi)-based host-induced gene silencing of critical A. flavus gene targets, are being evaluated to increase aflatoxin resistance in susceptible crops. Past studies have shown an important role of &#945;-amylase in A. flavus pathogenesis and aflatoxin production, suggesting this gene/enzyme is a potential target to reduce both A. flavus growth and aflatoxin production. In this regard, the current study was undertaken to evaluate heterologous expression (under control of the constitutive CaMV 35S promoter) of a Lablab purpureus L. &#945;-amylase inhibitor-like protein (AILP) in maize against A. flavus. AILP is a 36-kDa protein, which is a competitive inhibitor of A. flavus &#945;-amylase enzyme and belongs to the lectin&#8211;arcelin&#8211;&#945;-amylase inhibitor protein family in common bean. In vitro studies prior to the current work had demonstrated the role of AILP in inhibition of A. flavus &#945;-amylase activity and fungal growth. Fungal growth and aflatoxin production in mature kernels were monitored in real time using a GFP-expressing A. flavus strain. This kernel screening assay (KSA) is very simple to set up and provides reliable and reproducible data on infection and the extent of spread that could be quantified for evaluation of germplasm and transgenic lines. The fluorescence from the GFP strain is closely correlated to fungal growth and, by extension, it is well-correlated to aflatoxin values.&#160; The goal of the current work was to implement this previous knowledge in a commercially important crop like maize to increase aflatoxin resistance. Our results show a 35%&#8211;72% reduction in A. flavus growth in AILP-expressing transgenic maize kernels which, in turn, translated into a 62%&#8211;88% reduction in aflatoxin levels.

Inhibition of Aspergillus flavus Growth and Aflatoxin Production in Transgenic Maize Expressing the &#945;-amylase Inhibitor from Lablab purpureus L.

Here we present a protocol to analyze Aspergillus flavus growth and aflatoxin production in maize kernels expressing an antifungal protein.&#160; Using a GFP-expressing A. flavus strain we monitored the infection and spread of the fungus in mature kernels in real time. The assay is rapid, reliable, and reproducible.

تثبيط النمو رشاشيه الخنزير و "إنتاج الأفلاتوكسين" في "الإعراب عن الذرة المعدلة وراثيا" المانع α-الأميليز من purpureus لابلاب ل.

Aflatoxin contamination in food and feed crops is a major challenge worldwide. Aflatoxins, produced by the fungus Aspergillus flavus (A. flavus) are ...

Environment

Microarray Analysis for Saccharomyces cerevisiae

In this protocol, gene expression in yeast (Saccharomyces cerevisiae) is changed after exposure to oxidative stress induced by the addition of hydrogen peroxide (H2O2), an oxidizing agent. In the experiment, yeast is grown for 48 hours in 1/2X YPD broth containing 3X glucose. The culture is split into a control and treated group. The experiment culture is treated with 0.5 mM H2O2 in Hanks Buffered Saline (HBSS) for 1 hour. The control culture is treated with HBSS only. Total RNA is extracted from both cultures and is converted to a biotin-labeled cRNA product through a multistep process. The final synthesis product is taken back to the UVM Microarray Core Facility and hybridized to the Affymetrix yeast GeneChips. The resulting gene expression data are uploaded into bioinformatics data analysis software.

Microarray Analysis for Saccharomyces cerevisiae

In this protocol, gene expression in yeast (Saccharomyces cerevisiae) is changed after exposure to oxidative stress induced by the addition of hydrogen peroxide (H2O2), an oxidizing agent.

تحليل لميكروأري السكيراء الجعوية</em

In this protocol, gene expression in yeast (Saccharomyces cerevisiae) is changed after exposure to oxidative stress induced by the addition of hydrogen ...

Histological Quantification to Determine Lung Fungal Burden in Experimental Aspergillosis

The quantification of lung fungal burden is critical for the determination of the relative levels of immune protection and fungal virulence in mouse models of pulmonary fungal infection. Although multiple methods are used to assess fungal burden, quantitative polymerase chain reaction (qPCR) of fungal DNA has emerged as a technique with several advantages over previous culture-based methods. Currently, a comprehensive assessment of lung pathology, leukocyte recruitment, fungal burden, and gene expression in mice with invasive aspergillosis (IA) necessitates the use of a significant number of experimental and control animals. Here the quantification of lung histological staining to determine fungal burden using a reduced number of animals was examined in detail. Lung sections were stained to identify fungal structures with Gomori&#39;s modified methanamine silver (GMS) staining. Images were taken from the GMS-stained sections from 4 discrete fields of each formalin-fixed paraffin-embedded lung. The GMS stained areas within each image were quantified using an image analysis program, and from this quantification, the mean percentage of stained area was determined for each sample. Using this strategy, eosinophil-deficient mice exhibited decreased fungal burden and disease with caspofungin therapy, while wild-type mice with IA did not improve with caspofungin. Similarly, fungal burden in mice lacking &#947;&#948; T cells were also improved by caspofungin, as measured by qPCR and GMS quantification. GMS quantification is therefore introduced as a method for the determination of relative lung fungal burden that may ultimately reduce the quantity of experimental animals required for comprehensive studies of invasive aspergillosis.

Here we describe a protocol to determine pulmonary fungal burden in mice with invasive aspergillosis by quantification of Gomori&#39;s modified methanamine silver staining in histological sections. Use of this method resulted in comparable results with less animals compared to assessment of fungal burden by quantitative PCR of lung fungal DNA.

القياس الكمي نسيجية لتحديد عبء الرئة الفطرية في أسبيرجيلوسيس التجريبية

The quantification of lung fungal burden is critical for the determination of the relative levels of immune protection and fungal virulence in mouse ...

High Throughput Yeast Strain Phenotyping with Droplet-Based RNA Sequencing

The powerful tools available to edit yeast genomes have made this microbe a valuable platform for engineering. While it is now possible to construct libraries of millions of genetically distinct strains, screening for a desired phenotype remains a significant obstacle. With existing screening techniques, there is a tradeoff between information output and throughput, with high-throughput screening typically being performed on one product of interest. Therefore, we present an approach to accelerate strain screening by adapting single cell RNA sequencing to isogenic picoliter colonies of genetically engineered yeast strains. To address the unique challenges of performing RNA sequencing on yeast cells, we culture isogenic yeast colonies within hydrogels and spheroplast prior to performing RNA sequencing. The RNA sequencing data can be used to infer yeast phenotypes and sort out engineered pathways. The scalability of our method addresses a critical obstruction in microbial engineering.

A bottleneck in the &#8216;design-build-test&#8217; cycle of microbial engineering is the speed at which we can perform functional screens of strains. We describe a high-throughput method for strain screening applied to hundreds to thousands of yeast cells per experiment that utilizes droplet-based RNA sequencing.

ارتفاع الإنتاجية الخميرة سلالة Phenotyping مع تسلسل الحمض النووي الريبي القائم على دروبليت

The powerful tools available to edit yeast genomes have made this microbe a valuable platform for engineering. While it is now possible to construct ...

Bioengineering

Sexual Development and Ascospore Discharge in Fusarium graminearum

Fusarium graminearum has become a model system for studies in development and pathogenicity of filamentous fungi. F. graminearum most easily produces fruiting bodies, called perithecia, on carrot agar. Perithecia contain numerous tissue types, produced at specific stages of perithecium development. These include (in order of appearance) formation of the perithecium initials (which give rise to the ascogenous hyphae), the outer wall, paraphyses (sterile mycelia which occupy the center of the perithecium until the asci develop), the asci, and the ascospores within the asci14. The development of each of these tissues is separated by approximately 24 hours and has been the basis of transcriptomic studies during sexual development12,8. Refer to Hallen et al. (2007) for a more thorough description of development, including photographs of each stage. Here, we present the methods for generating and harvesting synchronously developing lawns of perithecia for temporal studies of gene regulation, development, and physiological processes. Although these methods are written specifically to be used with F. graminearum, the techniques can be used for a variety of other fungi, provided that fruiting can be induced in culture and there is some synchrony to development. We have recently adapted this protocol to study the sexual development of F. verticillioides. Although individual perithecia must be hand picked in this species, because a lawn of developing perithecia could not be induced, the process worked well for studying development (Sikhakolli and Trail, unpublished).
The most important function of fungal fruiting bodies is the dispersal of spores. In many of the species of Ascomycota (ascus producing fungi), spores are shot from the ascus, due to the generation of turgor pressure within the ascus, driving ejection of spores (and epiplasmic fluid) through the pore in the ascus tip2,7. Our studies of forcible ascospore discharge have resulted in development of a "spore discharge assay", which we use to screen for mutations in the process. Here we present the details of this assay.
F. graminearum is homothallic, and thus can form fruiting bodies in the absence of a compatible partner. The advantage of homothallism is that crossing is not necessary to generate offspring homozygous for a particular trait, a facet that has facilitated the study of sexual development in this species14,7. However, heterothallic strains have been generated that can be used for crossing5,9. It is also possible to cross homothallic strains to obtain mutants for several genes in one strain1. This is done by coinoculating one Petri dish with 2 strains. Along the meeting point, the majority of perithecia will be recombinant (provided a mutation in one of the parent strains does not inhibit outcrossing). As perithecia age, they exude ascospores en masse instead of forcibly discharging them. The resulting spore exudate (called a cirrhus) sits at the tip of the perithecium and can easily be removed for recovery of individual spores. Here we present a protocol to facilitate the identification of recombinant perithecia and the recovery of recombinant progeny.

Sexual Development and Ascospore Discharge in Fusarium graminearum

Sexual crosses and isolation of recombinant progeny are important research tools for the filamentous fungus, Fusarium graminearum, The techniques necessary successfully carry out these processes are presented.

الجنسي للتنمية والتفريغ في بوغ زقي فيوزاريوم graminearum</em

Fusarium graminearum has become a model system for studies in development and pathogenicity of filamentous fungi. F. graminearum most easily produces ...

Infection of Zebrafish Larvae with Aspergillus Spores for Analysis of Host-Pathogen Interactions

Invasive aspergillosis (IA) is one of&#160;the most common fungal infections among immunocompromised individuals. Despite the availability of antifungal drugs, IA can cause &#62;50% mortality in infected immunocompromised patients. It is crucial to determine both host and pathogen factors that contribute to infection susceptibility and low survival rates in infected patients in order to develop novel therapeutics. Innate immune responses play a pivotal role in recognition and clearance of Aspergillus spores, though little is known about the exact cellular and molecular mechanisms. Reliable models are required to investigate detailed mechanistic interactions between the host and pathogen. The optical clarity and genetic tractability of zebrafish larvae make them an intriguing model&#160;to study host-pathogen interactions of multiple human bacterial and fungal infections in a live and intact host. This protocol describes a larval zebrafish Aspergillus infection model. First, Aspergillus spores are isolated and injected into the zebrafish hindbrain ventricle via microinjection. Then, chemical inhibitors such as immunosuppressive drugs are added directly to the larval water. Two methods to monitor the infection in injected larvae are described, including the 1) homogenization of larvae for colony forming unit (CFU) enumeration and 2) a repeated, daily live imaging setup. Overall, these techniques can be used to mechanistically analyze the progression of Aspergillus infection in vivo and can be applied to different host backgrounds and Aspergillus strains to interrogate host-pathogen interactions.

Infection of Zebrafish Larvae with Aspergillus Spores for Analysis of Host-Pathogen Interactions

This protocol describes an Aspergillus infection model in zebrafish larvae. Aspergillus spores are microinjected into the hindbrain of larvae, and chemical treatment is used to induce immunosuppression. Infection progression is monitored via a daily imaging setup to monitor fungal growth and immune responses as well as enumeration of live spores by colony forming unit plating.

عدوى يرقات حمار وحشي مع جراثيم أسبيرغيلوس لتحليل التفاعلات المضيفة الممرضة

Invasive aspergillosis (IA) is one of&#160;the most common fungal infections among immunocompromised individuals. Despite the availability of antifungal ...

Quantitative Analysis of Aspergillus nidulans Growth Rate using Live Microscopy and Open-Source Software

It is well established that colony growth of filamentous fungi, mostly dependent on changes in hyphae/mycelia apical growth rate, is macroscopically estimated on solidified media by comparing colony size. However, to quantitatively measure the growth rate of genetically different fungal strains or strains under different environmental/growth conditions (pH, temperature, carbon and nitrogen sources, antibiotics, etc.) is challenging. Thus, the pursuit of complementary approaches to quantify growth kinetics becomes mandatory in order to better understand fungal cell growth. Furthermore, it is well-known that filamentous fungi, including Aspergillus spp., have distinct modes of growth and differentiation under sub-aerial conditions on solid media or submerged cultures. Here, we detail a quantitative microscopic method for analyzing growth kinetics of&#160;the model fungus Aspergillus nidulans, using live imaging in both submerged cultures and solid media. We capture images, analyze, and quantify growth rates of different fungal strains in a reproducible and reliable manner using an open source, free software for bio-images (e.g., Fiji), in a way that does not require any prior image analysis expertise from the user.

Quantitative Analysis of Aspergillus nidulans Growth Rate using Live Microscopy and Open-Source Software

We present a label-free live imaging protocol using transmitted light microscopy techniques to capture images, analyze and quantify growth kinetics of the filamentous fungus A. nidulans in both submerged cultures and solid media. This protocol can be used in conjunction with fluorescence microscopy.

التحليل الكمي لمعدل نمو النيدولانات Aspergillus باستخدام المجهر الحي والبرمجيات مفتوحة المصدر

It is well established that colony growth of filamentous fungi, mostly dependent on changes in hyphae/mycelia apical growth rate, is macroscopically ...

Isolation of Culturable Yeasts and Molds from Soils to Investigate Fungal Population Structure

Soil is host to an incredible amount of microbial life, with each gram containing up to billions of bacterial, archaeal, and fungal cells. Multicellular fungi such as molds and unicellular fungi, broadly defined as yeasts, fulfill essential roles in soil ecosystems as decomposers of organic material and as food sources for other soil dwellers. Fungal species diversity in soil is dependent on a multitude of climatic factors such as rainfall and temperature, as well as soil properties including organic matter, pH, and moisture. Lack of adequate environmental sampling, especially in regions of Asia, Africa, South America, and Central America, hinders the characterization of soil fungal communities and the discovery of novel species. 
We characterized soil fungal communities in nine countries across six continents using ~4,000 soil samples and a protocol developed in the laboratory for the isolation of yeasts and molds. This protocol begins with separate selective enrichment for yeasts and the medically relevant mold Aspergillus fumigatus, in liquid media while inhibiting bacterial growth. Resulting colonies are then transferred to solid media and further processed to obtain pure cultures, followed by downstream genetic characterization. Yeast species identity is established via sequencing of their internal transcribed spacer (ITS) region of the nuclear ribosomal RNA gene cluster, while global population structure of A. fumigatus is explored via microsatellite marker analysis. 
The protocol was successfully applied to isolate and characterize soil yeast and A. fumigatus populations in Cameroon, Canada, China, Costa Rica, Iceland, Peru, New Zealand, and Saudi Arabia. These findings revealed much-needed insights on global patterns in soil yeast diversity, as well as global population structure and antifungal resistance profiles of A. fumigatus. This paper presents the method of isolating both yeasts and A. fumigatus from international soil samples.

This protocol is an effective, speedy method of culturing yeasts and the mold Aspergillus fumigatus from large sets of soil samples in as little as 7 days. The methods can be easily modified to accommodate a range of incubation media and temperatures as needed for experiments.

عزل الخمائر والعفن القابل للزراعة عن التربة للتحقيق في التركيب السكاني الفطري

Soil is host to an incredible amount of microbial life, with each gram containing up to billions of bacterial, archaeal, and fungal cells. Multicellular ...

A zWEDGI Technique to Visualize Fungal Pathogen-Infected Zebrafish Larvae

Source: Thrikawala, S. et al., <a href="https://app.jove.com/v/61165">Infection of Zebrafish Larvae with Aspergillus Spores for Analysis of Host-Pathogen Interactions.</a>&#160;J. Vis. Exp.&#160;(2020)
The video demonstrates a zebrafish wounding and entrapment device technique for observing fluorescently labeled fungal pathogen infection in zebrafish larvae. Initial infection attracts macrophages, and as infection persists, neutrophil recruitment progresses at the injection site.

تقنية zWEDGI لتصور يرقات الزرد المصابة بممرضات الفطريات

1. Preparation of&#160;Aspergillus&#160;spores for injection

	From an&#160;Aspergillus&#160;spore suspension, calculate the volume needed to obtain 1 x ...

بوغ زقي zeae Gibberella الإنتاج والتحصيل للتجارب ميكروأري.

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