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Questions related to Fungal Biology
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The Ash dieback fungus Hymenoscyphus pseudoalbidus (or Chalara fraxinea) has been infecting European Ash (Fraxinus excelsior) across many areas of Europe, but seems to not have affected the Iberian peninsula. I have some questions about this:
1) Is the above statement true or are there reported cases on the Iberian Peninsula?
2) Dr Graham Rowe at Uni of Derby has suggested that genetic differences mean infection of F. excelsior may not be as virulent on the Iberian Peninsula: http://www.derby.ac.uk/news/dna-profile-of-british-ash-trees-could-make-them-at-less-risk-from-dieback-ecologist-claims What other data/publications are there on this suggestion?
3) Fraxinus angustifolia is more common than F. excelsior across much of the Iberian Peninsula. Is F. angustifolia as susceptible to infection as F. excelsior?
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Hi,
See the first report of Hf from Spain:
When it is moist and not too hot, the fungus will thrive well. The rather warm and dry conditions on the Iberian penninsula will likely limit the pathogen and thus disease intensity.
F. angustifolia is likely as susceptible as F. excelsior, but in Southern Europe the impact of ash dieback may be less, due to climatic conditions.
I hope that this helps.
With kind regards,
Thomas Kirisits
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sterile control of a mineral substrat for plants
grow medium: sabouraud agar
sample 2
tia
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In the picture above, the grayish brown mycelium represents the fungus Rhizopus. While the white and greenish blue colonies represent the fungus Penicillium.
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steril control of a mineral substrats for plants
grow medium: sabouraud agar
sample 8
tia
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The above micrograph shows penicillium spp.
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sterile control of a mineral substrate for plants
grow medium: sabouraud agar
tia
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The colony which limited with yellow marker with white mycelium may be belong to the fungus Beauveria bassiana, but not exactly. It needs more images to clearly identified.
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In order to study the optimization of moroccan algae biomass, we're monotoring the growth of 4 different green algae in a laboratory setting in liquid medium under different LED light intensities.
As the culture progressed, we noticed severe discoloration and loss of integrity of one of the macroalgae cultures. However, the rest of the algae species were not affected and remained green.
  • What could be the cause/causes of these changes? Is it a fungal contamination or a sign that this algae reacted negatively to the light intensities chosen?
  • What would be the best approach to investigate the cause and to determine it?
Any suggestion would be highly appreciated.
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Decoloration est dur aux rzyons solaire ou n'importe quels paramètres qui peuvent dégrader les couleurs
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I could find treatment strategy or guidelines on humans but not on fishes. Could you suggest me some related to fishes?
Thank you
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I have fungal contamination in my incubator and I want anything that may help me to get rid of this contamination because it's bothering me in cell culture or any work inside incubator.
I washed incubator by hot water and alcohol 75% then 99% but I don't know how to eradicate this fungi
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Mohamed S Ahmed yes when we are talking about fungal contaminations specially like aspergillus and penicillum which are the common source causing breakdown of the compounds such as gelatine and most of the fungi found in the environment cause infections. And the ways by which these can be avoided or reduced is specially the spores from the culture can contaminate the other cultures and we need to remove them and after removing the contaminated cultures clean the entire incubation centre with a cloth soaked in hypochlorite and then with ethanolto reomvoe the fungal contaminations
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I'm raising some D. magna and one of the cultures seems to be faltering. A lot of them have started to sink and turn a very bright white (not just translucent). Here is a quick picture I took under a dissecting scope of one of them.
They're in potable spring water (Crystal Springs which they seem to do very well in, at least better than Carolina's) and I feed them a mix of 50mg DIH2O/1g baker's yeast at around 2mL about every other day in a ~3-4L container. I started the culture on 2/1/21. We're trying to see how small/easy of a budget we can grow them in.
I have 5 cultures going right now and this is the only one showing issues. Is this some sort of disease and if so should I quickly quarantine/kill these in case it can spread to the other containers?
edit: I added a compound microscope picture as well. She looks... gunky. It looks like she's got a bunch of fuzz floating around inside.
edit 2: Now the water is certainly contaminated with either some sort of fungus or Vorticella that are attaching to her carapace. Other daphnia have had their eggs turn completely white as well
edit 3: After talking to a friend who has also raised daphnia, I realize now the source of my anguish is from parasitic copepods that lay eggs that then hatch and crawl inside other crustaceans and eat them from the inside out
I'm going to keep this question up and answered in case anyone else ever comes across this issue
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I figured it out. It was some freaking copepods contaminated in my samples from Carolina
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We have cultured Fusarium oxysporum on PDA medium and we want to store it for a long period of time. We would be thankful if someone can provide an easy method to store fungal strain especially Fusarium oxysporum using sterile water.
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Dear Abhik.
Most fungi can be kept alive on many of the agar media for 6 months or 1 year before transfers are necessary. Sporulating agar cultures may be frozen at -20 C. in the deep freeze and maintained for considerable periods of time. Another good method for keeping cultures for a number of years is by pouring sterilized mineral oil in the test tube until the culture is completely submerged . The best method for preservation of most fungi that sporulate is by the lyophil process or the freeze dry method . Cultures have been maintained for at least 20 years by this method.(Alexopoulos and Beneke, 1962). you can also use sterilized sand for preserve of F.oxysporum.
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Dear All,
The forward and reverse sequences obtained from the PCR products are not identical at all.
Can I still make the contigs for doing BLASTn?
Individually both sequences are showing the same species in BLASTn.
Together, by placing one sequence after another they are showing a different species.
Kindly suggest a way to identify the organism by using both forward and reverse sequences.
Best regards
Arush
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Matthew B Paddock is correct but I will add one more thing. If your amplimer has a small insert or deletion ( CA repeat or poly A or similar) near to one end then the sequence will look good from the other end but will be weak signalled and messy after the indel when sequenced from the indel end. You may have to trim off poorly sequenced bases at the ends of each sequence to get a good Blast result but if you are looking at gene coding sequences then genes are highly conserved through species and it may just be that your search finds many identical sequences and the one that you are interested in is way down a list of nearly identical sequences. You may have to increase the number of matches shown
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Hello,
Our project description page details the unusual origins of our research project/group- looking for the identity of a (likely novel) organism that may be causing a form of chronic illness.
We have successfully cultured an organism from blood donated by affected individuals, and failed to culture it from healthy control blood samples. The organism appears to grow preferentially in the deep matrix of the agar, and only the sporulation stage is visible as surface colonies.
Microscopically, the organism forms structures resembling pseudohyphae, fruiting bodies, and birefringent walled cysts. We hypothesize that it may be related to myxomycetes, oomycetes, apicomplexia and/or other Stramenopile organisms. This is based on morphology and also early metagenomics studies (done on tissue samples rather than the cultured colonies). We are pursuing additional sequencing studies as well asTEM, but have run into complications with DNA extraction (low yields) presumably due to the tendency for the spore nuclei to be encasked in a crystal-like capsule within the agar matrix, as well as the tiny size of the spores/zoospores (1-3 microns)
A recurring finding is that once the organism has infiltrated the agar matrix in the Petri dish, it begins to form spheres and pseudohyphae-like structures WITHIN the deep layers of the agar matrix. Grossly, the agar plate will look like there is no growth for three to six weeks (though samples of agar removed by sterile loop show infiltration of thousands of nuclei-which can be induced to form motile zoospores within 20 min of re-hydration) during this time. Contaminant growth is very rare during this time window. Then, as moisture is removed from the agar, sphere-like objects with a fibrous outer ring, and then fern-like patterns begin to become visible in the dessicated agar. This happens only in the samples cultured from affected individuals, and not in the sham or negative control samples.
I am wondering if anyone else has seen these patterns before in the deep layers of agar (and ideally confirmed presence of an organism by removing/staining blocks of the affected agar). If so, what organism was growing in the agar and, did you find any way of extracting good yields of DNA from the dried/crystallized agar?
PS- photos below are of surface of agar plate/Petri dish as viewed 100x-250x through dissecting microscope. Cultured tissues represented include blood, subcutaneous adipose collected by needle biopsy, and water from a hot tub that might have been a source of infection.
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PS- please excuse the typing and formatting errors in my replies, I am currently typing from a cell phone and cannot see or edit the area in which I am typing the reply.
Also, since it is difficult to add photo files to the “reply” section, I am including some links to videos we have taken of the microscopic appearance/behavior of the cells/spores retrieved from cultured growth in blocks of agar such as this, as well as what has been observed directly in tissue/fluid samples from some of the affected individuals.
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Dear all,
I am looking for recommendations for an efficient protocol for the extraction of Genomic DNA from several fungal cultures.
I have most of the chemicals and equipment used in the process, but I am not getting a complete procedure (Starting from Cell disruption) to purification and quantification of DNA.
Any kind of help will be appreciated.
Best Regards
Arush
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I can provide you a simple protocol for fungal DNA extraction.
Take 2-3 colonies directly from your culture plate using a pipette tip and dip it in a tube containing 100 microliters of nuclease free water. Vortex for a few seconds and heat in a water bath or incubator at 96 degree C for 10-15 min. Centrifuge at 13000 RPM and use 1 or 2 microliters of the supernatant for PCR.
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Actually these are two separate questions:
i. How to extract mycotoxins from the media PDA the fungal culture is growing on?
ii. Can we extract the toxin from the natural media it is growing on?
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Hello,yes you can extract mycotoxin from the substrate. You may culture the fung in a specific media preferred by this fungi .
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I had carried out an experiment to study plant growth response towards different music exposure. However, the results turn out differently when fungus appearance detected. For your information, the samples were exposed to music (rock and ballad genre) 4 hours every weekday. After the 10th day of the experiment, fungus spread in rock music treatment is vigorous on the planting media compared to the ballad music treatment. The germination is completely zero for both treatment.
May I ask what possibilities there are that make the fungus infection difference between the rock music treatment and ballad music treatment.
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Hello,
I know many studies on sound treatments for disease control for plants. It seems that certain sound waves triggers plants defense systems to protect themselves from insects, fungi or bacterias. I am informed that certain sound waves inhibit mycellium growth or germination of spores, however I am not speciliazed on microbiology. I recommend the studies below if you did not read yet.
Experimental Investigation on the Effects of Audible Sound
to the Growth of Aspergillus Spp.
Inhibition of Botrytis cinerea Spore Germination and Mycelia Growth by
Frequency-specific Sound
I would be so glad if you have more experiments or studies on fungi growth status via sound waves or different kind of music application.
Thanks,
Meltem
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Reactive oxygen species are produced during early plant-pathogen interactions as a plant defence response. Sometimes, necrotrophic pathogens also produce ROS to cause necrosis or plant cell death. Now the question arises, how can we distinguish between both the ROS, specially during infection? Is there any approach available to do so?
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Typically, superoxide (O2*-) is the first ROS being produced in most cells under aerobic conditions.
For bacterial cells, while it is true that a fraction of molecular oxygen consumed during respiration and ATP production is converted to superoxide, this amount of superoxide cannot be compared to the level of the host cells (which contains multiple mitochondria as a source of superoxide at resting state and during inflammation).
In modeling infection, typically host cell densities used are 1x10^4 to 1x10^5 cells /mL. In realistic terms, clinical infections involve infecting pathogens at densities at or below 10^7 microbes / mL at most (because the nutrient requirements including aeration in vivo won't allow them to grow to supraphysiological levels like 10^8 / mL) .
Also, typically, infection experiments use MOI (multiplicity of infection) ranging from 1 to 10. MOI like 100 (equivalent to 10^7/mL or more) again seems unrealistic for the reasons stated above.
Most bacteria possess multiple enzymatic and non-enzymatic antioxidant defenses to deal with superxide -- first by dismutation by SOD, then catalase or peroxiredoxins to terminate the ROS reactions. So, effectively, what you can expect to detect or see in resting bacteria for endogenous superoxide / ROS is very very low (we have some unpublished imaging data). Certainly not to be compared with a resting host cell.
***
Having said that, it means endogenous bacterial superoxide (or ROS) in host-pathogen interactions may not be a key concern in measuring the overall ROS turnover. It is expected to be a small contribution.
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If you have an appropriately selective chemical probe for ROS (say superoxide), then what you do is to use enzyme inhibitors (e.g. NOX inhbitors) or antioxidants (say mito-TEMOL for mitochondrial superoxide) to measure the effects of the intervention, compared to un-intervened controls.
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Also, if you are absolutely interested in finding about ROS in the bacteria/microbe, fluorescent protein based H2O2 reporting has been established (the HyPer series). Those can be integrated into your model strains by transgenesis. Then, you can image or quantify H2O2 changes during infection.
Currently with the known tools, precise investigation on ROS kinetics and dynamics remain quite challenging.
Good luck on your experiments!
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I am working with wood decay fungi. I want to make a homogenize mycelium suspension, I used waring blender for different times, But I have lumps of mycelium every time which don't want. I want to impregnate wood blocks with this mycelium suspension.
if any body have any suggestion, I will appreciate your answers.
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Hi Nasim
You can remove making a shaker for the broth that contain mycelium and spores, then you can do a filtration for it
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Dear collegues,
Can we use protocols of DNA extraction from plant tissues (from leaves) to extract DNA of fungal pathogen which lives inside of tissue? If yes, will the quantity of the pathogen's DNA will be sufficient for further amplification with specific primers?
Or we have to use special protocols to extract fungal DNA from plant leaves? In this case please send a link.
The pathogen is Taphrina deformans causing leaf curl disease in Peah tree.
Thanks a lot in advance,
Looking forward to your reply,
Lidia
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Dear colleagues,
Thank you for your comments,
we just used traditional CTAB method and our experience is that it works always much better than any kind of commercial kits.
Yes, we have lots od plant DNA together with the fungal DNA in the result. But if we have a fungal-specific primers PCR always wrks good. We just use higher concentration of DNA in PCR-mix:
DNA sample - 2 mkl (from the solution of 200-300 ng/mkl)
PCR mix volume - 15 mkl.
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Hello Colleagues
Recentrly, fungal infections cause an increase of morbidity and mortality in hospitalized patients and in immunocompromised persons. What are the most recent recommendations and guidelines for the control and prevention of nosocomial fungal infections.
Thanks
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The last two decades have shown an increase incidence of nosocomial fungal infections in hospital environment.The important fungi that are implicated in nosocomial infections are Candida albicans, Non-Candida albicans, Aspergillus fumigatus and Non-Aspergillus fumigatus, Fusarium species and others. We have isolated several fungal pathogens from burn wounds of patients admitted in burn ward of the hospital. Certain measures, such as personal protective wear, proper hand hygiene, respiratory hygiene, thorough cleaning and disinfection, safe injection practice, avoidance of sharp needle and scalpel injury, and waste disposal besides prompt medical attention to skin injury, and appropriate treatment with anti-fungal drugs (fluconazole, itraconazole, Amphotericin B, posaconazole etc) can be effective to prevent nosocomial mycoses.
One see the following paper on nosocomial mycotic infections.
Alangaden GJ. Nosocomial Fungal Infections: Epidemiology, Infection Control, and Prevention. Infectious Disease Clinics of North America 2011;25:201-25.
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Morbidity and mortality from invasive fungal infections remain unacceptably high, I really want to know why vaccines are not developed for fungal diseases, lack of scientific proficiency or ignorance?
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Michael Dare Asemoloye Factors which greatly limits the generation of fungal vaccines are,
1. Human commensal nature of fungi,
2. Capacity of fungi to establish clinical latency (Candida, Cryptococcus etc.),
3. Potential high costs in preparing the vaccines,
4. Mostly immunocompromised individuals are susceptible to fungal infections,
5. Vaccine against commensal organisms ( Candida, etc.) becomes a challenge as autoimmunity against the organism develops.
Despite these factors, many fungal vaccines are in development stages.
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i would like to know that i have isolated about 30 different kind of isolates from fruit peel at harvesting stage..and have pure colonies and i want to do Identification, so i have finished microscopy but for verification want to do molecular study. so
1. how to prepare sample for fungal isolate for DNA extraction (please explain from new culture grow to the end)?
2. Best method for unknown fungal isolates DNA extraction?
3. how can store fungal isolates for about 6 to 12 months =?
I shall be thankful to you
looking forward to hearing from you.
Regard. RIZWAN.HM
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The link experiment of mine will give you an outline. I extract fungal DNA isolates from infected wheat spike.
Regards,
Harun
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Fungi and animal manure.
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I want to extract the fungus isolate DNA for Sequencing and Identification as i have different isolates from fruit peel, so i want to know that is there some requirements for colony grow for DNA extraction ? or just grow the mycillium on PDA media and and then collect them after 7 to 10 days for DNA extraction ?
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Michael Dare Asemoloye Thanks a lot of you for sahrng :
i would like to know more that :
1: if some fungi grow slower ( took about 1 month for petri dish completion) and some faster (1 week for petri dish completion) so what should do in that case for sample collection for DNA extraction?
2: Can use more then 1 plug of same isolate in one petri dish for getting more quick material for DNA extraction?
3: Isolate sample collection for DNA extraction : one isolate per petri dish colony should collect in one 2 ml and put in liquid nitrogen and then grind and use CTAB or any kit ,,,,is ok? how will know that the sample is 30 mg for DNA extraction??
4: Spore calculation through haemocytometers procedure: can you please share a general formula for spore calculation ? as i have read many artic@les and questions but mostly labs used there own method and formula, ( pores/ml = (n) x 10^4, can explain this what is 10^4, ?
5: can you please share, that How can make the suspension solution for spore calculation through haemocytometers?
6: If i want to do inoculation of that isolates on fruit with different connections then how can make different concentrations?
I shall be thankful to you
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We saw during survey studies that there are mushrooms on pomegranate stems and branches, but we were unable to give it a meaning. Can someone explain this?
Thank you.
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Could you please tell the name of this mushroom??
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Endophytic Trichoderma species are rather aggresive in their colonization of a substrate; I understand that they do it faster and more effectively than mycorrhizal fungi (e.g. Glomus spp.). It seems Trichoderma spp. and Glomus spp. do not antagonize each other. However, given this faster colonizing process of Trichoderma over Glomus, would it be recommendable to inoculate an established crop first with the mycorrhizae inoculum and a few days after with Trichoderma? If so, how much time should one wait between applications?
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@ Jose, you have to inoculate separately as Tricoderma is growing very fast and they are not compatible with Glomus species. I think if you are very eager to see the combined effect you must inoculate first with Glomus and wait atleast for 2 weeks then go for Tricoderma inoculation.
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Thanks in advance for your replies.
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It is preferable to spray chemical pesticides at the beginning of flowering or adding biological fungi to the soil to increase the defenses of the host
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Has such a question been answered in a paper? Or does anyone have experience with that?
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While doing leaf litter decomposition experiment following Olsen (1963) we got k value (per year) 0.18 and half life 3.85. However, 54.01% weight has already lost at the end of first year. How can we interprete the value of half life and weight loss?
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I need know if Dicranidion is reported from America -mainly México.
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Please have a look at the following RG link.
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I want to culture Fusarium oxysporum. Can anyone provide me detailed protocol for Fusarium oxysporum culture?
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I face the same problem in growing fusarium oxysporum >> after one year of continuous sub- culturing on PDA medium the growth became transparent without apparent mycelium or spores . would you please tell me any solution for this case
thanks
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And is it possible to use that toxin against plant pathogen or pests?
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use the liquid media for the targeted fungus. and then put the flask in the shaker machine for some days. than you can use the centrifuge for extracting the fungus products.
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appeared after one week of culture ,  it is round and white at the at the bottom of a dish and once it appeared after one day
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Generally, this kind of contaminants are found when the culture plate is too old (≥6 months) or handled inappropiately. Normally yeast are found in our body and a part of healthy mix of normal flora. If I am not mistaken, it looks like yeast contamination; avoid unwanted movements, speaking while plating. Keep your work bench clean and wipe with 70% ethanol.
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Hi
As far as I know, ascorbate peroxidase enzyme (APX) is the major enzyme involved in H2O2 scavenging. It is specific to plants and algae.
Recently, I was shocked when I came across an article in which APX and guaiacol peroxidase (GPOX) from fungi were unusually assayed !
In fact, GPOX activity monitoring in fungal cells is not surprising since the famous class II peroxidases such as lignin peroxidase that is found in fungi can use guaiacol as electron donor. However, APX assay is well conducted and APX activity was even higher than catalase that is well reported in fungi !
Although ascorbate (or erythroascorbate) could be a natrural substrate for many peroxidases, what do you think about the presence of APX enzyme in non-photosynthetic organisms and especially in fungi ?
Your contributions will be welcomed.
Mohammad
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Hello, can't be evidence of ascorbate peroxidase in fungi. Peroxidase structures have significantly increased our understanding of the evolutionary and functional relationships within the plant peroxidase superfamily.
Three distantly related structural classes have emerged:
1. mitochondrial yeast cytochrome c peroxidase, chloroplast and cytosol ascorbate peroxidases, and gene duplicated bacterial peroxidase class I
2.secretory fungal peroxidases classII
3. classical, secretory plant peroxidases (class III).
So ascorbate peroxidase is class I peroxidase enzyme and catalyse the H2O2-dependent oxidation of ascorbate in plants, algae and certain cyanobacteria but not fungi. For this type of peroxidase your culture shuld be photosynthetic. My be we will see this activity in future in Radiotrophic fungus.
Good luck!
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I want to compile some information about the leading universities or research centers in the field. Feel free to contribute.
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The Mycology Unit of Universitat Rovira i Virgili (Reus, Catalonia, Spain) is an excellent research group working on systematics of clinical and environmental fungi. Some outstanding scientists who have worked (or currently work) there are Drs. Josep Guarro, Javier Pastor, Josepa Gené, Josep Cano, Alberto Stchigel and Maria José Figueras, among others.
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Hello! Im working on my thesis on Fungal Bioremediation and I noticed that my subcultured fungi took on a different color compared to our initial inoculation of it on PDA. I would like to ask if it is possible that fungi morphology could change if the fungi is exposed to different environments or media?
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Yes, it is possible. When fungus grow on different media or different environment exposure, their morphology could be change because of different substrate, environment parameters and medium condition.
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Dear all,
I’m looking for options to publish in Mycology (specific or non-specific Journals on Mycology). In 2011, Hyde and KoKo published a very interesting and applicable paper entitled “Where to publish in mycology?” where they provided information for the major journals that publish manuscripts entirely devoted to Mycology. However, were created new Journals in the last years and (unfortunately) open access has forced the author to choice Journals with very expensive publications fees and that yet charge for reads theirs access to our articles. Because of this, I would like to know where do you are publishing your papers? It is worth to publish without open access? Which Journals do you recommend most for fungal biology/ecology publication? I’m asking it just because I'm very worried with the elitism of scientific knowledge and how it can delay the science progress and the dissemination of “what we do in our labs/universities”.
Thanks for your response!
Hyde & KoKo (2011): https://bit.ly/2IuUQrQ.
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Other titles of peer-reviewed journals indexed in Scopus and Clarivate:
1-Current Research in Environmental and Applied Mycology
2-Sabouraudia Journal of Medical and Veterinary Mycology
3-Medical Mycology Case Report
Regards
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Dear Drs., We cordially invite you to contribute a manuscript in our proposed books on the major theme ‘Functional Operons in Secondary Metabolic Pathways of Fungal Origins Vol I-Fungi as Foes’ and ‘Functional Operons in Secondary Metabolic Pathways of Fungal Origins Vol II-Fungi as Friends’ to be published for the Fungal Biology Seriesled by Editors-in-Chief, Dr. Vijai Kumar Gupta and Dr. Maria Tuohy by Springer Science+Business Media (New York) by the end of 4 February 2019 .
We intend to include significant chapters exploring the major points/topics attached to this letter as Ahmad, TOC. However, other potential topics are also welcome. Please submit your response to the invitation by 31st October 2018, together with a provisional title, rough outline or abstract and a list of coauthors with affiliations (all of these may be provisional).
The full version of the manuscripts may be submitted by the end of 15th January 2018, although earlier submissions are of course appreciated. All files should be submitted as email attachments to malik4948@gmail.com
Manuscripts will be reviewed by the book Editors and/or by field Editors/Peer reviewers. We look forward to receiving first a confirmation email of your participation and eventually the provisional title with outline/summary/abstract and your actual manuscript.
Springer guidelines and a Chapter as Sample for the manuscript preparation is being attached herewith for your kind perusal. Please forward the same to other interested Researchers, Scientists and Professors. 
Should you have any queries whatsoever, please feel free to contact us.
Sincerest regards,
Editors
Dr. Malik M. Ahmad, PhD
Dr. Saba Siddiqui, PhD
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Thank you.
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Thanks for any contribution!
Francisco.
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Hi Francisco. Not really clear about your question, as the true coprophils are already in the dung when it is excreted - they require a passage through the herbivore gut to stimulate spore germination. If dung is dry when collected it can be kept in paper bags for months, even years, and will produce the normal growth of fungi when rehydrated and incubated. Coprophilous fungi alone are not responsible for decomposition - they will be interacting with the bacteria and fauna in the dung. I suppose you could do something along the lines of Webster and his students, e.g sterilise dung and then inoculate with different species, alone and in combination, and record the decomposition rate by weighing at intervals.
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Dear colleagues,
This mushroom sample is belonging to Coprinopsis genus but I'm a bit confused about the species to which it belongs, Would you please help??
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It seems to be Coprinus comatus
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Dear researchers,
What are the black structures that appear on Botrytis cinerea mycellium after two weeks of in vitro culture??
I tried to observe them under binocular microscope and I think that they are fruiting bodies but I am not sure.
Can anyone cofirm this ??
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Hi, These are sclerotia , The fungus produces highly resistant sclerotia as survival structures in older cultures.
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I'm trying to learn about phycobilin-related fluorescence. I've learned that there is a pathway of phycobilin biosynthesis, which consist of reduction of hemin through biliverdin to phycobilin using enzymes oxygenase-1 and phycocyanobilin:ferredoxin oxidoreductase. I know that phycocyanobilin in complex with protein do fluoresce. My question is: does phycocyanobilin alone, not complexed with any enzyme, exhibit any fluorescence? Also, would be great to get excitation/emission spectra.
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It is fluorescent, but can be made more or less fluorescent depending on the protein it binds to. Here is a link to the absorbance spectrum and other info: http://www.rsc.org/suppdata/cc/b8/b821687h/b821687h.pdf Hopefully after 3 years this helps someone :-)
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Primary diagnosis and treatment of onychomycosis (ugly toenails due to ONLY FUNGUS INFECTION) when dealing with ugly toenails has led to mostly failures in products, services, economics & EBM.
We need to admit that shoes, underpinning biomechanics and repetitive microtrauma (RMT) to toenails are the primary exogenous factors of onychodystrophy (ugly toenails due to unhealthy toenails for any/many reasons).
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It seems that there are at least three issues of importance besides dx. and tx.:
(1) prevention,
(1.1) who acquires them,
(1.2) do certain individuals/groups have a propensity to acquire them,
(1.3) do certain individuals/groups use successful preventative strategies already and, if so, what are the strategies and success rates.
Also, any RCTs out there and what are the results.
Dennis
Dennis Mazur
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i want to get gene deletion mutant in fungus, as the CRISPR/Cas9 is a rapid method, so i want to apply this method, can anyone tell me about using this method in Fungus?
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Kinda easy. Have been using it in my fungus for 2years now and it works perfectly well.
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Could it be useful to determine the size of a fungal organism?
Image for illustrative purposes only (Source: Webster & Weber 2007).
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Hola Cristian
VCG is a classification at the subspecies level. You are not going to be sure if two isolates, belonging to the same VCG, are the same, albeit they are more closely related than two isolates of two different VCGs. However, it is a well-known (and relatively easy) tool that is going to give you useful ecologic information about your population; for example, in general, and depending on your species, you population could sexually reproduce if you have isolates of different MAT within a given VCG.
Un saludo
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I want to use SSR marker to detect nuclear migration in Buller phenomenon in Agaricus bisporus but I want to use another method in my work (e.g. morphological and another molecular method except SSR marker ).
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Any co-dominant marker can be used
CAPS markers are easy to develop. Three markers linked to MAT are interesting: (1) PR6 used in the paper cited above by Aydin Hassanzadeh, (2) markers based on polymorphism in RPB2 (also linked to MAT), or (3) in gene mip (tightly linked to MAT). For the latter, the couple of primers used for A. subrufescens (marker mip) in the paper indicated below has been also successfully used with A. bisporus.
Evidence for amphithallism and broad geographical hybridization potential among Agaricus subrufescens isolates from Brazil, France and Thailand. Fungal Biology 118:1013-1023, DOI 10.1016/j.funbio.2014.10.004
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Currently, I am culturing fungus in petri dishes. Frequently, condensation of water occurred on the inner top of the petri dish. Although the water droplets didn't fall down it still blocks the view. Does anyone know why and how to prevent this?
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While pouring hot agar medium, stack petri plates in size of 10 or even more. It will prevent condensation in all plates except the top one.
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Is there any software which measure the capsule of cryptococcus neoformans? I have capture some pictures from mobile now i want to measure via software.
Thanks
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the media should encourage fungal growth and sporulation; should be simple and cheap to formulate. I have decided to formulate the V8 media [isolate fungus using the V8 media. A 100 mL of V8 juice will be combined with 900 ml of distilled water and stirred.1g CaCO3 and 0.05 g b-sitosterol are added and mix well. Leave stir bar in the flask for later mixing.15g of agar will then be added and autoclave at 15 psi for 20 minutes. A 1 g of Chloramphenical (before autoclave) or 1 mL of Streptomycin The medium is stirred while dispensing onto petri dishes at the Laminar Flow Cabinet to insure good mixing of CaCO3], however need further suggestion for alternate media otherwise
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The optimal media depend on the fungus being grown.  A good reference for several of the general media for fungal growth is Singleton et al. 1992. Methods for Research on Soilborne Phytopathogenic Fungi.  This mentions several of the types that have been widely used, such as malt extract agar, cornmeal agar, oatmeal  agar, etc.
Yes, V8 juice is most commonly clarified for use in media (e.g. Miller 1955, Ayers and Lumsden 1975, etc.) and then autoclaved.
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my research is about screening of mannanase producing fungi from leaf litter of Salaca zalacca var.pondoh. I have tried to do screening used selective media contain:
Urea : 0,3 g/L
NH42SO4 : 1,4 g/L
KH2PO4 :2 g/L
CaCl2 : 0,4 g/L
MgSO4.7H2O :0,3 g/L
Peptone :1 g/L
FeSO4.7H2O : 0,005 g/L
MnSO4.7H2O : 0,016 g/L
ZnSO4.7H2O : 0,0014 g/L
COCl2.6H2O : 0,002 g/L
Agar : 15 g/L
and adding 1% locust bean gum as carbon sources
the result is fungi can grow on the media, but when I add 1% congored dye, there are not any clear zone apppear. what should I do to make the clear zone appear?
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how long have you been staining the media? because in my experience, you need to flood it with congo red for about 15 minutes then wash it with aquadest (flood it also for 15 minutes) 2-3 times, then the clear zone that indicated enzyme activity will be shown
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Which fungus is in these pics ??
Kindly identify...
thanks
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Hello everybody, the fungus was isolated in PDA from the reed stems (Phragmites australis) near the soil surface, the colonies are fairly white as shown below, growing very fast, but no sporulation was obtained for couple of weeks. It seems belong to ....... I would be very grateful if you assist me to identify it. Thank you
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Dear & Simon, Saida and Łukasz
I attached new photos related to the blight reed fungus, Thank you
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Hi, I have made this video about fusarium pseudograminearum and wanted to make sure that it shows mating of heterothallic ascospores of fusarium pseudograminearum. Any comments?
Thanks Friederike
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You need to find the ascospores and photograph them. 
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I am currently establishing protocols for identification of plant pathogenic species by PCR. I tried primer pair BC108/BC563 published in 2006 by Rigottii and collegues, but it does not work with me (while I get excellent amplification with ITS1/ITS4 primer pair).  Any suggestions or any experiences with BC108/BC563 ?
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Hei,
To obtain a sensitive and specific assay, I would recommend the use of a primer/probe -based real-time PCR assay. It is probably very challenging to use the ITS rDNA gene cluster to design a specific assay for Botrytis cinerea. One option is to use betatubulin -see e.g. New Phytol (2005) 168: 465-474.
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Some fungus-like materials were found in the deep dermal inflammation tissue from a 76-year-old lady's right hand. She presented with soft tissue swelling with redness on her right hand after antibiotic treatment failure. She said she has high blood sugar when she tested herself with a blood glucose test kit. But she was not properly diagnosed or treated for that.
What kind of fungus should I include in the differential diagnosis?
(phaeohyphomycosis?) Is this definitely fungal infection? if not, what this would be?
Any ideas are welcome!
Attached pics are PAS stained slide taken at x1K
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Dear Yosep,
Not because I work on protist pathogens, but to me looks like trophozoites of some amoeba. Try to grow the swab assisted wound material on sabouraud's medium to rule out fungus. Could be Acanthamoebal trophozoites, deep eosinophilic cells??
Best,
Mannan
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Can anyone tell how to unbound/release the absorbed dye from the fungus. 
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Thank you Jean sir
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I've got some Corynespora cassiicola that I need to preserve for later use but am not really sure of the best storage method & condition.  Would really appreciate comments from those who have worked with this fungus.
Thank you
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You are welcome.
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I have finally got sclerotia of R. solani. I want to preserve for future work. so how could I preserve them and at which temperature? I don't like to want any contamination. Waiting for suggestions. 
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Hi Abbas, I used autoclaved mineral oil to preserve  Sclerotia at room temperature. it works good up to six month.
I hope it will help you.
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Please advice me about identity of this fungus produced spores on SDA media.
Regards
Shuvrah Rehman
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Hi Shuvrah,
You can also see this book "Seifert K, Morgan-Jones G, Gams W, Kendrick B. 2011. The Genera of Hyphomycetes. CBS Biodiversity Series no. 9: 1–997. CBS-KNAW Fungal Biodiversity Centre, Utrecht, Netherlands ".
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Trapping maize with mycorhhizae but not form vesicle in the root after 90 days. Why??
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Dear colleague Mostakim
There are not an exact time for find that structures in roots. It depends of diverse causes, among then, physiological processes, soil nutrient levels. For example it has been demonstrated that late embryogenesis abundant (LEA) proteins also accumulate in vegetative plant tissues during periods of water deficit, which reinforced a role for these proteins as desiccation protectant. It seems that during cellular dehydration LEA proteins play an important role in maintenance of the structure of other proteins, vesicles or endomembrane structures in the sequestration of ions such as calcium, in binding or replacement of water, and functioning as molecular chaperones. Any problem with that proteins could be one of the causes of the ausence of that vesicles.On the other hand, such vesicles, frequently hard to distinguish from intraradical spores, are more frequently formed on the most polluted locations and are seen as a part of the mycorrhizal survival strategy on metal-polluted sites (Pawlowska et al. 1996; Turnau et al. 1996; Regvar et al. 2006). Finally, with a management of nutrient solutions to the plant we can induce the formation of such structures also.
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1- Bacterial contamination is a major problem. Can I add antibacterial to prevent it?
2- At which temperature should I keep it and how long will it remain viable?
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I agree with Christoph Ottenheim  and all answers above ...good luck.
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I´m working with Solanum quitoense and Fusarium oxysporum f. sp quitoense. My intention is to determine if there are races within this special form. However, there are no previous studies that allow me to make comparisons or clear methodologies to perform this work
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Hi Duarte
I think, you can identify the races dependence on VCG, after you determine the race
you can do molecular biology
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Can anyone give a short explanation about the functional role of cheilocystidia? In several genera of the higher fungi, they have thick walls, or/and encrusted apex. Anyway, what is the function of this formations?
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First, things do not necessary have a function in Nature. Structures that do not impact the fitness of a genotype (neutral traits) will not be eliminated by natural selection and may thus get fixed and transmitted. 
This being said, the function of cystidia, whether cheilo or pleuro, is still unclear. They may play a role in maintaining a higher degree humidity around the gills (boundary layer effect). They may also play a defensive role in protecting  against gills feeding animals. See for instance:
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Dear colleagues,
Could you provide any information (publications or your own observations) on presence of geoglossoid fungi in mycorrhizal associations of any types? I`ve met a mention of its mycorrhizal state at some reviews long ago, but no references to research articles were provided.
According to our unpublished yet data Thuemenidium atropurpureum was detected in root system of Pyrola media and ascomata of Geoglossum sphagnophilum were observed in close proximity to bog orchid Hammarbya paludosa plants.
With great thanks for any help,
Elena
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HI, Elena
We published a small paper on Thuemenidium (http://www.mycologia.org/content/102/5/1089.full), and observed shared habitat between the fungus and crawberry. We have not collected any molecular evidence for any associations between earth tongues and other plants. However, I did observed tiny apothecia of Trichoglossum associated with mosses' rhizomes. Below was what we addressed about earth tongue ecology in that paper, hope this is helpful for your research.
"...The ecology of earth tongue fungi in Geoglossum, Trichoglossum, Microglossum and Thuemenidium was once considered homogenous, not only because all were found commonly in more or less damp lawns or pastureland (Nannfeldt 1942) but also because the ecology of these fungi, indeed of most Leotiomycetes, has been both understudied and overlooked for a long time. Although there is no available hard evidence many species of Geoglossum and Trichoglossum are believed to be associated in some way with bryophytes. The ecology of T. arenarium is unique because it grows in sand dunes near the seacoast. Of note it often grows with Clavaria argillacea (Ohenoja 1995, 2000) and has been confirmed to form mycorrhizae with the black crowberry Empetrum nigrum (Nitare 1982). In contrast T. atropurpureum usually is collected from acidic grasslands where diverse mosses are common. So far no relationships between T. atropurpureum and specific mosses have been proposed. Lumbsch and Huhndorf kept T. arenarium in Geoglossum following Nitrare (1982) and assigned only T. atropurpureum to Microglossum on the basis of molecular evidence (http://www8.umu.se/myconet)."
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I need the detail list of techniques used in the screening of fusarium wilt in Musk melon and even the scale required for evaluation.
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Hi Shridhar
You can follow this paper "Effect of root feeding by striped cucumber beetle larvae on the incidence and severity of Fusarium wilt of muskmelon"
Good Luck
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I have difficulties to sporulate the isolation of M. oryzae.
What factors can I modulate to increase plaque growth and sporulation?
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You can also write to Prof. Ning Zhang at Rutgers Un. She works with Magnaporthe.
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I rather confuse about factors that influence trichoderma (anamorph) to become Hypocrea (teleomorph).
can someone help to explain to me?
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Dear Fatia, 
As other colleagues have mentioned, not all conidial fungi have the ability to produce a sexual stage. Furthermore, those which do produce a sexual stage, can be either homothallic or heterothallic.
Homothallic fungi can produce sexual reproductive structures in the absence of a sexually compatible strain, but mating compatible strains is required to obtain a sexual phase in heterothallic species.
In general terms, it is easier to obtain a sexual stage on poor media which resemble the conditions in which the fungus grows in nature. For example, keratinophilic Onygenales commonly produce sexual stages on keratin-rich substrates such as sterilized feathers or hairs. If you have a fungus which is associated with decaying plant material, you can try water agar with sterilized fragments of straw, carnation leaves, etc., and poor media such as oatmeal agar or potato-carrot agar.
Success!
Hugo 
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Can anyone suggest me how to buy the fresh spores of Cordyceps sinensis and C. militaris fungi?Or please let me know any lab which is working with these fungi?
I am very interested in these species, since they are high value for medicine.
Thank you in advanced.
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i am working on it now. At our location Central Kalimantan Indonesia we have many of C, militaris infected on lavae and pupa of Setora nitens (Lepidoptera: Limacodidae)
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I want to know the methods and procedures for inoculating sawdust at a certain weight and the parameters to measure or take to know if degradation is enhanced.
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first make 5 to 6 sets of saw dust soaked with double distilled water ( including a control set without any inoculation). then add the microbes aseptically to each set after serial dilution with .01% MgSO4 solution ( upto 10-5/-6 level) and take your desired data after certain period of inoculation. 
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What are the crucial steps to take when one suspects a novel fungal isolate from soil which has continuously yielded in one hand "inconclusive result" and in the other "no result" through sequencing of its internal transcribed spacer (ITS) ribosomal DNA genes by two different recognized research labs in the US and Europe?
The "inconclusive result" arm produced a name down only to the genus level with a genetic distance so wide from the closest in their proprietary library, while the "no result" arm ran out continuously 'inability to produce visible bands'.
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Hi,
I aggree that you shouldn't base oyour decisions in molecular ID only. Some kind of preliminary phenotypic identification can sometimes help you define what type of fungus you are dealing with, and what other genes you should use for molecular ID.
Even though ITS is the current fungal barcode region, it is not fully discriminatory for numerous fungal genera of Ascomycota. βT1, TEF, calmodulin and other genes are not as standardised, but are in some cases more useful than ITS. You should have a multilocus approach.
I would advise you to go through publications of Studies in Mycology or others describing newly discovered species to have a clearer picture of the steps you should take when novelty is suspected. 
Good luck,
Paula Rodrigues
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We used the dual culture to detect the antagonism  of trichoderma , the methods are as follows :
Trichoderma and S. sclerotiorum were seeded in a same Petri dish containing PDA at opposite sides. Controls were performed using S. sclerotiorum against itself. Fungi were grown up to 7 days at 28 C.
The inhibition efficiency was calculated using the following formula:
 inhibition efficiency (%) = (C-T)/C*100
( C=growth radius of pathogen in control plates,T=growth radius of pathogen in dual culture plates).
My question is: if the Controls were performed using  S. sclerotiorum against itself, how to measure the growth radius of pathogen in control plates if the
S. sclerotiorum  hyphae overgrow/pass across each other ?
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I agree with Franci Aco Celar.  It's classical method where in control plate must be only one pathogen inoculum. In addition for Trichoderma strains with active growth is necessary to use the method of "deferred antagonism". In our studies, 7 days was too much. Trichoderma covers the surface of petri dish for 3-4 days.