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Dear Research Gate hivemind,
assuming, that liquid culture is not an option and you can not easily scrape mycelium off the surface without getting agar in your sample, because the fungus grows mainly in the medium.
Is there an established method for extraction of DNA for further PCR or even direct sequencing purposes?
Maybe using filtration with a mesh of a certain size, freeze drying, ultrasonic baths, centrifuges or a combination of that...
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You can use cellophane overlaid media plates. Once the fungus growth is completed on cellophane sheet, you can directly use to extract gDNA using liquid nitrogen and phenol chloroform extraction.
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Hello !
I would like to store spores of botrytis cinerea for long time , for future experiments. How can i process ! thank you !
someone have protocols?
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I am not getting authonticated information, is it Vegetable juice or ....
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V-8 Agar is a medium consists of 8 types of vegetables including carrot, spinach, tomato, celery, parsley, beets, watercresses and lettuce. They are mixing together with known percent and preparing a juice from them and adding CaCO3 (3gm). It's a good medium for isolating Pythium spp.
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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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I am trying to find some information on how the extraction of Active Hexose Correlated Compound (AHCC) from Shiitake mushroom can be done. I can't seem to find anything online. 
Thank you for the help. 
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What is the best method for quantifying arbuscular mycorrhizae in agriculture field soils? I have been attempting to extract spores from the soil by wet sieving and sucrose centrifugation, but have found very few spores. Root staining is another option, but are there any other suggestions?
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You can follow the Gerdemann and Nicolson (1963) method, see the
Baccharis incarum and fungus Arbuscular Mycorrhizal symbiotic relationship for land fallow in the Bolivian highland article (CienciAgro (2014) 3(1): 51- 58)
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Is there any report for purple pigment from fungal culture?
-It is extracellular
-water soluble and soluble in n-butanol.
Your suggestion and advice will be helpful.
Thank you
general 
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@Mattias Anderson. Sorry to say than orsellinic acid derivatives are colorless unless those are conjugated with other aromatic systems (dipole/quadrupoles preferably)
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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 have tried aniline blue and cotton blue staining for root sections. They're not effective, in this case.
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wheat germ agglutinin (WGA) staining
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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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We are conducting cultivation of Monascus purpureus on solid media in order to reduce citrinin and increase monakolin production. We are having quite some problems finding M. purpureus cultures. Is anyone willing to share/exchange? We have many basidiomicetes (especially lignivorous) - Ganoderma, Lentinula, Pleurotus, Grifola, Trametes etc. in our collection which we can offer to you.
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hi andrej did you ever find a source for Monascus purpreus . I've been looking for either live cultures or even spores just alittle harder to clean but not to hard.
thx steve
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Hi! Who can direct me by the culture medium the most advantageous for the isolation of fungi from the soil ( sabouraud or PDA)? 
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You can use PDA or SDA with chloramphenicol to recover common environmental fungi like Aspergillus, Fusarium, Chaetomium, Mucor, Rhizomucor, Alternaria, Rhizopus etc.If you wish to isolate dermatophytes such as Microsporum gypseum, you should use SDA with chloramphenicol and cycloheximide. However, for isolation of Cryptococcus neoformans, Candida, Geotrichum, Rhototorula from soil, avian droppings, bat guano, air, fruits and vegetables, we employed Pal sunflower seed medium with chloramphenicol.This medium was developed by us in 1980 and is used in many laboratories.We developed one more medium designated as "APRM" in 2015 and is helpful to recover common saprophytic fungi from soil, avian excreta,water, and air.I have uploaded several papers on Research Gate and Academia on the efficacy of Pal sunflower seed medium to isolate many fungi from environmental and clinical specimens.
Prof.Dr.Mahendra Pal
Founder Director of Narayan Consultancy on Veterinary Public Health and Microbiology, Anand, India
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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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I am inoculating bacteria and fungi onto separate wood blocks. I know how to do the fungi, the bacteria I am struggling with as there appears to be different ways. I was going to grow bacteria in nutrient broth, measure OD of cells, the spin it down, decant the broth then repeat using Ringers twice to wash them. The resuspend in Ringers and but 2 mL into a bijou tube and put the wood block in the bijou tube for a period of time to allow bacteria to colonise. Does that sound like a viable option and if I leave it for a month will it affect the wood or the bacteria regarding oxygen in the Ringers?
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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 objective of this work is to screen culture filtrates of different bioagent fungi, then obtaining adequate quantity from filtrate to determine their bioactivity. So it is quit necessary to learn about most suitable conditions for increasing the amount of fungal filtrate.
Many thanks
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For obtained sufficient fungal filtates is necessary temperature, aireation etc in optimal points whenyou selecting shake flasks in submerged fermentations.
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I want to inoculate large quantity of feed with fungus, which needs to be done out-laboratory . any suggestion about how can I do that?
What can I use to incubate the feed and fungus without having contamination with other fungus?
Is there specific containers for that kind of work?
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Dear Khalil
there are vats specified for that you can make yourself like cheese vats with double jacket in order to keep the temperature of your mix in the same time these vats made from stateless so the feed will be safe from metal contamination. in order to do your work it so easy to sterilize the vats, the feed mix the continua your experimental
shenana 
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I have isolated fungi from plant but not able to identify on microscopic level
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Hi!
It seems to be an Aspergillus but you must to look for species identification as Yehia told you. Colony morphology depends on culture media...useful for identification, also (i don't know what culture media you have used). 
I find this articles usefuls:
Best wishes!
Beatrice
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I want to perform a MIC test on filamentous fungi, but I can't find any good protocol for it. 
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Hi 
you can see this link its useful for you 
good luck
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I was trying to acclimatize a certain bacterial strain in Vat dyes. But I am unable to determine the concentration of the dye in the media at any particular time. After Sampling, I centrifuge 1ml of the culture medium at 8000 RPM, 10 mins. But after centrifugation, I find the dyes along with biomass in the pellet form at the bottom of the tube. Hence I find it difficult to quantify.
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We tried to inoculate penicillium in PDA ammeded with a fungicide for IC50 
the plate got a bacteria cover instead of penicillium... 
the technique used for inoculation is dicarding penicillium spores with sterile dis water 
is that the reason of our contamination ?
we tought to scratch the spore without using any water 
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Some bacteria live in close association with fungal colonies, and use hyphae as corridors.
In order to separate the bacteria from the fungus, take a sterile inoculating loop and touch over your Penicillium culture where you see colour (spores). Then perform a "streak" inoculation. Draw an s-shaped pattern with the loop (the streak). This technique allows for your sample to be progressively diluted accross the streaked surface. After culture, you should find individual Penicillium colonies, and probably, some bacterial colonies. Allow the single Penicillium colonues to spore and subculture from those.
If the contamination persissts, conduct the same experiment in agar containing chloramphenicol.
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 reverse side method  is used for detection of bacteriocin production by bacteria
wanna use it to detect the inhibitory effect of some endophytic fungi against some toxigenic fungi >>>instead of using agar disk diffusion assay
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Hello
Yes, you can use . It is standard method
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We know that the shaking incubator gives a large amount of the fungal growth due to agitation, mixing media with fungal elements but I would like to know :Please, does the shaking incubator effect to get the different chemical structures of the fungal secondary metabolites compared with incubator ( without shaking) for the same fungus in same conditions ( medium, pH, and temperature)?. By another hand, if anyone grows Penicillium notatum in shaking incubator by using potato dextrose broth at 25 ˚C for 7 days, and the same fungus is cultured by using the same conditions except using incubator (without shaking) instead of shaking incubator. Does the shaking incubator lead to get secondary metabolite differs in the chemical structure compared with those extracted by the incubator without shaking, and by using a same extracting solvent?
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The effect of the type of incubation just in the amount of metabolites not in the type of it.
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I need assistance with right protocol in prepare malt extract agar (MEA) with Spezieller Nahrstoffarmer media + PEG 8000 on a plate. the attached document methods describes the (Spezieller Nahrstoffarmer media + PEG 8000) liquid agar preparation as a guide.
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Crude extracts are widely used against fungi. So How I get crude extract of bacteria??
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Are you extracting the DNA or protein?
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I am working with a non-sporulating filamentous fungus, and I use blended mycelium as an inoculum for liquid cultures. Does anybody know approximately how long I can store a concentrated suspension of blended mycelium in demi-H2O in the fridge (<+7°) before it will lose significant viability?
Thanks in advance!
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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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There are variable methods for determination the composition of the culture filtrate of Trichoderma harzianum, I need, if possible, more reliable and accurate methods for this purpose. Thank you
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Hi Dr,
Hope you are doing well.
I think GC-MS is the best method.
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antifungal suceptibilty test
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molecular identification of Aspergillus falvus.
NS1 ,C18 primers is universal primer for identification of Aspergillus species?
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Please see the latest molecular ID procedures in  https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4260807/ 
We use the calmodulin (CaM) primers in this article as a DNA barcode.  Though ITS is also recommended, it rarely IDs aspergilli below the species group level.  To ID, blast the CaM sequence against the curated Aspergillus & Penicllium Database at http://www.westerdijkinstitute.nl/Aspergillus/DefaultInfo.aspx?Page=Home.
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How to storage fungus in a long time?
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Dear Nur,
In the following link you can find a proposed methodology to store R. solani:
Which contains the following protocol and a link to the original research paper:
A Method for Long-time Culture Storage of Rhizoctonia solani. Edward E. Butler, Professor, Department of Plant Pathology, University of California, Davis 95616; Phytopathology 70:820-821. Accepted for publication 22 February 1980. Copyright 1980 The American Phytopathological Society. DOI: 10.1094/Phyto-70-820.
A method is described for long-time preservation of cultures of Rhizoctonia solani in tubes in a dry soil containing 4% (w/w) wheat bran. The moisture level of the mixture at the time of inoculation with R. solani was 0.23 g water per gram (dry weight) of soil. About 1 mo at 23–27 C was required to allow the colonized mix to air dry; then it was stored at –25 C. Isolates from diverse sources survived 55 mo at –25 C. Virulence, morphology, and cultural characteristics of R. solani were not changed.
I guess it shouldn´t be difficult to find alternative protocols at the NCBI database.
Good luck!
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BC DSM1 seems to be pretty electroporation recalcitrant. 
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Dear Colleagues
I am trying to purify a FAD-binding protein. Is there a reliable method I can use to purity such proteins?
Best
Zha
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If your protein is very dilute, use a binding technique like ion exchange first to concentrate. The flavine has an absorbance maximum at 450 nm, so you are looking for something that absorbs there and at 280 nm (the aromatic aa in the protein). Since you have done 2DE, you know the pI of your protein, designing an IEC experiment should be straightforward.
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Candida albicans
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Thanks Oadi Matny thats very kind of you
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Yield of  mass culture is good. But after extraction of fungal mass with  ethyl alcohol or other solvent  like methanol, yield is very low.
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Methanol in my opinion is adecuate
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I growed Trichoderma harzianum from a product using a medium with a "special" aditive.
It growed in 3 to 4 days at room temperature 25ºC aprox, adn the medium  was done with:
  • 1000 ml water
  • 10g agar agar
  • 10g dextrose
  • Clavulanic acid + amoxicilin (generously, amount not recorded)
  • 0.01g of the product, that contains only T. harzianum: 10 8 ufc/g. 
  • And a drop of the aditive
I have been told that its not possible they are Trichoderma harzianum colonies: apparently they do not grow like that.
So I wonder if it would be possible for them to grow in a different way not yet seen, thank to that "additive" i am adding.
Can any of you identify Trichoderma harzianum in those Petri?
What interest could this have?
Thank you for your comments
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I have never seen it growing like that. Which strain is it? Cultivation time and temp?
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Which dimorphic fungi grow solely in yeast form in liquid media?
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Mohammed Fayyadh; Oadi Matny:
thank you for your help!!!
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I need to dissolve my extracts with a solvent that is non-toxic to ganoderma boninense. There are methanol, dichloromethane, chloroform and aqueous extracts. I've tried acetonitrile and DMSO but both can inhibit Ganoderma boninense.
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Thank you very much Mr. Oadi and Mr. Laith.
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Anyone can tell me about Adamek's Liquid Medium ? How to prepare it for the growth of fungus ?
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Good Luck
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For quality control of beauveria I need to try out spore germination percentage data too. Any method or any suggestion?
Regards
Shuvrah
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You welcome 
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please tell me selective medium for isolating fusarium udum?
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thank you all  for the answers
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i want to study and do research about mycovirus, but i don't know much about that. Anyone can help me? thanks
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You most welcome
Houda
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i want to know about the methodology for screening of Agaricus bisporus strains against Cladobotryum mycophilum fungus?
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@Laith Al-Ani but sir i cannot open these links....please guide me to open these.thanks sir.
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I have collected the diseased plant from field, and kept them at 0c now i want to isolate fungi from stem please tell what is appropriate procedure or protocol.
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Hi
you can disinfect with hypochlorite sodium %1 for 30 second and wash with autoclaved  distilled water two times and then dry samples with autoclaved paper and culture it on PDA or MEA or PCA
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we have commercial metarhizium acridum powder (talc) formulation and we would extract spores/propagules to calculate germinated spores
we first tested salin solution containing; NACl/ KCl and tween 80 but it gives us nothing on PDA 
 either with simple distilled dionised water 
I used to extract fungi from commercial (powder) products using simple tween 20 and NaCl solution but this did not work either in this case
would it be possible that the product is not actif anymore (one year conserved at 4°C) ?
 Is there another protocol for that fungi ?
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I have isolated fungi from a diseased fruit and cultured this fungi in Petri plates. Which protocol should I follow to extract the DNA of these unknown fungi? What strategy, I should use to do PCR? More importantly which primers I use for identification of species level as well as genus level?? 
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I think you can use the following primers, and  then sequencing and bioinformatics:
forward primer ITS1 (5’ TCC  GTA  GGT  GAA  CCT  GCG G 3’) and reverse primer ITS4 (5’ TCC  TCC  GCT  TAT  TGA  TAT  GC 3’)  (White et al., 1990)
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Proposed substrate is sawdust from sawmill. Target mushroom is oyster mushroom
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Dear Yehya Salih, 
Thank you for your response and attached
documents.
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Penicillium is a problem in Petri plates having PDA media. I have sterilized the PDA media but still, it is a potential threat. I want to work on other fungi but I face Penicillium and its spores. Kindly give me tips how to get rid of this notorious fungus. 
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Mr.  Abbas,  I  suggested that you can recheck the procedure and be highly aseptic in the operations especially during subculturing the isolates. 
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I am working on a fungi that grows best on solid media, but i need to assay the enzyme activities, find out the reducing sugar concentration, total protein and other assays that require a liquid medium, any suggestions?
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I have prepared the conidial suspension of fungi and now I want to adjust the spore concentration by hemocytometer> Your suggestions are needed.  
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Several good references here on ResearchGate.
Check this discussion on a 5x10(4) spore concentration:
Note the answer from: Susann Auer · - · Technische Universität Dresden:
"Since you obviously have to adjust your spores to an exact concentration I recommend that you count as many squares as possible and repeat your counting with another sample of your spore suspension. So you will assess proper mixing of the sample prior to counting.:
How to find the find the concentration of fungal spores? - ResearchGate. https://www.researchgate.net/post/How_to_find_the_find_the_concentration_of_fungal_spores2 [accessed Feb 7, 2017].
Also:
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I am growing Penicillium sp. in PDB.  With days the broth inoculated with the fungus is becoming gel like and thick.  I am not able to filter the inoculated broth for measuring dry weight.  Please suggest some ways of getting rid of this problem.  Also suggest how to filter this thick inoculated broth for carrying out extraction.
I have also diluted but the inoculate broth before filtering.  However even this approach failed.
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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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1. Mycelia is grown in PDB for 4 days, kept on a rotary shaker.
2. Mycelia is ground using liquid nitrogen.
3. About 50-80 mg of the mycelia is taken into eppendrof tubes and subjected to chloroform isoamyl extraction.
4. 600 ul of the DNA extraction buffer ( Nacl, tris HCl, EDTA, 10 % SDS). 20 ul of proteinase K is added to each tube and incubated at 56 C for 1 hour.
5. After 1 hour, 600 ul of chloroform : isomayl alcohol is added and centrifuged.
6. After this step, i added 4 ul of RNAse A to each and incubated at 37 C for 15 mins.
7. Further continued with the chloroform isoamyl extraction.
Here is the gel picture am not sure why i can still see some stuff at the end and their are no clear bands. Is the DNA degraded or its the RNA presence at the bottom?  
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67
Sometimes problem:
1) DNA concentration may be very little
2) Primer concentration is very high
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I am woking on TP53 (SW48) cell line. I noticed fungus in cells. I washed with PBS for two times. Hw can I remove fungus completely?
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Dear Karthik
Hi it's a TRY (It's worked for me)
Anyway 
Divide the entire cell culture in to small portions and observe under microscope for the fungal contamination: (there is a chance to get a portion of cells without fungi)
Gentle centrifugation  can also settles the mesh of fungi+cells
TRY 
Best of Luck
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mycologist, Fermenter specialist
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clean the filter with a 2 % formaldehyde and then autoclave by packing 
you can add 5ml formaldehyde to the fermenter containing water or remaining fungal culture and sterilize for 10mins keep a mask to your nose while sterilizing
and then clean thoroughly
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There are more than opinions for counting the number of spores . I want the optimal method. 
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Dear Dr. Yehya,
I hope you find the attached document useful.
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What are the most important manuals to study isolation, identification and taxonomy of rust fungi?
Is there any available online free to download?
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Ajay:
Have a look at this link for insights:
Best
Syed
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I had a fungal strain which i treated with ethidium bromide containing PDA media and after some alteration's, i got successful. Now for screening process i treated it with same media containing ET-BR but with different concentrations like 1 ml, 2ml, 3ml,4ml and 5ml of ET-BR media into Normal PDA media and I could clearly see that the PDA media with the most amount of ET-BR had the most growth and then the 4ml . Upon further screening with that ETBR- pure media on halfside of the Petri plate and half with Normal PDA the Results are quite hilarious. i can clearly see the growth on ET-BR media but no growth on the other side. So does that mean that this strain has either somehow become ET-BR dependent or there is some other logical explanation to it ?
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@ali
As far as i know, i haven't saw anyone using ETBR with PDA so it was on trial basis just to make sure that strain is susceptible to the carcinogenic media or not, and now as per your comment i am going to increase the amount of ETBR and so to check its Growth rate if enhanced over time or not. Secondly i ll be surely in contact with because this whole thing is my very own idea and i have pretty much no idea what parameters could effect the growth rate other then the ETBR concentrations and TEMP....
Looking Forward to talk with you 
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Hi, I am trying to get sporulation in Corynespora litchii so that I get good slide for photomicrograph of conidial characteristics. I have tried PDA as well as potato carrot agar with no satisfactory result.  Further, whether Corynespora and Solicorynespora are synonym? Which name is currently valid? Please suggest some solutions and discuss the issue.
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It has been Solicorynespora litchii since1990.
Both genera are valid but some species of Corynespora were transferred to Solicorynespora when this was created.
For this kind of questions check Mycobank and Index Fungorum
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I'm looking for the references regarding density of entomopathogenic fungi in relation to soil properties 
I have isolated entomopathogenic fungi from soil using selective medium (agar plate) which present the density of fungi. 
I found that most of the paper used insect-baiting method using the larvae of Galleria mellonella or Tenebrio molitor to isolate the entomopathogenic fungi from soil.
Some of the study used both method, which is the selective medium and insect-baiting method.
They described the occurence of entomopathogenic fungi in soil based on the occurence (presence & absence of entomopathogenic fungi in soil) rather than the density of entomopathogenic fungi.
But, I cannot find a published paper only determining the density of entomopathogenic fungi with soil properties. 
Therefore, I found it very hard to discuss my finding with the previous study. 
Hope insect pathologists can give me the references regarding density of entomopathogenic fungi if you do found it. 
But if not, can you suggest how to compare my study with previous study? 
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I'm trying to use sodium thiosulfate to arrest the reaction of sanitizers on my fungal spores after the holding period is over. As sodium thiosulfate is the best to take away the chemical, especially the peracetic acid but I found this could be antifungal too. Any suggestions? Does anyone know the concentration of sodium thiosulfate that could be safe while working with Mucor and Botrytis spores? Thank you.
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During antagonism process between Penicillium sp. and another different species of fungi, I found great antagonistic activity of Penicillium sp. against pathogenic fungi, but unfortunately this fungus (Penicillium sp.) has antagonistic activity against Trichoderma sp. and on some herbaceous plants as well. Thanks for any useful idea or references.
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Hi Dr Jawid it is known that Penicillium dangeadii(Telemorphic:Talaromyces flavus) reduce Verticillium wilt infection on potato, olive and eggplant.the mechanism by which P.dangeardii may be due to glucose oxidase releases from  Penicillium ,glucose oxidase metabolize glucose released from root plants and form hydrogen peroxide which is toxic to Verticillium(Fravel &Robert ,1991)
Fravel,D,Rand Roberts,D.P.(1991).In suito evidence for thr role of glucose oxidase in the biocontrol of verticillium wilt by Talaromyces flavus .Biocontrol Sci Tecnol,1:90-99
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I enclosed Microscopic photos for this fungus, Thank you
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Dear Liqaa,
Looking at the pics, it looks like you have Fusarium sp., definitely not T. rubrum.
Regards
Mohammed
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I am about to do artificial inoculation using Beauveria bassiana 
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You most welcome
Houda
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I got this fungal isolate from termite insects when I was trying to isolate entomopathogens.  Dear colleagues please let me know what is the identity of this fungi.
Regards
Shuvrah
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Dear Yehya A. Salih
No I did not test pathogenicity on termites. Till now I am completing isolation protocol. Yes I will upload plate culture.
Thanks.
Shuvrah
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I am currently work on endophytic fungi, using rice solid static cultures. One of my fungus shown blue methanol extract. Blue color appear after the molded media immerse using methanol over night and disappear after third day at room temperature. Any idea to isolate this compound? I think it is polar and unstable compound.
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Collect your blue extract. Filter it. Reduce it to dryness in vacuo under mild conditions to keep it under safer conditions for storage in the refrigerator than in solution.Take a small portion of your blue extract and give it on several TLC plates (but use enough material). Then develop it in small TLC chambers with pure solvents as eluents. These solvents should have different polarities, e.g. hexane, chloroform, ether or MTBE, ethyl acetate, methanol, acetone and others. But do not use mixtures. After the TLC ist developed have a look where the BLUE is. Upstairs, in the middle, downstairs?This will allow you to get an intention on the polarity of your compound and the qualitiy of your extract.
Then, it is possible to find a suitable eluent for column chromatography (CC) by mixing two of the solvents in an appropriate ratio. All of this can be done only by trial and error. But with small amounts it is not difficult to find a solution for the separation. I would recommend to try CC for separation of the blue.
Caution: Real dyestuffs with a high epsilon value may cause a colour already with tiny amounts of material. However, it cannot be seen at the begin if this is the case in your case or not.
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Trichoderma are promising biological bio-protectants agent against plant pathogenic fungi. Producing large number of spores and unaffected to environmental variabilities  and and have a long shelf life  are definitely required.
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Hi Jawad: There are a master thesis about formalization and spore production of Trichoderma  in the department of plant protection - University of Baghdad. I dont remember the title of the thesis but the name of the MS student is Mohammad Al-Hittar, his supervisor is Dr Farkad Al-Rawi. I think there is an electronic copy of the thesis, if you have any contact with any one in plant protection department - university of Baghdad, ask him to provide it to you. Unfortunately i am now in sabbatical leave, other whys I will send it to you.
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(a) Fungal spp. - Hypholoma spp., Phlebiopsis giganteaArmillaria ostoyae, Heterobasidion irregulare. (b) observing interaction pairings
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Hello! Could you suggest a proper analytical method used to quantify the number of each mycorrhizal species (Glomus intraradices, Glomus mosseae, Glomus aggregatum, Glomus clarum) in some product? Thanks!
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Dear Sergey
I coincide with Prof. Antoun. AMFspores can be present inside roots (if the product consist in a traditional inocula of spores reproduced at the host roots) or free in the proper product´s substrate. AMF can be extracted by wet sieving and decanting method, followed by centrifugation in sucrose solution (50%w/v) (Walker et al., 1982), be grouped initially by morphotypes, and finally identified the species under a microscope. By molecular methods, DNA extraction from the isolated can be performed using UltraCleanTM soil DNA kit (MO BIO Laboratories, Solana beach, CA, USA) following the manufacturer’s specifications. DNA extracted is checked by agarose gel electrophoresis (0.8%), stained with ethidium bromide and visualized under UV light. Then you can use use a nested-PCR DGGE analysis of the 18S rDNA followed by sequencing of excised bands.The products are analyzed by DGGE and the bands are excised for sequencing.
The fatty acid composition of lipids in AM fungi diferentiates them from other organisms. In most AM fungi, a large proportion of the total fatty acids is found as 16:1ω5c, 18:1ω7c, 20:3, 20:4 and 20:5. Furthermore, AM fungi have a rather high content of polyunsaturated 20-carbon fatty acids. The 16:1ω5c has been used as a marker fatty acid for AM fungi in controlled environments and the 18:1ω9c, 20:1ω9c, 20:2ω6c and 22:1ω9c could be used as possible markers for the detection of G. margarita, for example. You  can check also Madan et al. (2002).
Madan RC, Pankhurst BH, Smith H (2002) Use of fatty acids for identification of AM fungi and estimation of the biomass of AM spores in soil. Soil Biol Biochem 34:125–128. A big hug.
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I have done vegetative compatibility grouping for four isolates of Fusarium . from each isolate I selected 5 mutant sectors. however on determining their phenotypes, all of them turn to be nit 1 mutants. for compatibility mating or testing purpose i need a different isolate such as nit 3 or nit M. how should I proceed with
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Sure sir
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For: ITS, 18S or 28S?
Is ITS known to degrade with heat, and hence may not be best to use in practice?
Thankyou!!!
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The best option is to develop nested pcr model please refer this article Fungal-specific PCR primers developed for analysis of the ITS region of environmental DNA extracts
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From my laboratory experiment, A. versicolor is able to grow on PDA agar but the growth is really slow compared to A. flavus and A. niger
It takes 2 weeks to make the A. versicolor cover 2/3 of the agar surface (in petri dish) while it only take 6 days for A. flavus and A. niger to do so. Does anyone have suggestion on how to fasten A. versicolor growth?
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Dear Nizzal Syafiq
May be different requirement of temperature, Usually any slow growing rate in fungal we consider it as suspected viral infected fungal (mycoviruses), it may your isolate get infected with viruses
Pls. find the attached files
Page 206 in: Saul L. Neidleman . 1989. Advances in Applied Microbiology, Academic Press,Pp.320. ISBN 0080564488, 9780080564487
Ursula Kües, Reinhard Fischer. 2006. Growth, Differentiation and Sexuality Springer Science & Business Media,Pp.449. ISBN 3540281355, 9783540281351
Marcel Borgers, Roderick Hay, Michael G. Rinaldi. 2012. Current Topics in Medical Mycology, Springer Science & Business Media,Pp.274 ISBN 1461227623, 9781461227625
Hoping this will be helpful
Regards
Prof. Houda Kawas
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I have grown some pure colonies of mangrove endophytic fungi on potato dextrose agar media amended with150 μg/ml chloramphenicol. Now i want to test their antibacterial property against some test pathogenic bacteria by disk diffusion or well diffusion method by cutting the  block with cork borer  from the pure colonies. But i am in hesitation that weather the amended chloramphenicol of the media will interfere the test procedure or not. The age of each colony is about 1 month and storing them at 4° temperature. It will be very appreciated and greatness if somebody share his/her experience in this regard.
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Thank you so much you all to share your valuable suggestions and good wishes for you.
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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 need to extracted DNA from Aspergillus niger ..steps of method
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Hi Alaa
for isolation of total cellular DNA from plant cells and tissues, or fungi (or plant pathogen infected plants) I have good experience using the DNeasy Plant Mini Kit. Good luck