Beiträge von Steve_mt

Thank you Andreas, what % is the Ammonia, around 10% maybe?

I am preparing new solution from the powder now, that Congo Red is strange.

I take the long cut, I dissect and put the section in congo red. Leave for a minute or so, then I absorb the extra stain with a cotton bud or kitchen roll strips and add plenty of 3%KOH and wash the dissection in it (sometimes leave it to stand for another minute if the tissue is robust/hard). Then I pick the washed section and place it in a drop of KOH+Glycerol mixture, sometimes a dissect further in smaller pieces and space them out. I place the cover slip over, press with a rubber over tissue paper and ready to go. The glycerol mixture prevents the slide from drying.

Cheers

The one on the right is Congo Red Powder C.I. 22120 dissolved in water and filtered (I asked my friend who gave it to me) and remains red in KOH. Actually, when searching for my stuff, I just realised that I have bought a bottle and forgot about it - so at least this post was very beneficial !!! The one on the left was bought a few months ago from a commercial Lab Supplies and they provided it as a solution (1%). Well, I can make my own Congo Red solution from 22120... I guess 1% is a good concentration or a bit less is better? Since my Lab is home-based I prefer to avoid Ammonia.

P.s. Does it really expires 🤷‍♀️😁 ?

Hi guys, tomorrow I show you a video of what's happening. Thank you so much for your replies. I am a bit more relaxed with regards toxicity. The other (old) congo red do form some minute crystals and I know the notorious feeling under the microscope. Sometimes it goes away by heating it in water bath at 70C until water cools down gradually

I have had a stock of Congo Red donated by a friend and recently I bought a new stock from a local Lab supplies shop. The second one is a different kind!

In alkaline solution, it becomes purple and somewhat leaches out stain when a stained section is placed in clean KOH.

Are there different types of Congo Red because I may buy again this time from abroad.

Another point, it is really toxic and carcinogenic? Sometimes my fingers are stained and then I nibble my nails! : blow:

Dear Uwe, I just wrote this morning to Unite / Estonia and I see what they are gonna tell me. I will keep you informed with everything. Thank you for your help and I see no objection on sharing the sequence with serious colleagues like your kind self. I have no experience with the sequencing of fungi so I am glad to read your offer of helping.

Apart this I have other fungi that are crying for sequencing such as a Cosmospora (?) sp. [ Cosmospora species , Central Mediterranean region - Forum ASCOFrance] and a Talaromyces growing specifically on Washingtonia seeds. [ https://mushroomobserver.org/262509 ]

I would love to hug that fluffy brown Scottish 'buffalo / ox ' Thanks for sharing

Lovely!!! Thank you and I agree re sequencing - I check both and maybe start from the Estonian but 30 Eur is not much. I may submit a larger sample size (I have a Cosmospora that is really hard to determine) to the spanish Lab later.

Im happy in this forum

The stem is very hairy throughout, and I have most (the essential) micro data too (spores, paraphyses and asci). I read that latispora has a smooth / smoothish stem.

I recently found dozens of Helvella sp. which would correspond to a new record for the island of Gozo and most likely the whole archipelago. It was growing from very damp and shaded ground coved with Juglans leaves (and some twigs). I read the teeth on the lower surface is characteristic for the species, hence my determination to Helvella ephippium. Is it common in central and south Europe?

I see, so the infractus group can be confirmed at least! I wanted to add info (micro) perhaps it can lead to a narrower selection within the group. Ok then.

Who do you think would be interested to follow a DNA analysis?

Is a dry specimen enough to sequence or it has to be subcultured on special agar?

Or to put in different ways, what can I do to sequence it? (with my own budget)

Thanks.

Pileipellis: two layers (maybe only the upper is the true pellis). The outermost (suprapellis) a trichoderm of interwoven curved/ worm-shaped hyphae, 3-6um wide with rounded apices (very slightly inflated) and several clamp junctions.

Below lies the subpellis composed of wider and shorter hyphae, tubular or cylindrical in shape, with less frequent clamp junctions, 10-15(-17) um wide.

Spores: Pip shaped, subglobular to broadly ovate, heavily waited showing shadow-lines under the x100 oil immersion, apiculum distinct, central, elongated. Germ pore absent, but there is a discontinuous wall at the apiculum (I don't think it can be considered a sa germ pore ?!). Quite variable in size and sometimes shape , few are elongated and longer (+2.5 um above the average length). Inamyloid but wall seems to be dextrinoid.

Spores: Pip shaped, subglobular to broadly ovate, heavily warted showing shadow-lines under the x100 oil immersion, apiculum distinct, central, elongated. Germ pore absent, but there is a discontinuous wall at the apiculum (I don't think it can be considered a sa germ pore?!). Quite variable in size and sometimes shape (few are elongated and longer (+2.5 um above the average).

Cheilocystidia: If I have observed correctly, they are quite indistinct as expected for this genus, cylindrical, sometimes curved, do not stain very well in Congo red, rare. Same size of basidia (maybe I have observed basidiomes). Pleurocystidia not observed (or rare and missed)

Basidia: Rectangular-clavate, with a subtruncate apex, flexuose (curved), many with few to several oil bodies, in my opinion mostly 2-sterigmate, some 3- a few 4- (?), Sterigmata 3 um, horn- shaped, length 23-29 µm.

Basidia: Rectangular-clavate, with a subtruncate apex, flexuose (curved), many with few to several oil bodies, in my opinion mostly 2-sterigmate, some 3- a few 4-(?), sterigmata 3 um, horn-shaped, length 24-29um.

Lamella: Adnate to sub-decurrent, convex, cinnamon brown but it turns purplish-brown when bruised of wettened (eg in mount). I don't know if this is characteristic. Edge indifferent from face.

Dear all, I am very excited about this find. Many mycologist thrived to find the first Cortinarius from Malta. I spent the whole afternoon checking the microscopical features yesterday for a determination, and here i find some suggestions already.

T aste : It was incredibly bitter, I had to spit it out the small nibble I took.

Cuticle: The debris of soil and leaves stuck on the cuticle indicate that it was viscous when young. The stipe seem to be not. The colour was pale yellow, like straw changing to light cinnamon with age... so maybe it was paler when young.

Reactions: Iodine no reaction on cap or flesh; 4% KOH - slow reactions, slightly dark brown with a hint of olive-green hue on cap after 20 mins. Faster reaction with 10% KOH and somewhat more pronounced olive-green hue (tested on drying cap); 4% KOH on flesh gave a pale greyish-beige colour.

Zitat von boccaccio

Hi Stephen,

that smut sounds like an interesting find! I don't have any particular literature on smuts in the Mediterranean area, but a quick Google search revealed that there are at least three rusts on Lygeum. One species of Ustilago that seems to infect the flowers and two species of Tranzscheliella that form like a brown band of spores around the stem of the grass.

Björn

Hello Björn,

I have managed to find two photos of the infected plants but not of the spores and microscopical images. Must be Ustilago according to host specificity. If the infection is persistent, I can fetch samples this April and work on it together. I am under the impression (now I am not so sure!) that there was smuts on a nearby wheat or barley field but it can also be another species. Photos from Gnejna area (mt), 60-80m asl, April 2017.

Zitat von Bergwald

Hi Steve,

that's a "funny" idea. I must try as soon as possible.

Thanks for this information.

Regards

Thomas

Let us know if it works. The principle, although it may sound funny is a logical one - based on the assumption that there are hundreds of shed spores carpeting the hymenium surface. I have not read this technique anywhere, so maybe it does not work so well, but when I try it, I have success. The problem is for very small apothecia - you need a capillary tube then.

That would be the first Cortinarius sp. for the country 😍

Three specimens under Cistus and Thymus. Pileus 5-6cm, pale yellow, dry, though, glabrous-polished (when cleaned from debris), very bitter taste, no volva or ring or veil (at least not distinct), spore print light cinnamon brown, lamella adnate, stem shiny pale brown. Spores pip-shaped, surface rough (bumpy). IKI inamyloid (cell wall appears dextroid?!).

Am I on the right track if I consider Gymnopilus sp. ?

Hi, I often see questions on FB and online fora, asking how to get ascospores out of asci for proper measurement. I have this easy method to share with you which also works on tiny apothecia, even if dirty with soil or debris.

A droplet of water or cotton blue is placed on an upright cupulate or discoid apothecium with a fine-tip dropper. The water is agitated with a needle gently touching the surface. The spores are in suspension of this droplet. This is then transferred on a microscope glass slide and mounted for observation. If the apothecium was dirty with soil, let 10 seconds or more after agitation so that the heavy soil falls down and not sucked by the pipette.

How someone finds it useful

Zitat von mollisia

Hello Steve,

a warm welcome to the forum and hopefully we can contribute to your inventory of the Fungi of Malta. There are some people here familiar with the mediterranean fungus flora.

What concerns this "gall", I have some doubt that it really is fungal. And even more doubt that it originates from the gill edge of this Agaricales. May be this could be a myxomycete?

all the best,

Andreas

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I can confirm that the spherical body was attached and as if part / continuous from the gill. The idea of being a slime mould is possible but personally I don't know of any examples. Also, I would expect some verrucose-spinulose globular spores inside. The myserty continues... Thanks for the warm greeting.