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Orbilia?
Koszka Attila, 27-12-2019 04:11
On dead, still attached Sambucus-twig.

Spores 6,5-8 x 1,5 micrometers, with a head-shaped part towards the top of the ascus.
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Hans-Otto Baral, 27-12-2019 08:53
Hans-Otto Baral
Re : Orbilia?
Woww, this is incredible how you obtained these many groups of 8 spores!

This is an undescribed species, Orbilia ficicola, very close to O. abutilonis (a very badly described species from Hawaii) and O. siculispora, another novelty.

O. ficicola was repeatedly sampled from Ficus during the last years, but then also on other hosts. Which Sambucus is this and how sure is i Sambucus? Does it have a thick pith inside?

Zotto
Koszka Attila, 27-12-2019 10:48
Re : Orbilia?
Dear Zotto!
Many thanks for the information.
I saw the similar picture on Dragisa Savic's page, but there are no microphotograps.
A small piece of twig with fungus was placed in the microscope slide, and the
fungus was shot its spores on the slide in this 8-membered groups.
The substrate is Sambucus nigra, it is absolutely sure.
I send to you some dried material if you need it for your work.
When will it (the Orbilia-paper) complete?

Attila

Hans-Otto Baral, 27-12-2019 11:04
Hans-Otto Baral
Re : Orbilia?
Dear Attila

this is a wonderful collection but I do not need it for study, simply tell me please the collection data (with coordinates) and then I will take it up.

How much above ground was the twig? The diameter of the twig is below 5 mm?

What kind of ecology is there, which other trees?

At this occasion I can tell you that during the next days we will stop taking up any further records, as we are about to work on the layout. This is a pity but there must be an end, eventually.

Zotto
Koszka Attila, 27-12-2019 16:36
Re : Orbilia?
The collection data:

Hungary, Vértes mountains, near Pusztavám, on Sambucus nigra

47.411785 18.276400

The twig was 1,5 m high above the ground.

The diameter of the twig (at the mushroom-covered part) was 6-10mm.

This is an edge of an oak forest (Quercus cerris), with Sambucus nigra, Crataegus monogyna, and Pyrus pyraster.

I am writing a book as well, about the ascomycetes of Vértes mountains. The hardest thing is terminate (the best species will come after the finishing of the book anyway) :-)

With my huge respect:

Attila
Guy Marson, 27-12-2019 22:08
Re : Orbilia?
Dear Attila, 

Thanks for sharing your find of Orbilia ficicola growing on Sambucus.
As I just found out, I forgot to send the sequence from O. ficicola to GenBank. That will be done in the coming days.
As Orbilia ficicola already has a few SNPs ( https://en.wikipedia.org/wiki/Single-nucleotide_polymorphism ) within a radius of 150 km, I would be very interested to also sequence your Hungarian find.
If this is not a problem for you, I would ask you to send me a small part of your find - a little bit of wood with about 4-5 apothecia (usually, a 1 mm diameter apothecium should be sufficient for the PCR).
BTW: A PCR works best if the fungal material is dried as quickly as possible after the discovery and sent in a "ventilated" paper bag.  
I will send the remaining apothecia back to you when the sequencing is successfully completed.


Thanks a lot and best wishes for 2020.


Guy

-----

Guy Marson
45b, rue de Bettembourg
L-5810 Hesperange
Luxembourg

Hans-Otto Baral, 29-12-2019 09:07
Hans-Otto Baral
Re : Orbilia?
A problem wth this sample is the rather short spore size. Usually, O. ficicola has a bit longer spores than O. siculispora, i.e. 7.5-10.5 µm vs. 6.5-10.

I suggest to reexamine the sample, I am sure there will still be living spores of a larger size. Those on the pic are all dead.

To distinguish abutilonis is very complicated. Perhaps the hair cell size would help, but I am sceptical in this idea. Also the value of apothecial colour is far from settled. Cash said for abutilonis: "pale ochraceous buff to pinkish buff". Recently Adam Polhorsky found a mixture of two populations, beautiful pink and clear ochraceous. I imagined that two species grow together. Perhaps he will gain sequences in the future.

Here is the present key:

6. Disc orange-ochraceous; hairs broad till apex, upper cell *3–5.5 µm wide, thin-walled; conidia with (2–)4 parallel, 3.8–5.5 µm wide arms; spores *(6.5–)7–9(–10) µm long; bark & wood of Populus, Ulmus, cold-temperate humid Europe ...... O. siculispora
6. Disc ochraceous to often pinkish; hairs partly distinctly narrower in upper part, upper cell *2.5–3.3 µm wide, thin- to thick-walled; conidia with 2–4 strongly diverging, outwards curved, *3.5–4.3 µm wide arms; spores *(7.5–)8–10(–10.5) µm long ... 7

7. Basal inflation of */†spores max. 2× thicker than connecting part; hair cells 5–14 µm long; bark of Ficus, Populus, Tilia, Ulmus etc., submediterranean semihumid to cold-temperate humid Europe .... O. ficicola
7. Basal inflation of †spores > 4× thicker than connecting part; hair cells 10–18 µm long; bark of Abutilon, tropical subhumid Hawaii .... O. abutilonis

Attila, could you please tell me the date of collection?

Thanks
Koszka Attila, 30-12-2019 09:36
Re : Orbilia?
The collection date was 2019.12.25.
Hans-Otto Baral, 30-12-2019 11:17
Hans-Otto Baral
Re : Orbilia?
Thanks!
Hans-Otto Baral, 31-12-2019 08:47
Hans-Otto Baral
Re : Orbilia?
Falls du eine vierte Sequenz von ficicola hast, dann weiß ich leider nichts davon. Drei sind hochgeladen: MH221053, MK473415, MK473408
Adam Polhorský, 31-12-2019 13:58
Re : Orbilia?
Hello all,

I actually had 2 more collections of O. ficicola, one on Juglans wood and bark and one on Eounymus bark. All populations clearly pink, not ochraceous.

I can sequence it, but I do not mind sending it to Guy aswell. Whichever is more convenient.

I`m preparing the collection data for Zotto.
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Guy Marson, 31-12-2019 23:20
Re : Orbilia?
Ich hab nichts Neues ausser einem misslungenen Versuch die beiden IGS zu erfassen. Beim IGS2 (Upstream von der SSU) sagt Blast: CP039677 - Parastagonospora nodorum strain Sn79 chromosome 12. Der IGS1 ist Primer Dimer (Merde!). 
Die beiden letzten GB- Nummern (MK473415, MK473408) sind gesperrt, wenn die nicht von mir stammen, kannst du sie ja mal schicken. Die erste Nummer (MH221053) ist als Unveryfied deklariert aber das ist genau die Seq. die ich auch hab - bis auf das dritte nt im Intron der Domain 9 der LSU: das ist ein "A" (CTATGACTCAA). 

1 guten Rutsch.

Guy
Guy Marson, 31-12-2019 23:31
Re : Orbilia?
Hi Adam,

Congratulations for your finds!! 

You are welcome to send me some of the material (4-5 apothecia are sufficient) But it will take a while until I get results. Our ABI 3730 XL had to get new capillaries. Three weeks were planned but the exchange took almost 2 months and a lot f stuff has accumulated (...). I don't have the sequencer usage plan in mind, but in late January or early February I will be able to run one or two 96 well plates.

I wish you and all members of the forum all the best and many interesting finds for the new year 2020.

Cheers, 

Guy
Koszka Attila, 16-01-2020 12:38
Re : Orbilia?
Dear Guy!
Did the Orbilia ficicola sample reach you in order?
I sent it a week ago.
Attila
Hans-Otto Baral, 16-01-2020 12:40
Hans-Otto Baral
Re : Orbilia?
Yes it did. But I suppose Guy will answer soon.
Koszka Attila, 16-01-2020 12:45
Re : Orbilia?
Super! I was just worried, because nothing is going well in our country (especially the post office).
Guy Marson, 16-01-2020 22:05
Re : Orbilia?
Dear Attila, 


Your material arrived a week ago (09-Ja-2020).
The first thing I tried was to find anamorphic conidia (see below). 
Then I suspended an apothecium over corn meal agar (CMA) for spore discharge.
A second apothecium gave very good PCR results. The sequencing will be done later.
I sent the material back to you this afternoon. That should arrive in two working days.

Thank you very much for sending the material !

May I email the final results to your hu.g4s.com address?

Best regards,
Guy
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Koszka Attila, 17-01-2020 08:02
Re : Orbilia?
Dear Guy!

I am glad that the examination was successful.
It is not necessary to send the sample back,
because it was divided into two parts, I kept the rest.
If you get the results, please send a mail to the attila.koszka@hotmail.com.
Many thanks for your work!
Attila
Koszka Attila, 22-01-2020 09:27
Re : Orbilia?
Dear Guy!

The material has arrived back to me successfully.

Thank you for everything! :-)

Best regards
Attila
Guy Marson, 22-01-2020 16:25
Re : Orbilia?
Hi Attila,

I'm happy to read that the post worked well ;). Meanwhile, the culture grew quite abundantly and I hope to catch the anamorph with it. If not, you may could try the thicker part of the branch to catch some conidia? I've already examined the thin parts (the three parts you sent me). The method to find the conidia is not very complicated. If necessary (if the conidia don´t grow in culture), I will contact you by email.


Best regards,

Guy