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Lasiobelonium - which one?
Bharati Mandapati,
06-02-2025 06:25
Hi All,
I would love some help with this Lasiobelonium – based on Raitviir 1980 it could be L. subflavidum BUT there are some spores with no septation (perhaps younger ones), though there are 3-septate spores, some look like there are 1 or 2 septations. Picking the other choice at (9) in Raitviir leads me to species that do not match in other respects. I did note that the photographs for the L. subflavidum collections in Baral's excellent set of images also seems to have this type of variation in spores. My detailed notes (see notes field), photos (including photomicrography) and references consulted are at https://www.inaturalist.org/observations/260777677
Thanks in advance!
Bharati
Berkeley, CA, USA
Hans-Otto Baral,
06-02-2025 08:26
Re : Lasiobelonium - which one?
Hi
this is a very common error, apparently impossible to erase. Not less mature but alive!Â
You have plenty of ejected living spores, which are fully mature, as I defined the term mature (1992).
The septum is only not visible because of the many guttules. You must simply add MLZ or another killing agent that masks the LBs to the water mount, then you see pretty well the septum.
It seems to me that the mature spores are only 1-septate and get 3-septate only when overmature. But this you can find out when testing the numerous ejected spores with MLZ.
I am not sure why your first micro image shows dead elements only, although being in water. I suspect pressure or drying and rewetting´as reason?
Your ascus measurements sound like referring to dead asci, considering the low width. I see no living ascus in your pics.
The spores are quite large, so L. lonicerae is impossible.
Zotto
Bharati Mandapati,
06-02-2025 20:12
Re : Lasiobelonium - which one?
Hi Zotto,
>"this is a very common error, apparently impossible to erase. Not less mature but alive!
>You have plenty of ejected living spores, which are fully mature, as I defined the term mature (1992).
Thanks for the quick and helpful reply! I am an amateur beginner with asco microscopy and have a LOT to learn.
>"this is a very common error, apparently impossible to erase. Not less mature but alive!
>You have plenty of ejected living spores, which are fully mature, as I defined the term mature (1992).
>The septum is only not visible because of the many guttules. You must simply add MLZ or another killing agent that masks the LBs to the water mount, then you see pretty well the septum."
Ah! Just downloaded the 1992 paper – thanks for flagging it.
>"It seems to me that the mature spores are only 1-septate and get 3-septate only when overmature. But >this you can find out when testing the numerous ejected spores with MLZ."
Will do.
>"I am not sure why your first micro image shows dead elements only, although being in water. I suspect pressure or drying and rewetting´as reason?
>Your ascus measurements sound like referring to dead asci, considering the low width. I see no living ascus in your pics."
Pressure is what I suspect. I will review my current collection of photos (I have more and of different mounts) and better yet make a new set of slides after I read the 1992 paper.
>"The spores are quite large, so L. lonicerae is impossible."
 Noted. Based on the Raitviir key I get from 1-->2-->3-->9 which would rule out L. lonicerae – am I missing something again?
Bharati
Hans-Otto Baral,
06-02-2025 20:44
Re : Lasiobelonium - which one?
L. subflavidum is a really good option. I saw it only once, in Tenerife
Bharati Mandapati,
06-02-2025 20:54
Re : Lasiobelonium - which one?
>"L. subflavidum is a really good option. I saw it only once, in Tenerife"
Thanks again, Zotto! That is where I landed but for my doubts (see my first post here) but obviously I have MUCH to learn!
As I was checking to see whether there are records of L. subflavidum in the US, I found this wonderful correspondence between Lee Bonar and Dennis regarding specimens from California (in fact Bonar's collection was originally stored here in the Berkeley herbarium) - of course I don't know whether the species concept from that time is the same as now. https://www.gbif.org/tools/zoom/simple.html?src=//api.gbif.org/v1/image/cache/occurrence/4868707169/media/088eecbda47edd670796d23c61e06e00
Hans-Otto Baral,
06-02-2025 21:15
Re : Lasiobelonium - which one?
Ineresting. This is the question. I never went thoroughly into that species. But recently I collaborated on the new Outline (MYCOSPHERE 15(1): 5146–6239 (2024) www.mycosphere.org ISSN 2077 7019
Doi 10.5943/mycosphere/15/1/25) and copied an account by Peter Johnston which I found interesting:
Doi 10.5943/mycosphere/15/1/25) and copied an account by Peter Johnston which I found interesting:
Lasiobelonium (type L. subflavidum)
PRJ Outline: The type species of Lasiobelonium, L. subflavidum, and a species described from New Zealand as Dasyscyphus triseptatus, were placed in synonymy by Raitviir (1980) and this synonymy was accepted by Spooner (1987). Based on the description in Spooner (1987), a recent specimen from New Zealand (PDD 60106, https://scd.landcareresearch.co.nz/Specimen/PDD_60106) is morphologically typical of L. subflavidum. The multi-gene analysis of Johnston (2022) incorporates data from PDD 60106 and places Lasiobelonium in Solenopeziaceae sensu Johnston & Baschien (2020). This supports Johnston & Baschien (2020), an analysis where Lasiobelonium was represented by L. lonicerae rather than the type species.
Bharati Mandapati,
06-02-2025 21:49
Re : Lasiobelonium - which one?
Thanks Zotto. Sadly I don't have a copy of Spooner 1987 but I did note that all the records labeled L. subflavidum on Genbank seem to be from the Johnston collections and cultures and that the descriptions + photos are available here https://biotanz.landcareresearch.co.nz/scientific-names/1cb18cb4-36b9-11d5-9548-00d0592d548c
PS; what loci do you use when sequencing Solenopeziaceae?
Hans-Otto Baral,
06-02-2025 21:52
Re : Lasiobelonium - which one?
As in other Helotiales: ITS and LSU D1-D2 or D1-D4
Bharati Mandapati,
07-02-2025 01:42
Re : Lasiobelonium - which one?
Got it. Thanks again!
Bharati Mandapati,
17-02-2025 22:35
Re : Lasiobelonium - which one?
Hi Zotto,
I did not forget my homework assignment on this collection. :-) Life sometimes interferes with mycological pursuits.
I updated my inaturalist observation to include: an image of the dead spores showing septation (see photo 6; and one showing living asci with those guttulate spores (see photo 7); and relabeled photo 5 to reflect the fact that they are living spores.
Thanks again,
Bharati
I did not forget my homework assignment on this collection. :-) Life sometimes interferes with mycological pursuits.
I updated my inaturalist observation to include: an image of the dead spores showing septation (see photo 6; and one showing living asci with those guttulate spores (see photo 7); and relabeled photo 5 to reflect the fact that they are living spores.
Thanks again,
Bharati
Hans-Otto Baral,
17-02-2025 22:45
Re : Lasiobelonium - which one?
It requires some experience but I am sure from the MLZ photo that the spores get 1-septate. Wht could be taken as further septa is plasma.
Bharati Mandapati,
18-02-2025 00:25
Re : Lasiobelonium - which one?
Hi Zotto,
All this is very illuminating for me – what structures within spores are what and what is visible in what media - , so some more questions in my ongoing tutorial: :-)
1) What do you make of the septation in the version with KOH?
2) In your Google Drive collection for L. subflavidum
a) How would you interpret the MLZ image embedded in 100x_spo_koh+rc_24038_apomad0420.jpg?
b) In the image titled Lasiobelonium subflavidum cf., 23.VI.94-2a.jpg, (JM Castro 23-3-94), the note below the spore drawing says that 1 septum is clear, 2 (others) are less clear. The next drawing from this series, labeled 2b shows a variation in septa.
Thanks
Bharati
All this is very illuminating for me – what structures within spores are what and what is visible in what media - , so some more questions in my ongoing tutorial: :-)
1) What do you make of the septation in the version with KOH?
2) In your Google Drive collection for L. subflavidum
a) How would you interpret the MLZ image embedded in 100x_spo_koh+rc_24038_apomad0420.jpg?
b) In the image titled Lasiobelonium subflavidum cf., 23.VI.94-2a.jpg, (JM Castro 23-3-94), the note below the spore drawing says that 1 septum is clear, 2 (others) are less clear. The next drawing from this series, labeled 2b shows a variation in septa.
PS: I just noticed you added my previous inat images to the L. subflavidum Google Drive set (wow)! Perhaps you could update them to swap out/include the images I added today.
Thanks
Bharati
Hans-Otto Baral,
18-02-2025 08:34
Re : Lasiobelonium - which one?
Your images in MLZ and KOH are very instructive:
In MLZ you can best see the septum, while the plasma contents are rather masked.
In KOH you can see the lipid (LBs) while it is difficult or impossible to discern the septum. You need experience to interpret the "empty" space between the groups of LBs. In living spores this can be similarly difficult, so that I recommend to use a viscous and lethal medium in such cases to clarify the septa.
The collage you refer to is by Luis Quijada and shows in MLZ one spore with (at least) two septa. From his pics of living mature asci and living free spores I conclude that the spores are 1-septate before and during discharge. Whether 3-septate spores occur already inside living asci is unclear to me.
I added your two new images, but you must change the text in one: Living asci you need to replace by Dead asci. What concentration has your glycerol-water? Glycerol is not necessarily toxic but it causes water extraction, shrinkage, and masking of cell contents. You can test what happens when you wash the preparation with water afterwards.
Bharati Mandapati,
19-02-2025 00:49
Re : Lasiobelonium - which one?
Thanks! Will do re: changing the text on that image. Clearly I have much to learn re: identifying living asci. I thought these asci looked more like Figure 8a or 10a in your 1992 paper than their dead counterparts (8b and 10b).
I am using 20% glycerinated water, mainly to avoid the dreaded [typo was=Becke's] black line.
Hans-Otto Baral,
19-02-2025 20:25
Re : Lasiobelonium - which one?
Becke's line? Such a concentration immediately shrinks asci etc. You can make a test by adding it to a water mount of living asci.
Bharati Mandapati,
20-02-2025 03:01
Re : Lasiobelonium - which one?
Oops that was a typo - I meant the black line caused by the evaporation of the medium - apparently remedied by using a more viscous medium.
Alas this collection is past its prime, but will implement a with and without glycerin water test on my next asco.
Thanks!
Hans-Otto Baral,
20-02-2025 09:31
Re : Lasiobelonium - which one?
I think a small amount of a detergens (SDS) is better, but I am not familiar with that. Not evaporation but wettability is the problem you mean?
Bharati Mandapati,
20-02-2025 21:48
Re : Lasiobelonium - which one?
Marcel Lecomte has a nice description of the problem on p.77 of Microscopy & Fungi (English Ed, 2024) in the section re:Â Mishandling or Accidents in Microscopy: Areas of Retraction
I am attaching a photo of that paragraph from p. 77, in case others don't have a copy.
On pp. 41-42, he includes some options for viscous media including glycerinated water.
I am attaching a photo of that paragraph from p. 77, in case others don't have a copy.
On pp. 41-42, he includes some options for viscous media including glycerinated water.
Hans-Otto Baral,
20-02-2025 22:04
Re : Lasiobelonium - which one?
Ah, now I understand. Well, viscose media always cause damage to living cells. The easiest is to add water after a while.
Bharati Mandapati,
20-02-2025 22:29
Re : Lasiobelonium - which one?
You mean add some water to the edge of the coverslip once you observe the medium is evaporating in the initial prep?
Hans-Otto Baral,
21-02-2025 09:22
Re : Lasiobelonium - which one?
yes of course. The disadvantage is that free spores move away.
Bharati Mandapati,
21-02-2025 19:00
Re : Lasiobelonium - which one?
Thanks. Tradeoffs I guess - vital spores but escaping as fast as they can :-)