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PostPosted: Fri Mar 01, 2019 7:18 pm 
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Hi all,
The weather here in the U.K. is beginning to show signs of improvement and I've been wandering around the garden looking at many plants that are just beginning to 'come back to life'. A small rather tragic shrub has just started to open it's dormant buds to reveal smallish yellow flowers. I took a branch or two into the lab to have a look at the still-closed and just-opening buds, mainly to see if there was any interesting material for slide making or at least tissue sample collection....

I don't know the species or even the Genus I'm afraid; if anyone has any idea please let me know; it's a small ornamental cultivated garden shrub. Apologies for the following rather poor pictures but at the moment the plant is little more than twigs - no leaves yet...

Image

Image

The above said, my attention was drawn, during dissection under my stereo 'scope, to a number of rotting buds on the same branch as the clearly viable buds, that had died-off and become 'mouldy' for want of a better word.
Opening the tiny immature and mouldy buds I found that they had what almost looked like a tiny 'bracket fungus' within the folds of the leaves that protect the unopened bud. No fruiting bodies apparent but the commonly seen mycelium was evident.

The first thing I did was to place a few tiny pieces of the fungus and rotting-anther parts onto a slide and add a single drop of water. This is standard practice for me in preparation for the addition of a coverslip and examination with my Orthoplan (compound 'scope). Additionaly, as soon as any type of fungus enters the scene while dissecting I usually reach for one of the most well-known stains for this, 'Lactophenol Cotton-Blue' stain.
Adding a very lightly-staining quantity of this and rinsing I expected of course to see the mycelium strands and perhaps a few pollen grains, which I expected to survive the rotting process as pollen is extremely robust and able to withstand a huge variety of insults.

Both of these I saw, along with literally thousands of sickle-shaped objects that look a lot like diatoms - but having checked for birefringence with polarisation, and seeing none, I assumed that they were not Diatoms... They were not green either, as seen with those that remained unstained, as may be expected of the other that they resemble - algae. The question of how they get into the buds or indeed why they seem to thrive there in great numbers will have to wait without any even basic ID.

Some more images,
Here is some of the mycelium (the translucent woolly area on the left of the dark 'lump' which is a rotting anther) surrounded by many of the mystery objects that poured out of the mycelial tangle as water was added,
Image

Closer-in this is the mycelium loaded with the unidentified sickle-shaped 'things' - I chose a very lightly stained area here to try to maximise detail of both in one image,
Image

Here, backing away a little, are the objects en-masse, the round-ish darker objects are pollen grains,
Image

Apologies for the horrid image-blur....
Image

This is a little better, with pollen grain and scale-bar,
Image

So, I'm certain many here know exactly what these are, I personally have no real idea!

Amazing what pops-out when looking at plants, their charm and mystery never ceases to please and amaze me! :D :D

Ideas for ID please folks! John B. :)

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PostPosted: Fri Mar 01, 2019 8:08 pm 
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Hi John,
this should be a forsythia.

I also have started to look at buds and think about what to do with them.
They might be a suitable victim for my glycerin soap embedding tests.

Bob


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PostPosted: Fri Mar 01, 2019 8:08 pm 
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Location: NorthWest England
mrsonchus wrote:
I don't know the species or even the Genus I'm afraid; if anyone has any idea please let me know; it's a small ornamental cultivated garden shrub. Apologies for the following rather poor pictures but at the moment the plant is little more than twigs - no leaves yet...

First thought was 'Winter Jasmine' ...
https://www.rhs.org.uk/Plants/9452/i-Jasminum-nudiflorum-i/Details
... but your flowers appear to only have three petals.
Edit: or have I mis-read the image ?

Forsythia [of which there are many varieties] looks similar, but has four petals !

Mmm ...

MichaelG.

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PostPosted: Fri Mar 01, 2019 9:24 pm 
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MicroBob wrote:
Hi John,
this should be a forsythia.

I also have started to look at buds and think about what to do with them.
They might be a suitable victim for my glycerin soap embedding tests.

Bob


Bang-on Bob - I looked into it and the four petals, less trumpet-like flower and the appearance of the flowers before the leaves make it a Forsythia I think. Thanks for the ID - I couldn't for the life of me remember what it was!

John B.

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PostPosted: Fri Mar 01, 2019 9:25 pm 
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MichaelG. wrote:
First thought was 'Winter Jasmine' ...
https://www.rhs.org.uk/Plants/9452/i-Jasminum-nudiflorum-i/Details
... but your flowers appear to only have three petals.
Edit: or have I mis-read the image ?

Forsythia [of which there are many varieties] looks similar, but has four petals !

Mmm ...

MichaelG.


Aha Michael, yes, four petals rather than the five or six of Jasmine, and the less extensive fusion of those petals at their bases....
Thanks for the ID old chap.

John B.

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PostPosted: Fri Mar 01, 2019 9:31 pm 
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Here are a couple more images of the UMOs - these are taken in monochrome with the use of a green filter below the condenser, using a lovely x50 water-immersion objective I have, N.A. nominally 1, used with a dry 602 Achr 0.90 condenser.

They look a little more detailed but nothing special - anyone know what these are?

This one looks to have a joint in the middle... In the company of a pollen grain,
Image

Another group of the thousands present in the mycelium within a rotting bud, cropped,
Image


John B.

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PostPosted: Sat Mar 02, 2019 9:00 am 
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Hi John,
from look alone the closest to these mysterious objects I know are closterium desmidacae - probably not a valid identification.
I had a look through my biology books but didn't get any further.

Bob


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PostPosted: Sat Mar 02, 2019 11:26 am 
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MicroBob wrote:
Hi John,
from look alone the closest to these mysterious objects I know are closterium desmidacae - probably not a valid identification.
I had a look through my biology books but didn't get any further.

Bob


Hi Bob - you're a pal! Checked some images and they do indeed look like the closterium Desmids - a new area for me entirely but very intriguing! These ones have a couple of characters that may help with ID - the obvious banding that definitely seem to be external to the cytoplasm - maybe.... Also upon close examination under the 'scope they have numerouse closely-spaced longitudinal lines for want of a better term. Maybe these two characters together will help to home-in upon an ID?

If I get a chance I'll try to find some more today and maybe subject them to some oblique and darkfield examination,if I get the time....

A question that immediately occurs to me is "how do they get there?" - but I suppose they will have been transferred by birds, insects etc? Another question may be "do they thrive there?". They didn't look green but I suppose they may have been, or even that they may have died and lost the green....

I'd be very interested to identify them further if anyone can help.

Thanks for the lead Bob, you've wetted my appetite now to find out more! :D :D

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PostPosted: Sat Mar 02, 2019 6:09 pm 
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Hi John,

It certainly is not a Closterium. I think the answer can be found if you google "Fusarium conidia". You will find there pictures comparable to yours.

Roland


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PostPosted: Sat Mar 02, 2019 7:15 pm 
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Roland wrote:
Hi John,

It certainly is not a Closterium. I think the answer can be found if you google "Fusarium conidia". You will find there pictures comparable to yours.

Roland


Aha! Roland yes! (sorry Bob old chap ;) ).

That explains a few questions I had,
1) The size - if my scale-bar is correct these are (almost) an order of magnitude smaller than Closterium.
2) The affinity for LP-cotton-blue stain suggests fungi....maybe....
3) I couldn't find a Closterium with the 'bands' that the conidia have.
4) The context, i.e. that they were found literally among fungal hyphae.
5) They seemed to have no colour, certainly no green.

Now I have the leads from you kind fellows these thing make sense! :D :D

A really interesting mini-adventure from a side-track towards rotten buds whilst intending to examine the viable bud and developing flower! Microscopy is certainly a fascinating pursuit, and so accessible in terms of equipment, material for study and now we have t'internet, valuable and friendly help from kind folks such as those in this fine forum. :D

Thanks all, very good to have found the result, and learned quite a bit along the way.

John B. :)

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PostPosted: Sun Mar 03, 2019 12:39 pm 
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Congrats, John :( Ffor this is what we know about fusarium: "...cause fusarium rot in plants. They affect the roots that then blacken and die. Next the whole plant wilts and dies. There is no remedy. All affected plants must be destroyed asap to pevent further contamination. The soil must be sterilized (translated from wikipedia.nl)".


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PostPosted: Sun Mar 03, 2019 3:56 pm 
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Arrggghhhhh.... :o

My garden's doomed! :(

I'd better keep an eye on that plant..... :D

John B.

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