Trichoderma Harzianum & Vesicular Arbuscular Mycorrhizas

[oldtimer1]

None of this is really within my field, I suspect to do any work of real value a high powered compound microscope, oil immersion, stains etc would be needed.

I’m not sure how mycorrhizals produce spores, as to live in the vegative form it is a symbiont relying on a host to live, surely you have to have a vegative form complete its cycle to produce a fruiting body? I can’t see a life form reliant on a host to live and grow being cultured in the way you suggest, I suspect this type of fungi has to be cultured on the roots of a living plant as a host.

[Guest dr rockster]

I've just read some of the stuff in the link billious supplied(cheers dude! ) about trichoderma's.All fascinating stuff but I didn't realise the stonewash effect on denim jeans came from a process involving trichoderma's! :wink:

But I also didnt know they are a group of fungi and not a single strain.

[Felix Dzerzhinsky]

Agreed OT, I'd love to have a lab setup with dyes and a really good microscope but I'd be happy at the moment if my petri dishes looks like the one in the link Billious posted when they've spored.

Perhaps Arbuscule would like to explain for us as his username is the actual fruiting body of Mycorrhizal fungi. It's in the link he posted I believe, if not I'll try and dig out where I read it.

PDA is purposely designed for fruiting/sporulating fungi and provides the nutrients (complex carbohydrates) they would normally obtain from the plant roots and minerals they would get from the soil, apparently. This is the first time I've ever done this so it's a bit of a learning experience as I go. I just thought I'd try a few different things and see what happens.

Nothing ventured nothing gained :wink:

[distracted]

Hi all, I thought I would post an image of CANNA akTRIvator spores (10g). Even at the highest mag I have its impossible to resolve the spores themselves but thats not surprising as they are so small at 200th of a mm, just clusters of spores are visable. The image is polarised but with some side illumination. The very fine material is the spore custers and the more visable grains are additives maybe dessicants or organic detritus?

There are also grains that are transmitting light which is interesting:- with a polarised microscope you shine polarised light from below and then block the light past the image with a 90 degree polarisation. Glass and anything without a crystal structure looks black as no light from it gets through but crystaline material will twist the light so that the last filter doesn't block it, this may be only in one direction though so all the grains need to be revolved to see if they transmit light at different angle to the polarisation. This image shows that several grains are brightly coloured, and many more in the image would look the same if the image had been taken at a different rotation. This means that a lot of the larger grains have a crystal structure and therfore can't be spores. Most likely these are mineral grains. This also explains why my spray nozzle keeps blocking up as some grains were quite large maybe 300 microns or more.

Anyway thought that may interest felix at least!

[Guest dr rockster]

(little voice) Me too,I'm innerested too!

Do you think these mineral grains could be of the type found in Rock Dust and suchlike?

According to you clued up fungal chaps the minerals in Rock Dust when placed in the plants rhizosphere enable fungal beneficials to establish themselves very effectively,so maybe the inclusion of these particles at the time of application along with the fungals would be a logical and perhaps very beneficial thing to do?

Liked your explanation of polarisation/rotation for those not aware of dem tings dude.

Edited March 5, 2013 by distracted

[distracted]

V.hard to guess on the minerals as mineral grain ID is based on set thickness's of grains but I'd hazard that there are quartz grains, maybe a bit of calcite, some mica and what appeared to be zircon. Silt like really in appearance. However a full solubility test would be needed to exclude salts, initial tests suggests though that these are insoluble

[Guest dr rockster]

Which makes me think they are not there as a matrix or substrate to exist on but will aid the fungi to colonise on being applied mebbe?

[distracted]

Maybe Doc, or they could be a dessicant, a contaminant or even just a filler?

[Felix Dzerzhinsky]

Wow ,

That's an interesting picture, thanks for taking the time to post that. That's really cool with the polarised light I didn't know you could do that, it can reveal quite a bit more information that plain light, very interesting man.

So the really small particles are clusters of spores ? I wonder why there is so much other material ? Given that they are likely to be cultured on Potato Dextrose Agar or in a liquid broth I don't see what purpose the extra mineral could serve. I've also noticed blocking on my sprayer and wondered why, perhaps we have an explanation there. Could we possibly call this 'Grit Spores' ? Another reason to culture our own perhaps.

Nice one

Come on Doc, well get you fitted out with a Tefal forehead

Trichoderma Harzianum aren't the same as Mycorrhizal fungi as they do make symbiotic associations with plant roots but rather than absorbing minerals they parasitise pathogenic fungi and possibly encourage root expansion to increase their own environment ?

Edited March 5, 2013 by distracted

[Guest dr rockster]

Oi,fuck off Felix!

I've got a Tefal head larger than most star systems so please don't take me to task for summink that is usually no more interesting than stuff that grows between most peoples toes!

[Felix Dzerzhinsky]

[Felix Dzerzhinsky]

Not much happening in the petri dishes really. The little jelly like blobs have now extended to cover the entire surface of the agar. It's quite difficult to get a clear picture and it only really shows up well under sunlight and sadly there's not much of that around just before I leave for work in the morning.

The brown specks are just AKTrivtor powder where I sprinkled some on top to make sure it was populated. Next time round I'd use a really minute amount dissolved in sterilised water.

Just a quick word on the importance of sterilising your tools, errmmm I didn't so can't be totally sure I'm growing Trichoderma H until it produces spores

[Felix Dzerzhinsky]

Ok we have some major fungal growth occurring on the petri dishes, looks like I didn't screw up when I didn't sterilise everything. I apologise for these pictures being a little dark but it's not very light on my kitchen windowsill in the early morning.

Hopefully there will be some fruiting bodies visible in the next few days and may even see some spores by middle of next week

[withnail]

Wow they are really coming along felix

Can't see much bud formation though

[Felix Dzerzhinsky]

Do you think I should maybe cut down on the photoperiod then Withnail ?