Does anyone knows about the production of mycorrhiza inoculum from mycelium and spores?

Dear Donia. Inoculum preparation would depend on it's intended use. See if this helps...

http://www.springerlink.com/content/x155742041275164/

Best Wishes.

Hi Donia,

I'm agreeing with Ruhi, could you please be more specific? Like, what is supposed to be the use of the inocula you want to produce and what would you use as starting material for the inocula formulation?

Kind regards

hi everyone, thx for ur concern

i need to produce a suspension of spores of terfezia for direct inoculation to the roots of Helianthemum plants...

Hi! You can put directely the spore on distillated water but before you could testing the viability of spore

Dear Donia, I guess you will have to use a carrier, possibly assisted with a suitable nutrient for spores to germinate and then go for inoculation. Carrier based inoculation would ascertain better viability. You may mix growth medium with a sterilized carrier like charcoal, etc and dry before use. Regards.

Hi Donia, ... as per above advice, your request will depend on the plant types & mycorrhizal types you wish to propagate.

To start with, I suggest you check out an Australian text "Working with mycorrhizas in agriculture & forestry", see PDFs at http://aciar.gov.au/publication/MN032 .... Arbuscular mycorrhizal fungi (AMF) are obligate biotrophs - i.e. they can only be propagated together with a suitable & living plant symbionts or 'hosts'. In contrast, fungal spores of Ecto- & Ericoid mycorrhizal fungi can be propagated in axenic conditions, (e.g. in non-living nutrient media in a Petri dish etc).

There is a lot or published research & practical advice available for commercial propagation of mycorrhizal inocula, & or "on-farm" production & utilisation of mycorrhizas, according to cost restrictions etc, (e.g. see http://www.rodaleinstitute.org/20101206_a-complete-how-to-on-farm-am-fungus-inoculum-production )

Many commercial AMF inocula applications use aeroponic propagation methods which are ideal for the easy collection of fungal spores & hyphae. ( see http://invam.wvu.edu/methods/cultures/aeroponic.htm ) Most commercial AMF inocula production is focused on producing single genetic strains, ... however it is also possible to produce genetically diverse AMF, Ecto-M, Ericoid-M, etc inocula if required. From a practical & ecological perspective it is often optimal to 'eco-source' mycorrhizal inocula from soils, plants, climate & locations most similar to the site / crop etc for which it will be used. Conventional 'pot-culture' mycorrhizal inocula propagation methods are relatively cheap & quick, ... & importantly also allow ongoing screening for plant pathogens etc that may have been collected from these field soil sources.

In my opinion, fungal biological diversity is preferable for enabling diverse biological function & ecological 'fitness', (e..g. plant-fungal nutrient access, soil aggregation, drought resistance, disease resistance etc). This biological diversity can be thought of as the "scatter gun" approach, as compared to the single strains or "silver bullet" approach.

As per above comments re carrier media, it is possible to use gels such as Calcium Alginate ( a seaweed extract), expanded-clay media ( Vermiculite, Perlite & Zeolite), & lately biochar appears to offer a promising cheap & renewable alternative for at least some mycorrhizal fungal species. I am very keen to see the future advancement of affordable incocula production methods for use in direct seed drills, ... rather than the current methods which are largely restricted to nursery inoculation & propagation of seedlings for transplanting into field soils.

I`m agree with Don, you need to specify more about the plant and the symbionts assosciation.

Regards

Dear Donia,

As I understand from the comments above you just need a spore suspension. And after that you intend to use it fo infect a plant (correct me if I am wrong). I use Aspergillus niger to innoculate a bioreactor. And I just plate out spores on PDA (potato dextrose agar) plates from a cellbank, then wait for a week until there are enough spores on the plates and then get them of the plates with 0.1% v/v Tween80. Make sure you use something to scrape them of the plates so that you get a high concentration of spores. After that use a heamocytometer to determine the spore concentration

Hope this helps

Hi Ivo & Donia, ... et al

I wish it was always that easy... As per my above comments re Ectomycorrhizal, Ericoid & Orchid mycorrhizal fungi, it is possible to culture these Basidiomycete & Ascomycete fungi in artificial media & in the absence of a living plant 'host' (symbiont partner). These fungal groups reproduce sexually & may produce many 1,000,000s of spores from above ground or below ground fruiting bodies.

However, unfortunately Arbuscular mycorrhizal fungi (AMF) are asexual Zygomycete fungi & are also obligate biotrophic symbionts (i.e. .can't grow let alone reproduce without the presence of a suitable & living plant root partners); & they don't produce 'fruiting bodies' like mushrooms, puffballs, truffles etc ... AMF do however asexually produce relatively tiny sporocarps which contain very few spores. Evidence indicates the importance to AMF spores of bacterial communties living within AMF sporocarps, these bacteria are believed to be important co-symbionts of AMF spores. Some AMF species can contain 'vesicles' within plant roots, some of these may also contain AMF spores.

In aeroponically produced AMF inocula, it is relatively easy to collect 'extradical' spores, sporocarps & hyphae, & likewise root segments containing 'intra-radical' hyphae & spores. However, with soil-borne AMF & 'pot-cultures' it is usual to use 'wet-sieve' methods to separate & collect AMF inocula .... quite time consuming, but relatively low tech & very effective. As a general rule for AMF inocula collection, soil aggregates attached to host plant roots are more likely to contain AMF spores than in soils more distant from plant roots, thus there's a bit of an art involved in getting watering & soil moisture optimal for adhering to roots at harvest, rather than too dry & fragile.

AMF spore germination & root inoculation is most effective if spores are placed relatively close to roots of plants to be inoculated. Likewise viable AMF hyphae inocula needs to be fresh and placed in close proximity to potential symbiont plant roots. It is also possible inoculate plant roots with AMF hyphae placed on moist paper in a Petri dish, & to germinate AMF spores close to roots.

It is tempting to believe the marketing 'hype' about some commercial AMF & Ecto-M inocula, freeze dried fungal spores are very good / ok, (but are often mono-cultural strains), mycorrhizal fungal hyphae are best used fresh !!!

As per my previous message, for further details see free download PDFs from: http://aciar.gov.au/publication/MN032

Hi Donia,

If you want to propagate the isolated AMF, maybe you can place it in a pot with an sterlile mix of soil:vermiculite:perlite in a 1:1:1 proportion, with clamp or sorghum. Then (if the AMF colonized), you can collect the spores with a sieve.

Hope this helps!

hi everyone...

the spore suspension i need is from an ectomycorrhizal fungi (Terfezia) for pot-culture with helianthemum plants...

The spore suspension

 Please mention in vitro or in vivo inoculam production system.

https://bmcmicrobiol.biomedcentral.com/.../10.../s12866-019-1414-x