Hi Vipin,

what do you need to know exactly? I need more details: which type of substrate are you willing to use? which is the inculum formulation (pure spores of one or more isolates? a commercially available inoculum?) that you plan to inoculate? Maybe you want to quickly evaluate a particular symbiotic partnership between wheat and one AM fungus, in this case you could try to inoculate spores directly onto wheat seedling roots (the so called sandwich method). Or you want to inoculate wheat roots with a mixtures of AM fungi in order to evaluate any possible benefit on wheat growth, in this case you just add a quantity of the inoculum formulation to the pot (I normally add a layer of inoculum in between two thicker layers of substrate). If you give me a few more details maybe I can give you more precise suggestions...What's the inoculum made of?

bye

Andrea

Unfortunately, Amhishek Pyasi's proposed method is used far too often. It is similar to the so-called 'autocthonous' inoculum published in some of the work from Granada, Spain. This is no way to produce inoculum, unless you also want to include mites, nematodes, pathogenic fungi, oomycetes, chytrids, and any other kind of soil inhabitants. Not only that, but such work is scientifically unrepeatable. If you do the same thing the next time, there is no way of knowing if your 'inoculum' is the same as the previous time.

So, what you need to do is to isolate your fungi. If you feel the need to mix species, then you should isolate several species, and mix them yourself at the time of inoculation. That;s the scientist’s job – not to allow random acts of nature to operate. This way, what you do can be controlled and replicated, as should all scientific experiments.

So, you can begin with soil traps of the sort recommended by Abhishek, but not for use as inoculum. The traps will allow you to multiply up some (though unfortunately not all) of the AMF in your soil. Then you extract spores from these pots and use them to establish single species cultures, or better still, isolates (the term isolate should NEVER be used for anything other than a culture established from a single propagule). You can then bulk up these cultures or isolates, but you must ensure you use disinfested substrate and mycorrhiza-free host plants. Once bulked, you can use them as inoculum, preferably after establishing their colonisation potential with trials (most probably number).

As you live in India, I suggest you contact Prof. Alok Adholeya, in Delhi ([email protected]), who runs the Biotechnology and Management of Bioresources Division, The Energy and Resources Institute in New Delhi. His group can supply outstandingly good AMF inoculum: obtaining it in this way will save you much work and time.

Take a sterlized soil and sand (1:1) and mix it throughly, add this mixture of soil 3-4 layers and add AM inoculum of single type of spores, which you have already done multiplied by trap culture. this was added by 10g of inoculum of AM fungi by a layer then add 1 layer of mixture soil again in the top.

I think you must tell us which type of inoculum you have first, but generally the put some of the inoculum under the seeds, and some other inoculum mixed with soil above the seeds.

Thanks for kind suggestion