AMF Spore Isolation, Enumeration, and Identification
AMF spore isolation and enumeration was done following
the method of Uma et al. [35]. Suspension of 25 g soil
sample in water was passed through a series of 710 to
37 lm sieves. The residues on the sieves were washed into
beaker with water and filtered through filter papers. Each
filter paper was spread on petri dish and spores were
counted using a dissection microscope at 409 magnification.
Sporocarps and spore clusters were considered as one
unit. AMF spores were picked up using a needle, mounted
in polyvinyl alcohol-lactoglycerol with Meltzer’s reagent.
AMF spores were identified based on morphological
characteristics such as shape, size, colour, wall ornamentation,
etc. using identification keys of International Culture
Collection of Vesicular and Arbuscular Mycorrhizal
Fungi (http://www.invam.caf.wvu.edu) and AMF phylogeny
(www.amf-phylogeny.com). Spore density and species
richness were expressed as number of AM fungal spores
and numbers of AM fungal species in 25 g soil sample.