Method: How to count numbers of mycorrhizal species.
Counting a number of Glomus intraradices, Glomus mosseae, Glomus aggregatum, Glomus clarum) in a product like fertilizers, haylage, silage, hay etc.
1. Small samples (approximately 15 ml) from different stocks of examined product are suspended in water and thereafter sieved using 500 and 50 μm sieves.
2. Contents from each sieve are washed with water into a dish for examination under dissecting microscope.
3. Further microscopic studies of spore morphology are carried out (by a trained biologist) by examining features of spores mounted in PVLG (Polyvinyl alcohol-lactic acid-glycerol) (Omar et al. 1979) with and without the addition of Melzer’s reagent (5:1 v/v, Walker et al. 1993).
4. Numbers of spores of each mycorrhizal species are counted (by a trained biologist) in the examination dish under dissecting microscope. Alternatively it can be calculated using Neubauer's heamacytometer chamber as described in our protocol How to count fungi (at a glance).
5. The relative quantity or density of spores of each mycorrhizal species in the product stock is calculated using the following equation: d=n /Vs, where d - density of spores of each species in the product stock (spores/ml), n - quantity of spores of each species in the sample (spores), Vs - volume of the sample, 15 (ml).
Used literature (references)
1. Yuan Yuan Wang, Mauritz Vestberg, Christopher Walker, Timo Hurme, Xiaoping Zhang, Kristina Lindström. Diversity and infectivity of arbuscular mycorrhizal fungi in agricultural soils of the Sichuan Province of mainland China. Mycorrhiza (2008) 18:59–68, DOI: 10.1007/s00572-008-0161-x
2. Omar MB, Bolland L, Heather WA (1979) A permanent mounting medium for fungi. Bull Brit Mycol Soc 13:31–32
3. Walker C, Gianinazzi-Pearson V,Marion-Espinasse H (1993) Scutellospora castanea, a newly described arbuscular mycorrhizal species.Cryptogam Mycol 14:279–286