A comparison of methodologies for the staining and quantification of intracellular components of Arbuscular Mychorrizal (AM) fungi in the root cortex of two varieties of winter wheat

Arbuscular Mychorrizal (AM) fungi are one of the most common fungal organisms to exist in symbiosis with terrestrial plants facilitating the growth and maintenance of arable crops. Wheat has been studied extensively for AM fungal symbiosis using the carcinogen trypan blue as the identifying stain for fungal components, namely arbuscles, vesicles and hyphal structures. The present study uses Sheaffer® blue ink with a lower risk as an alternative to this carcinogenic stain. Justification for this is determined by stained wheat root sections (n = 120), with statistically significant increases in the observed abundance of intracellular root cortical fungal structures stained with Sheaffer® blue ink compared to trypan blue for both Zulu (P = 0.003) and Siskin (P = 0.0003) varieties of winter wheat. This new alternative combines an improved quantification of intracellular fungal components with a lower hazard risk at a lower cost.