Procedures and equipment for staining large numbers of plant root samples for endomycorrhizal assay

Abstract

A simplified method of clearing and staining large numbers of plant roots for vesicular-arbuscular (VA) mycorrhizal assay is presented. Equipment needed for handling multiple samples is described, and two formulations for the different chemical solutions are presented. Because one formulation contains phenol, its use should be limited to basic studies for which adequate laboratory exhaust hoods are available and great clarity of fungal structures is required. The second staining formulation, utilizing lactic acid instead of phenol, is less toxic, requires less elaborate laboratory facilities, and has proven to be completely satisfactory for VA assays.

Germination and growth of purple passion fruit seedlings under pre-germination treatments and mycorrhizal inoculation1

“…Leaf phosphate content was determined by non-destructive samples, using the blue molybdate method (Murphy & Riley 1962, Aziz & Habte 1987. Mycorrhizal colonization was determined by fuchsine acid staining (0.15 %), for 48 hours (Kormanik et al 1980). Roots were discolored with KOH (10 %), for 24 hours (Phillips & Hayman 1970).…”

Uptake of cadmium from an experimentally contaminated calcareous soil by arbuscular mycorrhizal maize ( Zea mays L.)

“…Root samples for assessing root colonization were prepared using the acid fuchsin staining-grid intersect method (Kormanik et al 1979;Giovannetti and Mosse 1980).…”

Influence of edapho-climatic factors on the sporulation and colonization of arbuscular mycorrhizal fungi in two Amazonian native fruit species

“…The segment roots were then cleared for 40-60 min (according to the species) in 10% KOH solution at 90 o C, bleached in alkaline H 2 O 2 (3 mL of NH 4 OH, 30 mL of H 2 O 2 at 10% and 567 mL of water), placed in 4% HCl solution for 3 min and then stained with glycerol-trypan blue solution (0.05%) at 90 o C for 60 min (Kormanick et al, 1980). The stained root samples were examined at x 40-100 magnification and AMF root colonization were estimated by the gridline intersection method (Giovanetti and Mosse, 1980).…”