Improved procedures for clearing roots and staining parasitic and vesicular-arbuscular mycorrhizal fungi for rapid assessment of infection

Chromium immobilization by extra- and intraradical fungal structures of arbuscular mycorrhizal symbioses

“…For evaluation of AM colonization, sub-samples of fresh roots were cut into 1 cm fragments, cleared in 10% KOH at 90 • C for 1 h, rinsed in 5% HCl for 5 min, and stained with 0.05% Trypan blue at 90 • C for 10 min following a modification of the methods of Phillips and Hayman [29], in which phenol was omitted from solutions. The intensity of mycorrhizal colonization in the whole root system (M%) was assessed by the method of Trouvelot et al [30] by using MYCOCALC software.…”

The arbuscular mycorrhizal fungus Glomus mosseae can enhance arsenic tolerance in Medicago truncatula by increasing plant phosphorus status and restricting arsenate uptake

“…Nodules were occasionally found on both mycorrhizal and nonmycorrhizal plant roots but their frequency of occurrence was very low (<5%). A randomly-selected portion of fresh roots from each pot was stained with Trypan blue (Phillips and Hayman, 1970) and both root length and the percentage of root length colonized were determined using the grid-line intersect method of Giovannetti and Mosse (1980). Shoots and roots were weighed after oven drying at 70 C for 72 h and ground sub-samples (0.1e0.2 g) were digested with 5 ml 4:1 (v/v) HNO 3 /HClO 4 for P and As analysis.…”

Growth and nutrient uptake of arbuscular mycorrhizal maize in different depths of soil overlying coal fly ash

“…A 1-g subsample of fresh roots was randomly selected from each pot and the roots were cleared overnight in 10% v/v KOH, acidified for 1 h in 1% v/v HCl, stained overnight with 0.05% w/v Trypan blue and de-stained with acidic glycerol. The proportion of root length infected was determined by examining the stained roots under a compound microscope using the gridline intersect method (Phillips and Hayman, 1970). The dry weights of bulk roots and shoots were determined after oven drying at 70°C for 48 h. Subsamples of oven-dried roots and shoots were milled to pass through a 0.5-mm sieve and dry ashed at 500°C for multi-element analysis by inductively coupled plasma-atomic emission spectroscopy (ICP-AES).…”