Symbiotic Fungi: Principles and Practice (Soil Biology)

102 H. Dupré de Boulois et al.
transport processes, (2) the difficulty of visualizing the development of the two
partners of the symbiosis and of recovering roots and extraradical fungal mycelium,
(3) the possible interaction between the element and the soil matrix, (4) the
potential direct uptake by the roots of the element under study due to tracer leakage
towards the root compartment caused by diffusion or mass-flow, and (5) the
complexity of performing physiological and molecular studies on extraradical
fungal mycelium and mycorrhizal roots (Schweiger and Jakobsen 2000; Rufyikiri
et al. 2005). For these reasons, compartmented in vitro (monoxenic) culture systems
became widely used (Rufyikiri et al. 2005).
The aim of this chapter is to provide methodologies to perform transport studies
using bi-compartmented in vitro culture systems, with root-organs or whole plants
as host, and to give a detailed description of the advantages and disadvantages of
these systems.
7.2 Equipment and Laboratory Material
7.2.1 Equipment
l Horizontal or vertical laminar flow hood
l Stereo-microscope (magnification 10– 40)
l Bunsen burner or bead sterilizer
l Cooled heated incubator
l Climatic chamber
l Rotating agitator
l Thermo-fusible glue applicator
7.2.2 Laboratory Material
l Bi-compartmented Petri plates (usual diameter: 90 mm)
l Polypropylene centrifugation tubes (50 ml)
l Scalpels and forceps
l Set of cork-borers
l Needles
l Manual or motorized pipettors adapted to 5–20 ml sterile pipettes
l 1–1,000 ml micropipettes with corresponding sterile tips
l Cellophane wrap (roll of 2 cm-large) and Parafilm (note that Seal View, Petri
Seal TM or other products can also be used)
l Thermo-fusible glue
l Silicon grease
l Black plastic bags (12 12 cm)
l Inclined (0–45 ) support for Petri plates