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The detection of arbuscular mycorrhizal fungi (AMf) in root
systems of different plants using fungal/mycorrhizal-specific
isozymes as molecular markers.
DODD1,J.C., B. TISSERANT2,
N. REQUENA3, V. BRENAC1,
J.M. BAREA3 & P. JEFFRIES2
1International Institute for Biotechnology
2Dept. of Biosciences, University of Kent,
Canterbury, Kent CT2 7YW, UK
3Estación Experimental del Zaidìn,
C.S.I.C., Profesor Albareda 1, E-18008 Granada, Spain
ICOM1 Abstract
Isozyme analysis is a simple and relatively inexpensive
technique which can discriminate closely-related organisms.
The use of FSI/MSIs as molecular markers has practical use in
the detection and identification of AMf in plant roots. Our
aim has been, therefore, to use non-denaturing polyacrylamide
gel electrophoresis to separate isozymes and to track well-
characterised AMf in young root systems. In a project funded
by the EU called REDEEM (Restoration of Environmental
Diversity by Effective Ecosymbiont Monitoring) we have used
this technique to identify individual AMf in the roots of
Anthyllis cytisoides (a desert shrub legume) and other test
plants. The results from a screening of various Glomus spp.
in laboratory microcosms using inert growth media
(Terragreen, durite sand), in soil from the study site in
Almeria (S. Spain), and ultimately in the field, have shown
that a cocktail of enzyme stains can be used to identify
individual AMf colonising roots at different stages of the
symbiosis. The use of an alkaline phosphatase (ALP) stain
showed that the non-indigenous symbiont, Glomus
microaggregatum (BEG 56), used as the fungal inoculant of A.
cytisoides seedlings could be detected 3 months after
outplanting into the field. The presence of the ALP marker
bands also implied that the fungus was probably still active
in the roots, with respect to P transfer, at a time when the
seasonal rains had finished. Staining of the roots for ALP
activity showed that most occurred in the arbuscules. The
results will be discussed with respect to the use of the
technique to study the functional ecology of AMf in natural
ecosystems.