The detection of Glomus spp. (arbuscular mycorrhizal fungi) forming mycorrhizas in three plants, at different stages of seedling development, using mycorrhiza‐specific isozymes

Abstract

A series of glasshouse experiments was used to determine mycorrhiza-specific isozymes (MSIs) produced by five species of Glomus colonizing roots of a desert shrub legume (Anthyllis cytisoides L.), Thymus vulgaris L. and Allium porrum L. over time. Extracts of colonized roots were electrophoresed on non-denaturing polyacrylamide gels (PAGE) and stained for 10 different enzymes. Staining protocols for esterase, glutamate oxaloacetate transaminase, alkaline phosphatase and malate dehydrogenase provided MSIs for the mycorrhizas formed by different arbuscular mycorrhizal (AM) fungi that had colonized roots of the three host plants. There was no apparent correlation between levels of colonization or arbuscular intensities, at or between each sampling, and the presence of MSIs. The development of colonization by the AM fungi differed little between the three plants when assessed with two methods of estimating fungal biomass. The variety of MSIs detected might reflect the diversity of metabolic activities of these Glomus species and, possibly, differing ecological functions. The high-level induction of two alkaline phosphatase MSIs in the mycorrhizas of Anthyllis cytisoides colonized by Glomus microaggregatum BEG56 was used to track the fate of this fungus when the same plant was inoculated and transplanted into a semi-arid site in south-east Spain. The probable fungal origin of the isozyme was indicated by detection of the same isozyme in the extraradical mycelium formed by Glomus microaggregatum BEG56 on Allium porrum. The use of MSIs to detect the mycorrhizas of species of Glomus in colonized roots is discussed.

Real‐time PCR to quantify composition of arbuscular mycorrhizal fungal communities—marker design, verification, calibration and field validation

“…Therefore, alternative approaches to microscopy have attracted more attention and investigation. A few of the following methods have been developed: AMF speciesspecific isoenzymes (Hepper et al 1988;Tisserant et al 1998), genera-or species-specific antibodies (Wilson et al 1983;Friese & Allen 1991;Treseder & Allen 2002) and various DNA-based molecular methods (Clapp et al 1995;van Tuinen et al 1998;Jansa et al 2003Jansa et al , 2008Lee et al 2008). Different molecular markers and approaches exist, and they have been proven to be unbeatable with respect to their capacity to discriminate between different sequence types used as biomarkers for AMF taxa such as genera, species or isolates (van Tuinen et al 1998;Jansa et al 2003;Bö rstler et al 2008;Sý korová et al 2011).…”

“…A few of the following methods have been developed: AMF species‐specific isoenzymes (Hepper et al. 1988; Tisserant et al. 1998), genera‐ or species‐specific antibodies (Wilson et al.…”

Evidence that differences in phosphate metabolism in mycorrhizas formed by species of Glomus and Gigaspora might be related to their life‐cycle strategies

“…On the basis of previous experimentation (e.g. Tisserant et al, 1998) the amount of inoculum added corresponded to a ' saturation dose ', i.e. an amount that resulted in the fastest colonization possible for each AM fungus on the host used under the prevailing environmental conditions.…”

Thyme travels: 15N isoscapes of Thymus vulgaris L. invasion in lightly grazed pastoral communities

“…Thus the observed difference between soil δ 15 N and foliar δ 15 N in this study could be a consequence of reliance on mycorrhizae as a source of N under N‐limited conditions and others have isolated AMF from thyme plant roots (Tisserant et al . ; del Val et al . ).…”