A quick and precise technique for identifying ectomycorrhizas by PCR.
A rapid procedure was developed to amplify ITS fragments directly from Tuber ectomycorrhizas either synthesized in a greenhouse or collected from the field. The addition of Bovine Serum Albumin (BSA) to the reaction mixtures overcame the presence of reaction inhibitors present in fungal and root cells, and enabled the amplification of the ITS regions directly from ectomycorrhizal tissues. This method is cheaper and less time-consuming than conventional procedures, and reduces the time required from 1-4 h to a few minutes. It is also much more sensitive, allowing the identification of just a small fragment of a mycorrhizal root tip. Because of this it is possible to select only the target fungal tissue and hence minimise the risk of contamination by saprobic or other mycorrhizal species. The method also avoids the use of toxic or hazardous substances. This method could have a wider application in other areas of applied mycology.
Iotti
Mirco
M
Dipartimento di Protezione e Valorizzazione Agroalimentare, Università degli Studi di Bologna, via Fanin 46, I-40127 Bologna, Italy.
Zambonelli
Alessandra
A
D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000032:analysis*; D004275:DNA, Ribosomal / Q000032:analysis*; D007887:Fabaceae / Q000033:anatomy & histology / Q000382:microbiology* / Q000502:physiology; D038821:Mycorrhizae / Q000235:genetics / Q000302:isolation & purification*; D018517:Plant Roots / Q000235:genetics / Q000382:microbiology; D016133:Polymerase Chain Reaction
D017931:DNA Primers; D004271:DNA, Fungal; D004275:DNA, Ribosomal