OVERVIEW OF SOME TECHNIQUES ON DETECTION ANDCHARACTERIZATION OF ARBUSCULAR MYCORRHIZAL FUNGI (A CASESTUDY IN ITALY)

Arbuscular mycorrhizal fungi are mutualistic symbionts widespread among both cultivatedand wild plants. They have different beneficial roles including stimulating plant growth andtolerance to wide varieties of biotic as well as abiotic stresses. So,the exact and optimizemethods for quantification and detection of these fungi in roots and soils are necessary.Thesetechniques are based on fungal sporesmorphological characterization as well as molecularcharacterization of fungi in soil or root samples.The work was carried out at the Institute forSustainable Plant Protection (IPSP) of the National Research Council of Italy (CNR) in Turin.For this purpose, the AM fungal root colonization patterns was measured based onmorphological assays proposed by Trouvelot et al in different roots samples of Mediterraneanendemic shrubs from Sicily. Also, the root genomic DNA was extracted using DNeasy® PlantMini kit (Qiagen) followed by quantification and amplification of fungal 18SrDNA withspecial primers for taxonomic and functional diversity of AM fungi. In another study, forcharacterization of AM fungal communities of soil samples from natural areas of Argentina,Sicily and Colombia, a soil core genomic DNA extracted using DNeasyPowerSoil® kitfollowed by amplification of 18S ( NS1- NS4, AML1-AML2, NS31- AM1) region usingspecialized primers designed to detect small differences in this region between closely relatedspecies. In addition, depending on the characteristics of the sample, the nested PCR techniquewas used, comprising two rounds of amplification with different pairs of primers in each one,in order to increase the sensitivity and specificity of the detection. Finally, to test the successof the process, the DNA concentration was measured by spectrophotometry in nanodrop®.