Overview of some techniques on detection and charaterization of arbuscular mycorrhizal fungi (A case study in Italy)

SAGUN, Ç.; ONTIVERO, R. E.; LOMBARDO, E.

Van

Conferencia; ISPEC 3rd INTERNATIONAL CONFERENCE ON AGRICULTURE AND RURAL DEVELOPMENT; 2019

Van University

Arbuscular mycorrhizal fungi are mutualistic symbionts widespread among both cultivated and wild plants. They have different beneficial roles including stimulating plant growth and tolerance to wide varieties of biotic as well as abiotic stresses. So, the exact and optimize methods for quantification and detection of these fungi in roots and soils are necessary. These techniques are based on fungal spores morphological characterization as well as molecular characterization of fungi in soil or root samples. The work was carried out at the Institute for Sustainable 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 on morphological assays proposed by Trouvelot et al in different roots samples of Mediterranean endemic shrubs from Sicily. Also, the root genomic DNA was extracted using DNeasy® Plant Mini kit (Qiagen) followed by quantification and amplification of fungal 18SrDNA with special primers for taxonomic and functional diversity of AM fungi. In another study, for characterization of AM fungal communities of soil samples from natural areas of Argentina, Sicily and Colombia, a soil core genomic DNA extracted using DNeasy PowerSoil® kit followed by amplification of 18S ( NS1- NS4, AML1-AML2, NS31- AM1) region using specialized primers designed to detect small differences in this region between closely related species.In addition, depending on the characteristics of the sample, the nested PCR technique was 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 success of the process, the DNA concentration was measured by spectrophotometry in nanodrop.