The aim of this study was to evaluate the performance of two of the most commonly used commercial kits for soil DNA extraction regarding the values of the taxonomic diversity of prokaryotes and community composition of saffron (Crocus sativus) cultivated fields. The impact of the QIAGEN-DNeasy PowerSoil Kit (MO) and Macherey-Nagel™ NucleoSpin™ Soil (MN) kit was tested on the soil of an Italian western alpine experimental site located in Saint Christophe (Aosta Valley, AO). Nine biological replicas of bulk soil were collected and analyzed independently with the two kits. 16S rRNA metabarcoding was applied to characterize soil microbial communities. We first noticed that both DNA extraction kits yielded nearly the same number of OTUs: 1284 and 1268 for MN and MO, respectively. Both kits did not differ in the alpha diversity of the samples, while they had an influence on the beta diversity. The comparative analysis of the microbial community composition displayed differences in microbial community structure depending on which kit was used. These differences were especially highlighted at Phylum and Class levels. On the other hand, the fact that, from a functional point of view, our approach did not highlight any differences allows us to state that the results obtained with the two extraction kits are comparable and interchangeable. Based on these results and those in the literature, we could undoubtedly recommend both commercial kits, especially if the soil target microorganisms are prokaryotes and the study focuses on agricultural sites.

Truth or Lie: Does the DNA Extraction Procedure Really Affect the Insight in Composition and Diversity of Microbial Communities in Saffron Cultivated Soils?

Voyron S;Bianciotto V;Ghignone S
;
Lumini E
2022

Abstract

The aim of this study was to evaluate the performance of two of the most commonly used commercial kits for soil DNA extraction regarding the values of the taxonomic diversity of prokaryotes and community composition of saffron (Crocus sativus) cultivated fields. The impact of the QIAGEN-DNeasy PowerSoil Kit (MO) and Macherey-Nagel™ NucleoSpin™ Soil (MN) kit was tested on the soil of an Italian western alpine experimental site located in Saint Christophe (Aosta Valley, AO). Nine biological replicas of bulk soil were collected and analyzed independently with the two kits. 16S rRNA metabarcoding was applied to characterize soil microbial communities. We first noticed that both DNA extraction kits yielded nearly the same number of OTUs: 1284 and 1268 for MN and MO, respectively. Both kits did not differ in the alpha diversity of the samples, while they had an influence on the beta diversity. The comparative analysis of the microbial community composition displayed differences in microbial community structure depending on which kit was used. These differences were especially highlighted at Phylum and Class levels. On the other hand, the fact that, from a functional point of view, our approach did not highlight any differences allows us to state that the results obtained with the two extraction kits are comparable and interchangeable. Based on these results and those in the literature, we could undoubtedly recommend both commercial kits, especially if the soil target microorganisms are prokaryotes and the study focuses on agricultural sites.
2022
Istituto per la Protezione Sostenibile delle Piante - IPSP
DNA extraction kits
environmental DNA
saffron cultivated soil
16S rRNA metabarcoding
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/20.500.14243/412877
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