Dataset Information

Pollen Monitoring by Optical Microscopy and DNA Metabarcoding: Comparative Study and New Insights.

ABSTRACT: Environmental samples collected in Brindisi (Italy) by a Hirst-type trap and in Lecce (Italy) by a PM10 sampler were analysed by optical microscopy and DNA-metabarcoding, respectively, to identify airborne pollen and perform an exploratory study, highlighting the benefits and limits of both sampling/detection systems. The Hirst-type trap/optical-microscopy system allowed detecting pollen on average over the full bloom season, since whole pollen grains, whose diameter vary within 10-100 μm, are required for morphological detection with optical microscopy. Conversely, pollen fragments with an aerodynamic diameter ≤10 μm were collected in Lecce by the PM10 sampler. Pollen grains and fragments are spread worldwide by wind/atmospheric turbulences and can age in the atmosphere, but aerial dispersal, aging, and long-range transport of pollen fragments are favoured over those of whole pollen grains because of their smaller size. Twenty-four Streptophyta families were detected in Lecce throughout the sampling year, but only nine out of them were in common with the 21 pollen families identified in Brindisi. Meteorological parameters and advection patterns were rather similar at both study sites, being only 37 km apart in a beeline, but their impact on the sample taxonomic structure was different, likely for the different pollen sampling/detection systems used in the two monitoring areas.

SUBMITTER: Fragola M

PROVIDER: S-EPMC8910172 | biostudies-literature | 2022 Feb

REPOSITORIES: biostudies-literature

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Pollen Monitoring by Optical Microscopy and DNA Metabarcoding: Comparative Study and New Insights.

Fragola Mattia M Arsieni Augusto A Carelli Nicola N Dattoli Sabrina S Maiellaro Sante S Perrone Maria Rita MR Romano Salvatore S

International journal of environmental research and public health 20220224 5

Environmental samples collected in Brindisi (Italy) by a Hirst-type trap and in Lecce (Italy) by a PM10 sampler were analysed by optical microscopy and DNA-metabarcoding, respectively, to identify airborne pollen and perform an exploratory study, highlighting the benefits and limits of both sampling/detection systems. The Hirst-type trap/optical-microscopy system allowed detecting pollen on average over the full bloom season, since whole pollen grains, whose diameter vary within 10-100 μm, are r ...[more]

PMID: 35270312

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Project description:Grass pollen is one of the leading causes of pollinosis, affecting 10-30% of the world's population. The allergenicity of pollen from different Poaceae species is not the same and is estimated from moderate to high. Aerobiological monitoring is a standard method that allows one to track and predict the dynamics of allergen concentration in the air. Poaceae is a stenopalynous family, and thus grass pollen can usually be identified only at the family level with optical microscopy. Molecular methods, in particular the DNA barcoding technique, can be used to conduct a more accurate analysis of aerobiological samples containing the DNA of various plant species. This study aimed to test the possibility of using the ITS1 and ITS2 nuclear loci for determining the presence of grass pollen from air samples via metabarcoding and to compare the analysis results with the results of phenological observations. Based on the high-throughput sequencing data, we analyzed the changes in the composition of aerobiological samples taken in the Moscow and Ryazan regions for three years during the period of active flowering of grasses. Ten genera of the Poaceae family were detected in airborne pollen samples. The representation for most of them for ITS1 and ITS2 barcodes was similar. At the same time, in some samples, the presence of specific genera was characterized by only one sequence: either ITS1 or ITS2. Based on the analysis of the abundance of both barcode reads in the samples, the following order could describe the change with time in the dominant species in the air: Poa, Alopecurus, and Arrhenatherum in early mid-June, Lolium, Bromus, Dactylis, and Briza in mid-late June, Phleum, Elymus in late June to early July, and Calamagrostis in early mid-July. In most samples, the number of taxa found via metabarcoding analysis was higher compared to that in the phenological observations. The semi-quantitative analysis of high-throughput sequencing data well reflects the abundance of only major grass species at the flowering stage.

Project description:BackgroundIntense conversion of tropical forests into agricultural systems contributes to habitat loss and the decline of ecosystem functions. Plant-pollinator interactions buffer the process of forest fragmentation, ensuring gene flow across isolated patches of forests by pollen transfer. In this study, we identified the composition of pollen grains stored in pot-pollen of stingless bees, Tetragonula laeviceps, via dual-locus DNA metabarcoding (ITS2 and rbcL) and light microscopy, and compared the taxonomic coverage of pollen sampled in distinct land-use systems categorized in four levels of management intensity (forest, shrub, rubber, and oil palm) for landscape characterization.ResultsPlant composition differed significantly between DNA metabarcoding and light microscopy. The overlap in the plant families identified via light microscopy and DNA metabarcoding techniques was low and ranged from 22.6 to 27.8%. Taxonomic assignments showed a dominance of pollen from bee-pollinated plants, including oil-bearing crops such as the introduced species Elaeis guineensis (Arecaceae) as one of the predominant taxa in the pollen samples across all four land-use types. Native plant families Moraceae, Euphorbiaceae, and Cannabaceae appeared in high proportion in the analyzed pollen material. One-way ANOVA (p > 0.05), PERMANOVA (R² values range from 0.14003 to 0.17684, for all tests p-value > 0.5), and NMDS (stress values ranging from 0.1515 to 0.1859) indicated a lack of differentiation between the species composition and diversity of pollen type in the four distinct land-use types, supporting the influx of pollen from adjacent areas.ConclusionsStingless bees collected pollen from a variety of agricultural crops, weeds, and wild plants. Plant composition detected at the family level from the pollen samples likely reflects the plant composition at the landscape level rather than the plot level. In our study, the plant diversity in pollen from colonies installed in land-use systems with distinct levels of forest transformation was highly homogeneous, reflecting a large influx of pollen transported by stingless bees through distinct land-use types. Dual-locus approach applied in metabarcoding studies and visual pollen identification showed great differences in the detection of the plant community, therefore a combination of both methods is recommended for performing biodiversity assessments via pollen identification.

Dataset Information

Pollen Monitoring by Optical Microscopy and DNA Metabarcoding: Comparative Study and New Insights.

Publications

Pollen Monitoring by Optical Microscopy and DNA Metabarcoding: Comparative Study and New Insights.

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