Using ITS2 metabarcoding and microscopy to analyse shifts in pollen diets of honey bees and bumble bees along a mass‐flowering crop gradient

Bänsch, Svenja; Tscharntke, Teja; Wünschiers, Röbbe; Netter, Leonie; Brenig, Bertram; Gabriel, Doreen; Westphal, Catrin

doi:10.1111/mec.15675

Cited by 29 publications

(23 citation statements)

References 82 publications

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“…Metabarcoding was able to detect many of the most abundant taxa identified with microscopy, often in similarly high proportions. Previous studies have found a positive correlation between the relative abundance of taxa identified in pollen samples by microscopy an metabarcoding, mostly for dominant taxa (Bänsch et al, 2020;Smart et al, 2017). The overlap in taxa identified by both methods was 21 families, representing 67.7% of the families detected by microscopy, and 12 genera, representing 44.4% of genera detected by microscopy.…”

Section: Pollen Metabarcoding Comparison With Microscopymentioning

confidence: 76%

Pollen DNA metabarcoding identifies regional provenance and high plant diversity in Australian honey

Milla

Sniderman

Lines

et al. 2021

Ecology and Evolution

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Accurate identification of the botanical components of honey can be used to establish its geographical provenance, while also providing insights into honeybee ( Apis mellifera L.) diet and foraging preferences. DNA metabarcoding has been demonstrated as a robust method to identify plant species from pollen and pollen‐based products, including honey. We investigated the use of pollen metabarcoding to identify the floral sources and local foraging preferences of honeybees using 15 honey samples from six bioregions from eastern and western Australia. We used two plant metabarcoding markers, ITS2 and the trnL P6 loop. Both markers combined identified a total of 55 plant families, 67 genera, and 43 species. The trnL P6 loop marker provided significantly higher detection of taxa, detecting an average of 15.6 taxa per sample, compared to 4.6 with ITS2. Most honeys were dominated by Eucalyptus and other Myrtaceae species, with a few honeys dominated by Macadamia (Proteaceae) and Fabaceae. Metabarcoding detected the nominal primary source provided by beekeepers among the top five most abundant taxa for 85% of samples. We found that eastern and western honeys could be clearly differentiated by their floral composition, and clustered into bioregions with the trnL marker. Comparison with previous results obtained from melissopalynology shows that metabarcoding can detect similar numbers of plant families and genera, but provides significantly higher resolution at species level. Our results show that pollen DNA metabarcoding is a powerful and robust method for detecting honey provenance and examining the diet of honeybees. This is particularly relevant for hives foraging on the unique and diverse flora of the Australian continent, with the potential to be used as a novel monitoring tool for honeybee floral resources.

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Section: Pollen Metabarcoding Comparison With Microscopymentioning

confidence: 76%

Pollen DNA metabarcoding identifies regional provenance and high plant diversity in Australian honey

Milla

Sniderman

Lines

et al. 2021

Ecology and Evolution

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“…Still, we claim that in the most important bumble bee forage plants, the proportional quantification of the reads produces a fairly good result. Bänsch et al [ 61 ] also used the DNA metabarcoding with reads from ITS2 region sequences to quantify the bumble bee-collected pollen but reminded that the interpretation must take the specific limitations of the outcome into account.…”

Section: Discussionmentioning

confidence: 99%

Bumble Bee Foraged Pollen Analyses in Spring Time in Southern Estonia Shows Abundant Food Sources

Bontšutšnaja

Karise

Mänd

et al. 2021

Insects

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Agricultural landscapes usually provide higher quantities of single-source food, which are noticeably lacking in diversity and might thus have low nutrient value for bumble bee colony development. Here, in this study, we analysed the pollen foraging preferences over a large territory of a heterogeneous agricultural landscape: southern Estonia. We aimed to assess the botanical diversity of bumble bee food plants in the spring time there. We looked for preferences for some food plants or signs of food shortage that could be associated with any particular landscape features. For this purpose, we took Bombus terrestris commercial hives to the landscape, performed microscopy analyses and improved the results with the innovative DNA metabarcoding technique to determine the botanical origin of bumble bee-collected pollen. We found high variability of forage plants with no strong relationship with any particular landscape features. Based on the low number of plant species in single flights, we deduce that the availability of main forage plants is sufficient indicating rich forage availabilities. Despite specific limitations, we saw strong correlations between microscopy and DNA metabarcoding data usable for quantification analyses. As a conclusion, we saw that the spring-time vegetation in southern Estonia can support bumble bee colony development regardless of the detailed landscape structure. The absence of clearly dominating food preference by the tested generalist bumble bee species B. terrestris makes us suggest that other bumble bee species, at least food generalists, should also find plenty of forage in their early development phase.

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“…Our primers further demonstrated unexpected detection of novel Encarsia and Eretmocerus parasitoid species in the African cassava and noncassava fields, however the true genetic and species diversity of these minute parasitoids will require independent assessment for confirmation, such as to include the standard barcode COI gene region using standard primers (e.g., C_LepFoIF/C_LepFoIR; Woodcock et al, 2013) as internal control. Nevertheless, understanding of trophic interactions between crop hosts, pest/beneficial insects and their potential biological control agents represents a molecular ecological network research area that could yield significant insights via metabarcoding approach (e.g., Bansch et al, 2020;Evans, Kitson, Lunt, Straw, & Pocock, 2016;Sow et al, 2019).…”

Section: Discussionmentioning

confidence: 99%

A high-throughput amplicon sequencing approach for population-wide species diversity and composition survey

Tay

Court²,

Jacomb³

et al. 2020

Preprint

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Management of agricultural pests requires an understanding of pest species diversity, their interactions with beneficial insects and spatial-temporal patterns of pest abundance. Invasive and agriculturally important insect pests can build up very high populations, especially in cropping landscapes. Traditionally, sampling effort for species identification involves small sample sizes and is labour intensive. Here, we describe a high throughput sequencing (HTS) PCR amplicon method and associated analytical workflow for a rapid, intensive, high-volume survey of pest species compositions. We demonstrate our method using the Bemisia pest cryptic species complex as examples. The whiteflies Bemisia including the ′tabaci′ species are agriculturally important capable of vectoring diverse plant viruses that cause diseases and crop losses. We use our HTS amplicon platform to simultaneously process high volumes of whitefly individuals, with efficiency to detect rare (i.e., 1%) test-species and beneficial hymenopteran parasitoid species. Field-testing our HTS amplicon method across the Tanzania, Uganda and Malawi cassava cultivation landscapes, we identified the sub-Saharan Africa 1 Bemisia putative species as the dominant pest species, with other cryptic Bemisia species being detected at various abundances. We also provide evidence that Bemisia species compositions can be affected by sampling techniques that target either nymphs or adults. Our method is particularly suitable to molecular diagnostic surveys of cryptic insect species with high population densities. Our approach can be adopted to understand species biodiversity across landscapes, with broad implications for improving trans-boundary biosecurity preparedness, thus contributing to molecular ecological knowledge and the development of control strategies for high-density, cryptic, pest-species complexes.

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Using ITS2 metabarcoding and microscopy to analyse shifts in pollen diets of honey bees and bumble bees along a mass‐flowering crop gradient

Abstract: This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

Cited by 29 publications

References 82 publications

Pollen DNA metabarcoding identifies regional provenance and high plant diversity in Australian honey

Pollen DNA metabarcoding identifies regional provenance and high plant diversity in Australian honey

Bumble Bee Foraged Pollen Analyses in Spring Time in Southern Estonia Shows Abundant Food Sources

A high-throughput amplicon sequencing approach for population-wide species diversity and composition survey

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