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Applying pollen DNA metabarcoding to the study of plant-pollinator interactions - PubMed

  • ️Sun Jan 01 2017

. 2017 Jun 12;5(6):apps.1600124.

doi: 10.3732/apps.1600124. eCollection 2017 Jun.

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Applying pollen DNA metabarcoding to the study of plant-pollinator interactions

Karen L Bell et al. Appl Plant Sci. 2017.

Abstract

Premise of the study: To study pollination networks in a changing environment, we need accurate, high-throughput methods. Previous studies have shown that more highly resolved networks can be constructed by studying pollen loads taken from bees, relative to field observations. DNA metabarcoding potentially allows for faster and finer-scale taxonomic resolution of pollen compared to traditional approaches (e.g., light microscopy), but has not been applied to pollination networks.

Methods: We sampled pollen from 38 bee species collected in Florida from sites differing in forest management. We isolated DNA from pollen mixtures and sequenced rbcL and ITS2 gene regions from all mixtures in a single run on the Illumina MiSeq platform. We identified species from sequence data using comprehensive rbcL and ITS2 databases.

Results: We successfully built a proof-of-concept quantitative pollination network using pollen metabarcoding.

Discussion: Our work underscores that pollen metabarcoding is not quantitative but that quantitative networks can be constructed based on the number of interacting individuals. Due to the frequency of contamination and false positive reads, isolation and PCR negative controls should be used in every reaction. DNA metabarcoding has advantages in efficiency and resolution over microscopic identification of pollen, and we expect that it will have broad utility for future studies of plant-pollinator interactions.

Keywords: DNA metabarcoding; ITS; palynology; plant–pollinator interactions; pollination networks; rbcL.

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Figures

Fig. 1.
Fig. 1.

Proportions of classified taxa (i.e., those identified at least to phylum) identifiable to each taxonomic rank from DNA metabarcoding with rbcL and ITS2.

Fig. 2.
Fig. 2.

Proportion of sequencing reads for family-level taxonomic identifications from samples FL179 (high DNA content), FL765 (moderate DNA content), and FL796 (low DNA content), with varying numbers of PCR cycles. Sequencing reads from rbcL are horizontally striped, sequencing reads from ITS2 are diagonally striped, and different families are represented in different colors.

Fig. 3.
Fig. 3.

Bipartite pollination network of 37 bee specimens (top nodes) to the 51 plant taxa present in their pollen loads based on ITS2 taxonomic classification (bottom nodes). Interactions were pooled within plant and bee taxa. Links between plants and pollinators are represented with lines whose width is proportional to the number of interactions while the width of the nodes represents total abundance of that taxon across all of its interactions.

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