Abstract Detail

Symbioses: Plant, Animal, and Microbe Interactions

Edwards, Christine [1], Lindsay, Denise [1], Minckley, Thomas [2], Lance, Richard [1].

Use of DNA metabarcoding to quantify the diversity of plant species utilized by a pollinator.

DNA metabarcoding combines the use of next-generation DNA sequencing, DNA barcodes, and bioinformatic analysis to identify and quantify diversity in a sample. Although this technique has been explored extensively in the field of microbiology, few studies have used this approach to quantify diversity in the plant species visited by a pollinator. In this study, we used DNA metabarcoding to identify the plant resources utilized by nectivorous bats, and compared the results to estimates based on visual identification of pollen. Scat and fur-swab samples were collected from two species of nectar-feeding bats, the Mexican long-tongued bat (Choeronycteris mexicana) and endangered lesser long-nosed bat (Leptonycteris curasoae), from Arizona, USA. Samples were divided, and plant diversity was quantified in one half of each sample by visual identification of pollen, with the other half analyzed using DNA metabarcoding. DNA metabarcoding was carried out by PCR enrichment for three plant DNA barcodes (rbcL, psbA-trnH, and trnL-F), followed by next-generation DNA sequencing of tagged, pooled samples. To identify the diversity of plant species in the sample, reads were assembled and subjected to BLAST searches of the NCBI nucleotide database and a database of DNA sequences generated from potential food plants collected from Arizona. Visual identification of pollen found that 99.6% was from Agave sp., which is thought to be the main food source utilized by the bats. DNA metabarcoding identified a much broader diversity of plant species, indicating that it is a very sensitive approach to identify the plant species visited by bats, potentially allowing for the identification of less frequently utilized food-plant species. However, many of the plant species identified by DNA metabarcoding may not be utilized as food resources by bats and instead may result from incidental transfer or pollen by-catch arising from the broader network of species interactions within the plant-pollinator community. These results suggest that metabarcoding may be a powerful tool to understand the ecology and interactions among species, but that careful analysis of results is necessary to derive accurate and robust ecological conclusions.

Broader Impacts:

1 - US Army Engineer Research and Development Center, Environmental Laboratory, 3909 Halls Ferry Rd, Vicksburg, MS, 39180, USA
2 - University of Wyoming, Department of Geography and Program in Ecology, 1000 E. University Avenue, Laramie, WY, 82071, USA

Keywords:
DNA metabarcoding
plant-pollinator interactions
nectar-feeding bats
Next-generation sequencing
environmental DNA
pollen identification.

Presentation Type: Oral Paper:Papers for Topics
Session: 8
Location: Prince of Wales/Riverside Hilton
Date: Monday, July 29th, 2013
Time: 10:00 AM
Number: 8008
Abstract ID:734
Candidate for Awards:None