Abstract
A comparative study on the use of traditional DNA barcoding and next-generation sequencing for determining the trophic interactions of herbivorous insects
Genome, Vol.60(11), pp.904-904
International Barcode of Life Conference, 7th (Skukuza, South Africa, 20-Nov-2017 - 24-Nov-2017)
2017
Abstract
Background: Measuring insect-herbivore trophic interactions is challenging, but the emerging field of ecological genomics is helping to overcome previous limitations with its increasing technological advances. Here, the efficiency of traditional single-species barcoding using Sanger sequencing is compared with novel next-generation sequencing (NGS) using an Illumina MiSeq. These two sequencing methods are trialled on the same selection of orthopterans to analyse their gut contents when host-plant species and diet breadth are predominantly unknown. Four standard plant DNA barcode markers were tested (rbcL, matK, psbA-trnH, and ITS), and a hybrid method was developed for NGS to allow for multiple marker PCR products to be sequenced simultaneously. Results: By sequencing more than one marker in the Illumina MiSeq cell, we increased the DNA fragment diversity which helped to reduce issues caused by low diversity that can cause lower quality reads and yield output. Overall, we found that the NGS method gave much more exhaustive results than the Sanger method in terms of number of sequences obtained. However, the Sanger sequences were longer in length and therefore could be matched more confidently to a plant species or genus level. One major drawback with Sanger sequencing for the use of diet analysis is that it often fails to deliver a readable sequence from a sample when there is DNA from multiple species within the gut-content, a limitation that is overcome by the NGS method. Significance: We suggest that only for known specialist species, the Sanger method is a more appropriate approach for measuring trophic interactions. Although both methods have valuable uses across many fields, NGS has a more extensive potential for developing a wider understanding of trophic interactions in ecological systems.
Details
- Title
- A comparative study on the use of traditional DNA barcoding and next-generation sequencing for determining the trophic interactions of herbivorous insects
- Authors
- Jane L Hardwick (Author) - Griffith UniversityAlison Shapcott (Author) - University of the Sunshine Coast, Queensland, GeneCology Research Centre - LegacySarah C Maunsell (Author) - Harvard UniversityNigel E Stork (Author) - Griffith UniversityRoger Kitching (Author) - Griffith University
- Publication details
- Genome, Vol.60(11), pp.904-904
- Conference details
- International Barcode of Life Conference, 7th (Skukuza, South Africa, 20-Nov-2017 - 24-Nov-2017)
- Audience
- Academic
- Publisher
- Canadian Science Publishing
- DOI
- 10.1139/gen-2017-0178
- ISSN
- 1480-3321
- Organisation Unit
- Centre for Bioinnovation; University of the Sunshine Coast, Queensland; School of Science, Technology and Engineering
- Language
- English
- Record Identifier
- 99669698602621
- Output Type
- Abstract
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