Abstract
Plants and their associated insect herbivores, represent more than 50% of all known species on earth. The first step in understanding the mechanisms generating and maintaining this important component of biodiversity is to identify plant-herbivore associations. In this study we determined insect-host plant associations for an entire guild of insect herbivores using plant DNA extracted from insect gut contents. Over two years, in a tropical rain forest in Costa Rica (La Selva Biological Station), we recorded the full diet breadth of rolled-leaf beetles, a group of herbivores that feed on plants in the order Zingiberales. Field observations were used to determine the accuracy of diet identifications using a three-locus DNA barcode (rbcL, trnH-psbA and ITS2). Using extraction techniques for ancient DNA, we obtained high-quality sequences for two of these loci from gut contents (rbcL and ITS2). Sequences were then compared to a comprehensive DNA barcode library of the Zingiberales. The rbcL locus identified host plants to family (success/sequence = 58.8%) and genus (success/sequence = 47%). For all Zingiberales except Heliconiaceae, ITS2 successfully identified host plants to genus (success/sequence = 67.1%) and species (success/sequence = 61.6%). Kindt’s sampling estimates suggest that by collecting ca. four individuals representing each plant-herbivore interaction, 99% of all host associations included in this study can be identified to genus. For plants that amplified ITS2, 99% of the hosts can be identified to species after collecting at least four individuals representing each interaction. Our study demonstrates that host plant identifications at the species-level using DNA barcodes are feasible, cost-effective, and reliable, and that reconstructing plant-herbivore networks with these methods will become the standard for a detailed understanding of these interactions.
Highlights
Plants, together with their associated insect herbivores, represent more than 50% of all known species on earth [1]
Over the two years of our field study, we recorded by direct observations at La Selva 20 species of rolled-leaf beetles, 33 species of Zingiberales, and 103 plant-herbivore interactions (Figure 2A)
Accumulation curves show that our efforts to record the diets of 7359 individual rolled-leaf beetles on 3092 host plants resulted in a comprehensive account of species and their interactions for this plant-herbivore system (Figure 2B)
Summary
Together with their associated insect herbivores, represent more than 50% of all known species on earth [1]. This overwhelming biological diversity is in part the product of the coevolutionary processes that have taken place between these two ecological partners [1,2,3,4]. The most common strategies have included: 1) direct observations of herbivory in the field; 2) laboratory feeding trials [5]; 3) morphological analyses of gut contents; and 4) stable isotope techniques [6,7]. For morphological and stable isotope analyses the resolution of the taxonomic and/or ecological information is often very limited
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