The use of DNA metabarcoding to analyse wild boar diets: Reproducibility of plant diet analyses and effective blocking of boar DNA amplification when evaluating animal dietary components

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  • メタバーコーディングに基づくイノシシの食性解析： 植物性食物の推定における再現性と動物性食物の推定における効果的な 解析対象種DNA の抑制方法の検討
  • メタバーコーディング ニ モトズク イノシシ ノ ショクセイ カイセキ : ショクブツセイ ショクモツ ノ スイテイ ニ オケル サイゲンセイ ト ドウブツセイ ショクモツ ノ スイテイ ニ オケル コウカテキ ナ カイセキ タイショウシュ DNA ノ ヨクセイ ホウホウ ノ ケントウ

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Abstract

We used a DNA metabarcoding method combined with high-throughput sequencing to analyse the diets of omnivorous wild boars. We extracted DNA from stomach contents. Three barcoded DNA regions (ITS-2, rbcL and P6 loop) were amplified and sequenced to identify plants consumed in different regions; the dietary plant genera differed significantly among the regions. To evaluate reproducibility, we analysed plant species compositions using three DNA samples from the same individuals; the samples did not differ significantly at the generic level, indicating that the stomach samples were well mixed. We next explored what the animals consumed. We used the polymerase chain reaction priming oligonucleotide and peptide nucleic acid methods to block the amplification of host DNA, mitochondrial cytochrome oxidase subunit 1 DNA. Both methods significantly reduced boar DNA amplification, but not DNA amplification from other animals.

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