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<p>When threatened by the giant hornet <i>Vespa mandarinia</i> Smith (Hymenoptera: Vespidae)<i>,</i> the Japanese honey bee <i>Apis cerana japonica</i> Radoszkowski (Hymenoptera: Apidae) collects substances from the surrounding environment and applies them to the entrance of its hive. This behavior is thought to help avoid mass attacks by hornets. To investigate the substances collected by the bees, we analyzed the DNA extracted from the applied spots using three methods. First, we placed glass slides at the hive entrance, extracted DNA from the spots mounted on the glass slides, and amplified the internal transcribed spacer (ITS) 1 using PCR. Sequencing of the cloned PCR products revealed the presence of both plant and fungal DNA. Second, we performed an amplicon analysis targeting ITS2 of plants and fungi using next-generation sequencing. A diverse array of fungi was detected; however, unexpectedly, no plant DNA was detected. The reason for this remains unclear; however, amplicon analysis appears to be biased toward the detection of fungi. Therefore, in the third experiment, we extracted DNA from individual spots on glass slides, amplified ITS1, and directly sequenced the PCR products. This experiment allowed us to quantify the utilization of plants and fungi based on the number of spots containing each type of material. We determined the sequences of 165 PCR products, including 56 obtained from plants, 68 from mushrooms, and 41 from non-mushroom fungi.</p>

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The Japanese honey bee Apis cerana japonica (Hymenoptera: Apidae) collects and applies plant and fungal materials to the hive entrance as a potential defense against the northern giant hornet Vespa mandarinia (Hymenoptera: Vespidae)

  • 期刊论文

  • 发表日期：

    2026年1月4日

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  • Yoshitaka Sumimiya,
  • Masato Ono,
  • Tetsuhiko Sasaki

摘要

When threatened by the giant hornet Vespa mandarinia Smith (Hymenoptera: Vespidae), the Japanese honey bee Apis cerana japonica Radoszkowski (Hymenoptera: Apidae) collects substances from the surrounding environment and applies them to the entrance of its hive. This behavior is thought to help avoid mass attacks by hornets. To investigate the substances collected by the bees, we analyzed the DNA extracted from the applied spots using three methods. First, we placed glass slides at the hive entrance, extracted DNA from the spots mounted on the glass slides, and amplified the internal transcribed spacer (ITS) 1 using PCR. Sequencing of the cloned PCR products revealed the presence of both plant and fungal DNA. Second, we performed an amplicon analysis targeting ITS2 of plants and fungi using next-generation sequencing. A diverse array of fungi was detected; however, unexpectedly, no plant DNA was detected. The reason for this remains unclear; however, amplicon analysis appears to be biased toward the detection of fungi. Therefore, in the third experiment, we extracted DNA from individual spots on glass slides, amplified ITS1, and directly sequenced the PCR products. This experiment allowed us to quantify the utilization of plants and fungi based on the number of spots containing each type of material. We determined the sequences of 165 PCR products, including 56 obtained from plants, 68 from mushrooms, and 41 from non-mushroom fungi.