Abstract
AbstractMany techniques have been developed to investigate the interactions between plants and herbivorous insects in natural environments and are generally used to determine either (a) which plant species are eaten by a specific herbivorous insect or (b) which herbivorous insect species are herbivores of a specific plant. The former problem is usually addressed by the direct observation of feeding and microscopic observation of gut contents and excrements, as well as the application of DNA‐barcoding techniques. However, the latter problem has typically been addressed using time‐consuming methods, such as direct observation and rearing. Therefore, more efficient techniques are needed for identifying and quantifying the interactions of plants with herbivorous insects. The present study demonstrates that the environmental DNA (eDNA) of herbivorous insects can be recovered from leaves with external foliage feeding marks. Mitochondrial DNA fragments of herbivorous insects were detected from insect‐exposed leaves using primer sets that amplified the DNA of target species. The amplification rate of the herbivorous insect DNA was positively associated with the rim length of feeding marks, which suggests that most of the insect DNA came from the feeding marks. Additionally, we showed that this method has the potential to detect eDNA from field‐collected leaves. This time‐efficient approach will contribute to the detection of plant–insect herbivore interactions.
Highlights
The present study demonstrates that the environmental DNA
(eDNA) of herbivorous insects can be recovered from leaves with external foliage feeding marks
Clean rubber gloves were used during ers and probe sets were tested by performing PCR or qPCR using leaf collection, and new rubber gloves were used when collecting the target and one or two related species that potentially occur at leaves from each plant to minimize cross-contamination
Summary
17H03735 and herbivorous insects in natural environments and are generally used to determine either (a) which plant species are eaten by a specific herbivorous insect or (b) which herbivorous insect species are herbivores of a specific plant. The present study demonstrates that the environmental DNA (eDNA) of herbivorous insects can be recovered from leaves with external foliage feeding marks. Allows efficient investigation after insect feeding behavior is necessary to understand interactions between plants and insect herbivores in a natural environment Such a method could help elucidate ecosystem structure, manage damage to cultivated crops, or conserve endangered species. Clean rubber gloves were used during ers and probe sets were tested by performing PCR or qPCR using leaf collection, and new rubber gloves were used when collecting the target and one or two related species that potentially occur at leaves from each plant to minimize cross-contamination. For the testing of B. mori primers, one DNA sample control leaf, without feeding marks, was collected to assess conextracted from the body of B. mori was used as the target species, tamination during sampling (i.e., sampling blank). The thermal cycle profile was as follows: 50°C for 2 min; 95°C for 10 min; 50 cycles of 95°C for
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