Plant‐derived eDNA metabarcoding is an effective approach for monitoring invertebrate rice pests

Abstract Traditional methods based on morphological taxonomy and visual identification of crop damage are increasingly challenged by the diversity and complexity of current and emerging pest occurrences. Although environmental DNA (eDNA) metabarcoding shows potential for improving agricultural pest management, comprehensive evaluation of its effectiveness in pest identification and monitoring remains scarce, particularly studies integrating controlled laboratory experiments with field monitoring. This study evaluated the potential of plant‐derived eDNA metabarcoding to identify and monitor invertebrate pests of rice through in silico polymerase chain reaction (PCR) analysis, controlled feeding experiments and field investigations. The results indicated that eDNA metabarcoding is effective in the detection of invertebrate rice pest species. In silico PCR analysis showed that the primer pair mlCOIintF/Fol‐degen‐rev can amplify a 313 bp fragment of the cytochrome c oxidase subunit I (COI) gene from 19 prevalent invertebrate rice pest species in China. The feeding experiment confirmed that the eDNA of Marasmia exigua larvae and adult Leptocorisa chinensis remains detectable for up to 16 days post‐feeding under natural conditions. The implementation of washing‐based extraction protocols, combined with increased amounts of substrate, showed to be effective in recovering target eDNA quantities. Regarding pest species richness, eDNA metabarcoding complements traditional morphological identification methods and facilitates plant part‐specific pest detection through targeted sampling. These results indicate that eDNA metabarcoding holds significant potential for monitoring invertebrate pests in rice cultivation systems. Findings support developing future eDNA‐based systems for precise pest identification and monitoring in agroecosystems.