A comprehensive and precise evaluation of Arthropoda diversity in agricultural landscapes can enhance biological pest control strategies. We used Malaise traps and sweep nets to collect insects from three double-cropping paddy fields. DNA was extracted from the ethanol preservative of the Malaise traps and from tissue samples of selected parasitoid wasps. This was followed by amplification using DNA barcoding primers to prepare high-throughput sequencing libraries. We annotated a total of 4956 operational taxonomic units (OTUs), encompassing 174 genera and 32 families of parasitoid wasps. The ethanol filter method efficiently captured a wide range of information. However, the method has low resolution and may result in a reduced estimate of species abundance. Additional insect species were also identified in the parasitoid samples. This suggests that high throughput sequencing from adult parasitoid wasps can also detect host species, enabling a better understanding of host species and providing insights into food webs.

The modifications in planting structure and continuous land use adjustments have significant implications for the structure, diversity, and functioning of arthropod communities in farmland landscapes, particularly regarding the biological control of pest insects by natural enemy species. Therefore, there is an urgent need to comprehensively and accurately assess insect community diversity in croplands. In this study conducted between 2022-2023, we utilized Malaise traps and sweep netting to collect insects from three different double-cropping paddy fields. We extracted DNA from preservative ethanol of Malaise traps and tissue samples of selected parasitoid wasps, followed by amplification using DNA barcoding primers to prepare high-throughput sequencing libraries. As a result, we annotated a total of 4956 operational taxonomic units (OTUs) belonging to 174 genera and 32 families of parasitoid wasps. Our findings demonstrate that the ethanol filter method efficiently captures a wide range of information diversity. However, it demonstrates a lower resolution and may result in a reduction of species abundance. Furthermore, we have identified additional insect species in the parasitoid samples. This demonstrates that high throughput sequencing from adult parasitoid wasps can effectively detect host specie, enabling a comprehensive understanding of host species and providing novel insights into food web construction.

This study reports the first complete mitochondrial genomes of three egg parasitoid wasps parasitizing Nilaparvata lugens—Pseudoligosita nephotetticum, Anagrus frequens, and Anagrus nilaparvatae. Genome sizes ranged from 15,429 to 15,889 bp, with all three mitogenomes displaying strong A + T bias, standard gene content, and characteristic strand asymmetries. While A. frequens and A. nilaparvatae exhibited conserved gene orders, extensive gene rearrangements, including multiple inversions in both protein-coding genes (PCGs) and tRNAs, were observed in P. nephotetticum. Codon usage analyses revealed a preference for codons ending in A or U. The non-synonymous (Ka) to synonymous (Ks) substitution ratio analysis identified signs of positive selection in multiple PCGs, particularly in atp8, nad6, and nad3, suggesting possible adaptive evolution related to host-searching behavior. Secondary structure analyses showed the loss of trnL1 in all Anagrus species, while trnS1 and trnR lacked the DHU arm, indicating possible derived traits in Mymaridae. Phylogenetic analysis was the first time to describe the relationship of the genus Anagrus within Mymaridae from the perspective of 13 protein genes. Furthermore, the grouping of (Pseudoligosita + Megaphragma) + Trichogramma was supporting the distinct evolutionary lineage of Pseudoligosita. This work provides new molecular resources and phylogenetic insight for Chalcidoidea, with implications for parasitoid evolution and biological control strategies.