Comparative Analysis of Gut Microbiota in Two Cucurbit Leaf Beetles Reveals Divergent Adaptation Strategies Linked to Host Plant Range

Insects’ gut microbiota and their hosts share a mutually dependent symbiotic relationship. However, how insect dietary breadth relates to microbial diversity remains unclear. This study compared the gut bacterial communities of the polyphagous Aulacophora indica and the oligophagous Aulacophora lewisii. Using an integrated approach of cultivation, 16S rRNA high-throughput sequencing, and bioinformatic analyses, we assessed their composition, diversity, and functional potential. Using cultivation-based methods revealed that A. indica showed a greater abundance and diversity of culturable bacteria, dominated by Proteobacteria and Actinobacteria, compared to A. lewisii (Proteobacteria and Firmicutes). In contrast, high-throughput sequencing revealed the opposite pattern: A. lewisii exhibited significantly higher overall species richness and diversity. This apparent paradox highlights the methodological complementarity between cultivation and sequencing. Furthermore, the community composition differed notably at the genus level. Functional prediction via PICRUSt2 v2.2.0 indicated that core metabolic pathways, including carbohydrate metabolism, amino acid metabolism, and energy metabolism, were more enriched in A. indica. In summary, this study reveals systematic multi-dimensional differences in the gut microbiomes of these beetles, providing a theoretical foundation and microbial resources for understanding ecological adaptation and developing targeted control strategies based on gut microbiota.