Molecular ontogeny of Bemisia tabaci life stages reveals miRNA-governed development and egg-stalk support for early embryogenesis

BACKGROUND: Deciphering how developmental gene-regulatory programs interface with lineage-specific reproductive novelties in Bemisia tabaci demands a temporally resolved, integrative molecular framework. Using deep miRNA sequencing, high-throughput transcriptomics, quantitative proteomics, two-dimensional proteo-mapping, and spatially multiplexed FISH, we delineate the circuitry governing whitefly ontogeny. RESULTS: This atlas uncovers a previously unrecognized morphological innovation: a transporter-enriched egg stalk operating as an active, metabolically competent nutrient-harvesting appendage. Integrated temporal profiling indicates that the strongest molecular reprogramming in B. tabaci occurs at hatching and adult emergence, consistent with conserved developmental transition points described in other insects. Conserved miRNAs (Btab-mir-34 and Btab-mir-2944b) were associated with early developmental transitions, whereas clade-restricted miRNAs (Btab-mir-307a, Btab-mir-352a, and Btab-mir-107a) were predicted to regulate metabolic, detoxification, and chemosensory networks. Spatial FISH supports that Btab-novel-mir-018a represses vitellogenin during late-nymphal stages, establishing a developmentally gated post-transcriptional module relieved at adult emergence. Comparison of egg versus isolated stalk tissue proteomics revealed a transporter-enriched molecular signature for the B. tabaci egg stalk, extending earlier physiological evidence for pedicel-mediated water and solute uptake. Localization of NaPi-III and RNAi-associated egg viability phenotypes further supports a transport-related role for this structure during embryogenesis. CONCLUSION: Collectively, this study provides a stage-wise molecular reconfiguration framework toward understanding the developmental and evolutionary architecture of B. tabaci. Furthermore, preliminary evidence for egg-stalk-associated transport identifies a candidate ontogeny-specific process that may inform future precision pest-management strategies.