No abstract

The Formosan subterranean termite, Coptotermes formosanus Shiraki, is a serious pest worldwide. Juvenile hormone analog (and its analogs such as methoprene) can induce the transformation of the worker caste into soldier caste in C. formosanus. However, several factors, such as feeding substrate and colony origin, influence the proportion of solider formation. The molecular mechanism of worker to soldier transformation of C. formosanus is still not clear. RT-qPCR is a powerful tool for molecular studies. Accurate gene quantification by the relative quantification method requires a stable expressed gene as the reference gene. However, no reference genes were available for this species in the methoprene bioassay. To study the problem of gene response to methoprene by RT-qPCR we have to first screen reference genes in C. formosanus. Workers were fed with methoprene. Termites were collected during the methoprene bioassay and separated into head and thorax+abdomen. Expression profiles of 10 candidate reference genes in the two body part types were investigated using RT-qPCR. The results were analyzed by a set of established methods (geNorm, NormFinder, BestKeeper, and RefFinder) as well as comparative ΔCt method. Our results suggest that RPS18 is the most stably-expressed gene both in the head and thorax+abdomen for expression analysis in the methoprene bioassay of C. formosanus. The screening of suitable reference genes in C. formosanus establishes the foundation for the molecular study of soldier caste differentiation in this species.

Termites have a distinct polyphenism controlled by concise hormonal and molecular mechanisms. Workers undergo double molts to transform into soldiers (worker–presoldier–soldier). Juvenile hormone analogs, such as methoprene, can induce workers to transform into presoldiers. However, the molecular mechanism underlying the worker-to-presoldier transformation in Coptotermes formosanus Shiraki is still not clear. We sequenced the transcriptome of workers four days after they had fed on methoprene-treated filter paper and control group workers, which fed on acetone-treated filter paper. The transcriptome of C. formosanus was assembled using the de novo assembly method. Expression levels of unigenes in the methoprene-treated group and the control group were compared. The differentially expressed genes were further analyzed by Gene Ontology (GO) term enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis. Tetrapyrrole binding, oxidoreductase activity, and metal ion binding were the only three enriched GO terms. Juvenile hormone synthesis was the first ranked enriched pathway. Carbohydrate, amino acid, and lipid metabolism pathways were also enriched. These three pathways may be related to fat body development, which is critical for presoldier formation. Our results have demonstrated the significance of JH synthesis pathways, and pathways related to fat body development in the artificial induction of presoldiers.