Screening of reference genes for expression analysis in the study of soldier caste differentiation of Formosan subterranean termite <i>Coptotermes formosanus</i> Shiraki

Du, He; Wu, Wenjing; Huang, Xueyi; Li, Zhiqiang

doi:10.7717/peerj.7981

Cited by 5 publications

(3 citation statements)

References 25 publications

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“…The qPCR was performed in CFX Connect Optics Module (Bio-Rad, Hercules, CA, USA). RPS18 was used as the reference gene [37]. The fold change was calculated for each of the eleven unigenes, and then log 2 transformed.…”

Section: Validation Of Rna-seq Resultsmentioning

confidence: 99%

Methoprene-Induced Genes in Workers of Formosan Subterranean Termites (Coptotermes formosanus Shiraki)

Du¹,

Tong

Huang³

et al. 2020

Insects

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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.

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Section: Validation Of Rna-seq Resultsmentioning

confidence: 99%

Methoprene-Induced Genes in Workers of Formosan Subterranean Termites (Coptotermes formosanus Shiraki)

Du¹,

Tong

Huang³

et al. 2020

Insects

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“…The resulting cDNA library was used for qRT‐PCR reaction with the ChamQ SYBY qPCR Master Mix (Vazyme, Nanjing, China) on a CFX96™ Real‐Time system (Bio‐Rad, Hercules, CA, USA). The reaction was performed as follows: 95°C for 30 s, followed by 39 cycles at 95°C for 10 and 60°C for 30 s. The housekeeping gene, termite RPL (Ribosomal protein L7), was used for normalisation (Du et al, 2019). Primer sets were designed using Primer‐Blast (https://www.ncbi.nlm.nih.gov/tools/primer-blast/) and are listed in Table S1.…”

Section: Methodsmentioning

confidence: 99%

The influence of first desaturase subfamily genes on fatty acid synthesis, desiccation tolerance and inter‐caste nutrient transfer in the termite Coptotermes formosanus

Xu,

Tong,

Chen

et al. 2023

Insect Molecular Biology

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Desaturase enzymes play an essential role in the biosynthesis of unsaturated fatty acids (UFAs). In this study, we identified seven “first desaturase” subfamily genes (Cfor‐desatA1, Cfor‐desatA2‐a, Cfor‐desatA2‐b, Cfor‐desatB‐a, Cfor‐desatB‐b, Cfor‐desatD and Cfor‐desatE) from the Formosan subterranean termite Coptotermes formosanus. These desaturases were highly expressed in the cuticle and fat body of C. formosanus. Inhibition of either the Cfor‐desatA2‐a or Cfor‐desatA2‐b gene resulted in a significant decrease in the contents of fatty acids (C16:0, C18:0, C18:1 and C18:2) in worker castes. Moreover, we observed that inhibition of most of desaturase genes identified in this study had a negative impact on the survival rate and desiccation tolerance of workers. Interestingly, when normal soldiers were reared together with dsCfor‐desatA2‐b‐treated workers, they exhibited higher mortality, suggesting that desaturase had an impact on trophallaxis among C. formosanus castes. Our findings shed light on the novel roles of desaturase family genes in the eusocial termite C. formosanus.

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“…The selection criteria of the best reference genes by this method consider the following factors: 1) genes with a standard deviation of raw Ct values higher than 1.0 are considered inconsistent, and 2) genes with expression 3-fold over or under their geometric mean Ct should be discarded. Primers for all reference genes were taken from Du, et al [ 47 ] ( S3 Table ), and primer specificity was determined by the melting curve analysis. Efficiency for each qRT-PCR reaction was determined utilizing the raw amplification data and the LinRegPCR software [ 48 , 49 ].…”

Section: Methodsmentioning

confidence: 99%

Molecular characterization and expression variation of the odorant receptor co-receptor in the Formosan subterranean termite

2022

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Subterranean termites live in underground colonies with a division of labor among castes (i.e., queens and kings, workers, and soldiers). The function of social colonies relies on sophisticated chemical communication. Olfaction, the sense of smell from food, pathogens, and colony members, plays an important role in their social life. Olfactory plasticity in insects can be induced by long- and short-term environmental perturbations, allowing adaptive responses to the chemical environment according to their physiological and behavioral state. However, there is a paucity of information on the molecular basis of olfaction in termites. In this study, we identified an ortholog encoding the odorant receptor co-receptor (Orco) in the Formosan subterranean termite, Coptotermes formosanus, and examined its expression variation across developmental stages and in response to social conditions. We found that C. formosanus Orco showed conserved sequence and structure compared with other insects. Spatial and temporal analyses showed that the Orco gene was primarily expressed in the antennae, and it was expressed in eggs and all postembryonic developmental stages. The antennal expression of Orco was upregulated in alates (winged reproductives) compared with workers and soldiers. Further, the expression of Orco decreased in workers after starvation for seven days, but it was not affected by the absence of soldiers or different group sizes. Our study reveals the molecular characteristics of Orco in a termite, and the results suggest a link between olfactory sensitivity and nutritional status. Further studies are warranted to better understand the role of Orco in olfactory plasticity and behavioral response.

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Screening of reference genes for expression analysis in the study of soldier caste differentiation of Formosan subterranean termite Coptotermes formosanus Shiraki

Cited by 5 publications

References 25 publications

Methoprene-Induced Genes in Workers of Formosan Subterranean Termites (Coptotermes formosanus Shiraki)

Methoprene-Induced Genes in Workers of Formosan Subterranean Termites (Coptotermes formosanus Shiraki)

The influence of first desaturase subfamily genes on fatty acid synthesis, desiccation tolerance and inter‐caste nutrient transfer in the termite Coptotermes formosanus

Molecular characterization and expression variation of the odorant receptor co-receptor in the Formosan subterranean termite

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