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Regulation of Hormone-Related Genes in Ericerus Pela (Hemiptera: Coccidae) for Dimorphic Metamorphosis

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Regulation of Hormone-Related Genes in Ericerus Pela (Hemiptera: Coccidae) for Dimorphic Metamorphosis

Liu Pengfei et al. J Insect Sci.

Abstract

Insect hormones regulate metamorphosis including that leading to sexual dimorphism. Using RNA-Seq, we discovered that the second-instar male larva (SM) of the white wax insect, Ericerus pela, have 5,968 and 8,620 differentially expressed transcripts compared with the second-instar female larva (SF) and the first-instar male larva (FM), respectively. The expression levels of genes involved in the apoptosis of old tissues and the reconstruction of new ones in the SM significantly enhanced, while the SF mainly has enhanced expression levels of anabolic genes such as chitin. We predicted that the second-instar larvae are the developmental origin of sexual dimorphic metamorphosis. Meanwhile, in the juvenile hormone (JH) metabolic pathway, CYP15A1 and JH esterase (JHE) are differentially expressed; and in the 20-hydroxyecdysone (20E) metabolic pathway, CYP307A1, CYP314A1, and CYP18A1 are differentially expressed. In the SM, the expression levels of CYP307A1 and CYP314A1 are significantly increased, whereas the expression level of CYP18A1 is significantly decreased; in the SF, the expression levels of the above genes are opposite to that of the SM. Expression trends of RNA-seq is consistent with the expression level of qRT-PCR, and seven of them are highly correlated (R ≥ 0.610) and four are moderately correlated (0.588 ≥ R ≥ 0.542).

Keywords: dimorphic; hormone; metamorphosis; transcriptome.

Figures

Fig. 1.
Fig. 1.
(a) The life history of male and female worms of E. pela, in which E represents egg, FF represents first-instar female nymph, SF represents second-instar female nymph, EA represents early female adult, and LA represents late female adult. FM represents the first-instar male nymph, SM represents the second-instar male nymph, PP represents the pre-pupa, P represents the pupa stage, and MA represents the male adult stage. (b) Results of database annotation of the transcriptome of E. pela; x-axis is the database, right-y-axis is the number of transcripts annotated and left-y-axis is percentage. (c) VN diagram analysis of transcripts of the expressed between the six stages of E. pela.
Fig. 2.
Fig. 2.
Transcriptional differential gene analysis. (a) Differential gene clustering analysis (FPKM ≥ 1). (b) Expression results of differentially expressed transcripts in different periods. The x-axis represents the comparison between the two periods, and the y-axis represents the number of transcripts. (c) VN diagram analysis of transcripts of the differential expression of the six stages of E. pela. (d) GO enrichment expression results between two stages. The x-axis represents the comparison between the two periods, and the y-axis represents the number of transcripts. (e) GO enrichment transcripts. The x-axis represents genes, the y-axis represents the number of transcripts, and the red bars represented biological processes and the green bars represented molecular functions.
Fig. 3.
Fig. 3.
Differential genes of KEGG metabolic pathways of terpene and insect hormone. (a) Differential transcripts of KEGG metabolic pathways of terpene and insect hormone. The x-axis represents the period contrast and the y-axis represents the number of transcripts. (b, c) The x-axis represents differential genes of terpene, and the y-axis represents the number of transcripts. (d, e) The x-axis represents the differential genes of insect hormone, and the y-axis represents the number of transcripts. (f) Brief diagram of JH and 20E metabolic pathways, left-JH and right-20E. Blue letters represent the differential gene, and the graph on the left of the letter represents the expression result of the gene differential transcript.
Fig. 4.
Fig. 4.
qPCR validation. (a) results of insect hormone metabolism pathway SM versus SF stage differentially expressed genes; (b) results of insect hormone metabolism pathway SM versus FM stage differentially expressed genes; (c) RT–PCR was used to verify the results of RNA-seq sequencing, the x-axis represents the different periods of insects and the y-axis represents the expression quantity.

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