doi: 10.3390/ijms241914797.
Ecdysteroid Biosynthesis Halloween GeneSpook Plays an Important Role in the Oviposition Process of Spider Mite,Tetranychus urticae
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- PMID:37834248
- PMCID: PMC10573261
- DOI: 10.3390/ijms241914797
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Ecdysteroid Biosynthesis Halloween GeneSpook Plays an Important Role in the Oviposition Process of Spider Mite,Tetranychus urticae
Liang Wang et al. Int J Mol Sci..
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Abstract
In insects, the ecdysteroid hormone regulates development and reproduction. However, its function in the reproduction process of spider mites is still unclear. In this study, we investigated the effect of the Halloween geneSpook on the oviposition of the reproduction process in a spider mite,Tetranychus urticae. The expression patterns of the ecdysteroid biosynthesis and signaling pathway genes, as analyzed by RT-qPCR, showed that the expression pattern of the Halloween genes was similar to the oviposition pattern of the female mite and the expression patterns of the vitellogenesis-related genesTuVg andTuVgR, suggesting that the Halloween genes are involved in the oviposition of spider mites. To investigate the function of the ecdysteroid hormone on the oviposition of the reproduction process, we carried out an RNAi assay against the Halloween geneSpook by injection in female mites. Effective silencing ofTuSpo led to a significant reduction of oviposition. In summary, these results provide an initial study on the effect of Halloween genes on the reproduction inT. urticae and may be a foundation for a new strategy to control spider mites.
Keywords: RNAi; Spook; Tetranychus urticae; ecdysteroid; oviposition.
Conflict of interest statement
The authors declare no conflict of interest.
Figures
Average daily egg production (number of eggs per day and per mite) of female adult mites ofT. urticae from the 1st day of the adult stage (=day 1) until death of the female mite at day 10. Every point represents a biological replicate and each replicate included 15 mites. In this experiment, we performed eight biological replicates per time point (day). The results are presented as mean (±SE) based on eight biological replicates per day. Lowercase letters above each column indicate significant differences amongst the ten days using one-way analysis of variance (ANOVA) followed by Tukey’s honestly significant difference (HSD) test (p < 0.05).
Expression pattern of the ecdysteroid biosynthesis Halloween genes (A–D) and the ecdysteroid signaling pathway genes (E–L) in female mites ofT. urticae as determined by quantitative real-time PCR (RT-qPCR). The relative expression levels were calculated using the method of 2−ΔΔCT and based on the value of the lowest expression level. Lowercase letters above each bar indicate significant differences amongst the ten days using one-way ANOVA followed by a Tukey’s HSD test (p < 0.05).
Expression pattern of the vitellogenesis-related genes,Vg (A–C) andVgR (D), in female mites ofT. urticae as determined RT-qPCR. The relative expression levels were calculated using the method of 2−ΔΔCT and based on the value of the lowest expression level. Lowercase letters above each bar indicate significant differences amongst the ten days using ANOVA followed by a Tukey’s HSD test (p < 0.05).
RNAi effects againstTuSpo by injection in female mites ofT. urticae. (A) The silencing efficiency ofTuSpo was detected by RT-qPCR at 24 h after the female mites were injected with 60–80 ng of dsTuSpo. The mean (±SE) expression level is based on four biological replicates, and each replicate consisted of 50 mites. The relative expression was calculated based on the value of dsefgp (control). The down arrow indicates a decrease of 64% in relative expression ofTuSpo. (B) Effects of dsTuSpo injection in female mites against the average daily egg production. Every point represented a small round leaf (2 cm diameter) with 5 mites on each leaf. The mean (±SE) average daily egg production is based on 18 biological replicates. (C) Effects of dsTuSpo injection in female mites on the total egg number. The mean (±SE) total egg number is based on four biological replicates, and we used 25 female mites per replicate. The down arrow indicates a decrease of 61% in total egg number. (D) Effects of dsTuSpo injection in female mites on the oviposition cycle. The mean (±SE) oviposition period is based on 18 biological replicates, and we used 5 mites per replicate. The significant difference between the two groups is indicated with ‘**’,p < 0.01, ‘***’,p < 0.001 and ‘****’,p < 0.0001 after a Student’st-test.
Relative expression levels of the ecdysteroid biosynthesis and signaling pathway genes and the vitellogenesis-related genes in female mites ofT. urticae, as determined by RT-qPCR at 24 h after the female mites were injected with 60–80 ng of dsTuSpo. (A) Relative expression levels of the other ecdysteroid biosynthesis Halloween genesTuDib,TuSad, andTuShd. (B) Relative expression levels of the ecdysteroid receptor complex genesTuEcR,TuRXR1, andTuRXR2. (C) Relative expression levels of the ecdysteroid signaling genesTuHR4,TuHR3,TuE75,TuE78, andTuFTZ-f1. (D) Relative expression the vitellogenin genes and their receptor gene,TuVg1,TuVg2,TuVg3, andTuVgR1. The mean (±SE) expression level is based on four biological replicates, and we used 50 mites per replicate. Significant differences between the treatment and control are indicated with ‘*’,p < 0.05 and ‘**’,p < 0.01 after a Student’st-test.
Scheme proposing the ecdysteroid biosynthesis and signaling cascade pathway regulating vitellogenesis and reproduction inT. urticae. Lowercase letters above the average daily egg production indicate significant differences during the first six days (day 1–6) in the adult stage using ANOVA followed by a Tukey’s HSD test (p < 0.05). ‘*’ indicates a significant difference of change during day 1–6 in the adult stage for the relative expression levels of the respective gene. Data on egg production and expression of genes are based on Figure 1, Figure 2 and Figure 3.
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This study was supported by the Guizhou Provincial Science and Technology Foundation (QianKeHeJiChu ZK [2022]-YiBan065), the Special Natural Science Foundation of Guizhou University (Gui Da Te Gang He Zi [2022]13), the National Natural Science Foundation of China (3175002), and the Project of Occurrence, Monitoring, and Investigation and Integrated Prevention and Control Technology of major pest mites from Ministry of Agriculture and Rural Affairs, China (No. 15226022).
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