The lethal giant larvae Gene in Tribolium castaneum:  Molecular Properties and Roles in Larval and  Pupal Development as Revealed by RNA Interference

Xiao, Da; Xiao, Liang; Gao, Xiwu; Yao, James T. P.; Zhu, Kun Yan

doi:10.3390/ijms15046880

Cited by 15 publications

(34 citation statements)

References 33 publications

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“…The peak effect of injected dsRNA is more likely dose and insect species-dependent. T. castaneum larvae injected with dsRNA of lethal giant larvae gene (TcLgl) showed significant suppression of the TcLg1 transcripts 2 days post-injection 53 . In the brown plant hopper, Nilaparvata lugens , knockdown of two trehalose-6-phosphate synthases caused inhibition of mRNA expression of twelve chitinase genes 48 h after treatments and up to 72 h for other genes 54 .…”

Section: Discussionmentioning

confidence: 99%

Characterization and RNAi-mediated knockdown of Chitin Synthase A in the potato tuber moth, Phthorimaea operculella

Mohammed

Diab

Abdelsattar

et al. 2017

Sci Rep

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Chitin is a major component of insect exoskeleton, tracheal system and gut where it is synthesized by chitin synthase (CHS) enzymes. In this paper, we report the isolation and RNAi of chitin synthase A (PhoCHSA) from the potato tuber moth Phthorimaea operculella. The full-length cDNA of PhoCHSA is 5,627 bp with 4,689 bp open reading frame coding for 1,563 amino acids. Structural analysis of conceptual amino acid translation showed three distinct regions found in all known insect CHS proteins; N-terminus region having 9 transmembrane helices, middle catalytic region containing several conserved domains identified in insect CHS enzymes, and C-terminus region containing seven transmembrane spans. Phylogenetic analysis showed that PhoCHSA protein clustered with CHSA enzymes identified from insects from different insect orders. RNAi targeting three different regions of the gene showed different efficacy against potato tuber moth larvae and dsRNA targeting the 5′ region has the highest efficacy. Results were verified by qRT-PCR which showed that dsRNA targeting the 5′ region caused the highest reduction in PhoCHSA mRNA level. Our results show the importance of selecting the RNAi target region and that chitin synthase A can be a suitable RNAi target for the potato tuber moth control.

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Section: Discussionmentioning

confidence: 99%

Characterization and RNAi-mediated knockdown of Chitin Synthase A in the potato tuber moth, Phthorimaea operculella

Mohammed

Diab

Abdelsattar

et al. 2017

Sci Rep

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“…Since preliminary studies with different doses showed no difference in either control mortality or expression of target gene 30 , subsequent controls were only carried out at a single (highest) dose for each treatment.…”

Section: Methodsmentioning

confidence: 99%

“…RNAi-based gene silencing thus has the potential to represent a novel insecticide technology, since it is theoretically possible to protect plants against insects by down regulating the expression of essential genes in the pest 20 29 30 31 . Furthermore, this technology should also allow non-conserved sequences to be specifically targeted, thus conferring a high degree of specificity.…”

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confidence: 99%

RNAi-mediated knockdown of the voltage gated sodium ion channel TcNav causes mortality in Tribolium castaneum

Halim

Alshukri

Ahmad

et al. 2016

Sci Rep

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The voltage-gated sodium ion channel (VGSC) belongs to the largest superfamily of ion channels. Since VGSCs play key roles in physiological processes they are major targets for effective insecticides. RNA interference (RNAi) is widely used to analyse gene function, but recently, it has shown potential to contribute to novel strategies for selectively controlling agricultural insect pests. The current study evaluates the delivery of dsRNA targeted to the sodium ion channel paralytic A (TcNav) gene in Tribolium castaneum as a viable means of controlling this insect pest. Delivery of TcNav dsRNA caused severe developmental arrest with larval mortalities up to 73% post injection of dsRNA. Injected larvae showed significant (p < 0.05) knockdown in gene expression between 30–60%. Expression was also significantly (p < 0.05) reduced in pupae following injection causing 30% and 42% knockdown for early and late pupal stages, respectively. Oral delivery of dsRNA caused dose-dependant mortalities of between 19 and 51.34%; this was accompanied by significant (p < 0.05) knockdown in gene expression following 3 days of continuous feeding. The majority of larvae injected with, or fed, dsRNA died during the final larval stage prior to pupation. This work provides evidence of a viable RNAi-based strategy for insect control.

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“…The RT-qPCR was performed with EvaGreen qPCR MasterMix-iCycler (Applied Biological Materials) by using the Bio-Rad iCycler iQTM multi-color real-time PCR detection system (Bio-Rad Laboratories, Hercules, CA, USA). A gene encoding ribosomal protein S3, TcRPS3 , was used as an internal reference [ 57 ]. Primers for RT-qPCR were designed by Beacon Designer™ ( Table 3 ).…”

Section: Methodsmentioning

confidence: 99%

Insecticide-Mediated Up-Regulation of Cytochrome P450 Genes in the Red Flour Beetle (Tribolium castaneum)

Liang

Xiao

et al. 2015

IJMS

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Some cytochrome P450 (CYP) genes are known for their rapid up-regulation in response to insecticide exposures in insects. To date, however, limited information is available with respect to the relationships among the insecticide type, insecticide concentration, exposure duration and the up-regulated CYP genes. In this study, we examined the transcriptional response of eight selected CYP genes, including CYP4G7, CYP4Q4, CYP4BR3, CYP12H1, CYP6BK11, CYP9D4, CYP9Z5 and CYP345A1, to each of four insecticides in the red flour beetle, Tribolium castaneum. Reverse transcription quantitative PCR (RT-qPCR) revealed that CYP4G7 and CYP345A1 can be significantly up-regulated by cypermethrin (1.97- and 2.06-fold, respectively), permethrin (2.00- and 2.03-fold) and lambda-cyhalothrin (1.73- and 1.81-fold), whereas CYP4BR3 and CYP345A1 can be significantly up-regulated by imidacloprid (1.99- and 1.83-fold) when 20-day larvae were exposed to each of these insecticides at the concentration of LC20 for 24 h. Our studies also showed that similar levels of up-regulation can be achieved for CYP4G7, CYP4BR3 and CYP345A1 by cypermethrin, permethrin, lambda-cyhalothrin or imidacloprid with approximately one fourth of LC20 in 6 h. Our study demonstrated that up-regulation of these CYP genes was rapid and only required low concentrations of insecticides, and the up-regulation not only depended on the CYP genes but also the type of insecticides. Our results along with those from previous studies also indicated that there were no specific patterns for predicting the up-regulation of specific CYP gene families based on the insecticide classification.

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The lethal giant larvae Gene in Tribolium castaneum: Molecular Properties and Roles in Larval and Pupal Development as Revealed by RNA Interference

Cited by 15 publications

References 33 publications

Characterization and RNAi-mediated knockdown of Chitin Synthase A in the potato tuber moth, Phthorimaea operculella

Characterization and RNAi-mediated knockdown of Chitin Synthase A in the potato tuber moth, Phthorimaea operculella

RNAi-mediated knockdown of the voltage gated sodium ion channel TcNav causes mortality in Tribolium castaneum

Insecticide-Mediated Up-Regulation of Cytochrome P450 Genes in the Red Flour Beetle (Tribolium castaneum)

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