Comparison of the <scp>RNA</scp> interference effects triggered by <scp>dsRNA</scp> and <scp>siRNA</scp> in <i>Tribolium</i><i>castaneum</i>

Wang, Jinda; Wu, Min; Wang, Baoju; Han, Zhaojun

doi:10.1002/ps.3432

Cited by 46 publications

(36 citation statements)

References 45 publications

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“…Injecting the same amount of corresponding siRNA, however, yielded only 45% silencing from days 2-4 with no observable phenotypic changes (Wang et al, 2013). These findings, coupled with the recent observation that clathrin-mediated endocytosis serves as the predominant cellular uptake mechanism for dsRNA in T. castaneum (Xiao et al, 2015), points to possible lengthdependent uptake of dsRNA in T. castaneum as has been observed in S2 cells.…”

Section: Nucleotide Sequence Lengthsupporting

confidence: 59%

Diet-delivered RNAi in Helicoverpa armigera – Progresses and challenges

Lim

Robinson

Jain

et al. 2016

Journal of Insect Physiology

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Section: Nucleotide Sequence Lengthsupporting

confidence: 59%

Diet-delivered RNAi in Helicoverpa armigera – Progresses and challenges

Lim

Robinson

Jain

et al. 2016

Journal of Insect Physiology

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“…The thermal cycling conditions were as follows: 40 cycles at 95 °C for 30 s, 58 °C for 30 s, and 72 °C for 45 s and a last extension at 72 °C for 10 min9. Purified TRE amplicons were in-vitro transcribed using T7 RiboMax Express RNAi System (Promega, Madison, USA)71. GFP amplicon was used to prepare control dsRNA.…”

Section: Methodsmentioning

confidence: 99%

Functional characterization of three trehalase genes regulating the chitin metabolism pathway in rice brown planthopper using RNA interference

Zhao

Yang

Shen

et al. 2016

Sci Rep

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RNA interference (RNAi) is an effective gene-silencing tool, and double stranded RNA (dsRNA) is considered a powerful strategy for gene function studies in insects. In the present study, we aimed to investigate the function of trehalase (TRE) genes (TRE 1-1, TRE 1-2, and TRE-2) isolated from the brown planthopper Nilaparvata lugens, a typical piercing-sucking insect in rice, and investigate their regulating roles in chitin synthesis by injecting larvae with dsRNA. The results showed that TRE1 and TRE2 had compensatory function, and the expression of each increased when the other was silenced. The total rate of insects with phenotypic deformities ranged from 19.83 to 24.36% after dsTRE injection, whereas the mortality rate ranged from 14.16 to 31.78%. The mRNA levels of genes involved in the chitin metabolism pathway in RNA-Seq and DGEP, namely hexokinase (HK), glucose-6-phosphate isomerase (G6PI) and chitinase (Cht), decreased significantly at 72 h after single dsTREs injection, whereas two transcripts of chitin synthase (CHS) genes decreased at 72 h after dsTRE1-1 and dsTREs injection. These results demonstrated that TRE silencing could affect the regulation of chitin biosynthesis and degradation, causing moulting deformities. Therefore, expression inhibitors of TREs might be effective tools for the control of planthoppers in rice.

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“…RNAi has also been employed for the silencing of vital genes in Tribolium castaneum , Nilaparvata lugens and other insects [4, 6]. Furthermore, RNAi could be used in novel methods of pest management and may be more effective than other pest control measures once the problems of the efficiency and specificity of gene silencing are solved [7]. …”

Section: Introductionmentioning

confidence: 99%

“…T. castaneum is a holometabolous insect in the larva, pupa and adult stages and represents a serious pest that damages grain in storage. It is thought to be an effective and reliable insect for conducting RNAi research [7, 31, 32]. It has also been reported that RNAi is highly effective in Tribolium , even in the postembryonic stages of Tribolium [9, 33].…”

Section: Introductionmentioning

confidence: 99%

Knockdown of five trehalase genes using RNA interference regulates the gene expression of the chitin biosynthesis pathway in Tribolium castaneum

et al. 2016

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BackgroundRNA interference is a very effective approach for studies on gene function and may be an efficient method for controlling pests. Trehalase is a key gene in the chitin biosynthesis pathway in insects. Five trehalase genes have been cloned in Tribolium castaneum, though it is not known whether the detailed functions of these trehalases can be targeted for pest control.ResultsThe functions of all five trehalase genes were studied using RNAi, and the most important results showed that the expression of all 12 genes decreased significantly from 12 to 72 h compared with the control groups, except GP1 at 72 h, when the expression of the TcTre2 gene was suppressed. The results also revealed different abnormal phenotypes, and the observed mortality rates ranged from 17 to 42 %. The qRT-PCR results showed that the expression of TPS, GS, two GP, CHS1a and CHS1b genes decreased significantly, while that of the CHS2 gene decreased or increased after RNAi after the five trehalases were silenced at 48 h. In addition, TPS gene expression decreased from 12 to 72 h after dsTcTre injection.ConclusionsThese results demonstrate that silencing of any individual trehalase gene, especially Tre1-4 and Tre2 gene can lead to moulting deformities and a high mortality rate through the regulation of gene expression in the chitin biosynthesis pathway and may be a potential approach for pest control in the future.Electronic supplementary materialThe online version of this article (doi:10.1186/s12896-016-0297-2) contains supplementary material, which is available to authorized users.

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Comparison of the RNA interference effects triggered by dsRNA and siRNA in Triboliumcastaneum

Cited by 46 publications

References 45 publications

Diet-delivered RNAi in Helicoverpa armigera – Progresses and challenges

Diet-delivered RNAi in Helicoverpa armigera – Progresses and challenges

Functional characterization of three trehalase genes regulating the chitin metabolism pathway in rice brown planthopper using RNA interference

Knockdown of five trehalase genes using RNA interference regulates the gene expression of the chitin biosynthesis pathway in Tribolium castaneum

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