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

Abstract

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.

Study on the Effect of Wing Bud Chitin Metabolism and Its Developmental Network Genes in the Brown Planthopper, Nilaparvata lugens, by Knockdown of TRE Gene

“…Trehalose is known as the blood sugar of insects and plays a key role in various physiological processes (Kikuta et al, 2012 ; Yasugi et al, 2017 ). Trehalase (TRE), in insects, is divided into two types- soluble TRE, also called TRE1 or Treh1, and membrane-bound TRE, also called TRE2 or Treh2 (Shukla et al, 2015 ; Tang et al, 2016 ; Zhao et al, 2016 ). TRE1 is involved in the hydrolysis of endogenous trehalose, and TRE2 functions in the assimilation of exogenous trehalose as a carbon source (Becker et al, 1996 ; Chen et al, 2010a ).…”

“…Most insect species, including Spodoptera exigua (Tang et al, 2008 ; Chen et al, 2010a ), Apis mellifera (Lee et al, 2007 ), Bombyx mori (Mitsumasu et al, 2005 ; Kamei et al, 2011 ), Laodelphax striatellus (Zhang et al, 2012 ), Omphisa fuscidentalis (Tatun et al, 2008a , b ), and Bemisia tabaci (Wang et al, 2014 ) have one soluble TRE gene as well as one membrane-bound TRE gene (Bansal et al, 2013 ). In addition, some insect have more than one soluble TRE; two Treh1 were found in Leptinotarsa decemlineata (Shi J. F. et al, 2016 ), five Treh1 were found in Harmonia axyridis (Tang et al, 2014 ; Shi Z. K. et al, 2016 ) and four Treh1 were found in Tribolium castaneum (Tang et al, 2016 ). In addition, two TRE1 and one TRE2 have been cloned and reported in Nilaparvata lugens (Zhao et al, 2016 ).…”

“…Previous studies have shown that target gene silencing via RNA interference (RNAi) or inhibition of TRE activities can lead to insect death or morphological defects because TRE plays an important regulatory role in chitin metabolism (Zhao et al, 2016 ; Tang et al, 2017 ). TRE regulates the expression of CHS and Cht in the insect cuticle and midgut, and inhibiting chitin biosynthesis kills pests when TRE gene expression is suppressed or knocked down (Chen et al, 2010a ; Zhang et al, 2012 ; Tang et al, 2016 , 2017 ; Zhao et al, 2016 ). When insect chitin synthesis or degradation malfunction or the chitin content are decreases, normal molting does not occur, and abnormalities are observed (Zhao et al, 2016 ; Zhang et al, 2017a ).…”

Regulatory functions of trehalose-6-phosphate synthase in the chitin biosynthesis pathway in Tribolium castaneum (Coleoptera: Tenebrionidae) revealed by RNA interference

“…Several studies have revealed that RNAi in T. castaneum is efficient in maintaining the silencing effect for a longer period and inducing a higher rate of deformations (Chen et al, 2010a;Xi et al, 2015), even in the postembryonic stages (Tomoyasu & Denell, 2004;Arakane et al, 2005;Minakuchi et al, 2008Minakuchi et al, , 2015. Previous studies have also shown that T. castaneum is an effective and reliable insect for conducting RNAi research (Noh et al, 2012;Tang et al, 2016).…”

Targeting the potassium ion channel genes SK and SH as a novel approach for control of insect pests: efficacy and biosafety

“…This technology can be seen as an 'environmentally friendly' approach for the control of insect pests, with a high degree of specificity. 4 Many insect species from different orders, including Coleoptera, [5][6][7] Hemiptera, 8 Orthoptera, 9 Diptera 5,10,11 and Lepidoptera, [12][13][14][15] have been shown to be sensitive to dsRNA, although some species are far more sensitive than others. Previous studies have revealed that the Coleoptera are more susceptible to dsRNA compared to other arthropod orders, 16 while the Lepidoptera require higher concentrations of dsRNA.…”