RNA interference of <i>chitin synthase</i> genes inhibits chitin biosynthesis and affects larval performance in <i>Leptinotarsa decemlineata</i> (Say)

Shi, Ji-Feng; Mu, Lili; Chen, Xu; Guo, Wen‐Chao; Li, Guoqing

doi:10.7150/ijbs.14464

Cited by 67 publications

(61 citation statements)

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“…RNAi technology has been successfully applied to the studies of gene function in insects (Hannon, 2002;Ghanim et al, 2007). There are also successful examples in Coleoptera (Arakane et al, 2005;Shi et al, 2016). In our study, the target genes Hsp70, Hsp21, and sHsp21 were silenced with high efficiency in A. hygrophila, and the expression of these three Hsps was significantly downregulated after RNA interference.…”

Section: Discussionmentioning

confidence: 60%

Induced Thermotolerance and Expression of Three Key Hsp Genes (Hsp70, Hsp21, and sHsp21) and Their Roles in the High Temperature Tolerance of Agasicles hygrophila

et al. 2020

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Thermal adaptation plays a fundamental role in the expansion and distribution of insects, and heat shock proteins (Hsps) play important roles in the temperature adaptation of various organisms. To determine the roles of Hsp genes (Hsp70, Hsp21, and sHsp21) on the high temperature tolerance of Agasicles hygrophila, we obtained complete cDNA (complementary DNA) sequences for Hsp70, Hsp21, and sHsp21 by rapid amplification of cDNA ends (RACE), analyzed their expression profiles under different high temperature treatments by quantitative reverse transcription polymerase chain reaction (RT-qPCR), and performed functional verification by RNA interference (RNAi). The open reading frames of Hsp70, Hsp21, and sHsp21 were 1940, 543, and 567 bp, encoding 650, 180, and 188 amino acids, respectively. Their molecular weights (MWs) were 71.757, 20.879, and 21.510 kDa, and the isoelectric points were 5.63, 6.45, and 6.24, respectively. Phylogenetic tree analysis showed that the Hsp70, Hsp21, and sHsp21 genes of A. hygrophila were relatively conserved in evolution. The Hsp70 and Hsp21 genes in A. hygrophila were homologous to those in Leptinotarsa decemlineata (87 and 79% similarity, respectively), and the sHsp21 gene in A. hygrophila was homologous to that in Lissorhoptrus oryzophilus (74% similarity). The amino acid polypeptide chain had highly conserved sequences of DLGGGTFD, VLVGGSTR, and GPTIEEVD. The sequence of EEVD was the characteristic motif of cytoplasmic Hsp70, and the highly conserved sequences of MALFR and MSLLP were characteristic sequences of Hsp2 and sHsp21, respectively. Relative quantitative real time PCR showed that the three Hsps could be induced by 4-h treatment at high temperatures. Significant upregulation of these Hsps was observed when the temperature was further increased. The RNAi results showed that the injection of the three Hsps' dsRNA could suppress the expression at the gene level significantly. Compared with the control group,

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Section: Discussionmentioning

confidence: 60%

Induced Thermotolerance and Expression of Three Key Hsp Genes (Hsp70, Hsp21, and sHsp21) and Their Roles in the High Temperature Tolerance of Agasicles hygrophila

et al. 2020

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“…Chitin synthase A (CHSA) was reported to affect tracheal aECM in two insects besides fruit fly. RNA interference of CHSA resulted in thinner taenidia and the disappearance of taenidial stripes in Leptinotarsa decemlineata larvae, while the reduced expression of SeCHSA in Spodoptera exigua reduced chitin accumulation in the tracheae, resulting in severe tracheal malformation (Chen et al ., ; Shi et al ., ). Here, the disrupted expression of NlCPAP1‐N , the homolog of mtg , resulted in typical defects of the chitin spiral covering the tube.…”

Section: Discussionmentioning

confidence: 97%

Involvement of mind the gap in the organization of the tracheal apical extracellular matrix in Drosophila and Nilaparvata lugens

Yue

et al. 2019

Insect Science

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The tracheal apical extracellular matrix (aECM) is vital for expansion of the tracheal lumen and supports the normal structure of the lumen to guarantee air entry and circulation in insects. Although it has been found that some cuticular proteins are involved in the organization of the aECM, unidentified factors still exist. Here, we found that mind the gap (Mtg), a predicted chitin-binding protein, is required for the normal formation of the apical chitin matrix of airway tubes in the model holometabolous insect Drosophila melanogaster. Similar to chitin, the Mtg protein was linearly arranged in the tracheal dorsal trunk of the tracheae in Drosophila. Decreased mtg expression in the tracheae seriously affected the viability of larvae and caused tracheal chitin spiral defects in some larvae. Analysis of mtg mutant showed that mtg was required for normal development of tracheae in embryos. Irregular taenidial folds of some mtg mutant embryos were found on either lateral view of tracheal dorsal trunk or internal view of transmission electron microscopy analysis. These abnormal tracheae were not fully filled with gas and accompanied by a reduction in tracheal width, which are characteristic phenotypes of tracheal aECM defects. Furthermore, in the hemimetabolous brown planthopper (BPH) Nilaparvata lugens, downregulation of NlCPAP1-N (a homolog of mtg) also led to the formation of abnormal tracheal chitin spirals and death. These results suggest that mtg and its homolog are involved in the proper organization of the tracheal aECMs in flies and BPH, and that this function may be conserved in insects.

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“…In our experiments, we tested a standardized bacteria concentration by growing the culture at defined OD 600 nm , followed by resuspension in a constant volume, and we did not explore additional bacteria concentrations. In L. decemlineata, a reduction in chitin synthase (ChS) transcript is dependent on the larval phase, and silencing of ChS increased larval mortality after continuous feeding for 3 days with the dsRNA ChS . In H. armigera , larvae fed on tobacco plants with transformed chloroplast expressing dsRNA of chitin synthase showed a significant mortality of 53.3% compared to larvae fed on wild‐type plants .…”

Section: Discussionmentioning

confidence: 99%

“…In L. decemlineata, a reduction in chitin synthase (ChS) transcript is dependent on the larval phase, and silencing of ChS increased larval mortality after continuous feeding for 3 days with the dsRNA ChS. 41 In H. armigera, larvae fed on tobacco plants with transformed chloroplast expressing dsRNA of chitin synthase showed a significant mortality of 53.3% compared to larvae fed on wild-type plants. 42 The efficiency reported in silencing CHI, with the subsequent increase in mortality of insects from several orders, together with the fact that chitin synthase is not present in vertebrates and plants, emphasizes the potential of using this gene in RNAi pest control.…”

Section: Wileyonlinelibrarycom/journal/psmentioning

confidence: 99%

Gene silencing by RNAi via oral delivery of dsRNA by bacteria in the South American tomato pinworm, Tuta absoluta

Bento

Marques

Campana

et al. 2019

Pest Management Science

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BACKGROUND RNA interference (RNAi) has been evaluated in several insect pests as a novel strategy to be included in integrated pest management. Lepidopterans are recognized to be recalcitrant to gene silencing by RNAi. As such, double‐stranded RNA (dsRNA) delivery needs to be adjusted to assure its stability until it reaches the target gene transcript for silencing. Gene silencing by RNAi offers the potential to be used in the control of Tuta absoluta (Meyrick), one of the main insect pests of tomato (Solanum lycopersicum) worldwide. Here, we tested the delivery of dsRNA expressed in Escherichia coli HT115(DE3) and supplied to larvae in an artificial diet by screening target genes for silencing. We tested six target genes: juvenile hormone inducible protein (JHP); juvenile hormone epoxide hydrolase protein (JHEH); ecdysteroid 25‐hydroxylase (PHM); chitin synthase A (CHI); carboxylesterase (COE); and arginine kinase (AK). RESULTS Based on larval mortality, the duration of the larval stage in days, pupal weight, and the accumulation of the target gene transcript, we demonstrated the efficacy of bacterial dsRNA delivery for the functional effects on larval development. Providing dsRNA targeted to JHP, CHI, COE and AK by bacteria led to a significant decrease in transcript accumulation and an increase in larval mortality. CONCLUSION Bacteria expressing dsRNA targeting essential T. absoluta genes supplied in artificial diet are efficient to screen RNAi target‐genes. The oral delivery of dsRNA by bacteria is a novel potential alternative for the control of T. absoluta based on RNAi. © 2019 Society of Chemical Industry

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RNA interference of chitin synthase genes inhibits chitin biosynthesis and affects larval performance in Leptinotarsa decemlineata (Say)

Cited by 67 publications

References 50 publications

Induced Thermotolerance and Expression of Three Key Hsp Genes (Hsp70, Hsp21, and sHsp21) and Their Roles in the High Temperature Tolerance of Agasicles hygrophila

Induced Thermotolerance and Expression of Three Key Hsp Genes (Hsp70, Hsp21, and sHsp21) and Their Roles in the High Temperature Tolerance of Agasicles hygrophila

Involvement of mind the gap in the organization of the tracheal apical extracellular matrix in Drosophila and Nilaparvata lugens

Gene silencing by RNAi via oral delivery of dsRNA by bacteria in the South American tomato pinworm, Tuta absoluta

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