Identification and Functional Analysis of Chitin Synthase A in Oriental Armyworm, <i>Mythimna separata</i>

Zhai, Yifan; Fan, Xiaolong; Yin, Zhenjuan; Yue, Xiangzhao; Men, Xingyuan; Zheng, Li; Zhang, Wenqing

doi:10.1002/pmic.201700165

Cited by 20 publications

(23 citation statements)

References 47 publications

(68 reference statements)

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“…Also, we had detected a relatively high level of expression in the ovary. Similar results have been observed in Mythimna separata 54 , where MsCHS1 was highly expressed in the ovary. A previous study using the fluorescently labeled lectin technique had also documented that chitin was present in A .…”

Section: Discussionsupporting

confidence: 89%

Molecular cloning, expression, and functional analysis of the chitin synthase 1 gene and its two alternative splicing variants in the white-backed planthopper, Sogatella furcifera (Hemiptera: Delphacidae)

Wang

Yang

Zhou

et al. 2019

Sci Rep

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Chitin synthase is responsible for chitin synthesis in the cuticles and cuticular linings of other tissues in insects. We cloned two alternative splicing variants of the chitin synthase 1 gene (SfCHS1) from the white-backed planthopper, Sogatella furcifera. The full-length cDNA of the two variants (SfCHS1a and SfCHS1b) consists of 6408 bp, contains a 4719-bp open reading frame encoding 1572 amino acids, and has 5′ and 3′ non-coding regions of 283 and 1406 bp, respectively. The two splicing variants occur at the same position in the cDNA sequence between base pairs 4115 and 4291, and consist of 177 nucleotides that encode 59 amino acids but show 74.6% identity at the amino acid level. Analysis in different developmental stages showed that expression of SfCHS1 and SfCHS1a were highest just after molting, whereas SfCHS1b reached its highest expression level 2 days after molting. Further, SfCHS1 and SfCHS1a were mainly expressed in the integument, whereas SfCHS1b was predominately expressed in the gut and fat body. RNAi-based gene silencing inhibited transcript levels of the corresponding mRNAs in S. furcifera nymphs injected with double-stranded RNA of SfCHS1, SfCHS1a, and SfCHS1b, resulted in malformed phenotypes, and killed most of the treated nymphs. Our results indicate that SfCHS1 may be a potential target gene for RNAi-based S. furcifera control.

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Section: Discussionsupporting

confidence: 89%

Molecular cloning, expression, and functional analysis of the chitin synthase 1 gene and its two alternative splicing variants in the white-backed planthopper, Sogatella furcifera (Hemiptera: Delphacidae)

Wang

Yang

Zhou

et al. 2019

Sci Rep

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“…For instance, knockdown of chitin synthase A (CHSA) gene induced severe phenotypic alterations in the normal course of ecdysis and oogenesis in M. separata. 47 M. separata fed with Polyoxin AL, a fungicide that inhibits chitin synthase, died at the molting stage. 48 Knockdown of Chitinase genes MseChi1 and Mse-Chi2 resulted in increased mortality and reduced body weight of M. separata larvae.…”

Section: Discussionmentioning

confidence: 99%

Expression and functional analysis of cytochrome P450 genes in the integument of the oriental armyworm, Mythimna separata (Walker)

Huang

Yin

Zhu

et al. 2020

Pest Management Science

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BACKGROUND: Mythimna separata is a devastating agricultural pest that has recently developed insecticide resistance. Integument-specific cytochrome P450s were reported to participate in cuticle formation and could be potential targets for pesticide synthesis. RESULTS: The transcriptome of integuments of M. separata larvae was constructed, generating a total of 38 058 unigenes with an average length of 1243 bp. These unigenes are enriched in functional categories such as lipid transport and metabolism, and secondary metabolites biosynthesis, transport and catabolism. Amongst unigenes, cytochrome P450s were identified and 66 unique P450s with complete open reading frames were named. These P450s were divided into 17 families and 32 subfamilies, containing conserved motifs such as helix C, helix I, helix K, and the heme-binding region. RNA-Seq and RT-qPCR analyses showed different expression levels of P450s in integuments of M. separata larvae. Further RT-qPCR analysis of P450s among different tissues showed that five P450s, especially CYP4G199, were specifically highly expressed in integuments. Moreover, knockdown of CYP4G199 disturbed cuticle formation, leading to imperfection in larval cuticle, and prevented pupation of M. separata. CONCLUSION: Transcriptome of larval integuments provided sequence and expression of genes in M. separata. CYP4G199 is specifically highly expressed in larval integuments and is important for cuticle formation in M. separata.

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“…RNAi technology can be used to eliminate or knock down the specific expression of specific genes, so this technique has been widely used to explore gene function in physiological processes of different insect species. Improved administration and formulation of dsRNAs for insects coupled with the current progress in RNAi technology enables RNAi to be an alternative strategy for insect pest control [31–34]. Several methods of RNAi have been successfully used by researchers with their own advantages and limitations [24, 35, 36].…”

Section: Introductionmentioning

confidence: 99%

RNA interference-mediated knockdown of 3, 4-dihydroxyphenylacetaldehyde synthase affects larval development and adult survival in the mosquito Aedes aegypti

Jing

Zhang

et al. 2019

Parasites Vectors

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Background The cuticle is an indispensable structure that protects the mosquito against adverse environmental conditions and prevents pathogen entry. While most cuticles are hard and rigid, some parts of cuticle are soft and flexible to allow movement and blood-feeding. It has been reported that 3, 4-dihydroxyphenylacetaldehyde (DOPAL) synthase is associated with flexible cuticle formation in Aedes aegypti . However, the molecular function of DOPAL synthase in the ontogenesis of mosquito remains largely unknown. In this study, we characterized gene expression profiles of DOPAL synthase and investigated its functions in larvae and female adults of Aedes agypti by RNAi. Results Our results suggest that the expression of DOPAL synthase is different during development and the transcriptional level reached its peak at the female white pupal stage, and DOPAL synthase was more highly expressed in the cuticle and midgut than other tissues in the adult. The development process from larva to pupa was slowed down strikingly by feeding the first-instar larvae with chitosan/ DOPAL synthase dsRNA nanoparticles. A qRT-PCR analysis confirmed that the dsRNA-mediated transcription of the DOPAL synthase was reduced > 50% in fourth-instar larvae. Meanwhile, larval molt was abnormal during development. Transmission electron microscopy results indicated that the formation of endocuticle and exocuticle was blocked. In addition, we detected that the ds DOPAL synthase RNA caused significant mortality when injected into the female adult mosquitoes. Conclusions Our findings demonstrate that DOPAL synthase plays a critical role in mosquito larval development and adult survival and suggest that DOPAL synthase could be a good candidate gene in RNAi intervention strategies in mosquito control. Electronic supplementary material The online version of this article (10.1186/s13071-019-3568-7) contains supplementary material, which is available to authorized users.

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Identification and Functional Analysis of Chitin Synthase A in Oriental Armyworm, Mythimna separata

Cited by 20 publications

References 47 publications

Molecular cloning, expression, and functional analysis of the chitin synthase 1 gene and its two alternative splicing variants in the white-backed planthopper, Sogatella furcifera (Hemiptera: Delphacidae)

Molecular cloning, expression, and functional analysis of the chitin synthase 1 gene and its two alternative splicing variants in the white-backed planthopper, Sogatella furcifera (Hemiptera: Delphacidae)

Expression and functional analysis of cytochrome P450 genes in the integument of the oriental armyworm, Mythimna separata (Walker)

RNA interference-mediated knockdown of 3, 4-dihydroxyphenylacetaldehyde synthase affects larval development and adult survival in the mosquito Aedes aegypti

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