RNA interference of trehalose-6-phosphate synthase and trehalase genes regulates chitin metabolism in two color morphs of Acyrthosiphon pisum Harris

AbstractTrehalose-6-phosphate synthase (TPS) and trehalase (TRE) directly regulate trehalose metabolism and indirectly regulate chitin metabolism in insects. Real-time quantitative PCR (RT-qPCR) and RNA interference (RNAi) were used to detect the expressions and functions of the ApTPS and ApTRE genes. Abnormal phenotypes were found after RNAi of ApTRE in the Acyrthosiphon pisum. The molting deformities were observed in two color morphs, while wing deformities were only observed in the red morphs. The RNAi of ApTPS significantly down-regulated the expression of chitin metabolism-related genes, UDP-N-acetyglucosamine pyrophosphorylase (ApUAP), chitin synthase 2 (Apchs-2), Chitinase 2, 5 (ApCht2, 5), endo-beta-N-acetylglucosaminidase (ApENGase) and chitin deacetylase (ApCDA) genes at 24 h and 48 h; The RNAi of ApTRE significantly down-regulated the expression of ApUAP, ApCht1, 2, 8 and ApCDA at 24 h and 48 h, and up-regulated the expression of glucose-6-phosphate isomerase (ApGPI) and Knickkopf protein (ApKNK) genes at 48 h. The RNAi of ApTRE and ApTPS not only altered the expression of chitin metabolism-related genes but also decreased the content of chitin. These results demonstrated that ApTPS and ApTRE can regulate the chitin metabolism, deepen our understanding of the biological functions, and provide a foundation for better understanding the molecular mechanism of insect metamorphosis.

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Trehalose and glucose levels regulate feeding behaviour in two color morphs of Acyrthosiphon pisum Harris

10.21203/rs.3.rs-198698/v1 ◽

2021 ◽

Author(s):

Guang Wang ◽

Jing-Jiang Zhou ◽

Yan Li ◽

Yuping Gou ◽

Peter Quandahor ◽

...

Keyword(s):

Rna Interference ◽

Feeding Behavior ◽

Nonlinear Regression ◽

Strong Evidence ◽

Growth And Development ◽

Feeding Behaviour ◽

Acyrthosiphon Pisum ◽

Glucose Levels ◽

Color Morphs ◽

Phosphate Synthase

Abstract Trehalose serves multifarious roles in growth and development in insects. We have previously shown that trehalose regulates Acyrthosiphon pisum chitin metabolism. Accordingly, we hypothesize here that trehalose-regulated A. pisum responses in chitin metabolism might also include trehalose-regulated feeding behaviour to involve in chitin metabolism. When RNA interference (RNAi) of trehalose-6-phosphate synthase gene increased the percentage of E2 (i.e. phloem ingestion) waveform and decreased the percentage of F (i.e. stylet work) and G (i.e. xylem ingestion) waveforms compared with the control A. pisum. RNAi of trehalase gene did not affect the percentage of each waveform compared with the control A. pisum. The high trehalose and glucose diets increased the percentage of E2 waveform of red A. pisum. The multiple nonlinear regression shown that the both low trehalose and glucose levels increased the percentage of np (i.e. non-probing), E1 (i.e. phloem salivation), and E2 waveforms. The high or low trehalose:glucose (T:G) ratio decreased the percentage of np, E1, and E2. Interestingly, the percentage of C (i.e. probing), F and, G waveforms were increased at low, low, and high T:G ratio, respectively. The results provided strong evidence that the trehalose and glucose levels regulate A. pisum feeding behavior.

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Functional Characterization of a Trehalose-6-Phosphate Synthase in Diaphorina citri Revealed by RNA Interference and Transcriptome Sequencing

Insects ◽

10.3390/insects12121074 ◽

2021 ◽

Vol 12 (12) ◽

pp. 1074

Author(s):

Jian-Chun Song ◽

Zhan-Jun Lu ◽

Long Yi ◽

Hai-Zhong Yu

Keyword(s):

Fatty Acid ◽

Rna Interference ◽

Functional Characterization ◽

Instar Nymph ◽

Amino Acid Residues ◽

Diaphorina Citri ◽

Real Time Quantitative Pcr ◽

Qpcr Analysis ◽

Phosphate Synthase

Trehalose-6-phosphate synthase (TPS) plays an important role in the synthesis of trehalose. In the current study, a TPS gene was obtained from Diaphorina citri, and named as DcTPS1 which encoded a protein of 833 amino acid residues. Real-time quantitative PCR (qPCR) analysis revealed that DcTPS1 had the highest expression level in the midgut and fifth-instar nymph stage. Knockdown of DcTPS1 by RNA interference (RNAi) induced an abnormal phenotype and increased mortality and malformation rate with a decreased molting rate. In addition, silencing of DcTPS1 significantly inhibited D. citri chitin metabolism and fatty acid metabolism, while the expression levels of fatty acid decomposition-related genes were downregulated. Furthermore, comparative transcriptomics analysis revealed that 791 differentially expressed genes (DEGs) were upregulated and 678 DEGs were downregulated when comparing dsDcTPS1 groups with dsGFP groups. Bioinformatics analysis showed that upregulated DEGs were mainly involved in oxidative phosphorylation, whereas downregulated DEGs were mainly attributed to the lysosome and ribosome. These results indicated that DcTPS1 played an important role in the growth and development of D. citri.

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