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

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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Identification of Halloween Genes and RNA Interference-Mediated Functional Characterization of a Halloween Gene shadow in Plutella xylostella

Frontiers in Physiology ◽

10.3389/fphys.2019.01120 ◽

2019 ◽

Vol 10 ◽

Cited By ~ 5

Author(s):

Lu Peng ◽

Lei Wang ◽

Ming-Min Zou ◽

Liette Vasseur ◽

Li-Na Chu ◽

...

Keyword(s):

Rna Interference ◽

Plutella Xylostella ◽

Functional Characterization

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Paving the way for transgenic schistosomes

Parasitology ◽

10.1017/s0031182011001466 ◽

2011 ◽

Vol 139 (5) ◽

pp. 651-668 ◽

Cited By ~ 21

Author(s):

S. BECKMANN ◽

C. G. GREVELDING

Keyword(s):

Rna Interference ◽

Initial Phase ◽

Functional Characterization ◽

Reporter Genes ◽

Significant Progress ◽

Gene Insertion ◽

Remarkable Progress ◽

Made In ◽

Silence Gene

SUMMARYIn parasitological research, significant progress has been made with respect to genomics and transcriptomics but transgenic systems for functional gene analyses are mainly restricted to the protozoan field. Gene insertion and knockout strategies can be applied to parasitic protozoa as well as gene silencing by RNA interference (RNAi). By contrast, research on parasitic helminthes still lags behind. Along with the major advances in genome and transcriptome analyses e.g. for schistosomes, methods for the functional characterization of genes of interest are still in their initial phase and have to be elaborated now, at the beginning of the post-genomic era. In this review we will summarize attempts made in the last decade regarding the establishment of protocols to transiently and stably transform or transfect schistosomes. Besides approaches using particle bombardment, electroporation or virus-based infection strateies to introduce DNA constructs into adult and larval schistosome stages to express reporter genes, first approaches have also been made in establishing protocols based on soaking, lipofection, and/or electroporation for RNA interference to silence gene activity. Although in these cases remarkable progress can be seen, the schistosome community eagerly awaits major breakthroughs especially with respect to stable transformation, but also for silencing or knock-down strategies for every schistosome gene of interest.

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