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Different functions of the insect soluble and membrane-bound trehalase genes in chitin biosynthesis revealed by RNA interference
- Source :
- PLoS ONE, PLoS ONE, Vol 5, Iss 4, p e10133 (2010)
- Publication Year :
- 2010
-
Abstract
- BACKGROUND: Trehalase, an enzyme that hydrolyzes trehalose to yield two glucose molecules, plays a pivotal role in various physiological processes. In recent years, trehalase proteins have been purified from several insect species and are divided into soluble (Tre-1) and membrane-bound (Tre-2) trehalases. However, no functions of the two trehalases in chitin biosynthesis in insects have yet been reported. PRINCIPAL FINDINGS: The membrane-bound trehalase of Spodoptera exigua (SeTre-2) was characterized in our laboratory previously. In this study, we cloned the soluble trehalase gene (SeTre-1) and investigated the tissue distribution and developmental expression pattern of the two trehalase genes. SeTre-1 was expressed highly in cuticle and Malpighian tubules, while SeTre-2 was expressed in tracheae and fat body. In the midgut, the two trehalase genes were expressed in different locations. Additionally, the expression profiles of both trehalase mRNAs and their enzyme activities suggest that they may play different roles in chitin biosynthesis. The RNA interference (RNAi) of either SeTre-1 or SeTre-2 was gene-specific and effective, with efficiency rates up to 83% at 72 h post injection. After RNAi of SeTre-1 and SeTre-2, significant higher mortality rates were observed during the larva-pupa stage and pupa-adult stage, and the lethal phenotypes were classified and analyzed. Additionally, the change trends of concentration of trehalose and glucose appeared reciprocally in RNAi-mutants. Moreover, knockdown of SeTre-1 gene largely inhibited the expression of chitin synthase gene A (CHSA) and reduced the chitin content in the cuticle to two-thirds relative to the control insects. The chitin synthase gene B (CHSB) expression, however, was inhibited more by the injection of dsRNA for SeTre-2, and the chitin content in the midgut decreased by about 25%. CONCLUSIONS: SeTre-1 plays a major role in CHSA expression and chitin synthesis in the cuticle, and SeTre-2 has an important role in CHSB expression and chitin synthesis in the midgut.
- Subjects :
- Insecta
lcsh:Medicine
Chitin
Spodoptera
Biochemistry
chemistry.chemical_compound
Biosynthesis
Molecular Biology/Translational Regulation
Gene expression
Animals
Tissue Distribution
Trehalase
RNA, Small Interfering
lcsh:Science
Molecular Biology
chemistry.chemical_classification
Chitin Synthase
Multidisciplinary
biology
Gene Expression Profiling
lcsh:R
fungi
Chitin synthase
Trehalose
Enzyme assay
Enzyme
chemistry
Solubility
Molecular Biology/Post-Translational Regulation of Gene Expression
biology.protein
lcsh:Q
RNA Interference
Biochemistry/Transcription and Translation
Research Article
Developmental Biology
Subjects
Details
- ISSN :
- 19326203
- Volume :
- 5
- Issue :
- 4
- Database :
- OpenAIRE
- Journal :
- PloS one
- Accession number :
- edsair.doi.dedup.....8ab2de45502f809701f24c96361ac04e