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Insect Sci. 2013 Feb;20(1):109-19. doi: 10.1111/j.1744-7917.2012.01573.x. Epub 2012 Nov 22.

RNA interference to reveal roles of β-N-acetylglucosaminidase gene during molting process in Locusta migratoria.

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  • 1Research Institute of Applied Biology, Shanxi University, Taiyuan.

Abstract

β-N-acetylglucosaminidases are crucial enzymes involved in chitin degradation in insects. We identified a β-N-acetylglucosaminidase gene (LmNAG1) from Locusta migratoria. The full-length complementary DNA (cDNA) of LmNAG1 consists of 2 667 nucleotides, including an open reading frame (ORF) of 1 845 nucleotides encoding 614 amino acid residues, and 233- and 589-nucleotide non-coding regions at the 5'- and 3'-ends, respectively. Phylogenetic analysis grouped the cDNA-deduced LmNAG1 protein with the enzymatically characterized β-N-acetylglucosaminidases in group I. Analyses of stage- and tissue-dependent expression patterns of LmNAG1 were carried out by real-time quantitative polymerase chain reaction. Our results showed that LmNAG1 transcript level in the integument was significantly high in the last 2 days of the fourth and fifth instar nymphs. LmNAG1 was highly expressed in foregut and hindgut. RNA interference of LmNAG1 resulted in an effective silence of the gene and a significantly reduced total LmNAG enzyme activity at 48 and 72 h after the injection of LmNAG1 double-stranded RNA (dsRNA). As compared with the control nymphs injected with GFP dsRNA, 50% of the dsLmNAG1-injected nymphs were not able to molt successfully and eventually died. Our results suggest that LmNAG1 plays an essential role in molting process of L. migratoria.

© 2012 Institute of Zoology, Chinese Academy of Sciences.

KEYWORDS:

Locusta migratoria; RNA interference; chitin degradation; β-N-acetylglucosaminidase

PMID:
23955831
[PubMed - indexed for MEDLINE]
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