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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.

Different Functions of the Insect Soluble and Membrane-Bound Trehalase Genes in Chitin Biosynthesis Revealed by RNA Interference