Lysozyme of the beet armyworm, Spodoptera exigua: activity induction and cDNA structure

Lysozyme of the beet armyworm, Spodoptera exigua, was characterized in its up-regulation pattern, and its cDNA was cloned by RT-PCR using degenerate primers designed from some conserved amino acid regions shared with related lepidopteran species. Lysozyme activity of the non-immunized S. exigua had developmental variation, with the highest level in the fifth instar larvae. The basal level of the lysozyme activity was significantly enhanced by the injection of laminarin or lipopolysaccharide (LPS). Among different LPSs tested, the extract from an entomopathogenic bacterium, Xenorhabdus nematophilus, proved to be the most potent. Fat body was the major tissue to express the lysozyme in S. exigua. Even though there was a significantly elevated level of lysozyme in the hemolymph at 12 h after laminarin injection, the transcript (∼1.1 kbp) was found in the fat body as early as 6 h after injection. The cDNA of the lysozyme was cloned as 602 bp with a deduced 141-amino-acid residue open reading frame containing two introns. Except for a signal peptide with 20 amino acid residues, the estimated molecular weight and isoelectric point of the lysozyme was 14 313.83 Da and 8.59, respectively. Only a single copy gene of the lysozyme was found in S. exigua genome from Southern analysis. The amino acid sequence of S. exigua lysozyme showed higher similarity (88.7%) with noctuid species compared to other lepidopteran species.