Formed of proteins, glycoproteins, and chitin microfibrils in a proteoglycan matrix, the peritrophic matrix (PM) separates the food from the midgut epithelium in most but not all insects. A PM occurs in two forms. A type I PM is delaminated from the entire midgut epithelium and, in some cases, may only be formed in response to feeding and the type of meal ingested. A type II PM is produced by a specialized region of the anterior midgut called the cardia and forms a continuous sleeve (or sleeves) that is always present. As it is positioned between food and midgut epithelium, the PM plays key roles in the intestinal biology of the insect. The PM may protect the midgut epithelium from mechanical damage and insult from pathogens and toxins; it must act as a semipermeable membrane regulating passage of molecules between the different midgut compartments; and it may separate the midgut lumen into different, physiologically significant compartments.

Peritrophic matrix structure and function.

Peritrophic matrix proteins.

Chitin is a major component of fungal walls and insect exoskeletons. Plants produce chitinases upon pathogen attack and chito-oligomers induce defense responses in plants, though the exact mechanism behind this response is unknown. Using the ATH1 Affymetrix microarrays consisting of about 23,000 genes, we examined the response of Arabidopsis (Arabidopsis thaliana) seedlings to chito-octamers and hydrolyzed chitin after 30 min of treatment. The expression patterns elicited by the chito-octamer and hydrolyzed chitin were similar. Microarray expression profiles for several genes were verified via northern analysis or quantitative reverse transcription-PCR. We characterized T-DNA insertion mutants for nine chito-oligomer responsive genes. Three of the mutants were more susceptible to the fungal pathogen, powdery mildew, than wild type as measured by conidiophore production. These three mutants included mutants of genes for two disease resistance-like proteins and a putative E3 ligase. The isolation of loss-of-function mutants with enhanced disease susceptibility provides direct evidence that the chito-octamer is an important oligosaccharide elicitor of plant defenses. Also, this study demonstrates the value of microarray data for identifying new components of uncharacterized signaling pathways.

Loss-of-function mutations in chitin responsive genes show increased susceptibility to the powdery mildew pathogen Erysiphe cichoracearum.

Peritrophic membranes (PMs) are an invertebrate-unique structure that lines the digestive tract, playing important roles in facilitating food digestion and providing protection to the gut epithelium. The importance of PMs in insects has been recognized ever since its presence was identified 200 years ago. In the last 5 years, significant progress towards understanding the PM molecular structure and the mechanism for PM formation has been made. Recent studies on Type 1 PMs from lepidopteran larvae have suggested a model for the PM molecular structure and formation. The important physiological functions of the PM suggest that PMs can be a significant structural target for insect control and the current understanding of the structure of lepidopteran larval PMs has provided us with potential opportunities for targeting the PM by various mechanisms. Arch. Insect Biochem. Physiol. 47:110–118, 2001. © 2001 Wiley-Liss, Inc.

Molecular structure of the peritrophic membrane (PM): identification of potential PM target sites for insect control.

A transient luminal chitinous matrix is required to model epithelial tube diameter in the Drosophila trachea.

The lectin wheat germ agglutinin (WGA) has a binding site which is able to bind a sequence of three N-acetyl-glucosamine residues. Therefore, it has a very strong affinity for the polymers of this sugar, especially chitin. Colloidal gold can be labelled with WGA and used as a specific electron-dense marker for the electron-microseopic localization of chitin. The specificity of the WGA-gold binding can be checked by competitive inhibition with 5–10 mM triacetyl chitotriose. The reliability of this method was tested in three species. In the formation zone of the radula of the snail, Biomphalaria glabrata Say, chitin or chitin precursors were localized in vesicles of the odontoblasts, outside the extremely long microvilli of odontoblasts and in the newly formed teeth. The inner peritrophic envelope of the earwig, Forficula auricularia L., is characterized by an orthognal texture of bundles of microfibrils that are thought to contain chitin. The pesence of chitin was proved using the present method. In the peritrophic membranes of the blowfly, Calliphora erythrocephala Meigen, it was possible to differentiate between chitin and glycoproteins which have N-acetylglucosamine residues.

I. Latka

Papers

Electron microscopic localization of chitin using colloidal gold labelled with wheat germ agglutinin.