Mulberry leaves have been selected as a food source for the silkworm (Bombyx mori) for over 5000 years. However, the interaction mechanisms of mulberry-silkworm remain largely unknown. We explore the interaction between mulberry and silkworm at the protein level. Total proteins were extracted from mulberry leaves and silkworm feces on day 5 of the fifth larval instar and analyzed on shotgun liquid chromatography-tandem mass spectrometry, respectively. In total, 2076 and 210 foliar proteins were identified from mulberry leaves and silkworm feces, respectively. These proteins were classified into four categories according to their subcellular location: chloroplast proteins, mitochondrial proteins, secretory-pathway proteins, and proteins of other locations. Chloroplast proteins accounted for 68.3% in mulberry leaves but only 23.2% in the feces. In contrast, secretory-pathway proteins had low abundance in mulberry leaves (7.3%) but were greatly enriched to the largest component in the feces (60.1%). Most of the foliar secretory-pathway proteins in the feces were found to be resistant to silkworm feeding by becoming involved in primary metabolite, proteinase inhibition, cell-wall remodeling, redox regulation, and pathogen-resistant processes. On the contrary, only six defensive proteins were identified in the fecal chloroplast proteins including two key proteins responsible for synthesizing jasmonic acid, although chloroplast proteins were the second largest component in the feces. Collectively, the comparative proteomics analyses indicate that mulberry leaves not only provide amino acids to the silkworm but also display defense against silkworm feeding, although the silkworm grows very well by feeding on mulberry leaves, which provides new insights into the interactions between host-plant and insect herbivores.
Keywords: defensive activities; mulberry leaves; proteomics; silkworm feces.