Comparative Proteomic Analysis between the Domesticated Silkworm (Bombyx mori) Reared on Fresh Mulberry Leaves and on Artificial Diet

Abstract

To gain an insight into the effects of different diets on growth and development of the domesticated silkworm at protein level, we employed comparative proteomic approach to investigate the proteomic differences of midgut, hemolymph, fat body and posterior silk gland of the silkworms reared on fresh mulberry leaves and on artificial diet. Seventy-six differentially expressed proteins were identified by MALDI TOF/TOF MS, and among them, 41 proteins were up-regulated, and 35 proteins were downregulated. Database searches, combined with GO analysis and KEGG pathway analysis revealed that some hemolymph proteins such as Nuecin, Gloverin-like proteins, PGRP, P50 and beta/-N-acetylglucosamidase were related to innate immunity of the silkworm, and some proteins identified in silkworm midgut including Myosin 1 light chain, Tropomyosin 1, Profilin, Serpin-2 and GSH-Px were involved in digestion and nutrition absorption. Moreover, two up-regulated enzymes in fat body of larvae reared on artificial diet were identified as V-ATPase subunit B and Arginine kinase which participate in energy metabolism. Furthermore, 6 down-regulated proteins identified in posterior silk gland of silkworm larvae reared on artificial diet including Ribosomal protein SA, EF-2, EF-1gamma, AspAT, ERp57 and PHB were related to silk synthesis. Our results suggested that the different diets could alter the expression of proteins related to immune system, digestion and absorption of nutrient, energy metabolism and silk synthesis poor nutrition and absorption of nutrition in silkworm. The results also confirmed that the poor nutrient absorption, weakened innate immunity, decreased energy metabolism and reduced silk synthesis are the main reasons for low cocoons yield, inferior filament quality, low survival rate of young larvae and insufficient resistance against specific pathogens in the silkworms fed on artificial diet.

Protein Patterns of Silkworm Bombyx mori L. Fed on Leaves of Wild and Cultivated Mulberry Varieties

“…One-dimensional sodium dodecyl sulfatepolyacrylamide electrophoresis (1D SDS-PAGE) were carried out according to (Zhou et al, 2008;Li et al, 2009) for haemolymph protein analysis, while the procedure established by Vasudha et al (2006) Qualitative analysis of proteins exhibited obvious variations in the number and position of bands between the larvae reared on each of the two investigated mulberry varieties. Most protein molecular weights were dominant in the two varieties except 91.23 and 72.92 kDa which appeared in M. alba var.…”

Proteomic analysis of <italic>Bm</italic>N cells (<italic>Bombyx mori</italic>) in response to infection with <italic>Nosema bombycis</italic>

“…Total protein extraction and quantification Each sample was homogenized in 200 ml of lysis buffer containing 8 M urea, 2 M thiourea, 4% (v/v) 3-[(3-Cholamidopropyl) dimethyl ammonio] propanesulfonate (CHAPS), 2% (v/v) immobilized pH gradient (IPG) buffer ( pH 3-10), and 0.1 M dithiothreitol for 10 min on ice and then kept for 10 min at room temperature, sonicated in an ice bath for 2 min, and centrifuged twice at 8000 g for 20 min at 188C [18]. The protein concentration in the supernatant was determined using BCA kit as described in manufacturer's instructions (SANGON, Shanghai, China).…”

The immune response of silkworm, Bombyx mori

“…Six belong to the short (S) subfamily . Comparative proteomic approach identified that PGRP was up-regulated when reared on fresh mulberry leaves when compared with on artificial diet (Zhou et al, 2008). B. mori GNBP was found to constitutively express in fat body and rapidly induced following a cuticular or hemoceolien bacterial challenge (Lee et al, 1996).…”