Gas Chromatography-Mass Spectrometry Based Midgut Metabolomics Reveals the Metabolic Perturbations under NaF Stress in Bombyx mori

Abstract

Fluoride tolerance is an important economic trait in sericulture, especially in some industrial development regions. Analyses of physiological changes involving structural damage to the insect body and molecular analyses of some related genes have focused on this area; however, the changes that occur at the metabolic level of silkworms after eating fluoride-contaminated mulberry leaves remain unclear. Here, metabonomic analysis was conducted using gas chromatography-mass spectrometry (GC-MS) to analyze the changes in midgut tissue after NaF stress using silkworm strains 733xin (susceptible stain) and T6 (strain resistant to fluoride), which were previously reported by our laboratory. Differential metabolomics analysis showed that both T6 and 733xin strains displayed complex responses after exposure to 200 mg/kg NaF. The purine metabolism and arginine and proline metabolic pathways of fluoride-tolerant strains reached significant levels, among which 3′-adenylic acid and hypoxanthine were significantly upregulated, whereas guanine, allantoic acid, xanthine, N-acetyl-L-glutamic acid, and pyruvate were significantly downregulated. These metabolic pathways may be related to the fluoride tolerance mechanism of NaF poisoning and tolerant strains.

Metabolic Characterisation of the Midgut of Bombyx mori Varieties after BmNPV Infection Using GC-MS-Based Metabolite Profiling

“…GC-MS analysis was conducted with a 7890-5977A GC/MSD (Agilent Technologies, Inc., CA, USA). The chromatographic conditions followed the methods of Li et al [ 13 ] as follows: DB-5MS capillary column (30 m × 0.25 mm × 0.25 um, Agilent J&W Science, Folsom, CA, USA); carrier gas was high purity helium (purity not less than 99.999%); flow rate was 1.0 mL/min; and temperature of the inlet was 260 °C. The injection volume was 1 μL not diverted and the solvent was delayed for 5 min.…”

“…Therefore, the midgut is the first barrier against the BmNPV in silkworms, and conducting a metabonomic study of the midgut tissues could reveal the silkworm’s anti-BmNPV mechanisms. With the advent of the current era of big data, the research and analysis of genomes [ 10 ], transcriptomes [ 11 ], proteomes [ 12 ] and metabonomes [ 13 ] of highly resistant and sensitive strains of silkworms will facilitate the analysis of antiviral mechanisms in silkworms. Recently, many investigators have integrated metabonomics and transcriptomics to study metabolic pathways, disease resistance, and stress resistance mechanisms of organisms under threat [ 14 , 15 ].…”