Metabolic Analysis of the Development of the Plant-Parasitic Cyst Nematodes Heterodera schachtii and Heterodera trifolii by Capillary Electrophoresis Time-of-Flight Mass Spectrometry

Abstract

The cyst nematodes Heterodera schachtii and Heterodera trifolii, whose major hosts are sugar beet and clover, respectively, damage a broad range of plants, resulting in significant economic losses. Nematodes synthesize metabolites for organismal development and social communication. We performed metabolic profiling of H. schachtii and H. trifolii in the egg, juvenile 2 (J2), and female stages. In all, 392 peaks were analyzed by capillary electrophoresis time-of-flight mass spectrometry, which revealed a lot of similarities among metabolomes. Aromatic amino acid metabolism, carbohydrate metabolism, choline metabolism, methionine salvage pathway, glutamate metabolism, urea cycle, glycolysis, gluconeogenesis, coenzyme metabolism, purine metabolism, pyrimidine metabolism, and tricarboxylic acid (TCA) cycle for energy conversion (β-oxidation and branched-chain amino acid metabolism) energy storage were involved in all stages studied. The egg and female stages synthesized higher levels of metabolites compared to the J2 stage. The key metabolites detected were glycerol, guanosine, hydroxyproline, citric acid, phosphorylcholine, and the essential amino acids Phe, Leu, Ser, and Val. Metabolites, such as hydroxyproline, acetylcholine, serotonin, glutathione, and glutathione disulfide, which are associated with growth and reproduction, mobility, and neurotransmission, predominated in the J2 stage. Other metabolites, such as SAM, 3PSer, 3-ureidopropionic acid, CTP, UDP, UTP, 3-hydroxy-3-methylglutaric acid, 2-amino-2-(hydroxymethyl-1,3-propanediol, 2-hydroxy-4-methylvaleric acid, Gly Asp, glucuronic acid-3 + galacturonic acid-3 Ser-Glu, citrulline, and γ-Glu-Asn, were highly detected in the egg stage. Meanwhile, nicotinamide, 3-PG, F6P, Cys, ADP-Ribose, Ru5P, S7P, IMP, DAP, diethanolamine, p-Hydroxybenzoic acid, and γ-Glu-Arg_divalent were unique to the J2 stage. Formiminoglutamic acid, nicotinaminde riboside + XC0089, putrescine, thiamine 2,3-dihydroxybenzoic acid, 3-methyladenine, caffeic acid, ferulic acid, m-hydrobenzoic acid, o- and p-coumaric acid, and shikimic acid were specific to the female stage. Overall, highly similar identities and quantities of metabolites between the corresponding stages of the two species of nematode were observed. Our results will be a valuable resource for further studies of physiological changes related to the development of nematodes and nematode–plant interactions.

An improved medium for in vitro studies of female reproduction and oviposition in Schistosoma japonicum

“…Six supplements, including l -asparagine, l -hydroxyproline, para -aminobenzoic acid ( p ABA), vitamin B12, glutathione, and casein hydrolysate, were absent from AB169 (Additional file 9 : Table S3). Previous studies have shown that l -hydroxyproline and glutathione are associated with nematode growth and reproduction, mobility, and neurotransmission [ 45 ]. As a component essential for the synthesis of folate, p ABA is required for the development of parasites, such as Brugia malayi larvae and malaria [ 46 , 47 ].…”

Capillary Electrophoresis: A Three-Year Literature Review

“…It should be noted that MS detection is usually crucial in metabolite analysis as many metabolites are not detectable optically without prior derivatization. Several CE-TOF methods have been successfully developed for various applications such as studying the metabolic profiling of cyst nematodes, investigating the metabolomic characteristics of macrophages infected with Leishmania amazonensis , exploring novel putative biomarkers of longevity in Caenorhabditis elegans, and examining skeletal muscle metabolites in lipoprotein lipase transgenic rabbits . Recent studies have also contributed to elucidating the metabolic pathways in human bodies associated with specific food intake and their connections to particular health conditions. , …”