Metabolomic Analysis of Combretum lanceolatum Plants Interaction with Diaporthe phaseolorum and Trichoderma spirale Endophytic Fungi through ¹H‐NMR

Abstract

Endophytic fungi are an important class of microorganisms, able to interact with a host plant via a mutualistic mechanism without visible symptoms of the fungal colonization. The synergy between endophytic fungi and their host plant can promote morphological, physiological and biochemical changes through the expression of bioactive metabolites. This work aims to correlate metabolic changes in the Combretum lanceolatum plant metabolome with its endophytic fungi Diaporthe phaseolorum (Dp) and Trichoderma spirale (Ts), and to discover corresponding metabolite‐biomarkers, with the principal focus being on its primary metabolism. The 1H‐NMR metabolomic analysis of qualitative and quantitative changes was performed through multivariate statistical analysis and the identification of primary metabolites was achieved on the Madison Metabolomics Consortium Database. The presence of Dp significantly impacted the plant's metabolic pathways, improving the biosynthesis of primary metabolites such as threonine, malic acid and N‐acetyl‐mannosamine, which are precursors of special metabolites involved in plant self‐defence. This work represents a valuable contribution to advanced studies on the metabolic profiles of the interaction of plants with endophytes.

Metabolomics Analysis of C. lanceolatum Roots in the Presence of Its Endophytic Fungi

“…23 The C. lanceolatum seeds disinfection, germination and cultivation as well as the endophytic fungi Dp and Ts inoculation were performed as previously described. 22 The experiment was divided into three fungal interaction times, immediately after inoculation (0 days), after 1 day and 7 days. For each fungal interaction time, the uninoculated C. lanceolatum plant was used as a negative control.…”

“…Furthermore, the fungal association induces different metabolomic variations towards the plant aerial parts and roots, since the interaction of Dp fungus with C. lanceolatum plants significantly affected the metabolic profile, increasing the primary metabolites such as threonine, malic acid and N-acetyl-mannosamine. 22 Based on these results, the Ts fungus interaction represents a more relevant influence on the biosynthesis of primary compounds in the plant roots, improving the plant's development and secondary metabolism pathways.…”

“…The 1 H NMR spectra were acquired at 25 ºC with 128 transients of 64 K data points, 90° pulse angle, 4.35 s acquisition time and 4 s recycle delay. 22,26 Using a general procedure, 27 1 H NMR spectra pre-processing and multivariate analyses of data for metabolite identification were performed with a few modifications. The spectra were phased and the baseline were corrected using MestReNova software.…”

“…[18][19][20] Although C. lanceolatum plant has been studied regarding its bioactivity properties, only a little attention has been given towards its phytochemical investigation in the literature. 21,22 In a previous work, 22 our research group described the metabolomic changes in the aerial parts of C. lanceolatum plants inoculated with its endophytic fungi Diaporthe phaseolorum (Dp) and Trichoderma spirale (Ts). Accordingly, the study demonstrated that Dp fungus has significantly affected the plant aerial parts metabolic pathways, improving the biosynthesis of primary metabolites such as threonine, malic acid and N-acetyl-mannosamine.…”

“…Accordingly, the study demonstrated that Dp fungus has significantly affected the plant aerial parts metabolic pathways, improving the biosynthesis of primary metabolites such as threonine, malic acid and N-acetyl-mannosamine. 22 Based on those results, the aim of our current work is to analyze the effects of Dp and Ts fungi on the chemical composition of C. lanceolatum roots in order to extend the metabolomics knowledge over the symbiotic interaction between these species.…”