Multimodal Spectroscopic Studies to Evaluate the Effect of Nod-Factor-Based Fertilizer on the Maize (Zea mays) Stem.

Maize (Zea mays) is one of the most cultivated plants in the world. Due to the large area, the scale of its production, and the demand to increase the yield, there is a need for new environmentally friendly fertilizers. One group of such candidates is bacteria-produced nodulation (or nod) factors. Limited research has explored the impact of nodulation, factors on maize within field conditions, with most studies restricted to greenhouse settings and early developmental stages. Additionally, there is a scarcity of investigations that elucidate the metabolic alterations in the maize stem due to nod-factor exposure. It was therefore the aim of this study. Maize stem's metabolites and fibers were analyzed with various imaging analytical techniques: matrix assisted laser desorption ionization-mass spectrometry imaging (MALDI-MSI), Raman spectroscopy, attenuated total reflection Fourier transform infrared spectroscopy (ATR FT-IR), and diffuse reflectance infrared Fourier transform spectroscopy. Moreover, the biochemical analyses were used to evaluate the proteins and soluble carbohydrates concentration and total phenolic content. These techniques were used to evaluate the influence of nod factor-based biofertilizer on the growth of a non-symbiotic plant, maize. The biofertilizer increased the grain yield and the stem mass. Moreover, the spectroscopic and biochemical investigation proved the appreciable biochemical changes in the stems of the maize in biofertilizer-treated plants. Noticeable changes were found in the spatial distribution and the increase in the concentration of flavonoids such as maysin, quercetin, and rutin. Moreover, the concentration of cell wall components (fibers) increased. Furthermore, it was shown that the use of untargeted analyses (such as Raman and ATR FT-IR, spectroscopic imaging, and MALDI-MSI) is useful for the investigation of the biochemical changes in plants.

MALDI MSI and Raman Spectroscopy Application in the Analysis of the Structural Components and Flavonoids in Brassica napus Stem.

Nod factors among the signaling molecules produced by rhizobia in response to flavonoids to induce root nodule formation in the legumes. It is, however, hypothesized that they might increase the yield and positively impact the growth of non-legumes. To evaluate this statement, rapeseed treated with Nod factor-based biofertilizers were cultivated, their stems was collected, and the metabolic changes were investigated using Raman spectroscopy and MALDI mass spectrometry imaging. Biofertilizer proved to increase the concentration of lignin in the cortex, as well as hemicellulose, pectin, and cellulose in the pith. Moreover, the concentration of quercetin derivatives and kaempferol derivatives increased, while the concentration of isorhamnetin dihexoside decreased. The increase in the concentration of the structural components in the stem might therefore increase the lodging resistance, while the increase in concentration of the flavonoids might increase their resistance to fungal infection and herbivorous insects.

Application of Vibrational Spectroscopic Techniques in the Study of the Natural Polysaccharides and Their Cross-Linking Process.

Polysaccharides are one of the most abundant natural polymers and their molecular structure influences many crucial characteristics-inter alia hydrophobicity, mechanical, and physicochemical properties. Vibrational spectroscopic techniques, such as infrared (IR) and Raman spectroscopies are excellent tools to study their arrangement during polymerization and cross-linking processes. This review paper summarizes the application of the above-mentioned analytical methods to track the structure of natural polysaccharides, such as cellulose, hemicellulose, glucan, starch, chitosan, dextran, and their derivatives, which affects their industrial and medical use.

Spectroscopic Evaluation of the Potential Neurotoxic Effects of a New Candidate for Anti-Seizure Medication-TP-315 during Chronic Administration (In Vivo).

The aim of this study was to investigate the potential neurotoxic effect of the new anti-seizure medication candidate-5-(3-chlorophenyl)-4-hexyl-2,4-dihydro-3H-1,2,4-triazole-3-thione (TP-315), after chronic administration to mice. TP-315 was administered to mice intraperitoneally for 14 days. At 24 h post the last injection, animals were decapitated, their brains were acquired, flash-frozen in liquid nitrogen and cut into 10 µm slices. The FT-IR chemical imaging technique was used for the investigation of the potential neurotoxic effect in the cerebral cortex and hippocampus. The effect on the lipidomic and proteomic profile and on oxidative stress was investigated. The results showed no statistically significant changes in the above-mentioned parameters. TP-315 seems to pose no neurotoxic effect on the mouse brain after chronic use, therefore, its use should be safe.

Multimodal Spectroscopic Imaging of Pea Root Nodules to Assess the Nitrogen Fixation in the Presence of Biofertilizer Based on Nod-Factors.

Multimodal spectroscopic imaging methods such as Matrix Assisted Laser Desorption/Ionization Mass Spectrometry Imaging (MALDI MSI), Fourier Transform Infrared spectroscopy (FT-IR) and Raman spectroscopy were used to monitor the changes in distribution and to determine semi quantitatively selected metabolites involved in nitrogen fixation in pea root nodules. These approaches were used to evaluate the effectiveness of nitrogen fixation by pea plants treated with biofertilizer preparations containing Nod factors. To assess the effectiveness of biofertilizer, the fresh and dry masses of plants were determined. The biofertilizer was shown to be effective in enhancing the growth of the pea plants. In case of metabolic changes, the biofertilizer caused a change in the apparent distribution of the leghaemoglobin from the edges of the nodule to its centre (the active zone of nodule). Moreover, the enhanced nitrogen fixation and presumably the accelerated maturation form of the nodules were observed with the use of a biofertilizer.

Biological activity of Nod factors.

Chemically, the Nod factors (NFs) are lipochitooligosaccharides, produced mainly by bacteria of the Rhizobium genus. They are the main signaling molecules involved in the initiation of symbiosis between rhizobia and legume plants. Nod factors affect plant tissues at very low concentrations, even as low as 10-12 mol/L. They induce root hair deformation, cortical cell division, and root nodules' formation in the host plant. At the molecular level, the cytoskeleton is reorganized and expression of genes encoding proteins called nodulins is induced in response to Nod factors in the cell. Action of Nod factors is highly specific because it depends on the structure of a particular Nod factor involved, as well as the plant receptor reacting with it.

Recent developments of MALDI MSI application in plant tissues analysis.

Mass spectrometry imaging (MSI) combined with matrix-assisted laser desorption/ionization (MALDI) is an efficient technology applied in plant metabolomics research. This technique allows for visualization of spatial distribution of metabolites such as: lipids, proteins, peptides and DNA sequences, by determining the x, y coordinates of the compounds exactly in plant tissue. Simplicity of the tissue preparation without the need of prior exact knowledge about the analytes is a great advantage of this method. In this review, we provide an overview of experimental workflow including sample preparation, data acquisition and analysis, methodology, and some recent applications of MALDI MS imaging in plant metabolomics research.

The FT-IR and Raman Spectroscopies as Tools for Biofilm Characterization Created by Cariogenic Streptococci.

Fourier transform infrared (FT-IR) and Raman spectroscopy and mapping were applied to the analysis of biofilms produced by bacteria of the genus Streptococcus. Bacterial biofilm, also called dental plaque, is the main cause of periodontal disease and tooth decay. It consists of a complex microbial community embedded in an extracellular matrix composed of highly hydrated extracellular polymeric substances and is a combination of salivary and bacterial proteins, lipids, polysaccharides, nucleic acids, and inorganic ions. This study confirms the value of Raman and FT-IR spectroscopies in biology, medicine, and pharmacy as effective tools for bacterial product characterization.