Exploring the cellular surface polysaccharide and root nodule symbiosis characteristics of the rpoN mutants of Bradyrhizobium sp. DOA9 using synchrotron-based Fourier transform infrared microspectroscopy in conjunction with X-ray absorption spectroscopy.

The functional significance of rpoN genes that encode two sigma factors in the Bradyrhizobium sp. strain DOA9 has been reported to affect colony formation, root nodulation characteristics, and symbiotic interactions with Aeschynomene americana. rpoN mutant strains are defective in cellular surface polysaccharide (CSP) production compared with the wild-type (WT) strain, and they accordingly exhibit smaller colonies and diminished symbiotic effectiveness. To gain deeper insights into the changes in CSP composition and the nodules of rpoN mutants, we employed synchrotron-based Fourier transform infrared (SR-FTIR) microspectroscopy and X-ray absorption spectroscopy. FTIR analysis of the CSP revealed the absence of specific components in the rpoN mutants, including lipids, carboxylic groups, polysaccharide-pyranose rings, and ß-galactopyranosyl residues. Nodules formed by DOA9WT exhibited a uniform distribution of lipids, proteins, and carbohydrates; mutant strains, particularly DOA9∆rpoNp:ΩrpoNc, exhibited decreased distribution uniformity and a lower concentration of C=O groups. Furthermore, Fe K-edge X-ray absorption near-edge structure and extended X-ray absorption fine structure analyses revealed deficiencies in the nitrogenase enzyme in the nodules of DOA9∆rpoNc and DOA9∆rpoNp:ΩrpoNc mutants; nodules from DOA9WT and DOA9∆rpoNp exhibited both leghemoglobin and the nitrogenase enzyme. IMPORTANCE This work provides valuable insights into how two rpoN genes affect the composition of cellular surface polysaccharides (CSPs) in Bradyrhizobium sp., which subsequently dictates root nodule chemical characteristics and nitrogenase production. We used advanced synchrotron methods, including synchrotron-based Fourier transform infrared (SR-FTIR) microspectroscopy and X-ray absorption spectroscopy (XAS), for the first time in this field to analyze CSP components and reveal the biochemical changes occurring within nodules. These cutting-edge techniques confer significant advantages by providing detailed molecular information, enabling the identification of specific functional groups, chemical bonds, and biomolecule changes. This research not only contributes to our understanding of plant-microbe interactions but also establishes a foundation for future investigations and potential applications in this field. The combined use of the synchrotron-based FTIR and XAS techniques represents a significant advancement in facilitating a comprehensive exploration of bacterial CSPs and their implications in plant-microbe interactions.

Molecular structure and linear-non linear rheology relation of rice starch during milky, dough, and mature stages.

Immature rice has potential to be used as healthy food. The relation between molecular structure and rheological properties was investigated. The lamellar repeating distance (8.42-8.63 nm) and crystalline thickness (4.60-4.72 nm) were not different among stages indicating a complete lamellar structure even at early stage. The relative crystallinity was higher in dough (39.62 %) than milky (36.69 %) and mature starch (35.22 %) caused by molecular structure, amylose, and amylose-lipid complex. The short amylopectin branched chains (A and B1) in dough starch were easily entangled resulted in higher Payne effect and elastic dominant. Dough starch paste exhibited higher G'Max (738 Pa) than milky (685 Pa) and mature (645 Pa) starch. In a non-linear viscoelastic regime, small strain hardening was found in milky and dough starch. Mature starch showed the highest plasticity and shear thinning at high-shear strains as the long-branched chains (B3) microstructure was disrupted, disentangled, followed by chain orientation along shear.

Development of a Sericin Hydrogel to Deliver Anthocyanins from Purple Waxy Corn Cob (Zea mays L.) Extract and In Vitro Evaluation of Anti-Inflammatory Effects.

Sericin-alginate hydrogel formulations with purple waxy corn (Zea mays L.) cob extract (PWCC) for topical anti-inflammatory application are developed and evaluated. The physical properties such as viscosity, pH, and anthocyanin release are examined and in vitro anti-inflammatory activities, such as NO inhibition and IL-6, IL-1ß, TNF-α, iNOS, and COX-2 expression, are evaluated in LPS-stimulated RAW 264.7 murine macrophages. The sericin-alginate hydrogel is prepared by physical crosslinking through the ionic interaction of the polymers combined with anthocyanin from PWCC at pH 6.5. The hydrogel formulation with 2.00% w/v sericin, 0.20% w/v alginate, and 0.15% w/v PWCC (SN6) shows a suitable viscosity for topical treatment, the highest nitric oxide inhibition (79.43%), no cytotoxicity, and reduced expression of IL-6, IL-1ß, and TNF-α mediators. Moreover, the SN6 formulation displays a sustained anthocyanin release over 8-12 h, which correlates with the Korsmeyer-Peppas model. The FT-IR spectrum of SN6 confirmed interaction of the sericin polymer with anthocyanins from PWCC via H-bonding by the shifted peak of amide I and amide III. In addition, the anthocyanin is stable in sericin hydrogels under heating-cooling storage conditions. Therefore, we suggest that this hydrogel formulation has potential as an anti-inflammatory agent. The formulation will be further investigated for in vivo studies and clinical trials in the future.

Retrogradation and Digestibility of Rice Starch Gels: The Joint Effect of Degree of Gelatinization and Storage.

Retrogradation affects acceptability of starchy foods; however, it is preferred in some products such as rice noodles. Amylose content, gelatinization temperature, and storage condition were reported to affect retrogradation but with disputed data. The joint effects of these parameters were interested in this study. Rice starch was gelatinized using different temperatures (77, 81, 95, and 121 °C) and stored isothermally with temperature cycles for 10 days. Results showed that the most important parameter that affected retrogradation was storage time followed by storage condition and gelatinization temperature. The recrystallization rate constant (k) and Avrami exponent (n) of retrograded starches stored under temperature cycle were higher than isothermal storage. All samples showed similar polymorphs of a mixture of B and V types. High-temperature gelatinized starch gel stored under temperature cycle condition produced higher yield of resistant starch. The study provided useful information on how to apply these parameters to control the retrogradation of starchy foods, especially rice noodle. PRACTICAL APPLICATION: Retrogradation is found to be more prominent at higher gelatinization temperature and longer storage time. Resistant starch produced from retrograded starch depended largely on storage time than storage condition. This finding can be applied to improve rice noodle qualities (by increasing retrogradation) with lower digestibility (by producing higher resistant starch).

Crystallization and chain reorganization of debranched rice starches in relation to resistant starch formation.

The effects of chain distribution, concentration, temperature and hydrothermal treatments on the recrystallization behavior and formation of resistant starch (RS) were investigated. Waxy and normal rice starches were debranched at 10 and 21% w/w solid concentrations, incubated at 25 or 50 °C, and further subjected to annealing or heat moisture treatment (HMT) to enhance RS formation. The crystallization at 25 °C favored the formation of the B-type structure, whereas crystallization at 50 °C led to the A-type structure with a higher melting temperature (100-120 °C) and a higher RS content (52%). All incubated samples showed an increase in RS content after subsequent hydrothermal treatments. The sample incubated at a high temperature contained the highest RS content (74.5%) after HMT with larger/perfect crystallites. These results suggested that the RS formation could be manipulated by crystallization conditions and improved by hydrothermal treatments which are dependent on the initial crystalline perfection.