Formate dehydrogenase in rice plant: growth stimulation effect of formate in rice plant

It was previously suggested that a population of phototrophic bacteria, Rhodopseudomonas palustris strain RN1 and Blastochloris sulfoviridis strain GN1 could be induced from granules in a lighted upflow anaerobic sludge blanket (LUASB) reactor. The present study showed that both strains RN1 and GN1 could use acetate, propionate, butyrate, and lactate as electron donors under anaerobic light conditions. The composition of organic acids in the effluent from the LUASB reactor was studied to investigate competitive consumption between acetogenic bacteria, methanogens, and phototrophic bacteria in the reactor. When acetate, propionate and lactate were supplied to the reactor, a small amount of acetate and propionate was observed in the effluent under light conditions. The concentrations of acetate and propionate increased under dark conditions compared with those under light conditions using organic acid and peptone media as the influent. When starch was supplied to the reactor, the concentrations of formate, acetate, propionate, butyrate, and lactate in the effluent were less than 0.5 mg C.l(-1) during operation under light and dark conditions. The concentrations of ammonium and phosphate in the effluent under dark conditions were higher than those under light conditions. These results suggested that phototrophic bacteria in the LUASB reactor consumed acetate and propionate as well as ammonium and phosphate in competition with methanogens and acetogenic bacteria.     

A novel gene delivery system in plants with calcium alginate micro-beads

We have produced micrometer-sized calcium alginate beads referred to as "bio-beads" that encapsulate plasmid DNA molecules carrying a reporter gene. In order to evaluate the efficiency of the bio-beads in mediating genetic transfection, protoplasts isolated from cultured tobacco cells (BY-2) were transfected with bio-beads containing a plasmid that carries the modified green fluorescent protein gene CaMV35S-sGFP. With the bio-beads treatment, approximately ten-fold higher GFP expression was observed after 24 h incubation compared to that with the conventional method using a naked plasmid solution. Transfection was up to 0.22% efficient. These results indicate that bio-beads have a possibility for efficient transformation in plants.     

Formate protects photosynthetic machinery from photoinhibition

In plants, environmental adversity often leads to the formation of reactive oxygen species (ROS) because of excess light energy. We found that incubation of rice leaves under high light intensity together with CO2-depleted air seriously damaged photosynthesis; however, the application of potassium formate (2 mM) before the photoinhibitory treatment protected the photosynthesis of the plant from photoinhibition. Further analysis revealed that formate protects PSII, RuBisCO, and FBPase from photoinhibition. Formate is thought to be involved in endogenous radical scavenging and/or in the supply of CO2, derived from the formate, thereby reducing oxidative damage to the photosystems under photoinhibitory conditions.     

The occurrence of geometric polyprenol isomers in the rubber-producing plant, Eucommia ulmoides Oliver

The chain length and geometric isomerism of polyprenols from Eucommia ulmoides Oliver were analyzed using supercritical fluid chromatography. After intensive effort to establish separation conditions for geometric isomers, a phenyl-bonded silica gel-packed column was found that cleanly separated poly-trans and-cis prenols. The presence of longchain poly-trans prenols (>9 mers) was confirmed for the first time in plants. Trans isomers were found in the leaf, seed coat, and root, but not in the bark and seed. Poly-trans prenols in this plant may act as intermediates for trans-polyisoprene biosynthesis.     

Formate dehydrogenase gene of Arabidopsis thaliana is induced by formaldehyde and not by formic acid

Variability of expression of formate dehydrogenase (FDH) caused by uptake of C-1 compounds was examined by using Arabidopsis thaliana as a model plant. Effects of uptake of several C-1 compounds were evaluated by Northern blot analysis using cDNA of A. thaliana FDH prepared by cloning on the basis of known sequence. As a result, expression of the FDH gene in A. thaliana was not intensely influenced by formic acid, an inherent substrate for FDH, but strongly induced by its reduced form, formaldehyde.     

Single cell-based analysis of torenia petal pigments by a combination of ArF excimer laser micro sampling and nano-high performance liquid chromatography (HPLC)-mass spectrometry

The molecular constituents of the petal pigments of the Torenia plant (Torenia hybrida) were analyzed on a single-cell basis by a combination of newly developed laser-microsampling and nano-flow liquid chromatography-electro spray ionization mass spectrometry (LC-ESIMS) techniques. Our method should provide a facile method for obtaining precise metabolic profiles of each cell in a single plant tissue.     

Metabolomics-based component profiling of hard and semi-hard natural cheeses with gas chromatography/time-of-flight-mass spectrometry, and its application to sensory predictive modeling

Gas chromatography/time-of-flight mass spectrometry (GC/TOF-MS) was used to analyze hydrophilic low molecular weight components, including amino acids, fatty acids, amines, organic acids, and saccharides, in cheese, and the sensometric application for practical metabolomic studies in the food industry is described. Derivatization of target analytes was conducted prior to the GC/TOF-MS analysis. Data on 13 cheeses, six Cheddar cheeses, six Gouda cheeses and one Parmigiano-Reggiano cheese, were analyzed by multivariate analysis. The uniqueness of the Parmigiano-Reggiano cheese metabolome was revealed. Principal component analysis (PCA) showed no grouping of the Cheddar cheeses and Gouda cheeses according to production method or country of origin. The PCA loading plot confirms that many amino acids contribute positively to PC1, suggesting that PC1 is closely related to degradation of proteins, and that lactic acid contributed positively to PC2, whereas glycerol contributed negatively to PC2, suggesting that factors regarding degradation of carbohydrates and fats were expressed in PC2. Partial least squares (PLS) regression models were constructed to predict the relationship between the metabolite profile and two sensory attributes, "Rich flavor" and "Sour flavor", which were related to maturation. The compounds that play an important role in constructing each sensory prediction model were identified as 12 amino acids and lactose for "Rich flavor", and 4-aminobutyric acid, ornithine, succinic acid, lactic acid, proline and lactose for "Sour flavor". The present study revealed that metabolomics-based component profiling, focusing on hydrophilic low molecular weight components, was able to predict the sensory characteristics related to ripening.     

Gallic acid oxidizes Met residues in peptides released from bovine β-lactoglobulin by in vitro digestion

Phenolic compounds (PCs) are frequently present in foods. However, little is known about the effect of PCs on enzymatic digestion process of food proteins and their products. In this study, the effect of gallic acid (GA) on in vitro digestion of β-lactoglobulin (β-LG) was investigated as a model system for analysis of the interaction between PCs and food proteins. GA showed no effect on the initial rate of β-LG digestion. However, after 1.5 h of digestion, the observed degree of hydrolysis of β-LG was lower in the presence than in the absence of GA. The peptides released from β-LG were characterized by LC/IT-TOF-MS and thirty peptides were identified. In particular, four new peaks were obtained following in vitro digestion of β-LG in the presence of GA. Met(7), Met(24) and Met(145) in the peptides corresponding to these peaks were oxidized to methionine sulfoxide residues.     

Application of electrospray ionization ion trap/time-of-flight mass spectrometry for chemically-synthesized small RNAs

In this study, we have demonstrated an accurate and rapid small RNA analytical method with both sequence determination and detailed modification analysis by electrospray ionization-ion trap/time-of-flight mass spectrometry (ESI-IT/TOFMS). To develop this ideal method, we have examined the performance of ESI-IT/TOFMS using various chemically-synthesized model sequences of modified or unmodified microRNAs (miRNAs). The deconvoluted mass of a 22-nucleotide (nt) miRNA was obtained from a multiply charged precursor ion (MS(1)). The ion exhibited high mass accuracy (< 7 ppm) and high mass resolution (a value of m/Δm=10,000) and was therefore very useful in RNA composition assignment. The optimized MS(2) method using ion trap collision-induced dissociation, as well as automatic annotation analysis of product ions based on the accurate mass information, enabled the precise sequencing determination of intact miRNAs. Further, the detailed structural analysis of 3'-terminal modified nucleic acid in intact methylated miRNA was carried out using the MS(3) capability of the hybrid IT/TOFMS. The direct infusion method also provided a high throughput and good sensitivity because the analytical time and sample concentration needed in a series of experiments with reliable data were only 3 min and 100 nM, respectively. This study provides a novel approach for characterizing the intact chemically-synthesized small RNA without chemical and enzymatic digestions and would be widely applicable for the structural analysis of complicated modified small RNAs.