Livia Simon-Sarkadi

Determination of biogenic amines by capillary electrophoresis

A method for determining biogenic amines in food using micellar electrokinetic capillary chromatography has been developed. Derivatization of the amines was performed with AccQ (6-aminoquinolyl-N-hydroxysuccinimidyl carbamate; Waters, Milford, MA, USA) reagent. The influence of buffer composition on the separation (including pH, SDS concentration and various additives) was investigated. The separation of seven biogenic amines (histamine, tyramine, tryptamine, spermine, spermidine, cadaverine and putrescine) could be achieved within 25-30 min with good repeatability. The biogenic amine profiles in three different food samples (wine, salami and chive) were determined and quantitated.

Stress-induced changes in the free amino acid composition in transgenic soybean plants having increased proline content

Following drought stress at supraoptimal temperature the increase in proline (Pro) content in transgenic (T) soybean [Glycine max (L.) Merr. cv. Ibis] plants overexpressing the gene coding for the last enzyme of Pro biosynthesis, L-Δ1-pyrroline-5-carboxylate reductase, was much greater than in wild type (W) plants (105-fold versus 19-fold after 7 d). Under control conditions arginine accounted for nearly 60 % of the total free amino acid content. After stress treatment the content of Pro was more than 50 % in both T and W genotypes, and at the end of recovery the γ-aminobutyrate content reached 27 and 53 % in the W and T plants, respectively. Without stress treatment there was only a 2-fold difference between T and W in the tyrosine content. However, during the stress period and the subsequent recovery a similar difference was found for many amino acids. The present results indicate that manipulating of the content of a single amino acid influences the whole free amino acid composition in soybean.

Differential effects of cold acclimation and abscisic acid on free amino acid composition in wheat

The effect of cold acclimation and abscisic acid (ABA) treatment on the free amino acid composition was compared in Chinese Spring chromosome 5A substitution lines with different levels of freezing tolerance. The total amino acid content gradually increased during the 3-week cold acclimation period, while the effect of ABA became visible only after 7 d. The ratio of members of the glutamate family increased during cold acclimation and the ratio of amino acids belonging to the aspartate family decreased. Opposite changes were observed after treatment with ABA. Consistently with these results, ABA only induced a major increase in the Asn content, while the Asp, Glu, Gln and Pro levels were greatly induced by cold. A corresponding alteration at the gene expression level was only found for Pro and Glu. With the exception of Pro, cold- or ABA-induced changes in the amino acid levelsor Pro, did not correlate with the freezing tolerance of the three genotypes examined and were not affected by chromosome 5A. Since cold acclimation induced the accumulation of most of the amino acids, while ABA had a significant effect only on Asn, the cold-induced changes in free amino acid levels were probably not mediated by ABA.

Effect of cadmium and titanium-ascorbate stress on biological active compounds in wheat seedlings

ABSTRACT The effect of cadmium (Cd) and combined Cd and titanium (Ti)-ascorbate was examined on wheat seedlings (Triticum aestivum L.). The one-week-old hydroponically-grown seedlings of wheat were exposed to Cd2+(10−7 M, 10−3 M) and Cd2+(10−7 M, 10−3 M)+Ti-ascorbate (5 μg L−1) stresses. The stress effects were followed by changes in free amino acid and polyamine contents and the peroxidase enzyme activity in roots and shoots. The results showed significant differences between the two plant organs (root and shoot) with respect to the biochemical response to the applied stress. Titanium-ascorbate treatment seemed to reduce the negative effects of heavy metal damage in wheat seedlings.

Different Accumulation of Free Amino Acids during Short- and Long-Term Osmotic Stress in Wheat

The effect of wheat chromosome 5A on free amino acid accumulation induced by osmotic stress was compared in chromosome 5A substitution lines with different freezing tolerance. Treatment with 15% polyethylene glycol (PEG) resulted in greater total free amino acid content even after 3 days compared to the controls. The ratio of amino acids belonging to various amino acid families differed after 3-week treatment in the control and PEG-treated plants only in the case of the freezing-sensitive substitution line. There was a transient increase with a maximum after 3 days in the amounts of several amino acids, after which their concentrations exhibited a more gradual increase. During the first days of osmotic stress the Glu, Gln, Asp, Asn, Thr, Ser, Leu, and His concentrations were greater in the tolerant substitution line than in the sensitive one, while the opposite relationship was observed at the end of the PEG treatment. The coordinated changes in the levels of individual amino acids indicated that they are involved in both the short- and long-term responses to osmotic stress. The alterations differed in the two chromosome 5A substitution lines, depending on the stress tolerance of the chromosome donor genotype.

Effect of Drought Stress at Supraoptimal Temperature on Polyamine Concentrations in Transgenic Soybean with Increased Proline Levels

Abstract The effect of drought stress at supraoptimal temperature on free proline and polyamine levels was compared in wild type and transgenic soybean (Glycine max cv. Ibis) plants having increased proline levels. Since glutamate and arginine are precursors of both proline and polyamines, it was assumed that the genetic manipulation of proline levels would also affect the polyamine levels. The proline and spermine concentrations increased, while the putrescine concentration generally decreased or did not change after the treatments in both genotypes. Following drought higher proline and lower spermine levels were detected in the transgenic plants compared to the wild type ones, which could be explained by the increased use of their common precursors for proline biosynthesis in the transgenic plants.

Nitric oxide affects salt-induced changes in free amino acid levels in maize

It was assumed that salt-induced redox changes affect amino acid metabolism in maize (Zea mays L.), and this influence may be modified by NO. The applied NaCl treatment reduced the fresh weight of shoots and roots. This decrease was smaller after the combined application of NaCl and an NO-donor ((Z)-1-[N-(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate, DETA/NO) in the shoots, while it was greater after simultaneous treatment with NaCl and nitro-L-arginine (L-NNA, inhibitor of NO synthesis) in the roots. The quantum yield efficiency of photosystem II was not influenced by the treatments. NaCl had a significant effect on the redox environment in the leaves as it was shown by the increase in the amount of glutathione disulphide and in the redox potential of the glutathione/glutathione disulphide redox pair. This influence of NaCl was modified by DETA/NO and L-NNA. Pharmacological modification of NO levels affected salt-induced changes in both the total free amino acid content and in the free amino acid composition. NaCl alone increased the concentration of almost all amino acids which effect was strengthened by DETA/NO in the case of Pro. L-NNA treatment resulted in a significant increase in the Ala, Val, Gly and Tyr contents. The Ile, Lys and Val concentrations rose considerably after the combined application of NaCl and DETA/NO compared to NaCl treatment alone in the recovery phase. NaCl also increased the expression of several genes related to the amino acid and antioxidant metabolism, and this effect was modified by DETA/NO. In conclusion, modification of NO levels affected salt-induced, glutathione-dependent redox changes and simultaneously the free amino acid composition and the level of several free amino acids. The observed much higher Pro content in plants treated with both NaCl and DETA/NO during recovery may contribute to the protective effect of NO against salt stress.

Modification of cadaverine content by NO in salt-stressed maize

NO has an important role in the control of plant development, growth, and the response to abiotic stress. In our recent paper it was demonstrated that NO affected the salt-induced changes in free amino acid levels in maize.1 Since polyamines are synthesized from lysine and arginine, it was supposed that their concentrations are also influenced by NO. Cadaverine levels were increased by a NO donor and decreased by an inhibitor of NO synthesis in salt-stressed maize. These findings indicate that NO participates in the mediation of the effect of salt on cadaverine content. The coordinated changes in the NO and cadaverine levels may be involved in regulating of the response to salt stress in maize.

Comparison of protein quality and mineral element concentrations in grain of spelt (Triticum spelta L.) and common wheat (Triticum aestivum L.)

The cultivation of Triticum spelta (spelt) has no tradition in Hungary. In recent years the interest towards this old species renewed in many countries. This high‐nutritional cereal, which has a high ash and fibre content, can be used in many health‐oriented grain‐based food products. Therefore, field experiments have been conducted for some years to test the performance of this species under home growing conditions. Here we report the results of analyses for some important quality parameters of grain samples from the 1996/97 season in comparison with those of older and new home‐grown bread wheat cultivars. Three common wheat cultivars and one advanced spelt line were grown on small plots fertilised with an NPK dose necessary to reach the highest yield and quality. Spikes were sampled weekly from the time of 70–77% grain moisture to full ripening. The grains were analysed for ash, N, P and K content and amino acid composition. Concentrations of 16 other macro + micro elements and in the ripe grains, baking quality parameters were also assessed. The grain development of spelt showed a remarkable time‐lag compared to that of the common wheat cultivars. However, the highest thousand‐grain‐masses, ash, N, and P concentrations were measured in this cultivar after milk ripening. The grains of spelt contained the macro‐nutrient Mg and four micro‐nutrients (Zn, Mn, Fe, Cu) in higher concentrations compared to those of the common wheat varieties. The total and essential amino acid concentrations measured in the ripe grains of spelt were also remarkably higher. Although its wet gluten content (47.5%) was considerably higher than that of the bread wheat cultivars, its breadmaking quality was poor.

OPLC Analysis of Polyamines in Wheat Seedlings under Cadmium Stress

Two wheat cultivars (Triticum aestivum L. cv. Chinese Spring and Cappelle Desprez) differing in drought stress tolerance were treated with two concentrations of cadmium (10-7 and 10-3 M) to investigate whether there is also any difference in their heavy-metal tolerance. Overpressured layer chromatography (OPLC) with off-line fluorescence detection was used to monitor cadmium stress-induced changes in the polyamine content of wheat seedlings. Significant differences were found between the spermidine (Spd) and putrescine (Put) content of the two wheat cultivars. The Spd content of the moderately drought-tolerant Chinese Spring increased both in shoots and roots at the higher Cd concentration compared with the control whereas in the drought-sensitive Cappelle Desprez the Spd level was reduced by both Cd treatments. Put content increased with the amount of Cd to which the roots of Chinese Spring wheat were exposed. In the Cappelle Desprez cultivar the Put content of both organs was higher at both Cd concentrations compared with the control.