The Effect of Combined Elicitation with Light and Temperature on the Chlorogenic Acid Content, Total Phenolic Content and Antioxidant Activity of Berula erecta in Tissue Culture.

Chlorogenic acid is one of the most prominent bioactive phenolic acids with great pharmacological, cosmetic and nutritional value. The potential of Berula erecta in tissue culture was investigated for the production of chlorogenic acid and its elicitation combined with light of different wavelengths and low temperature. The content of chlorogenic acid in the samples was determined by HPLC-UV, while the content of total phenolic compounds and the antioxidant activity of their ethanol extracts were evaluated spectrophotometrically. The highest fresh and dry biomasses were obtained in plants grown at 23 °C. This is the first study in which chlorogenic acid has been identified and quantified in Berula erecta. The highest chlorogenic acid content was 4.049 mg/g DW. It was determined in a culture grown for 28 days after the beginning of the experiment at 12 °C and under blue light. The latter also contained the highest content of total phenolic compounds, and its extracts showed the highest antioxidant activity. Berula erecta could, potentially, be suitable for the in vitro production of chlorogenic acid, although many other studies should be conducted before implementation on an industrial scale.

Use of Emulsion-Templated, Highly Porous Polyelectrolytes for In Vitro Germination of Chickpea Embryos: a New Substrate for Soilless Cultivation.

The application of highly porous and 3D interconnected microcellular polyelectrolyte polyHIPE (PE-PH) monoliths based on (3-acrylamidopropyl)-trimethylammonium chloride as soilless cultivation substrates for in vitro embryo culture is discussed. The embryo axes isolated from chickpea seeds are inoculated onto the surface of the monoliths and allowed to germinate. Germination study show that the newly disclosed PE-PH substrate performs much better than the conventionally used agar as the germination percentage, shoot and root length, fresh and dry weight as well as the number of leaves are enhanced. The PE-PHs exhibit a higher absorption capacity of the plant growth medium, that is, 36 g·g-1 compared to agar, that is, 20 g·g-1, and also survive autoclaving conditions without failing. The key advantage over standard agar substrates is that they can be reused several times and also without prior sterilization. These results suggest that PE-PHs with exceptional absorption/retention properties and robustness have great potential as soilless substrates for in vitro plant cultivation.

Diversity and Content of Carotenoids and Other Pigments in the Transition from the Green to the Red Stage of Haematococcus pluvialis Microalgae Identified by HPLC-DAD and LC-QTOF-MS.

H. pluvialis is a unicellular freshwater alga containing many bioactive compounds, especially carotenoids, which are the strongest antioxidants among the pigments. This study evaluates the composition and content of carotenoids and other pigments in both stages of algae life cycle, especially in the green vegetative stage, less studied in comparison to the red stage. To determine the composition and content of carotenoids, a combination of HPLC-DAD and LC-QTOF-MS was used. The content of carotenoids in the green vegetative stage was significantly lower than in the red vegetative stage. In the green vegetative stage, 16 different carotenoids and other pigments were identified. Among the total 8.86 mg g-1 DW of pigments, 5.24 mg g-1 DW or 59% of them were chlorophyll a with its derivatives, and 3.62 mg g-1 DW or 41% of them were free carotenoids. After the transition from the green to the red stage, the carotenoid composition was replaced by secondary carotenoids, astaxanthin and its esters, which predominated in the whole carotenoid composition. In addition to free astaxanthin, 12 astaxanthin monoesters, 6 diesters and 13 other carotenoids were determined. The majority of 37.86 mg g-1 DW pigments were monoesters. They represented 82% of all pigments, and their content was about 5 times higher than both, diesters (5.91 mg g-1 DW or 12% of all) and free carotenoids (2.4 mg g-1 DW or 6% of all). The results of the study contribute to the data on the overall pigment composition and content of H. pluvialis algae and provide the basis for further improvement of cultivation of the H. pluvialis algae.

Identification and Content of Astaxanthin and Its Esters from Microalgae Haematococcus pluvialis by HPLC-DAD and LC-QTOF-MS after Extraction with Various Solvents.

Haematococcus pluvialis, a unicellular green microalga that produces a secondary metabolite under stress conditions, bears one of the most potent antioxidants, namely xanthophyll astaxanthin. The aim of our study was to determine the content of astaxanthin and its esterified forms using three different solvents-methyl tert-butyl ether (MTBE), hexane isopropanol (HEX -IPA) and acetone (ACE)-and to identify them by using high performance liquid chromatography coupled with diode array detection and the quadrupole time-of-flight mass spectrometry (HPLC-DAD and LC-QTOF-MS) technique. We identified eleven astaxanthin monoesters, which accounted for 78.8% of the total astaxanthin pool, six astaxanthin diesters (20.5% of total), while free astaxanthin represented the smallest fraction (0.7%). Astaxanthin monoesters (C16:2, C16:1, C16:0), which were the major bioactive compounds in the H. pluvialis samples studied, ranged from 10.2 to 11.8 mg g-1 DW. Astaxanthin diesters (C18:4/C18:3, C18:1/C18:3) were detected in the range between 2.3 and 2.6 mg g-1 DW. All three solvents were found to be effective for extraction, but MTBE and hexane-isopropanol extracted the greatest amount of free bioactive astaxanthin. Furthermore, MTBE extracted more low-chain astaxanthin monoesters (C16), and hexane-isopropanol extracted more long-chain monoesters (C18 and above) and more diesters. We can conclude that MTBE is the solvent of choice for the extraction of monoesters and hexane-isopropanol for diesters.

Response of Berula erecta to Lead in Combination with Selenium.

Tissue culture of Berula erecta is suitable cultivation system for research purposes connected with contamination and phytoremediation studies. In previous investigation we determined the optimal dose concentration at which Se stimulates plant growth and positively affects the antioxidative status in this experimental system. In current study, we investigate its response to exposure to lead (Pb) and further the possible protective effect of Se(IV) against Pb exposure. Plants were grown in 10 and 50 mg Pb L-1 solution without and with added Se (0.1 mg L-1) for six weeks. Plants possessed a high affinity to uptake Pb and Se in roots. Addition of Pb inhibited roots elongations and the plant height. In contrast, the combined effect of Se + Pb treatment was reflected in increased weight of plants when compared to Pb treatment alone. Pb decreased the amount of chlorophylls and consequently photochemical efficiency was lowered, whereas in Pb + Se treatment the photochemical efficiency was higher. Furthermore, Pb treatment caused a gradual increase in glutathione in both roots and shoots, however, to a greater percentage in shoots when compared to controls. Exposure to both Pb and Se did not cause any significant changes in root's glutathione level when compared to Pb treatment alone. In shoots, the combined treatment lowered the glutathione significantly, but the levels remained 50% above those of untreated control samples, reflecting that this might be related with the antioxidative effects of Se treatment.

Knowledge about and acceptance of genetically modified organisms among pre-service teachers: a comparative study of turkey and Slovenia

Genetically modified organisms cannot be regarded as merely a topic for academic debate, since these have serious implications as a research field and for production based on genetic engineering. Public debates rarely base their arguments on elements rooted in scientific arguments and knowledge but are heavily loaded with emotions, opinions and informal reasoning. This study aimed to investigate the knowledge and acceptance of genetically modified organisms among prospective teachers in Slovenia and Turkey. Knowledge of genetic modification was measured with a two-tier instrument. The level of acceptance of genetic modification was measured with a 17-item instrument. Findings revealed that knowledge of genetics and biotechnology barely influenced the acceptability of genetic modification, and correlations are low. The relationship between knowledge and acceptance was not significant among Slovenian students and while significant for the Turkish or combined groups, the r values were only 0.179 and 0.244. It was found that differences in the acceptability of clusters of different kinds of genetically modified organisms do exist between the two countries. In both countries, participants recognized microorganisms and plants that produce something useful as the most acceptable organisms, while at the other end were animals used for consumption or as donors of organs. Practical implications for teaching are discussed and implications for further studies are drawn.

Knowlege of, attitudes toward, and acceptance of genetically modified organisms among prospective teachers of biology, home economics, and grade school in Slovenia.

The objective of this study was to investigate knowledge, opinions, and attitudes toward, as well as readiness to accept genetically modified organisms (GMOs) among prospective primary and secondary Slovene teachers. Our findings are that prospective teachers want to take an active role in rejecting or supporting individual GMOs and are aware of the importance of education about genetically modified organism (GMO) items and their potential significance for society. Through cluster analysis, we recognized four clusters of GMOs, separated by degree of genetically modified acceptability. GM plants and microorganisms which are recognized as useful are accepted. They are undecided about organisms used in research or medicine and reject organisms used for food consumption and for fun. There are only weak correlations between knowledge and attitudes and knowledge and acceptance of GMOs, and a strong correlation between attitudes and acceptance. The appropriate strategies and actions for improving university courses in biotechnology are discussed.