Degradation of aromatic compounds in plants grown under aseptic conditions

The aim of the work is to investigate the ability of higher plants to absorb and detoxify environmental pollutants - aromatic compounds via aromatic ring cleavage. Transformation of 14C specifically labelled benzene derivatives, [1-6-14C]-nitrobenzene, [1-6-(14)C]-aniline, [1-(14)C]- and [7-(14)C]-benzoic acid, in axenic seedlings of maize (Zea mays L.), kidney bean (Phaseolus vulgaris L.), pea (Pisum sativum L.) and pumpkin (Cucurbita pepo L.) were studied. After penetration in plants, the above xenobiotics are transformed by oxidative or reductive reactions, conjugation with cell endogenous compounds, and binding to biopolymers. The initial stage of oxidative degradation consists in hydroxylation reactions. The aromatic ring can then be cleaved and degraded into organic acids of the Krebs cycle. Ring cleavage is accompanied by 14CO2 evolution. Aromatic ring cleavage in plants has thus been demonstrated for different xenobiotics carrying different substitutions on their benzene ring. Conjugation with low molecular peptides is the main pathway of aromatic xenobiotics detoxification. Peptide conjugates are formed both by the initial xenobiotics (except nitrobenzene) and by intermediate transformation products. The chemical nature of the radioactive fragment and the amino acid composition of peptides participating in conjugation were identified.     

Molecular and mineral responses of corals grown under artificial Calcite Sea conditions

The formation of skeletal structures composed of different calcium carbonate polymorphs (e.g. aragonite and calcite) appears to be both biologically and environmentally regulated. Among environmental factors influencing aragonite and calcite precipitation, changes in seawater conditions—primarily in the molar ratio of magnesium and calcium during so-called ‘Calcite’ (mMg:mCa below 2) or ‘Aragonite’ seas (mMg:mCa above 2)—have had profound impacts on the distribution and performance of marine calcifiers throughout Earth's history. Nonetheless, the fossil record shows that some species appear to have counteracted such changes and kept their skeleton polymorph unaltered. Here, the aragonitic octocoral Heliopora coerulea and the aragonitic scleractinian Montipora digitata were exposed to Calcite Sea-like mMg:mCa with various levels of magnesium and calcium concentration, and changes in both the mineralogy (i.e. CaCO₃ polymorph) and gene expression were monitored. Both species maintained aragonite deposition at lower mMg:mCa ratios, while concurrent calcite presence was only detected in M. digitata. Despite a strong variability between independent experimental replicates for both species, the expression for a set of putative calcification-related genes, including known components of the M. digitata skeleton organic matrix (SkOM), was found to consistently change at lower mMg:mCa. These results support the previously proposed involvements of the SkOM in counteracting decreases in seawater mMg:mCa. Although no consistent expression changes in calcium and magnesium transporters were observed, down-regulation calcium channels in H. coerulea in one experimental replicate and at an mMg:mCa of 2.5, pointing to a possible active calcium uptake regulation by the corals under altered mMg:mCa.     

Proteolytic enzymes and trypsin inhibitors of higher plants under stress conditions

The response of the components of a protease-inhibitor system of legume and cereal crops to stress factors was studied. It was found that salinization, heavy metal ions, and phytopathogenic flora inhibit the activity of neutral, acidic, and alkaline proteases at early stages of seed germination, the degree of the inhibition of the endoprotease activity being dependent on the index of tolerance of legume and cereal crops. It was shown that, in response to unfavorable conditions, accumulation of trypsin inhibitors occurs, which is accompanied by the appearance of new protein components, as indicated by electrophoresis. The results confirm the presumption that serine protease inhibitors are involved in the response of plants to stress factors.     

Proteolytic enzymes and trypsin inhibitors of higher plants under stress conditions

The response of the components of a protease-inhibitor system of legume and cereal crops to stress factors was studied. It was found that salinization, heavy metal ions, and phytopathogenic flora inhibit the activity of neutral, acidic, and alkaline proteases at early stages of seed germination, the degree of the inhibition of the endoprotease activity being dependent on the index of tolerance of legume and cereal crops. It was shown that, in response to unfavorable conditions, accumulation of trypsin inhibitors occurs, which is accompanied by the appearance of new protein components, as indicated by electrophoresis. The results confirm the presumption that serine protease inhibitors are involved in the response of plants to stress factors.     

Effects of nitrogen limitation on the growth and composition of unicellular algae in continuous culture

Since fat accumulation takes place in many algae as a response to exhaustion of the nitrogen supply, it has been suggested that this may provide a means of enhancing the potential food value of algae. To test this possibility, chemostatic continuous cultures of Chlorella sorokiniana and Oocystis polymorpha were subjected to successive reductions in influent nitrogen. As cellular nitrogen content decreased from about 10 to 4%, oxygen evolution, carbon dioxide uptake, chlorophyll content, and tissue production were drastically reduced, but total lipid content was essentially unchanged. Caloric values and C, H, and N analyses suggested a moderate increase in carbohydrate content, but gas chromatographic analyses revealed no significant qualitative or quantitative changes in the fatty acid fraction. In batch-cultured cells, nitrogen could be reduced to 3% of dry weight, causing a concomitant increase in total fatty acids and pronounced changes in the composition of the fatty acid fraction. These results suggest that cellular nitrogen must fall to approximately 3% of dry weight before appreciable increases in lipid synthesis can occur. Cell nitrogen is then apparently completely bound in essential cell constituents, and carbon subsequently fixed is converted into lipid products. The findings indicate that nitrogen limitation may be useful in increasing the food quality of batch-cultured cells, but the technique has little value for continuous culture systems per se.     

Fatty acid composition of Spirulina strains grown under various environmental conditions

The fatty acid distribution in 19 strains of Spirulina was studied. All but one contained γ-linolenic acid (GLA). No GLA was found in S. subsalsa, which had a very high content of palmitoleic acid. The fatty acid content of all but one of the tested strains increased with cultivation temperature and the relative amount of polyunsaturated fatty acid decreased. The highest content of GLA was found at 30–35° for most strains. High light intensities at a high temperature (38°), while not affecting the fatty acid composition, had a drastic effect on the fatty acid content, reducing it by as much as 46 %.     

几种高等水生植物的克藻效应研究

研究了五种高等水生植物(黑藻、金鱼藻、水花生、茭白、空心菜)对蛋白核小球藻、斜生栅藻生长的抑制作用。试验结果显示,各种受试植物均有不同程度的克藻效果,金鱼藻与黑藻的克藻作用最强,水花生的作用次之。克藻效果以受试植物与藻共培养最佳,其次为连续添加种植水,表明在植物的生长过程中其化感物质是连续释放且能迅速降解的,而黑藻干物质的浸提液所具有的克藻效应说明有些化感物质对一定温度和光照是较稳定的。     

Comparative chemical composition of the Barents Sea brown algae

Comparative study of phytochemical compositions of the most widespread brown algae species (one laminarian and four fucoid algae) from Barents Sea has been performed. A modified technique for mannitol determination in brown algae is proposed. It was revealed that fucus algae (fam. Fucaceae) contain 3% (of total dry weight) less mannitol than laminaria (Laminaria saccharina). The contents of alginic acid and laminaran in the Barents Sea fucoids are more than 10% less compared to laminaria. The alga L. saccharina contains almost two times more iodine than the species of fam. Fucaceae. The amounts of fucoidan and sum lipids in the Barents Sea fucoid algae is higher than in Laminaria saccharina (4-7% and 1-3%, respectively). In terms of contents of main biologically active compounds, fucus and laminarian algae from Barents Sea are inferior to none of the Far-Eastern species. The Barents Sea algae may become an important source of biologically active compounds.     

Changes in the acyl lipid composition of photosynthetic bacteria grown under photosynthetic and non-photosynthetic conditions

The acyl lipids and their constituent fatty acids were studied in the photosynthetic bacteria Rhodospirillum rubrum, Rhodopseudomonas capsulata and Rhodopseudomonas sphaeroides, which were grown under photosynthetic and non-photosynthetic conditions. The major lipids were found to be phosphatidylethanolamine, phosphatidylglycerol and cardiolipin in each bacterium. The two Rhodopseudomonas species also contained significant quantities of phosphatidylcholine. Other acyl lipids accounted for less than 10% of the total. On changing growth conditions from non-photosynthetic to photosynthetic a large increase in the relative proportion of phosphatidylglycerol was seen at the expense of phosphatidyl-ethanolamine. In Rhodospirillum rubrum the fatty acids of the major phospholipids showed an increase in the proportion of palmitate and stearate and a decrease in palmitoleate and vaccenate on changing growth conditions to photosynthetic. In contrast, the exceptionally high levels (>80%) of vaccenate in individual phospholipids of Rhodopseudomonas capsulata and Rhodopseudomonas sphaeroides were unaffected by changing growth conditions to photosynthetic. Analysis of the lipids of chromatophores, isolated from the three bacteria, showed that these preparations were enriched in phosphatidylglycerol. The large increase in this phospholipid, seen during growth under photosynthetic conditions, appeared, therefore, to be due to a proliferation of chromatophore membranes. Possible roles for acyl lipids in the formation and function of the photosynthetic apparatus of bacteria are discussed.     

Comparative chemical composition of the Barents Sea brown algae

Comparative study of phytochemical compositions of the most widespread brown algae species (one laminarian and four fucoid algae) from Barents Sea has been performed. A modified technique for mannitol determination in brown algae is proposed. It was revealed that fucus algae (fam. Fucaceae) contain 3% (of total dry weight) less mannitol than laminaria (Laminaria saccharina). The contents of alginic acid and laminaran in the Barents Sea fucoids are more than 10% less compared to laminaria. The alga L. saccharina contains almost two times more iodine than the species of fam. Fucaceae. The amounts of fucoidan and sum lipids in the Barents Sea fucoid algae is higher than in Laminaria saccharina (4–7% and 1–3%, respectively). In terms of contents of main biologically active compounds, fucus and laminarian algae from Barents Sea are inferior to none of the Far-Eastern species. The Barents Sea algae may become an important source of biologically active compounds.     

Comparative proteomic studies in Rhodospirillum rubrum grown under different nitrogen conditions

Forty-four differentially expressed proteins have been identified in the photosynthetic diazotroph Rhodospirillum rubrum grown anaerobic and photoheterotrophically, with different nitrogen sources, using 2D-PAGE and MALDI-TOF, from gels containing an average of 679 +/- 52 (in N(+)) and 619 +/- 37 (in N(-)) protein spots for each gel. A higher level of expression was found under nitrogen-rich growth, for proteins involved in carbon metabolism (reductive tricarboxylic acid cycle, CO(2) fixation, and poly-beta-hydroxybutyrate metabolism) and amino acid metabolism. The key enzymes RuBisCO and alpha-ketoglutarate synthase were found to be present in higher amounts in nitrogen-rich conditions. Ntr and Nif regulated proteins, such as glutamine synthetase and nitrogenase, were, as expected, induced under nitrogen-fixing conditions and glutamate dehydrogenase was down regulated. A novel 2Fe-2S ferredoxin with unknown function was identified from nitrogen-fixing cultures. In addition to differential expression, two of the identified proteins revealed variable p I values in response to the nitrogen source used.     

Comparative analysis of plastocyanin–cytochrome f complex formation in higher plants,green algae and cyanobacteria

Mechanisms of the complex formation between plastocyanin and cytochrome f in higher plants (Spinacia oleracea and Brassica rapa), green microalgae Chlamydomonas reinhardtii and two species of cyanobacteria (Phormidium laminosum and Nostoc sp.) were investigated using combined Brownian and molecular dynamics simulations and hierarchical cluster analysis. In higher plants and green algae, electrostatic interactions force plastocyanin molecule close to the heme of cytochrome f. In the subsequent rotation of plastocyanin molecule around the point of electrostatic contact in the vicinity of cytochrome f, copper (Cu) atom approaches cytochrome heme forming a stable configuration where cytochrome f molecule behaves as a rather rigid body without conformational changes. In Nostoc plastocyanin molecule approaches cytochrome f in a different orientation (head‐on) where the stabilization of the plastocyanin–cytochrome f complex is accompanied by the conformational changes of the G188E189D190 loop that stabilizes the whole complex. In cyanobacterium P. laminosum, electrostatic preorientation of the approaching molecules was not detected, thus indicating that random motions rather than long‐range electrostatic interactions are responsible for the proper mutual orientation. We demonstrated that despite the structural similarity of the investigated electron transport proteins in different photosynthetic organisms, the complexity of molecular mechanisms of the complex formation increases in the following sequence: non‐heterocystous cyanobacteria – heterocystous cyanobacteria – green algae – flowering plants.