PH-DEPENDENCE OF CHLORTETRACYCLINE(CTC)-INDUCED PLUG FORMATION IN NITELLA FLEXILIS (CHARACEAE)1

The formation of chlortetracycline(CTC)-induced wall appositions (callose plugs) in Nitella flexilis (L.)Ag. was pH-dependent in the range between 4.3-8.3. Plug number and plug diameter increased with the pH of the CTC solution. At pH 4.3 plug formation was light-dependent and occurred below the alkaline regions of the cell surface which form during photo synthetic assimilation of HCO₃^?. Inhibition of photosynthesis by 3–(3′,4′-dichlorophenyl)-1, 1-dimethylurea prevented plug formation in the light. Dark-treated cells could be induced to form plugs by raising the pH of the CTC solution. The formation of large but incomplete plugs in the presence of cytochalasin B is explained by the formation of numerous weak alkaline sites. I suggest that CTC enhances locally the Ca²⁺content at the cytoplasm near the plasmamembrane. The ionophoric character of CTC is probably more pronounced at high pH mainly because of a weaker binding with cations and a closer contact with the membrane.     

Lignin genetic engineering

Although lignins play important roles in plants, they often represent an obstacle to the utilization of plant biomass in different areas: pulp industry, forage digestibility. The recent characterization of different lignification genes has stimulated research programmes aimed at modifying the lignin profiles of plants through genetic engineering (antisense and sense suppression of gene expression). The first transgenic plants with a modification of monomeric composition of lignins and lignin content have been recently obtained. Down regulation of the OMT gene induces dramatic reduction of syringyl units. CAD down regulated plants exhibit a unusual red phenotype associated with the developing xylem and several chemical modifications of their lignins including an increase in cinnamaldehydes in the polymer structure. Interestingly this novel lignin is removed more easily during the pulping process. In both OMT and CAD down regulated plants no changes in phenotypic characteristics such as growth architecture and morphology were observed. More recent experiments have shown that a reduction of CCR activity determines specific changes in the coloration of the xylem area suggesting significant chemical modifications which are currently being studied.These different results show that it is possible to manipulate lignins through targeted genetic transformation of plants and that lignins exhibit a relative flexibility of their chemical structure. Future developments should probe the impact of down regulating the expression of other recently characterized lignification genes such as F5H and CCoAOMT and also of a combination of genes in order to tailor lignins more adapted to specific purposes. In addition to biotechnological applications which should provide important economical benefits for utilization of wood in the pulp industry, genetic engineering of lignins offer very promising perspectives for the understanding of lignin synthesis, structure and properties.     

Cadmium replaces calcium in the cell wall ofUlva lactuca

Electron microscopy, in conjunction with X-ray microanalysis, was used to investigate the effects of exposure to cadmium on the elemental composition of the macroalgaUlva lactuca. The cell wall was the only region of the cell to show any marked change in chemical composition as a result of exposure to cadmium, with less calcium evident in cadmium-treated thallus compared with untreated thalli. The cell wall ofU. lactuca is a complex structure made up of polysaccharides consisting of many-branched chains composed mostly of rhamnose and galactose subunits. Some of the hydroxyl groups on the subunits are substituted by sulphate groups. Borate is associated with the rhamnose subunits, which contain no sulphate groups, and calcium binds to borate, cross-linking the rhamnose groups. The borate-calcium complex adds rigidity to the cell wall; the replacement of calcium by cadmium will, therefore, influence the rigidity of the thallus. The ecological significance of this work is discussed with respect to the ability of the alga to withstand grazing or emersion.     

Pollen allergens: development and function

Pollen allergens interact with the human immune system and the resulting IgE antibodies provide specific probes for their identification and characterisation. In one case, grass allergenic proteins are expressed late in pollen development coincident with the laying down of reserves. Sequence similarity of allergens has indicated possible functions for some allergens. The major birch pollen allergen shows sequence similarity with pathogenesis-related proteins, which form a secondary response in plant host-pathogen interactions and show anti-microbial activity. Some allergens of unknown function are cysteine-rich proteins, while some others have cysteine-rich regions; for example, the major allergen from rye-grass pollen, Lol p 1, has a cysteine-rich N-terminal region, while at the C-terminal region four tryptophan residues together with tyrosine and phenylalanine residues resemble those of cellulose- or sugar-binding domains of other proteins. Several pollen allergens show sequence similarity to cell wall-associated enzymes, while others show hydrolytic enzyme activity often associated with cell walls.     

壳多糖酶研究的概况及最新进展

壳多糖酶专一性水解壳多糖中的β－1,4糖苷键,在自然界的碳循环中具有极其重要的意义．壳多糖酶分布广泛,功能多样,在真菌生长发育、植物抗真菌感染等生理过程中壳多糖酶均发挥重要作用．文章概述了壳多糖酶研究的现状及最新进展,介绍了壳多糖酶的分布、理化性质、催化性质、在细胞中的定位及其调控;简介了近年来真菌和植物壳多糖酶研究的动态及最新进展.     

Ultrastructural immunolocalization of periodic pectin depositions in the cell wall ofNicotiana tabacum pollen tubes

Summary The monoclonal antibody (MAb) JIM5, marking acidic pectins, was used to localize ultrastructurally pectin molecules in the pollen tube wall ofNicotiana tabacum. Longitudinal sections of LR-White embedded pollen tubes were exposed to antibody treatment; accumulations of pectins were identified by counting the density of the gold particles representing the pectin epitopes along the pollen tube wall. Significant accumulations of gold grains were marked and the distances between them were measured. In many pollen tubes a more or less regular distribution of the accumulations was observed along the tube indicating a periodical deposition of pectin. The distances between the accumulations were 4–6 m. Most of the label was found in the inner part of the outer layer of the bilayered cell wall. These findings correspond to and confirm the earlier observation by our group reporting ring-shaped periodical deposits in pollen tubes after immunofluorescence labelling with the MAb JIM5 under the confocal laser scanning microscope.Abbreviations Ab antibody - MAb monoclonal antibody     

Activity of cell wall-associated enzymes in ripening olive fruit

Enzymatically active cell wall isolaled from olive (Olea europaea) fruit was employed Hi investigate some hydrolytic enzymes bound to the cell wall and the changes in these during ripening. Seven glycosidases. β-glucosidase (EC 3.2.1.21) α-galactosidase (EC 3.2.1.22). β-galactosidase (EC 3.2.1.23). α-arabinosidase (EC 3.2.1.55), α-mannosidase (EC 3.2.1,24). β-xylosidase (EC 3.2.1.37) and β-N-acetylglucosamidase (EC 3.2.1.30). as well as Cx-cellulase (EC 3.2.1.4) and endo-polygalacturonase (EC 3.2.1.15). were identified in the cell wall preparation, at four stages of ripeness (mature green. changing colour, black and black-ripe). Activities of all these cell wall-associated enzymes fionicallv and covalently linked) were determined either by cell wall incubation with artificial substrate or after extraction from the cell wall with buffers of high salt concentration (Cx-cellulase). and were compared to those of forms solubilized from acetone powders with 500 nM citrate buffer (cytoplasmic and/or apoplastic plus ionically hound to cell wall) In general, the activities of low ionic strength buffer-soluble enzymes were found to be much higher than those of the bound enzymes. The bound enzymes are present in the fruit at the green colour stage, whereas the activities of the soluble enzymes only increased from the changing colour stage onwards. The tenacity of binding of enzymes to the wall was investigated by treating the walls with high salt and measuring residual activity. The nature of the ionic and covalent binding and the changes during ripening were also established for wall-hound glycosidase During ripening there was a marked change in the percentages of covalently- and tonically linked activities of β-glucosidase and β-galaclosidase: al the changing colour stages about 75–80% of the bound active in was present in high ionic strength buffer while al the black-ripe stage it was only 15–20. A possible role for these cell wall degradative enzymes in olive softening is discussed.     

Effects of UV-B on flavonoids, ferulic acid, growth and photosynthesis in barley primary leaves

Barley (Hordeum vulgare L.) was grown with UV-B (280–320 nm) at levels simulating 25 nr 5% ozone depletion on the date of the summer solstice al 40°N latitude, with UV-A (320–400 nm), or with no supplemental irradiation. In plant growth chambers providing 300 μmol m^?2 s^?1 photosynthetically active radiation (PAR). UV-B-grown leaves elongated more slowly than controls but reached the same final length 1 day later. Leal specific fresh weight (mass leaf area^?1) was significantly increased by UV-B after the 7th day of growth. IV-B did not significantly affect leaf area, fresh weight, dry weight, total chlorophylls, total carotenoids or photosynthetic quantum efficiency. CO₂ assimilation was decreased by UV-B only at internal CO₂ levels above 250 μl l^?1. By the 8th day of growth, UV-B increased flavonoid (saponarin and lutonarin) accumulation in both the lower epidermis and the mesophyll: about 40% of the saponarin and 20% of the lutonarin were in the lower epidermis under all experimental conditions. Glasshouse conditions proved too variable for reproducible determination of growth and photosynthesis but were reliable for determining developmental changes in flavonoid (saponarin and lutonarin) accumulation and provided up to 800 μmol m^?2 s^?1 PAR. In the glasshouse UV-B-grown leaves had more flavonoids than controls al all stages from 5 to 30 days after planting: ca 509 more saponarin and 100% more lutonarin. Levels of soluble (vacuolar) ferulic acid esters were similar under all conditions on day 5. and on day 20 or later, but were significantly higher in UV-B-grown plants on days 10 and 15. UV-B decreased insoluble (cell-wall-bound) ferulic acid esters on a whole leaf basis but significantly increased this fraction in the lower epidermis. UV-A had no significant effects on growth, photosynthesis or ferulic acid, but it slightly increased flavonoid accumulation. The results are discussed in terms of secondary phenolics as a tissue-specific, developmentally regulated adaptive response to UV-B.