植物体内的木质素

木质素是植物体内重要的大分子物质,它是一类在化学结构上密切相关、高分子量的多聚体。主要论述了木质素单体的类型,它们在植物体内的合成过程,单体间的聚合机制及其在植物体内的分布和作用。     

Lignin composition and structure in young versus adult Eucalyptus globulus plants

Lignin changes during plant growth were investigated in a selected Eucalyptus globulus clone. The lignin composition and structure were studied in situ by a new procedure enabling the acquisition of two-dimensional nuclear magnetic resonance (2D-NMR) spectra on wood gels formed in the NMR tube as well as by analytical pyrolysis-gas chromatography-mass spectrometry. In addition, milled-wood lignins were isolated and analyzed by 2D-NMR, pyrolysis-gas chromatography-mass spectrometry, and thioacidolysis. The data indicated that p-hydroxyphenyl and guaiacyl units are deposited at the earlier stages, whereas the woods are enriched in syringyl (S) lignin during late lignification. Wood 2D-NMR showed that β-O-4' and resinol linkages were predominant in the eucalypt lignin, whereas other substructures were present in much lower amounts. Interestingly, open β-1' structures could be detected in the isolated lignins. Phenylcoumarans and cinnamyl end groups were depleted with age, spirodienone abundance increased, and the main substructures (β-O-4' and resinols) were scarcely modified. Thioacidolysis revealed a higher predominance of S units in the ether-linked lignin than in the total lignin and, in agreement with NMR, also indicated that resinols are the most important nonether linkages. Dimer analysis showed that most of the resinol-type structures comprised two S units (syringaresinol), the crossed guaiacyl-S resinol appearing as a minor substructure and pinoresinol being totally absent. Changes in hemicelluloses were also shown by the 2D-NMR spectra of the wood gels without polysaccharide isolation. These include decreases of methyl galacturonosyl, arabinosyl, and galactosyl (anomeric) signals, assigned to pectin and related neutral polysaccharides, and increases of xylosyl (which are approximately 50% acetylated) and 4-O-methylglucuronosyl signals.     

Lignin deficiency in transgenic flax resulted in plants with improved mechanical properties

Flax (Linum usitatissimum L.) is a very important source of natural fibres used by the textile industry. Flax fibres are called lignocellulosic, because they contain mainly cellulose (about 70%), with hemicellulose, pectin and lignin. Lignin is a three-dimensional polymer with a high molecular weight, and it gives rigidity and mechanical resistance to the fibre and plant. Its presence means the fibres have worse elastic properties than non-lignocellulosic fibres, e.g. cotton fibres, which contain no lignin. The main aim of this study was to produce low-lignin flax plants with fibres with modified elastic properties. An improvement in the mechanical properties was expected. The used strategy for CAD down-regulation was based on gene silencing RNAi technology. Manipulation of the CAD gene caused changes in enzyme activity, lignin content and in the composition of the cell wall in the transgenic plants. The detected reduction in the lignin level in the CAD-deficient plants resulted in improved mechanical properties. Young's modulus was up to 75% higher in the generated transgenic plants (CAD33) relative to the control plants. A significant increase in the lignin precursor contents and a reduction in the pectin and hemicellulose constituents was also detected. A decrease in pectin and hemicellulose, as well as a lower lignin content, might lead to improved extractability of the fibres. However, the resistance of the transgenic lines to Fusarium oxysporum was over two-fold lower than for the non-transformed plants. Since Fusarium species are used as retting organisms and had been isolated from retted flax, the increased sensitivity of the CAD-deficient plant to F. oxysporum infection might lead to improved flax retting.     

Tolerance to waterlogging in young Euterpe oleracea plants

This study investigated whether gas exchange and the present content of antioxidant compounds can contribute to the survival of Euterpe oleracea plants in environments of frequent waterlogging. A factorial randomised, experimental design included two distinct water conditions (waterlogging and control) and five evaluation times (0, 6, 12, 18, and 24 d). Gasexchange parameters, leaf temperature, electrolyte leakage, and contents of antioxidant compounds were measured. Waterlogging did not promote significant alterations in net photosynthetic rate and transpiration, and stomatal conductance was reduced only after 18 d. Malondialdehyde and glutathione contents did not significantly change during waterlogging. Additionally, electrolyte leakage was significant only after 18 d of waterlogging. Thus, this study revealed that maintenance in gas exchange and antioxidant compounds might contribute to the survival of E. oleracea plants in environments exposed to waterlogging.     

Chilling injury and nucleotide changes in young cotton plants

The effects of chilling at 3 to 5 C on the nucleotide composition of roots and leaves of cotton (Gossypium hirsutum L.) seedlings were determined. Chilling decreased the concentration of nucleotides, especially di- and triphosphates, in both leaves and roots. Chilling also caused an increase in free nucleosides. The results are interpreted to mean that general phosphorolytic activity is associated with chilling injury rather than damage to the phosphorylating mechanisms alone. Hardening at 10 to 20 C prior to chilling protected the seedlings against subsequent chilling injury and prevented nucleotide losses.     

Silicon mitigates cadmium inhibitory effects in young maize plants

The influence of silicon on the growth of maize plants cultivated in hydroponics in the presence of cadmium (5 μM) was investigated. Four different treatments were used: Control (C), Cadmium (Cd), Silicon (Si) and Cadmium plus Silicon (Cd + Si). The Si concentration was 35 mM. Thirteen-day-old plants were harvested. Growth parameters (length of primary seminal root, leaf area of first and second fully developed leaves, fresh and dry weight of below- and above-ground parts of the plants), and Cd concentration and total amount of Cd in the below- and above-ground parts were determined. In roots, the development of the endodermal barrier was observed by fluorescent staining with Fluorol yellow 088.Inhibitory effects of Cd on plant growth were observed. Silicon treatment in the absence of Cd had positive effects on most of observed growth parameters compared with the control. Moreover, Si in the Cd + Si treatment improved all growth parameters compared with the cadmium treatment. Silicon increased the cell-wall extensibility both in Si and Cd + Si treatments when compared with the control. Alleviation of the Cd-inhibitory effect on maize plants by Si was not due to exclusion of Cd from the plant; in contrast, Cd concentration in below- and above-ground plant parts and the total amount of Cd per plant were significantly higher in the Cd + Si plants than in the Cd treatment. The increased Cd content in Cd + Si plants was correlated with the development of the endodermis; during the second stage of endodermal development, suberin lamellae were formed at a greater distance from the root apex in the Cd + Si than in the Cd treatment. Silicon itself did not influence the development of suberin lamellae in the maize roots compared with the control.     

Distribution of wheat germ agglutinin in young wheat plants

A liquid phase, competition-binding radioimmunoassay for wheat germ agglutinin, with a detection limit of 10 nanograms, was developed in order to determine the distribution of this lectin in young wheat plants. Affinity columns for wheat germ agglutinin removed all antigenically detectable activity from crude extracts of wheat tissue; thus, the antigenic cross-reactivity detected by the assay possesses sugar-binding specificity similar to the wheat germ-derived lectin. The amount of lectin per dry grain is approximately 1 microgram, all associated with the embryo. At 34 days of growth, the level of lectin per plant was reduced by about 50%, with approximately one-third in the roots and two-thirds in the shoot. The data also indicate that actively growing regions of the plant (the bases of the leaves and rapidly growing adventitious roots) contain the highest levels of lectin. Half of the lectin associated with the roots could be solubilized by washing intact roots in buffer containing oligomers of N-acetylglucosamine, whereas the remainder is liberated only upon homogenization of the tissue.     

Physiological responses to salt stress in young umbu plants

Soil salinity affects plant growth and development due to harmful ion effects and water stress caused by reduced osmotic potential in the soil solution. In order to evaluate the effects of salt stress in young umbu plants, research was performed in green house conditions at the Laboratory of Plant Physiology at Federal Rural University of Pernambuco, Brazil. Growth, stomatal behaviour, water relations, and both inorganic and organic solutes were studied aiming for a better understanding of the responses of umbu plants to increasing salinity. Plants were grown in washed sand with Hoagland and Arnon nutrient solution with 0, 25, 50, 75, and 100 mM NaCl. Growth, leaf water potential, transpiration, and diffusive resistance were evaluated. Na⁺, K⁺, Cl⁻, soluble carbohydrates, and free amino acid contents were measured in several plant organs. Most variables were affected with salinity above 50 mM NaCl showing decreases in: number of leaves, plant height, stems diameter, and dry masses, and increases in root-to-shoot ratio. Reductions in ψ_pd were observed in plants grown under 75 and 100 mM NaCl. All salt levels above zero increased Na⁺ and Cl⁻ contents in leaves. However, K⁺ content was not affected. Na⁺ and Cl⁻ in stems and roots reached saturation in treatments above 50 mM NaCl. Organic solute accumulation in response to salt stress was not observed in umbu plants. These results suggest that umbu plants tolerate salt levels up to 50 mM NaCl without showing significant physio-morphological alterations.     

Source-sink coupling in young barley plants and control of phloem loading

Phloem loading of carbohydrate within a mature exporting leaf of a barley seedling is shown to respond quickly to a change in the temperature of the root and the shoot meristem. This is interpreted as a close coupling between source supply and sink demand for carbohydrate, through the hydrostatic pressure gradient linking source and sink generated by the solute concentration within the sieve tubes. This interpretation was tested by using anoxia to alter solute concentration within the sieve tubes of one region of a leaf while observing phloem loading in an adjacent region. Responses to anoxia could not be explained by the above model, suggesting that either this model is incorrect or other signalling pathways are involved. There is evidence in the literature for coarse control of phloem loading but no evidence was found of fine control by solute content of the loaded sieve elements.     

Levels of oxygenated Fatty acids in young corn and sunflower plants

Three oxygenated unsaturated fatty acids were investigated to determine whether they were present in seedlings of corn (Zea mays L. cv. NK PX443) and sunflower (Helianthus annuus L. cv. Sundak). The three compounds, 13-hydroxy-12-oxo-cis-9-octadecenoic acid (I), 13-hydroxy-12-oxo-cis,cis-9, 15-octadecadienoic acid (II), and 12-oxo-cis,cis-10, 15-phy-todienoic acid (III), were detected and estimated by gas chromatography-mass spectrometry selected ion monitoring of their trimethylsilyloxy, methyloxime derivatives with 20-carbon analogs added as internal standards. In corn, the concentration of III increased between 5 and 10 days, while I and II remained relatively constant. A higher concentration of II was observed in corn seedlings grown in the light than those grown in the dark. Wounding increased the levels of all three compounds. In sunflower seedlings, the concentrations of I, II, and III increased between 6 and 10 days. The intracellular concentration of III in 10-day-old light-grown seedlings was estimated to be 200 nm in corn and 40 nm in sunflower.     

Tetramethylputrescine from young plants of Ruellia rosea

Tetramethylputrescine was isolated from the roots and aerial parts of Ruellia rosea. The compound was characterized by MS, ¹³C-NMR-spectroscopy and comparison with a synthetic specimen.     

Response of young barley plants to CO2 enrichment

Barley (Hordeum vulgare L. cv. Digger) was grown for 22 d inenclosed chambers with a CO₂ enrichment of 35, 155, 400 or 675µmol CO₂ mol1. CO₂ enrichment increased photosyntheticcapacity in the plants grown at either of the two highest levelsof pCO₂. A CO₂ enrichment of 675µmol CO₂ caused a significantincrement of shoot dry weight, whereas no changes were observedin fresh weight, chlorophyll or protein levels. At a light intensityof 860µmol m^–2s^–1 CO₂ enrichment caused photosyntheticcapacity to increase by 250%, whereas no effect was observedat 80 µmol m^–2 s^–1. Over time, photosynthesisdecreased by 70% independent of CO₂. A time-dependent increasein the level of extractable fructose was observed whereas totalextractable carbohydrate only changed slightly. Key words: Carbohydrates, CO₂ enrichment, Hordeum vulgare, photosynthesis, respiration     

Long-term chilling of young tomato plants under low light

The properties of two Calvin-cycle key enzymes, i.e. stromal fructose-1,6-bisphosphatase (sFBPase) and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) were studied in the cultivated tomato (Lycopersicon esculentum Mill.) and in four lines of a wild tomato (L. peruvianum Mill.) from different altitudes. During chilling for 14 d at 10°C and low light, the activation energy (E_A) of the reaction catalyzed by sFBPase decreased by 5–10 kJ·mol^–1 inL. esculentum and the threeL. peruvianum lines from high altitudes. InL. peruvianum, no loss or only small losses of enzyme activity were observed during the chilling. Together with the change in E_A, this indicates that the latter species is able to acclimate its Calvin-cycle enzymes to low temperatures. InL. esculentum, the chilling stress resulted in the irreversible loss of 57% of the initial sFBPase activity. Under moderately photoinhibiting chilling conditions for 3 d, theL. peruvianum line from an intermediate altitude showed the largest decreases in both the ratio of variable to maximum chlorophyll fluorescence (F_v/F_m) and the in-vivo activation state of sFBPase, while the otherL. peruvianum lines showed no inhibition of sFBPase activation. Ribulose-1,5-bisphosphate carboxylase/oxygenase was isolated by differential ammonium-sulfate precipitation and gel filtration and characterized by two-dimensional electrophoresis. The enzyme fromL. esculentum had three isoforms of the small subunit of Rubisco, each with different isoelectric points. Of these, theL. peruvianum enzyme contained only the two more-acidic isoforms. Arrhenius plots of the specific activity of purified Rubisco showed breakpoints at approx. 17°C. Upon chilling, the specific activity of the enzyme fromL. esculentum decreased by 51%, while E_A below the breakpoint temperature increased from 129 to 189 kJ·mol^–1. In contrast, Rubisco from theL. peruvianum lines from high altitudes was unaffected by chilling. We tested several possibile explanations for Rubisco inactivation, using two-dimensional electrophoresis, analytical ultracentrifugation, gel filtration and inhibitor tests. No indications were found for differential expression of the subunit isoforms, proteolysis, aggregation, subunit disassembly, or inhibitor accumulation in the enzyme from chilledL. esculentum. We suggest that the activity loss in theL. esculentum enzyme upon chilling is the result of a modification of sulfhydryl groups or other sidechains of the protein.Abbreviations a.s.l. above sea level - Chl chlorophyll - DTT dithiothreitol - E_A activation energy - FBP fructose-1,6-bisphosphate - F_v/F_m ratio of variable to maximum chlorophyll fluorescence - HL high light (500 mol photons·m^–2·s^–1) - LSU large subunit of Rubisco - ME 2-mercaptoethanol - Rubisco ribulose-1,5-bisphosphate carboxylase/oxygenase - RuBP ribulose-1,5-bisphosphate - sFBPase stromal fructose-1,6-bisphosphatase - SSU small subunit of Rubisco