Dihydroflavonol Reductase Activity in Relation to Differential Anthocyanin Accumulation in Juvenile and Mature Phase Hedera helix L

Juvenile phase English ivy (Hedera helix L.) plants accumulate anthocyanin pigment in the hypodermis of stems and petioles, whereas genetically identical plants of the mature phase do not. The objective of this work was to assess which enzyme(s) might limit anthocyanin accumulation in mature phase ivy. Leaf discs of both juvenile and mature phase ivy accumulated comparable levels of the flavonols kaempferol and quercetin, whereas only juvenile phase discs accumulated anthocyanin. The accumulation of quercetin, but lack of accumulation of leucocyanidin or anthocyanin in mature phase discs, suggested that mature discs lacked dihydroflavonol reductase activity. There was no detectable dihydroflavonol reductase activity in mature phase discs, whereas there was an induction of activity in juvenile phase discs in response to sucrose, or photosynthetically fixed carbon, and light as a photomorphogenic signal. Phenylalanine ammonia-lyase, an enzyme early in the anthocyanin biosynthetic pathway, was induced above its basal level by sucrose and light in discs of both phases of ivy, with greater activity in mature phase discs. Phenylpropanoids, a class of compounds that are precursors to flavonoids, accumulated in leaf discs of both phases, with greater levels in mature phase discs. These results indicate that the lack of dihydroflavonol reductase activity limits the accumulation of anthocyanin in mature phase tissue.     

Adventitious Root Initiation In Reciprocally Grafted Leaf Cuttings from the Juvenile and Mature Phase of Hedera helix L

Reciprocal grafts involving leaf petioles and lamina of thejuvenile and mature phase of Hedera helix were prepared to determinethe relative importance of petiole and lamina on root initiationin leaf cuttings. The results indicated that root initiationwas mainly a function of the potential of cells in the petioleto respond in a specific morphogenetic pattern. Initially, rootinitiation was unaffected by the type of lamina. However, overtime, a factor translocated from the juvenile lamina promotedroot initiation in the mature petiole. This factor decreasedthe time taken for root initiation and increased the numberof roots per mature petiole. There was no evidence for an inhibitorfrom the mature lamina affecting root initiation in the juvenilepetiole. Key words: Rejuvenation, root initiation, rooting cofactors     

Evidence for Ideal and Non-Ideal Equilibrium Freezing of Leaf Water in Frosthardy Ivy (Hedera helix) and Winter Barley (Hordeum vulgare)

Temperature dependences of leaf water potentials (ψ_leaf) of frozen leaves of frosthardy ivy and winter barley were determined psychrometrically and found to coincide with the respective water potentials of ice which were obtained using the same technique. The water potentials of ice showed good agreement with theoretically established data. Analysis of the components of ψ of frozen leaves of Hedera helix revealed ideal equilibrium freezing, i.e. the governing of the relative content of liquid (or frozen) water solely by the osmotic potential. In winter barley, by contrast, a negative pressure potential was demonstrated to contribute to ψ_leaf. even under conditions of moderate frost. This reduced the degree of protoplast dehydration and the extent to which the concentrations of the cellular solutes rose. Such a freezing behavior is termed non-ideal equilibrium freezing. Depending on the original content of leaf water, the volume increments of liquid water due to the negative pressure potential amounted up to 10% at ?6 °C and even more at a lower temperature. In addition to the experimental data, a theoretical treatment of psychrometry at subzero temperatures is presented.     

Polyamines and Adventitious Root Formation in the Juvenile and Mature Phase of English Ivy

The involvement of polyamines during adventitious root formationwas evaluated using a de-bladed petiole rooting assay for theeasy-to-root juvenile and difficult-to-root mature phase ofEnglish ivy (Hedera helix L.). Auxin (NAA 0.1 mM) stimulatedroot formation in juvenile phase cuttings, but failed to promoterooting in the mature phase. The addition of putrescine, spermineor spennidine (1.0 mM) with or without NAA (0.1 mM) did notaffect the rooting response in either the juvenile or maturephase cuttings. There was a significant increase in endogenouslevels of putrescine and spermidine in NAA-treated cuttings,but the only significant difference between the root formingjuvenile and the non-root forming mature phase cuttings wasan increase in putrescine levels. In NAA-treated juvenile cuttings,the polyamine biosynthesis inhibitor DFMA (1.0 mM) promotedroot formation from 9.2 to 14.5 roots per cutting, while DFMO(1.0 mM) reduced root formation from 9.1 to 1.4 roots per cutting.The promotion of rooting by DFMA was completely reversed byputrescine (1.0 mM), but putrescine, spermine or spermidine(1.0 mM) could not reverse the inhibitory effect of DFMO. NeitherDFMA nor DFMO promoted root formation in mature phase cuttings.DFMA was also added to NAA-treated juvenile petioles at variousstages during the root formation process. DFMA promoted rootingwhen applied during the early stages of root induction (0–3d), but became inhibitory to root formation when applied duringthe organization (6–9 d) or root elongation stages (9–12d). Key words: Hedera helix, organogenesis, root initiation, polyamines, DFMA, DFMO     

Nanoparticle biofabrication using English ivy (Hedera helix)

Background

English ivy (Hedera helix) is well known for its adhesive properties and climbing ability. Essential to its ability to adhere to vertical surfaces is the secretion of a nanocomposite adhesive containing spherical nanoparticles, 60?C85 nm in diameter, produced exclusively by root hairs present on adventitious roots. These organic nanoparticles have shown promise in biomedical and cosmetic applications, and represent a safer alternative to metal oxide nanoparticles currently available.

Results

It was discovered that the maximum adventitious root production was achieved by a 4 h application of 1 mg/ml indole-3 butyric acid (IBA) to juvenile English ivy shoot segments cultured in custom vessels. After incubation of the shoots under continuous light at 83 ??mol/m² s at 20°C for 2 weeks, the adventitious roots were harvested from the culture system and it was possible to isolate 90 mg of dry weight nanoparticles per 12 g of roots. The nanoparticle morphology was characterized by atomic force microscopy, and found to be similar to previous studies.

Conclusions

An enhanced system for the production of English ivy adventitious roots and their nanoparticles by modifying GA7 Magenta boxes and identifying the optimal concentration of IBA for adventitious root growth was developed. This system is the first such platform for growing and harvesting organic nanoparticles from plants, and represents an important step in the development of plant-based nanomanufacturing. It is a significant improvement on the exploitation of plant systems for the formation of metallic nanoparticles, and represents a pathway for the generation of bulk ivy nanoparticles for translation into biomedical applications.     

Changes in the Amount of Complex Lipids in the Seeds and in the Pericarp during the Development of Ivy Fruit (Hedera helix)

By a combination of thin-layer chromatography and gas liquid chromatography, a complete study of the development of the different lipid classes and of their fatty acids, during the development of the fruit of Hedera helix L., the English Ivy, has been achieved. In any part of the fruit observed, at any particular stage, the phospholipids and the neutral lipids are the most abundant lipid classes. They accumulate during the entire process of maturation, whereas significant changes occur in their relative proportions, phospholipids being largely dominant until fruit blackening. The accumulation of fatty acids during maturation is characterized by large amounts of C_18:1 in the neutral lipids, especially in the seed, where petroselinic acid (C_18:1Δ⁶) reaches 86% of the total fatty acids. To a smaller extent, the phospholipids also accumulate and thus have the character of reserve molecules. However, their composition remains more stable, which relates them to the “structural lipids” such as galactolipids that maintain their characteristic fatty acid composition, despite the radical changes occurring in the fatty acid metabolism during fruit ripening.     

Intraspecific Variation of Ribosomal Gene Redundancy in ZEA MAYS

Ribosomal genes in eukaryotes are highly redundant. Considerable variation in the level of redundancy among species, especially in higher plants, has been reported; but except for deletion and duplication mutants, it is generally accepted that intraspecific variability in redundancy level is small. We have examined the level of redundancy in several lines of maize by DNA-rRNA saturation hybridization. The amount of nuclear DNA which hybridizes with rRNA in the ten lines examined varied from 0.24% to 0.50%. The number of rRNA genes per diploid genome thus ranges from 1.12 x 10⁴ to 2.32 x 10⁴. Results also indicate that the level of redundancy is genetically transmitted.     

Phase Change in Hedera helix: Induction of the Mature to Juvenile Phase Change by Gibberellin A3

A sensitive and reproducible method to obtain GA₃ induced morphological reversion of mature Hedera helix to the juvenile form has been developed. Dose response experiments indicate that GA₃ stimulates reversion over a 50–100 fold range with a half maximal response at approximately 0.5 μg GA₃ per plant. The individual characteristics involved in phase change revert to the juvenile form in a sequential manner as GA₃ dose is increased. Variations in light intensity from 1.2–3.6 × 10⁴ lux and temperature from 15 to 26°C do not affect this hormonal response. Other growth regulators including indoleacetic acid, kinetin, abscisic acid and (2-chloroethyl)phosphonic acid (Ethephon) are inactive but other gibberellins (GA₁ and a mixture of A₄–A₇) are active in stimulating reversion. Therefore, the response is specific for gibberellins as a class of hormones but non-specific for a particular form of gibberellin. The significance of this response in relation to juvenility in woody plants is discussed.     

Mineral nutrient stoichiometric variability in Hedera helix (Araliaceae) seeds

Background and Aims

The analysis of variability in mineral allocation to seeds has rarely been considered in relation to allometric patterns and deviations from the allometric trajectory. Here, I examine the scaling of carbon (C), nitrogen (N), phosphorus (P) and sulfur (S) with seed mass in field-collected seeds of Hedera helix, taking into account that brood size might influence the allocation patterns.

Methods

C, N and S contents of 56 individual seeds of different sizes were analysed by combustion using a standard automated CNHS procedure. P content was determined for 200 individual seeds using spectrophotometry after acid digestion of ashed samples. This sample included seeds from different brood sizes.

Key Results

C and N content both varied isometrically with seed mass and this variable explained most of the variation in their content in seeds. P and S, however, varied allometrically with seed mass. Additionally, seed mass only explained 37·3 % and 37·6 % of the total variance in P and S content, respectively. Seeds with higher mass contained proportionately more P and, furthermore, the seeds from small broods contained proportionally more P than those from large broods.

Conclusions

Although seed mass in this species can be used as a surrogate of investment in C and N, it does not account for variability in allocation of nutrients such as P and S. The fact that larger seeds increased their P concentration when found in smaller broods might be a consequence of intense competition for this nutrient among developing seeds. Then, brood size may influence the mineral nutrient concentration of seeds.     

Photosynthesis of Ivy Leaves (Hedera helix) after Heat Stress I. CO2-Gas Exchange and Diffusion Resistances

For an analysis of the inhibition of the photosynthetic CO₂-uptake after heat stress attached leaves of Hedera helix L. were heat-stressed for 30 min at various temperatures. Subsequently their photosynthetic CO₂-uptake, transpiration, respiration in light and darkness, and CO₂-compensation concentration were measured under optimal conditions. After heat stress the stomatal resistance increased only corresponding to the raised CO₂-concentration inside the leaves (due to the reduced CO₂-uptake). The physical resistance between the mesophyll cell walls and the chloroplasts remained unchanged after heat stress. A non-stomatal inhibition of the CO₂-uptake is indicated by a strong increase of the CO₂-compensation concentration after heat stress. This is hardly due to a stimulation of the respiration in light, as the CO₂-evolution into CO₂-free air in light was even reduced. Therefore, it must be concluded that the photosynthetic process itself is impaired after heat stress.     

Phase Change in Hedera helix: Stabilization of the Mature Form with Abscisic Acid and Growth Retardants

Applications of ABA to the mature form of Hedera helix stabilize its morphological characteristics and prevent GA₃ induced reversion to the juvenile form. Plants treated with GA₃ reverted to the juvenile form whereas those supplied with ABA in conjunction with GA₃ remained mature. When mature plants were treated with 5 nanomoles of GA₃ and 5 micromoles of ABA, reversion did not occur, but when the GA₃ dose was raised to 25 nanomoles with the same level of ABA, reversion did occur. This implies that the relative amounts of GA₃ and ABA applied are important in controlling growth form and not the absolute levels of these hormones. Applications of growth retardants (Chlormequat, Ancymidol, and SADH) stabilize the mature form by preventing spontaneous reversions induced under low light intensity. These two lines of evidence support the hypothesis that the mature morphological form can be stabilized by regulating the effective level of gibberellins in the plant and this can be accomplished by inhibition of gibberellin action or gibberellin biosynthesis.     

Pollinator effectiveness and fruit set in common ivy, Hedera helix (Araliaceae)

Flowers of common ivy (Hedera helix L.) provide late season pollen and nectar for several insect groups, and its fruits are a winter and spring food source for frugivorous birds. Ivy benefits from insect pollination in order to set fruit, but it is unknown which flower-visiting insects are the most effective pollinators. Our observations suggest that Vespula wasps are potentially the most effective pollinators since they were frequent visitors, had relatively fast foraging rates, carried large numbers of pollen grains on their bodies and had the highest ‘Pollination potential PP index’ score (a measure of pollinator effectiveness) of all the insect groups examined. There was also a positive linear relationship between the proportion of ivy flowers that set fruit and wasp foraging activity in 0.5 m² quadrats. Visits by Vespula wasps may therefore be important for ensuring a supply of ivy fruits for birds.     

Photosynthesis in leaves of the juvenile and adult phase of ivy (Hedera helix)

Abstract Photosynthetic and anatomical parameters of leaves from the juvenile and adult part of an ivy plant (Hedera helix L.) have been determined and compared with each other. Light-saturated net photosynthesis (per unit leaf area) was about 1.5 times higher in adult leaves than in juvenile ones. The lower photosynthetic capacity of juvenile leaves was caused by a lower stomatal and especially a lower residual conductance to the CO₂-transfer. This corresponds with anatomical features of the leaves, i.e. lower stomatal frequency, fewer chloroplasts per cell, and – most important – thinner leaves, as well as with a less efficient photosynthetic apparatus measured as Hill reaction of isolated broken chloroplasts and activity of ribulose bisphosphate carboxylase. No differences in the respiration in light (relative to net photosynthesis) and in the CO₂-compensation concentration could be detected between the two leaf types. These observed anatomical and photosynthetic parameters of the juvenile and adult ivy leaves resemble those reported for shade and sun leaves, respectively, although the leaves investigated originated from the same light environment.     

Coordination of Ribosomal Protein and Ribosomal RNA Gene Expression in Response to TOR Signaling

Cells grow in response to nutrients or growth factors, whose presence is detected and communicated by elaborate signaling pathways. Protein kinases play crucial roles in processes such as cell cycle progression and gene expression, and misregulation of such pathways has been correlated with various diseased states. Signals intended to promote cell growth converge on ribosome biogenesis, as the ability to produce cellular proteins is intimately tied to cell growth. Part of the response to growth signals is therefore the coordinate expression of genes encoding ribosomal RNA (rRNA) and ribosomal proteins (RP). A key player in regulating cell growth is the Target of Rapamycin (TOR) kinase, one of the gatekeepers that prevent cell cycle progression from G1 to S under conditions of nutritional stress. TOR is structurally and functionally conserved in all eukaryotes. Under favorable growth conditions, TOR is active and cells maintain a robust rate of ribosome biogenesis, translation initiation and nutrient import. Under stress conditions, TOR signaling is suppressed, leading to cell cycle arrest, while the failure of TOR to respond appropriately to environmental or nutritional signals leads to uncontrolled cell growth. Emerging evidence from Saccharomyces cerevisiae indicates that High Mobility Group (HMGB) proteins, non-sequence-specific chromosomal proteins, participate in mediating responses to growth signals. As HMGB proteins are distinguished by their ability to alter DNA topology, they frequently function in the assembly of higher-order nucleoprotein complexes. We review here recent evidence, which suggests that HMGB proteins may function to coordinate TOR-dependent regulation of rRNA and RP gene expression.Key Words: Rapamycin, TORC1, HMO1, high mobility group, yeast, RP gene, rDNA.     

洋常春藤的组织培养与快速繁殖

1植物名称洋常春藤(Hedera helix cv.aureovariegata). 2材料类别带芽茎段. 3培养条件(1)诱导休眠芽萌动培养基:MS 6-BA 0.5 mg·L-1(单位下同) NAA 0.2;(2)芽增殖培养基:MS 6-BA 1.0 IBA 0.2;(3)生根培养基:1/2MS IBA 0.5.上述培养基中均加入30 g.L-1蔗糖、6.0 g·L-1琼脂,pH 5.8.培养温度为25～27℃,光照度2 000～2 500 lx,光照时间12 h.d-1.     

Light acclimation in leaves of the juvenile and adult life phases of ivy (Hedera helix)

Hoflacher, H. and Bauer, H. 1982. Light acclimation in leaves of the juvenile and adult life phases of ivy (Hedera helix). – Physiol. Plant. 56: 177–182. Light acclimation was investigated during the juvenile and adult life phases of the whole-plant-development in Hedera helix L. For this purpose, cuttings of the juvenile and adult parts of one single parent plant were grown under low-light (PAR 30–50 μmol photons m^?2 s^?1) and high-light (PAR 300–500 μmol m^?2 s^?1) conditions: CO₂ exchange, chloroplast functions, and specific anatomy of fully developed leaves differentiated under these conditions were determined. In juvenile plants the leaves formed under low and high light had light-saturated rates of net photosynthesis of 6.5 and 11.1 mg CO₂ (dm leaf area)^?2 h^?1, respectively. In adult plants the rates were 9.4 and 22.2 mg dm^?2 h^?1, indicating a more pronounced capacity for acclimation to strong light in the adult life phase. Higher photosynthetic capacities were accompanied by higher conductances for the CO₂ transfer through the stomata, leading to almost the same CO₂ concentration in the intercellular spaces. Thus, stomatal conductances were not primarily responsible for the different photo-synthetic capacities. The higher rates in adult and high-light grown leaves were mainly the result of formation of thicker leaves with more chloroplasts per unit leaf area. Expressed per chloroplast, the photosynthetic capacity, the Hill reaction, and the activity of ribulose bisphosphate carboxylase were almost identical in plants grown in low-light and high-light. Measurements of photosynthetic capacity and thickness of leaves of Hedera sampled from field habitats with contrasting light regimes confirm the results of growth chamber studies. It is, therefore, concluded that both life phases of Hedera are capable of acclimating to strong light, but that during the juvenile phase this capacity is not fully developed.     

Regulation of Ribosomal RNA Gene Multiplicity in DROSOPHILA MELANOGASTER

The ribosomal RNA (rRNA) genes of Drosophila melanogaster can undergo a disproportionate replication of their number. This occurs when the cluster of rRNA genes (rDNA) of one chromosome is maintained with a homologous chromosome that is completely or partially deficient in its rDNA. Under appropriate genetic conditions, it appears that disproportionate rDNA replication can be generated at the level of both somatic and germ line cells. In the latter case, mutants partially deficient for rDNA can increase their rRNA gene number to the wild type level and transmit this new genotype to successive generations.     

Ultrastructure and Radiolabelling of Leaf Cuticles from Ivy (Hedera helixL.) Plantsin vitroand duringex vitroAcclimatization

Cuticle ultrastructure and radiolabelling of isolated cuticlesafter incorporation of [¹⁴C] acetate in foliar discs were investigatedwith ivy plants grownin vitrothenex vitro. Results show an increasein thickness, mass and wax content, between young and expandedleaves, for bothin vitroandex vitrocuticles. The cuticle ofinvitrounexpanded leaves was very thin and only constituted alamellate zone. The ultrastructure ofin vitroyoung and expandedleaf cuticles showed characteristics similar toin situcuticles.The thickness of the lamellate zone remained fairly constantand represented 33% of the cuticle thickness in young leaves,but only 11.4% in expanded leaves. The number of lamellar unitsdecreased from 14 to nine between these two growth stages. Themain difference between young leaves developedin vitroorex vitrowasa thinner lamellate zone forex vitrocuticles. However, theselatter cuticles had an intermediary zone between the lamellateand reticulate zones. The cuticle thickness of expanded leaveswas greater forin vitrocuticles suggesting a temporary decreasein cuticle biosynthesis after transfer of the plant fromin vitrotoexvitro.Results from cuticle radiolabelling show higher radioactivityincorporation in cuticles isolated from leaves developedex vitrocomparedtoin vitro. This radiolabelling was particularly marked forexvitroyoung leaf cuticles and depended on the duration of theexvitrogrowth period revealing a progressive activation of cuticlebiosynthesis in response to new environmental conditions. Hedera helix; ivy leaf cuticle; in vitroplants; electron microscopy; radiolabelling; isolated cuticles     

RNA Polymerase I Activators Count and Adjust Ribosomal RNA Gene Copy Number

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