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  • nitrogen  (54)
  • Microtubules
  • Nicotiana
  • phosphorus
  • Springer  (161)
  • Blackwell Publishing Ltd
  • National Academy of Sciences
  • 1990-1994  (108)
  • 1980-1984  (53)
  • 1925-1929
  • 1992  (108)
  • 1980  (53)
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  • Springer  (161)
  • Blackwell Publishing Ltd
  • National Academy of Sciences
  • Wiley-Blackwell  (1)
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  • 1990-1994  (108)
  • 1980-1984  (53)
  • 1925-1929
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  • 1
    Electronic Resource
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    Springer
    Entomologia experimentalis et applicata 64 (1992), S. 225-238 
    ISSN: 1570-7458
    Keywords: Lymantria dispar (L.) ; diet switching ; nitrogen ; specialization ; plant-animal interactions ; insect behavior
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Gypsy moth (Lymantria dispar (L.) (Lepidoptera: Lymantriidae)) larvae were reared from hatch on 1.25% N or 3.5% N artificial diet (previous diet) and switched reciprocally to the other diet (current diet) after molting into the second, third, fourth, or fifth instar. The nitrogen concentration of food consumed during previous instars had a strong residual effect on the growth rate in subsequent instars when a diet switch was made during instars two through four, but did not affect growth rate of fifth-instar larvae despite effects on food consumption and utilization. In early instars, larvae reared on 1.25% N artificial diet and then switched to 3.75% N diet had lower mass-adjusted growth rates than larvae continuously reared on 3.75% N diet. Conversely, larvae reared on 3.75% N diet and switched to 1.25% N had higher mass-adjusted growth rates than larvae reared continuously on 1.25% N diet. Relative to larvae previously reared on 1.25% N diet, fifth-instar male larvae previously reared on 3.75% N diet had slightly lower consumption rates, higher net growth efficiency (ECD), and higher gross growth efficiency (ECI). Larvae previously reared on 3.75% N diet tended to have lower food assimilation efficiency (AD) and lower nitrogen assimilation efficiency (AD(N)). Although both previous and current diet nitrogen concentration strongly affected larval growth and food utilization, the interaction term between these was not significant for any response variables except ECD and ECI. Because the interaction term reflects the effect of switching per se, the results indicate that there was a metabolic cost associated with switching, but no inherent net cost or benefit of diet-switching to growth.
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  • 2
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    Catalysis letters 13 (1992), S. 45-53 
    ISSN: 1572-879X
    Keywords: Ammonia ; synthesis ; nitrogen ; rate-determining step ; potassium promotion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The simple concept of a single rate-determining step, the dissociation of N2(a), in ammonia synthesis is inconsistent with experimental results. It is proposed that three reaction steps, N2(a) → 2N(a) (1) N(a) + H(a)→NH(a) (2) NH3(a)→NH3(g) (3) are dominant in determining the rate of synthesis. Under industrial conditions reactions (2) and (3) are more important than (1). The role of K+ as promoter in these reactions is discussed, especially as an “anchor” for adsorbed oxygen.
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  • 3
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    Catalysis letters 16 (1992), S. 159-164 
    ISSN: 1572-879X
    Keywords: Chemisorption ; nitrogen ; carbon dioxide ; platinum ; water gas shift reaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The relationship between sites responsible for nitrogen chemisorption and sites responsible for stronger adsorption of carbon dioxide on platinum black is reported. A 2 to 1 ratio has been found between molecules of more strongly adsorbed carbon dioxide and molecules of nitrogen chemisorbed on individual samples. This relationship has allowed us to deduce the structure of chemisorbed carbon dioxide. Carbon dioxide is relatively weakly chemisorbed on platinum. Reasons for the weakness of this chemisorption are discussed.
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  • 4
    ISSN: 1420-9055
    Keywords: Sediment ; interstitial water ; phosphorus ; iron ; persulfate digestion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract It is shown that sorption of orthophosphate to iron compounds, formed during persulfate digestion, can cause a significant underestimation of total dissolved phosphorus in interstitial waters rich in iron and poor in phosphorus. Labelling the samples with carrier free32PO4 before digestion allows to correct for these losses.
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  • 5
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    Entomologia experimentalis et applicata 62 (1992), S. 29-36 
    ISSN: 1570-7458
    Keywords: Alkaloids ; cinnabar moth ; insect genetics ; larval performance ; Lepidoptera ; nutritional ecology ; nitrogen ; sugar ; Tyria jacobaeae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In a laboratory experiment using full-sibs, 60% of the variation in pupal weight of the monophage Tyria jacobaeae L. (Lepidoptera, Arctiidae) could be explained by variation in the nitrogen concentration of the food plant, Senecio jacobaea L. and only 4% by variation in sugar concentration. Larval weight and growth rates of young and old larvae were also positively correlated with nitrogen and sugar concentration. Developmental time was negatively correlated with nitrogen concentration. In a second experiment full-sib families differed significantly in larval weight at day 7, mortality, growth rate and developmental time. Pupal weight did not differ significantly among families, but was positively correlated with nitrogen concentration of Senecio. Larval performance was not significantly influenced by concentrations of sugars or alkaloids. We conclude that larval performance of Tyria during most of the larval period is mainly determined by genetic factors, but pupal weights are primarily determined by nitrogen concentration of the food plant.
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  • 6
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    Microchimica acta 109 (1992), S. 201-209 
    ISSN: 1436-5073
    Keywords: phosphonate base scale inhibitor ; brine ; direct current plasma ; inductively coupled plasma atomic emission spectrometry ; phosphorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The determination of phosphonate base scale inhibitors in brines by direct current plasma (DCP) and inductively coupled plasma atomic emission spectrometry (ICP-AES) is described. The first method is based on a direct nebulization of the brine samples and plasma using the phosphorus line at 213.618 nm. The second method involves extraction of phosphorus as phospho-antimonyl molybdate complex into methylisobutyl ketone (MIBK) phase and analysis of the extract for molybdenum using the Mo 313.260 nm line. Comparison between the proposed methods and an established recommended method [1] shows excellent agreement between the results in addition to the sensitivity and ease of automation provided by AES.
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  • 7
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    Nutrient cycling in agroecosystems 1 (1980), S. 95-101 
    ISSN: 1573-0867
    Keywords: soil fertility ; phosphorus ; potassium ; green manure ; crop rotation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Changes in the soil phosphorus and potassium status on three farming systems in the Northeast Polder during 25 years are related to the fertilizer regime over this period. The changes in the nutrient status of the soil are reflected in fertilizer policy. High rates of phosphate dressings markedly increased the phosphate status of the soil and built up a reserve of phosphate. In contrast, the potassium status decreased by about half, because only potatoes in the rotation were dressed with potassium. Methods of fertilization are discussed and suggestions are made with regard to possible improvements.
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  • 8
    ISSN: 1573-0867
    Keywords: Wheat ; nitrogen ; grain protein content ; grain filling ; urea spraying
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The objective of this work was to determine the effect of foliar urea spraying during or after anthesis, on dry matter and nitrogen accumulation in the wheat grain of two cultivars, differing in grain protein content. The experiment was carried out in the field, and 20 kg N ha−1 as urea (46-0-0) was sprayed at anthesis, 7 or 14 days after anthesis. Dry weight, N content and N percentage of the grain were determined at several stages of its development. Neither the rate nor the duration of dry matter accumulation in the grain was significantly modified by N sprayings at anthesis or later and, consequently, not its final weight. Nitrogen sprayings at anthesis or later increased the grain N content and N percentage at ripeness in both cultivars, but to a greater extent in the low grain protein cultivar. Greater N content in the grain was attributed to a rapid uptake of the N applied and not to a longer duration of the period of N accumulation. The different responses of the cultivars to N sprayings can be related to differences in N demand, and is probably associated, also, to a genetic ability to accumulate N in the grain.
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  • 9
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    Nutrient cycling in agroecosystems 32 (1992), S. 55-59 
    ISSN: 1573-0867
    Keywords: 15N ; nitrogen ; rice ; soil N ; N fertilizer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In the southern U.S. rice belt it is recommended that rice (Oryza sativa L.) grown in the dry-seeded, delayed flood cultural system have the preflood N fertilizer applied and the field flooded at the fourth to fifth leaf stage of plant development. The objective of this field study was to determine if delaying the flood and preflood N application past the fifth leaf stage was detrimental to rice total N and fertilizer15N uptake, total dry matter, and grain yield. This study was conducted on a Crowley silt loam (Typic Albaqualfs) and a Perry clay (Vertic Haplaquepts). The preflood N fertilizer and flood were delayed 0, 7, 14, or 21 d past the fourth to fifth leaf stage, after which time a permanent flood was established and maintained until maturity. All treatments received 20.5 g N m−2 as15N-labeled urea in three topdress applications. All plant and soil samples were taken at maturity. Harvest index increased as the preflood N and flood were delayed past the 4 to 5 leaf stage. Total N in the grain + straw either decreased or showed a decreasing trend as the N and flood were delayed. Similarly, uptake of native soil N decreased as flood was delayed. Conversely, percent recovery of fertilizer N in the rice plant and the plant-soil system increased as the preflood N and flood were delayed. Rice grain yield was not significantly affected by delaying the preflood N and flood up to 21 d.
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  • 10
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    Nutrient cycling in agroecosystems 32 (1992), S. 223-227 
    ISSN: 1573-0867
    Keywords: Soil testing ; phosphorus ; relative yield
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A greenhouse experiment, with Okra (Abelmoschus esculentus L.) as the test crop, was conducted on twenty-one soils ranging in Olsen's extractable phosphorus from 1.8 to 15.5µg Pg−1 soil. The experiment was conducted at Punjab Agricultural University, Ludhiana, India. The soils were nonsaline with pH ranging from 7.7 to 8.6. A critical level of 2.55µg Pg−1 soil was predicted by Cate and Nelson's (1971) statistical procedure. Because of a wide range in relative yields, this value did not accurately predict response to applied P. An approach to compute minimum response to applied fertilizer, which is likely to be obtained at a particular Olsen P level, has been presented. It involves calculation of lower 60 percent confidence limits for relative yield and fitting loge-linear regression to the transformed data. The regression was tested on a published data set and was found to hold well.
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  • 11
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    Nutrient cycling in agroecosystems 32 (1992), S. 259-267 
    ISSN: 1573-0867
    Keywords: Nitrogen ; phosphorus ; timber increment ; fertilization ; Pinus radiata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Two trials inPinus radiata growing on different sites in N.S.W. allowed consideration of fertilizer applications after 2nd or 3rd thinning. The trials included factorial applications of N and P at a single thinning intensity plus a further treatment which allowed assessment of different thinning intensities. The most significant growth responses were obtained by application of N and P in combination. The largest response (additional productivity compared with the unfertilized control) occurred 4 years after application and after 7 years there was no additional absolute response for either of the two sites. The largest fertilizer response was 70 m3 ha−1 over 7 years on one site and 36 m3 ha−1 on the other, indicating differences in absolute responses between sites. It was concluded that in planning treatments the most responsive sites near the end of the rotation should be selected to maximise economic returns. Foliage analyses indicated differences between sites at the commencement of the study. It was concluded that either a single year of foliage analyses at study commencement is of value, or sampling every year of the study should be used to analyse responses, but a single year of analysis during or at the end of the study would not be of value.
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  • 12
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    Aquatic sciences 54 (1992), S. 58-76 
    ISSN: 1420-9055
    Keywords: Eutrophication ; lake management ; phosphorus ; nitrogen ; chlorophyll-a ; slope estimator
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We estimate the response of chl-a (mg · m−3) to changes in concentrations of total phosphorus (TP) by calculating the slopeS = Δchl-a/ΔTP in chl-a =f(TP) graphs. Results show that in years where algae are P-limited oligotrophic lakes respond less (median slope 0.21) to changes in nutrient concentrations than eutrophic lakes, (median slope 0.31) and these again less than hypereutrophic lakes, (median slope 1.02). We find no saturation value for the slope within the TP range considered (6–480 mg · m−3). Chl-a in eutrophic lakes responds more frequently to non-nutrient factors than oligotrophic and hypereutrophic lakes. Results obtained by replacing TP with a new nutrient parameter, TP′ = 0.056 · TP · IN0.226, in which inorganic nitrogen, IN, is factored in, suggest that nitrogen has an influence on chl-a in oligotrophic lakes. Blue-green algae respond less to changes in TP than other algal species, e.g., diatoms.
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  • 13
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    Aquatic sciences 54 (1992), S. 381-390 
    ISSN: 1420-9055
    Keywords: Eutrophication ; phosphorus ; lake restoration ; internal restoration measures for lakes ; Swiss lakes ; Lake Lugano (Lago di Lugano)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In most lakes eutrophication is linked to an excessive input of phosphorus. Lake restoration by reduction of P-input (external measure) has led to a considerable drop of the P-concentration in all major Swiss lakes as well as in many other lakes. Internal restoration measures such as artificial mixing, drainage of hypolimnetic water, flushing, aeration, biomanipulation and others serve to improve and accelerate the response of a lake to external measures. For the case of Lago di Lugano, a simple two-box model is employed to demonstrate that a reduction of the P-input to about 25% of the present values is necessary to reach the “P-criterion” (P-concentration below 30 µg/l). Internal measures could possibly accelerate the extremely slow response of the northern basin.
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  • 14
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    Development genes and evolution 188 (1980), S. 65-73 
    ISSN: 1432-041X
    Keywords: Nuclear migration ; Cleavage ; Microtubules ; Ultrastructure ; Gall midge
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In the eggs ofWachtliella persicariae the cleavage nuclei move relative to the surrounding ooplasm. This ‘active’ migration is caused by an organelle whose ultrastructure was studied throughout the mitotic cycle. It consists of a greatly enlarged polar cytaster derived from the mitotic apparatus, linked to the nucleus by 100 Å filaments. The microtubules of the cytaster were found only during periods of active nuclear migration, i.e., from the onset of anaphase to the early prophase of the next mitotic cycle. They are always solitary and follow the course of the astral rays, which are known to temporarily adhere to peripheral structures of the egg cell and to exert tractive forces. In contrast to the cytaster microtubules, the microtubules in the spindle are bundled and persist from early metaphase through late telophase. During ontogenesis the first migration cytaster is built up between 3 and 12 min after oviposition near the anterior egg pole, in the vicinity of the sperm nucleus. In non-inseminated eggs time lapse films show a migration cytaster to develop autonomously in a region free from nuclei, but it does not follow the normal path of the male pronucleus. In several cases the female pronucleus, which remains without a cytaster of its own, was observed to move to the cytaster generated in the absence of the male pronucleus. Whether or not it is adhering to a nucleus, the cytaster divides into two at the correct time, i.e, corresponding to the first cleavage division in fertilized eggs. In some non-inseminated eggs this type of ‘pseudocleavage’ has been observed to occur repeatedly, giving rise to an increasing number of anucleate cytasters.
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  • 15
    ISSN: 1432-2048
    Keywords: mRNA ; Protein synthesis ; Protoplasts ; Nicotiana ; Translation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Studies of proteins synthesized in vitro by messenger RNA (mRNA) extracted from tobacco protoplasts showed that the changes in protein synthesis and especially the lack of certain proteins observed previously in isolated protoplasts did not result from a failure of translation.
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  • 16
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    Planta 148 (1980), S. 462-467 
    ISSN: 1432-2048
    Keywords: Flower formation ; Leaves (in flower formation) ; Meristem (shoot) ; Nicotiana ; Roots (and flower formation)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The terminal, apical shoot meristem ofN. tabacum cv. Wisconsin 38 normally differentiates into a flower after producing 30 to 40 nodes. The influence of leaves and roots on the regulation of flowering was evaluated by counting the number of nodes produced after removal of leaves or the induction of adventitious roots. Leaf removal has no effect on the number of nodes produced before flower formation. Root induction significantly increases the number of nodes produced before flower formation. The plant behaves as if it were measuring the number of nodes between the meristem and the roots as a means of regulating meristem conversion from vegetative to floral differentiation.
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  • 17
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    Planta 148 (1980), S. 491-497 
    ISSN: 1432-2048
    Keywords: Leaves (polysomes) ; Nicotiana ; Polysomes ; Poly(A)+ RNA ; Protein synthesis ; RNA (polysomal, polyA+)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The isolation of intact polysomes from leaves of tobacco (Nicotiana tabacum L.) is dependent on the age and state of development of leaves. Undegraded polysomes from young leaves in the early stages of expansion can be isolated easily by extracting the leaves in ice-cold extraction buffer (200 mM tris(hydroxymethyl)aminomethylmethane(Tris)-HCl, pH 9.0; 400 mM KCl; 200 mM sucrose; 35 mM MgCl2). Medium-size leaves give best yields of undegraded polysomes when extractions are carried out in the above buffer and in the presence of ethyleneglycol-bis-(β-amino-ethyl ether)-N,N′-tetracetic acid (EGTA) and mercaptoethanol. Isolation of polysomes from large, nearly fully expanded (mature) leaves requires all of the above plus diethyldithiocarbamate (DIECA) in the extraction medium. An extraction medium consisting of 25 mM EGTA, 0.01 M mercaptoethanol, 25 mM DIECA and 0.5% of the nonionic detergent, Nonidet-P40 (NP 40) was found to be very suitable for extraction of polysomes from all developmental stages of leaves. The polysomes extracted in the above medium showed active translation of protein in the wheat-germ in-vitro protein-synthesizing system. The translational products were similar when translations were carried out directly with polysomes or polysomal RNA, or polysomal poly(A)+ RNA from tobacco leaves. Poly(A)− polysomal RNA was a poor template in the in-vitro wheat-germ system.
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  • 18
    ISSN: 1432-2048
    Keywords: Auxin metabolism ; Cell suspensions (low density) ; Medium conditioning ; Nicotiana ; Nitrogen metabolism ; Protoplasts ; Vitamin requirements
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Preliminary attempts to define a completely synthetic medium able to support divisions of haploid tobacco mesophyll protoplasts at low initial densities have failed. High protoplast concentrations together with large amounts of naphtaleneacetic acid in the medium (3 mg l-1 NAA) were required for maximal induction of protoplast division. However, cell suspensions derived from haploid protoplasts after four days of preculture at high initial cell densities could be diluted to densities as low as 1–4 cells ml-1, provided the concentration of NAA in the medium was lowered to below 0.3 mg l-1. The optimal NAA supply for low cell density growth was affected by the nature of the nitrogen source. A simple minimal medium which supports the growth of these haploid cells with a plating efficiency of 30–40%, independent of the cell density in the range of 1–4 to 3·104 cells ml-1, has been established. In this medium inositol was the only vitamin stringently required for growth. Growth of cells at low densities was also possible in a medium initially containing 3 mg l-1 NAA, provided it was conditioned by the growth of protoplasts at high densities. Preliminary experiments with [14C]NAA showed that the amount of free NAA remaining in the medium after preincubation at high densities was drastically reduced. Simultaneously, NAA conjugates accumulated in the medium. The implications of these results are discussed.
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  • 19
    ISSN: 1432-2048
    Keywords: Auxin receptor ; Callus ; Nicotiana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cultured tobacco-pith tissue contains a cytoplasmic receptor for indoleacetic acid (IAA). The concentration of binding sites is very low in comparison to that of several auxin receptors found by other investigators. A few obvious possible causes (degradation or inactivation) were investigated. From the results we conclude that the low number of binding sites is real. The receptor binds IAA optimally at pH 7.5–7.8 and at a temperature of 24–30°C, when incubated for 25–30 min. The binding is very specific, as is shown by competition experiments. The concentration of the receptor in the callus tissue changes dramatically during each culture period, which suggests a possible role in development. The receptor was partly purified by gel filtration on Sepharose 6B followed by ion-exchange chromatography on DEAE-cellulose.
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  • 20
    ISSN: 1432-2048
    Keywords: Cell walls ; Cellulose ; Graptopetalum ; Microtubules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In the “regeneration” of a shoot from a leaf of the succulent, Graptopetalum paraguayense E. Walther the first new organs are leaf primordia. The original arrangement of cellulose microfibrils and of microtubules (MTs) in the epidermis of the leaf-forming site is one of parallel, straight lines. In the new primordium both structures still have a congruent arrangement but it is roughly in the form of concentric circles that surround the new cylindrical organ. The regions which undergo the greatest shift in orientation (90°) were studied in detail. Departures from the original cellulose alignment are detected in changes in the polarized-light image. Departures from the original cortical MT arrangement are detected using electron microscopy. The over-all reorganization of the MT pattern is followed by the tally of MT profiles, the various regions being studied in two perpendicular planes of section. This corrects for the difference in efficiency in counting transverse versus longitudinal profiles of MTs. Reorientation takes place sporadically, cell by cell, for both the cellulose microfibrils and the MTs, indicating a coordinated reorientation of the two structures. That MTs and cellulose microfibrils reorient jointly in individual cells was shown by reconstruction of the arrays of cortical MTs in paradermal sections of individual cells whose recent change in the orientation of cellulose deposition had been detected with polarized light. Closeness of the two alignments was also indicated by images where the MT and microfibril alignments co-varied within a single cell. The change-over in alignment of the MTs appears to involve stages where arrays of contrasting orientation co-exist to give a criss-cross image. During this critical reorganization, the frequency of the MTs is high. It falls during subsequent enlargement of the organ. It was found that the rearrangement of the cortical MTs to approximate a series of concentric circles on the residual meristem occurred before the emergence of leaf primordia. Through their apparent influence on microfibril alignments, the changes in MT disposition, described here, have the potential to generate major biophysical changes that accompany organogenesis.
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  • 21
    ISSN: 1432-2048
    Keywords: 1-Aminocyclopropane-1-carboxylic acid ; Avena ; Ethylene ; Etiolation ; Light (ethylene production) ; Nicotiana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract White light inhibits the conversion of 1-amino-cyclopropane-1-carboxylic acid (ACC) in discs of green leaves of tobacco (Nicotiana tabacum L.) and segments of oat (Avena sativa L.) leaves by from 60 to 90%. Etiolated oat leaves do not show this effect. The general nature of the effect is shown by its presence in both a mono- and a dicotyledon. Since the leaves have been grown and pre-incubated in light, yet can produce from 2 to 9 times as much ethylene in the dark as in the light, it follows that the light inhibition is fully reversible. The inhibition by light is about equal to that exerted in the dark by CoCl2; it can be partly reversed by dithiothreitol and completely by mercaptoethanol. Thus the light is probably acting, via the photosynthetic system, on the SH group(s) of the enzyme system converting ACC to ethylene.
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  • 22
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    Planta 149 (1980), S. 205-206 
    ISSN: 1432-2048
    Keywords: Callus cultures ; Nicotiana ; Nicotine ; Tissue cultures
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Callus cultures of two low-alkaloid lines of Nicotiana tabacum L. had considerably lower nicotine contents than cultures from the respective highalkaloid cultivars which were isogenic except for the two loci for alkaloid accumulation. Thus, there was a strong correlation between the nicotine content of callus cultures and the plants from which they were derived.
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  • 23
    ISSN: 1432-2048
    Keywords: Ca2+ transport ; Fungicides ; Herbicides ; Microtubules ; Mitochondria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The herbicides amiprophosmethyl (APM) trifluralin, and oryzalin as well as the fungicides methylbenzimidazolyl carbamate (MBC), O-isopropyl N-phenyl carbamate (IPC), and chlorisopropyl N-phenyl carbamate (CIPC), which are known to cause the destruction of microtubules in vivo but do not interfere with tubulin polymerization in vitro, have been examined with respect to their ability to affect Ca2+ transport in isolated cell organelles. In contrast to colchicine which has no effect on Ca2+ transport in isolated mitochondrial and microsomal fractions, all of the substances investigated caused considerable reduction of ca2+ net uptake into mitochondrial but not into microsomal fractions. This reduction has been shown to be due to an increase in passive Ca2+ efflux. These results have been extrapolated to in vivo situations where they are postulated to act by raising cytoplasmic Ca2+ levels.
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  • 24
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    Planta 150 (1980), S. 9-12 
    ISSN: 1432-2048
    Keywords: Auxin binding ; Nicotiana ; Plasma membrane ; Protoplast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In vitro binding of 1-naphthaleneacetic acid (NAA) to particulate fractions from tobacco leaf protoplasts was studied. In freshly isolated protoplasts no specific binding could be detected, whereas it was present in particulate fractions from tobacco leaves. It is concluded that the NAA-binding-sites are probably located at the external face of the plasma membrane; they are destroyed during protoplast isolation by proteolytic enzymes in the cellulase and macerozyme preparations. After culturing the protoplasts for 3–4 d, the first cell divisions were observed and at the same time specific NAA-binding became detectable. The affinity constant for NAA was approx. 2·106 mol-1 and the number of binding sites increased during further culture.
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  • 25
    ISSN: 1432-2048
    Keywords: Cytokinin habituation ; Habituation, cytokinin ; Nicotiana ; Position effects (habituation) ; Size effects (habituation) ; Tissue culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Pith tissue of Nicotiana tabacum L. cv. “Havana 425” exhibits a gradient in its tendency to habituate for cytokinin on an auxin-containing medium at 35° C, about 10° C above the standard culture temperature. Explants of pith from below the 8th to 11th internode, counting from the bottom of the plant, rarely habituate for cytokinin; explants from above this threshold habituate rapidly. The explants must also be above a critical size, about 20–30 mg, to habituate. There was a pronounced interaction between size and position effects; the threshold position for cytokinin habituation shifted upward with decreasing explant size.
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  • 26
    ISSN: 1432-2048
    Keywords: Abscisic acid ; Nicotiana ; Ribonucleic acid synthesis ; Tobacco mosaic virus
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    Topics: Biology
    Notes: Abstract Uptake of abscisic acid from the culture medium by discs of healthy and tobacco mosaic virus-infected tobacco leaves was measured. Small (two to five-fold) increases in abscisic acid concentration in discs caused increases in rates of [3H]uridine and [3H]adenine incorporation into total nucleic acid, virus RNA and host ribosomal RNA. Net accumulation of virus RNA was also enhanced by abscisic acid. This evidence for stimulation of RNA synthesis is compared with previous reports showing inhibition of RNA synthesis in other tissues. It is suggested that the increase in endogenous abscisic acid caused by tobacco mosaic virus infection may be at least partly responsible for observed increases in rates of RNA synthesis after infection.
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  • 27
    ISSN: 1432-2048
    Keywords: Arabinogalactan epitope (immunofluores-cence localization) ; Cell wall ; Monoclonal antibody (JIM 8, MAC 207) ; Nicotiana ; Pollen tube
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The monoclonal antibodies MAC 207 and JIM 8, recognizing arabinogalactan epitopes, were used to localize the corresponding antigenic sites in pollen tubes of Nicotiana tabacum L. grown in vitro or semi in vivo. The analysis of the immunofluorescence labelling was performed by means of confocal laser scanning microscopy. Most pollen tubes were labelled along their length, with the exception of the tip region, in a ring-like pattern with remarkable periodicity. The diameter of the rings was approx. 12 μm and the distance between two rings was about 6 μm. No labelling of the cytoplasm, the vegetative nucleus or the generative cell was observed. The presence of labelling in the non-apical tube wall after pectinase and cellulase digestion indicates that the epitopes for MAC 207 and JIM 8 are located in the inner callosic sheath of the pollen-tube wall.
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  • 28
    ISSN: 1432-2048
    Keywords: Hydrogenperoxide ; Lignification ; Nicotiana ; Peroxidase
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    Topics: Biology
    Notes: Abstract Three peroxidase isoenzyme-groups found in cell walls of tobacco were tested for their capacity to form H2O2. Isoenzyme-group GI, located only in cell walls (GII and GIII are also found in protoplasts) showed the highest Kapp-value for H2O2-formation. The lowest Kapp-value, i.e., maximal H2O2-formation was received for group GIII which is ionically bound to the cell wall. As shown before, GI yields maximal polymerization rates for coniferyl- and p-coumarylalcohol. These facts indicate that each of the peroxidase isoenzyme groups of the cell wall is involved with different catalytic functions within the same pathways of H2O2-formation and succeeding lignification. H2O2-formation catalyzed by all 3 groups was increased by very low concentrations of Mn2+-ions. The required amount of Mn2+ leading to maximal stimulation was in each case dependent on the basic rate of H2O2-formation. Maximal stimulation of H2O2-formation by phenolic compounds was achieved by coniferylalcohol at a concentration of 10-4M for all groups. Stimulation by p-coumaryl-and by sinapylalcohol was not as significant.
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  • 29
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    Planta 149 (1980), S. 402-407 
    ISSN: 1432-2048
    Keywords: Cytokinin ; Habituation rates ; Nicotiana ; Temperature and habituation ; Tissue culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Pith parenchyma tissue ofNicotiana tabacum L. cv. “Havana 425” becomes cytokinin habituated when incubated at 35°C on an auxin-containing medium. Under these conditions, habituated, hyperplastic nodules appear on the tissues. We used these nodules to estimate the incidence of habituation by a statistical method. The rate of habituation varied with the season. Tissue isolated from plants in the spring habituated approx. 7 times faster than did tissue isolated from plants in winter. The fact that the average rate, 〉4×10−3 per cell generation, was 100–1,000 times faster than the rate of somatic mutation inNicotiana species and depended on the physiological state of the tissue provides further evidence that habituation involves epigenetic changes rather than rare, random genetic mutations. We also found that kinetin (6-furfurylaminopurine) induced habituation and that the concentration required depended on the duration of cytokinin treatment. For long incubation times, approx. 6×10−10 M kinetin, which is about 1,000-fold lower than the concentration optimal for growth of cytokinin-requiring pith tissue, was sufficient to induce habituation. These results support the hypothesis that the habituated state is maintained by a positive feedback loop in which cytokinins either induce their own synthesis or inhibit their own degradation.
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  • 30
    ISSN: 1432-2048
    Keywords: Acetate incorporation ; Catharanthus ; Cell suspension cultures ; Fatty acids ; Glycihe ; Nicotiana ; Temperature and fatty-acid synthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The fatty-acid composition of C. roseus and N. tabacum cell suspension cultures was unaffected by subculture on Wood and Braun, Murashige and Skoog, or Gamborg B5C media. However, placing the cultures — which were normally grown at 25° C — at 15° C reduced growth but resulted in enhanced formation of oleic and linolenic acids in C. roseus cultures and increased levels of linoleic and linolenic acids in cultures of G. max and N. tabacum, respectively. The incorporation of [14C]acetate into [14C]linoleic acid was more rapid in N. tabacum cells than in G. max cells, but was very poor in C. roseus where the [14C] label was distributed mainly between palmitic and oleic acids.
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  • 31
    ISSN: 1432-2048
    Keywords: Fraction-1 protein ; Hybrid, somatic ; Nicotiana ; Protein (Fraction 1) ; Ribulose-1,5-bisphosphate carboxylase ; Somatic hybrid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In the sexual interspecific cross, Nicotiana rustica L.xN. tabacum L., N. rustica can serve as the female but not as the male parent. By fusion of protoplasts, the barrier to fertilization was overcome and somatic hybrids containing N. tabacum cytoplasm were produced as shown by isoelectric focusing of the Fraction-1 protein (F-1-protein). All somatic hybrids displayed polypeptides of the large subunit of F-1 protein (which is coded by the chloroplast genome) characteristic of only one or the other parental species. Two hybrids had large subunits of the N. tabacum type and two hybrids had those of the N. rustica type. Three hybrids contained three smallsubunit polypeptides (coded by the nuclear genome), one being characteristic of N. rustica, one characteristic of N. tabacum, and one with an isoelectric point common to both species. A fourth hybrid contained only two small-subunit polypeptides of the N. tabacum type but in a F-1 protein macromolecule whose large subunits were of the N. rustica type. One somatic hybrid was self-fertile and its F2 progeny contained large- and small-subunit polypeptides indistinguishable in their isoelectric points from those in the parent F1 hybrid. All somatic hybrids showed an aneuploid chromosome number and morphological characteristics intermediate between those of N. rustica and N. tabacum.
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  • 32
    ISSN: 1432-2048
    Keywords: Catharanthus ; Cell suspension cultures ; Glycine ; Fatty acids (synthesis, desaturation) ; Nicotiana ; Linoleic, linolenic, oleic, acids ; Temperature and fatty acids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cell suspension cultures of Catharanthus roseus G. Don, Glycine max (L.) Merr. and Nicotiana tabacum L. were incubated with [14C]acetate, [14C]oleic acid and [14C]linoleic acid at five different temperatures ranging from 15 to 35° C. When the incubation temperature was increased, [14C]acetate was incorporated preferentially into [14C]palmitate, with a concomitant drop in [14C]oleate formation. Between 15 and 20° C, [14C]oleic acid accumulated in C. roseus cells. In all cultures, optimum desaturation of [14C]oleic acid to [14C]linoleic acid occurred between 20 and 25° C, and in G. max this was also the optimal range for desaturation of [14C]linoleic acid to [14C]linolenic acid. Elongation of [14C]palmitic acid was inhibited when cultures grown at 15° C for 25 h were subsequently incubated with [14C]acetate at 25° C. [14C]oleic acid accumulated in G. max and C. roseus cultures grown at 35° C for 25 h and subsequently incubated at 25° C. Desaturation of [14C]oleic acid increased up to 25° C, but then decreased or leveled off depending on the cell line and on the temperature prior to incubation.
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  • 33
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    Planta 147 (1980), S. 405-413 
    ISSN: 1432-2048
    Keywords: Caulonema ; Cell growth (tip) ; Funaria ; Microtubules ; Organelle modification ; Polarity
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    Topics: Biology
    Notes: Abstract In the caulonema tip cells of Funaria hygrometrica, chloroplasts, mitochondria, and dictyosomes have differences in structure which are determined by cell polarity. In contrast to the slowly growing chloronema tip cells the apical cell of the caulonema contains a tip body. Colchicine stops tip growth; it causes the formation of subapical cell protrusions, redistribution of the plastids, and a loss of their polar differentiation. Cytochalasin B inhibits growth and affects the position of cell organelles. After treatment with ionophore A23 187, growth is slower and shorter and wider cells are formed. D2O causes a transient reversion of organelle distribution but premitotic nuclei are not dislocated. In some tip cells the reversion of polarity persists; they continue to grow with a new tip at their base. During centrifugation, colchicine has only a slight influence on the stability of organelle anchorage. The former polar organization of most cells is restored within a few hours after centrifugation, and the cells resume normal growth. In premitotic cells the nucleus and other organelles cannot be retransported, they often continue to grow with reversed polarity. Colchicine retards the redistribution of organelles generally and increases the number of cells that form a basal outgrowth. The interrelationship between the peripheral cytoplasm and the nucleus and the role of microtubules in maintaining and reestablishing cell polarity are discussed.
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  • 34
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    Planta 148 (1980), S. 491-497 
    ISSN: 1432-2048
    Keywords: Leaves (polysomes) ; Nicotiana ; Polysomes ; Poly(A)+ RNA ; Protein synthesis ; RNA (polysomal, polyA+)
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    Topics: Biology
    Notes: Abstract The isolation of intact polysomes from leaves of tobacco (Nicotiana tabacum L.) is dependent on the age and state of development of leaves. Undegraded polysomes from young leaves in the early stages of expansion can be isolated easily by extracting the leaves in ice-cold extraction buffer (200 mM tris(hydroxymethyl)aminomethylmethane(Tris)-HCl, pH 9.0; 400 mM KCl; 200 mM sucrose; 35 mM MgCl2). Medium-size leaves give best yields of undegraded polysomes when extractions are carried out in the above buffer and in the presence of ethyleneglycol-bis-(β-amino-ethyl ether)-N,N′-tetracetic acid (EGTA) and mercaptoethanol. Isolation of polysomes from large, nearly fully expanded (mature) leaves requires all of the above plus diethyldithiocarbamate (DIECA) in the extraction medium. An extraction medium consisting of 25 mM EGTA, 0.01 M mercaptoethanol, 25 mM DIECA and 0.5% of the nonionic detergent, Nonidet-P40 (NP 40) was found to be very suitable for extraction of polysomes from all developmental stages of leaves. The polysomes extracted in the above medium showed active translation of protein in the wheat-germ in-vitro protein-synthesizing system. The translational products were similar when translations were carried out directly with polysomes or polysomal RNA, or polysomal poly(A)+ RNA from tobacco leaves. Poly(A)− polysomal RNA was a poor template in the in-vitro wheat-germ system.
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  • 35
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    Planta 149 (1980), S. 389-401 
    ISSN: 1432-2048
    Keywords: Allium ; Cell wall Coated vesicles ; Guard cells ; Microtubules ; Protoplasts
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    Topics: Biology
    Notes: Abstract Protoplasts were prepared from the guard cells ofA. cepa. Epidermal peels taken from expanding green leaves and largely free of mesophyll were treated with Cellulysin, and protoplasts were harvested after 18 h of digestion. That the protoplasts were derived from guard cells was ascertained from their characteristic vacuolar autofluorescence and from observations showing that all other epidermal cells are killed in the peeling procedure. The protoplasts proved to be a good system with which to view the cell cortex and inner surface of the plasmalemma. The lysis of cells adhering to polylysine-treated, Formvar-coated grids, followed by negative staining in uranyl acetate, showed that many microtubules normally present in ordered arrays in situ remain closely applied to the inner surface of the plasmalemma in protoplasts. In addition, numerous vesiculate elements including coated vesicles and/or pits are present amongst the microtubules. Similar vesicles are evident in thin sections of fixed, embedded guard cells and protoplasts. The significance of these structures in the cell cortex is discussed.
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  • 36
    ISSN: 1432-2048
    Keywords: mRNA ; Nicotiana ; Protein synthesis ; Ribulose-1,5-bisphosphate carboxylase ; RNA (messenger) ; Translation
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    Topics: Biology
    Notes: Abstract The products synthesized in vitro by messenger RNA (mRNA) extracted from Nicotiana sylvestris were analyzed by electrophoresis on polyacrylamide slab gels. Only three of the major polypeptides synthesized are considered here: P55, P32, and P20. P55 and P32 were translated from chloroplast mRNA. P55 corresponds to the large subunit of ribulose-1,5-bisphosphate (RuP2) carboxylase; P32 is probably a chloroplast membrane protein. P20, the polypeptide synthesized from cytoplasmic poly(A)+ RNA, is the precursor of the small subunit of RuP2 carboxylase. The balance between P20 and P32, in which their relative proportions varied inversely, was regulated by the age of the leaves and the time of illumination; we took advantage of this phenomenon to isolate the mRNA from the small subunit in relatively large amounts. This mRNA has a molecular weight of 350,000.
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  • 37
    ISSN: 1432-2048
    Keywords: Adenosine 5′-phosphosulfate sulfotransferase ; Cell suspension culture ; Cysteine ; Enzyme regulation ; Nicotiana
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    Topics: Biology
    Notes: Abstract The properties and the regulation of adenosine 5′-phosphosulfate sulfotransferase extracted from cell suspension cultures ofNicotiana sylvestris was investigated. Optimal adenosine 5′-phosphosulfate sulfotransferase activity was obtained from the cells by extraction with 0.1 M tris-HCl, pH8.0, containing 2 M MgSO4 and 10 mM dithioerythritol. The K m for adenosine 5′-phosphosulfate in the sulfotransferase reaction was about 11 μM. Adenosine 5′-phosphosulfate in concentrations above 50 μM were inhibitory. The extratable adenosine 5′-phosphosulfate sulfotransferase activity decreased during cultivation with sulfate as the sole sulfur source, but after about 3 days it reached a constant level (50 to 100 nmol activated sulfate transferred h-1 mg-1 protein) which was maintained for at least 24 h. Addition of 0.5 mM cysteine to the culture medium decreased the extractable adenosine 5′-phosphosulfate sulfotransferase activity and blocked growth completely. With 0.1 mM cysteine an enzyme level of about 10% of the initial value was reached within 6 to 12 h without significant inhibition of growth. The added cysteine was absorbed rapidly and after 24 h cysteine could no longer be detected in the medium. Before the cysteine was completely depleted, the activity of adenosine 5′-phosphosulfate sulfotransferase started to increase, reaching ultimately a level which was comparable to the initial value.
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  • 38
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    Planta 147 (1980), S. 269-273 
    ISSN: 1432-2048
    Keywords: Crown gall ; Cytokinins ; Nicotiana
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    Topics: Biology
    Notes: Abstract Cytokinins were extracted from two cultures of tobacco crown gall tumor tissue: an unorganized tissue and a teratoma which produced leafy shoots. On Sephadex LH-20 column chromatography, extracts of both types of tissue yielded two peaks of cytokinin activity with elution volumes similar to ribosylzeatin and zeatin. Ribosylzeatin and zeatin were detected and quantified by coupled gas chromatography — mass spectrometry selected ion monitoring (GC/MS SIM), comparable quantities being found in the two extracts. Full mass spectral evidence for the presence of ribosylzeatin in both tissues was obtained. No evidence was found for the presence of N6-(Δ2-isopentenyl)adenosine (i6Ade) or N6-(Δ2-isopentenyl)adenine (i6Ade) although these compounds have been reported to occur in cytokinin-habituated tobacco callus tissues.
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  • 39
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    Planta 147 (1980), S. 274-276 
    ISSN: 1432-2048
    Keywords: Cell division ; Dikegulac ; Lysis ; Nicotiana ; Plasmalemma ; Protoplasts ; Tonoplast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Dikegulac (2,3:4,6 di-o-isopropylidine-2-keto-I-gulonate) is a growth regulator used to differentially kill terminal apices, and it analogously inhibits basic metabolic functions in dividing cells, but not stationary cells, in suspension culture. This report demonstrates an analogous situation in isolated tobacco protoplasts. At the lowest concentrations, dikegulac partially suppresses division of the protoplasts. Higher concentrations are required to produce visual cytoplasmic damage to the protoplasts, which probably first occurs at the level of the plasmalemma, as the vacuoles can be released intact. Later, tonoplast disruption occurs.
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  • 40
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    Planta 147 (1980), S. 500-506 
    ISSN: 1432-2048
    Keywords: Cell shape ; Colchicine ; Daucus ; Immunofluorescence ; Microtubules ; Protoplasts
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    Topics: Biology
    Notes: Abstract Indirect immunofluorescence has been used to study the function of cytoplasmic microtubules in controlling the shape of elongated carrot cells in culture. Using a purified wall-degrading preparation, the elongated cells are converted to spherical protoplasts and the transverse hoops of bundled microtubules are disorganised but not depolymerised in the process. Since microtubules remain attached to fragments of protoplast membrane adhering to coverslips and are still seen to be organised laterally in bundles, it would appear that re-orientation of the transverse bundles is due to loss of cell wall and not to the cleavage of microtubule bridges. After 24 h treatment in 10-3 M colchicine, microtubules are depolymerised in elongated cells but, at this time, the cells retain their elongated shape. This suggests that wall which was organised in the presence of transverse microtubule bundles can retain asymmetric shape for short periods in the absence of those tubules. However, after longer periods of time the cells become spherical in colchicine. Neither wall nor tubules therefore exert individual control on continued cellular elongation and so we emphasize the fundamental nature of wall/microtubule interactions in shape control. It is concluded that the observations are best explained by a model in which hooped bundles of microtubules—which are directly or indirectly associated with molecules involved with cellulose biosynthesis at the cell surface—act as an essential template or scaffolding for the orientated deposition of cellulose.
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  • 41
    ISSN: 1432-2048
    Keywords: Colchicine ; Membrane structure ; Microtubules ; Osmotic treatment ; Plasmalemma ; Poterioochromonas
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    Topics: Biology
    Notes: Abstract Changes in membrane topography in the flagellate Poterioochromonas malhamensis, as a result of colchicine and osmotic-stress treatments, have been studied using freeze-fracturing and thin sectioning. Ridges, but not rows of intramembrane particles, in the PF-face which denote the position of underlying cortical microtubules, together with the ridge associated with their point of origin (flagellar root fibre 1), dissappear after colchicine or short-term (5 min) osmotic treatments. Cortical microtubules are destroyed as a result of the former, but not the latter treatment. Longer periods in osmoticum allow a recovery of the microtubule — associated membrane ridges. Despite careful isosmotic fixations distinct cross-bridges between microtubules and the plasmalemma were not discernible in thin section.
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  • 42
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    Planta 148 (1980), S. 97-102 
    ISSN: 1432-2048
    Keywords: Abscisic acid ; Cell cultures ; Nicotiana ; Variants, resistant ; Water stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The goal of this work was to begin a genetic study of the molecular mode of action of abscisic acid (ABA), by isolating variant cultured cells resistant to the hormone, or to a factor which induces ABA synthesis, namely water stress. Cell cultures of Nicotiana tabacum L. cv. Wisconsin 38 and N. silvestris Speg. and Comes were chosen as the experimental materials. Studies of the effects of the two stresses on the growth of the cultures demonstrated that ABA or water stress imposed by mannitol could completely inhibit growth. These effects arose in both cases from a constant reduction of the growth rate of the cells throughout the culture period. Mannitol also induced an increase in ABA content of the cells and media of suspension cultures, although not to the concentrations required to achieve the same degree of growth inhibition when the hormone was applied exogenously.
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  • 43
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    Planta 148 (1980), S. 103-107 
    ISSN: 1432-2048
    Keywords: Abscisic acid ; Cell cultures ; Nicotiana ; Variants, resistant ; Water stress
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    Topics: Biology
    Notes: Abstract Variant clones were isolated from Nicotiana silvestris Speg. et Comes cell cultures at low frequencies following severe abscisic-acid (ABA) or mannitol-induced water-stress treatments of plated cells. N. tabacum L. variants were not recovered. Variants from the ABA selection experiments exhibited a 10-fold increase in resistance to the hormone. This trait was stable in non-selective conditions for as long as was tested (200 days), but did not alter the response of the cells to water stress. Cell lines from the waterstress selection were not more resistant to mannitol than the parent line, and had a wide range of response to ABA.
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  • 44
    ISSN: 1432-2048
    Keywords: mRNA ; Protein synthesis ; Protoplasts ; Nicotiana ; Translation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Studies of proteins synthesized in vitro by messenger RNA (mRNA) extracted from tobacco protoplasts showed that the changes in protein synthesis and especially the lack of certain proteins observed previously in isolated protoplasts did not result from a failure of translation.
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  • 45
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    Planta 148 (1980), S. 462-467 
    ISSN: 1432-2048
    Keywords: Flower formation ; Leaves (in flower formation) ; Meristem (shoot) ; Nicotiana ; Roots (and flower formation)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The terminal, apical shoot meristem ofN. tabacum cv. Wisconsin 38 normally differentiates into a flower after producing 30 to 40 nodes. The influence of leaves and roots on the regulation of flowering was evaluated by counting the number of nodes produced after removal of leaves or the induction of adventitious roots. Leaf removal has no effect on the number of nodes produced before flower formation. Root induction significantly increases the number of nodes produced before flower formation. The plant behaves as if it were measuring the number of nodes between the meristem and the roots as a means of regulating meristem conversion from vegetative to floral differentiation.
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  • 46
    ISSN: 1432-2145
    Keywords: Nicotiana ; Cytoplasmic male sterility ; Flower morphogenesis ; mtDNA organization ; In organello protein synthesis ; Mitochondrial transcripts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A detailed description of in planta floral ontogeny based on scanning electron microscopy, developmental histology and morphology is presented for three different alloplasmic gene-cytoplasmic male-sterile (CMS)Nicotiana tabacum isonuclear lines with cytoplasms ofN. bigelovii, N. debneyi and N. suaveolens and compared to the corresponding nuclear donorN. tabacum genotype. This allowed the precise determination of the developmental stages affected in the mutant forms as well as a thorough phenotypic characterization of them. The organization of the mitochondrial genome and expression of mitochondrial DNA (mtDNA) was investigated in the three different alloplasmic CMS tobacco analogs and compared to the corresponding malefertile parentalNicotiana species. Southern hybridizations of total cellular DNA and mtDNA from the different sets of lines with probes specific for mitochondrial genes coding for cytochrome oxidase subunits I, II and III, apocytochrome b, ATPase subunits α and 9 as well as 18S-5S ribosomal RNA indicated that: (a) mtDNA organization is different between mitochondrial genomes of fertile and sterile lines but identical in two different fertile tobacco lines; however genetic similarity among different mitochondrial genome types can be revealed by restriction fragment patterns; (b) although several differences were detected between the male-sterile and male-fertile plants, most of these were related to the origin of the mitochondria (cytoplasm donorNicotiana species); (c) identical mtDNA rearrangements — distinct to the cytoplasm donor — occur in cytoplasmic malesterile tobacco analogs bearing cytoplasm fromN. bigelovii in two differentN. tabacum nuclear backgrounds, indicating that mitochondrial genome structure inNicotiana is altered by substitution of the nuclear back-ground, since (d) inter- and intraspecific mitochondrial genome diversity among differentNicotiana species and the corresponding alloplasmic CMS tobacco analogs can be determined by hybridization with mtDNA specific probes. Analysis of in organello translation products in the three CMS-systems described confirmed the presence of variant proteins synthesized by mitochondria from CMS and male-fertileNicotiana isonuclear lines. In addition, differences due to the origin of both the nucleus and the cytoplasm, which involve changes in the presence or size of particular polypeptides, are apparent. A common feature of all three systems — including two different nuclear backgrounds — is the enhanced synthesis of a 31-kDa polypeptide in the strictly isonuclear CMS lines compared to the male-fertile tobacco. In addition, organellar gene expression was studied by Northern blot analysis of transcripts homologous to mitochondrial gene probes, revealing variant mtRNA species associated with some CMS lines.
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  • 47
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    Sexual plant reproduction 5 (1992), S. 64-71 
    ISSN: 1432-2145
    Keywords: Generative cell ; Isolation ; Microtubules ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Upon squashing of the pollen grain, the isolated generative cell ofNicotiana tabacum looses its spindle shape to become spherical; this phenomenon is independent of the sucrose concentration used. The time necessary for this change can vary from 1 min (0% sucrose) to 20 min (30% sucrose). The microtubular cytoskeleton was studied by means of immunofluorescence and electron microscopy. Just after isolation, 5 to 15 clearly visible bundles in microtubules organized in a basket-like structure are present. After 15 min in medium with 15% sucrose, the microtubular cytoskeleton disappears, and a diffusely spread tubulin can be observed. Neither the addition of 10–20 μM taxol to the medium, nor the omission of Ca2+ to the medium has any effect on the changes in cell shape and loss of microtubular bundles after isolation.
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  • 48
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    Sexual plant reproduction 5 (1992), S. 151-155 
    ISSN: 1432-2145
    Keywords: Calcium ; Chlorotetracycline ; Fertilization ; Nicotiana ; Synergid degeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Synergid degeneration was examined in the isolated embryo sac and egg apparatus of Nicotiana tabacum using quantitative cytology, fluorochromatic reaction (FCR) and chlorotetracycline (CTC). Most synergid degeneration occurs after pollen tubes (PT) arrive in the ovarian chamber between 42 and 48 h after pollination; synergid degeneration was precluded when PT were prevented from entering the ovary by stylar excision indicating that the signal that triggers synergid degeneration travels only relatively short distances in this plant. There was no evidence for any preferentiality between right or left synergids with regard to cell size or degeneration pattern. FCR staining confirms that synergid degeneration involves the loss of membrane integrity and is a reliable indicator of the onset of degeneration. CTC labeling of the degenerated synergid reveals that a concentrated reserve of membrane-bound calcium is present in the receptive synergid, possibly aiding in the attraction, arrest and discharge of the PT, releasing the sperms into the receptive ES.
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  • 49
    ISSN: 1432-2145
    Keywords: Female germ unit ; Nicotiana ; Megagametophyte ; Organelle movement ; Video microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Living embryo sacs and megagametophytic cells of Nicotiana alata and Nicotiana tabacum were obtained using enzymatic maceration and microdissection. The yields of isolated embryo sacs, egg apparatus and central cells were up to 35%, 40% and 35%, respectively. Vectorial movement of organelles and undulations of tubular structures, presumably endoplasmic reticulum, were observed in eggs, synergids and central cells using video-enhanced microscopy. Despite evident viability using the fluorochromatic reaction, the egg displays much less organelle movement and therefore appears to be quiescent. The large vacuole of the central cell is traversed by mobile strands of cytoplasm through which organelles migrate. A polygonal network is located at the periphery of the central cell, which may contribute to anchorage of the cell with the embryo-sac wall. The observation of organelle movement provides direct evidence of the condition of the cell and may be a useful approach for assessing cell vigor.
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  • 50
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    Sexual plant reproduction 5 (1992), S. 182-188 
    ISSN: 1432-2145
    Keywords: Isolated generative cells ; Microtubules ; Mitosis ; Immunofluorescence ; Confocal microscope ; Allemanda neriifolia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Organizational changes in the microtubules of isolated generative cells of Allemanda neriifolia during mitosis were examined using anti-α-tubulin and confocal laser scanning microscopy. Due to an improved resolution and a lack of out-of-focus interference, the images of the mitotic cytoskeleton obtained using the confocal microscope are much clearer than those obtained using the non-confocal fluorescence systems. In the confocal microscope one can see clearly that the spindle-shaped interphase cells contain a cage-like cytoskeleton consisting of numerous longitudinally oriented microtubule bundles and some associated smaller bundles. At prophase, the shape of the cells invariably becomes spherical. The microtubule cytoskeleton inside the cells concomitantly changes into a less organized form — consisting of thick bundles, patches, and dots. This structural form is not very stable, and soon afterwards the cytoskeleton changes into a reticulate network. Then the nuclear envelope breaks down, and the microtubules become randomly dispersed throughout the cell. Afterwards, the microtubules reorganize themselves into a number of half-spindle-like structures, each possessing a microtubule-nucleating center. The locations of these centres mark out the positions of the presumptive spindle poles. Numerous microtubules radiate from these centres toward the opposite pole. At metaphase, the microtubules form a number of bipolar spindles. Each spindle has two half-spindles, and each half-spindle has a sharply focused microtubule centre at the pole region. From the centres, kinetochore and non-kinetochore microtubules radiate toward the opposite half-spindle. At anaphase A, sister chromatids separate, the cells elongate, and the kinetochore microtubules disappear; the non-kinetochore microtubules, however, remain, and a new array of microtubules, in the form of a cage, appears. The peripheral cage bundles and the non-kinetochore bundles coverge into a sharp point at the pole region. Later, at anaphase B the microtubule cytoskeleton undergoes reorganization giving rise to a new array of longitudinally oriented microtubule bundles in the cell centre and a cage-like cytoskeleton in the periphery. At telophase, some of the cells elongate further, but some become spherical. The microtubules in the central region of the elongated cell become partially disrupted due to the formation of a phragmoplast-junction-like structure in the mid-interzone region. The microtubule bundles at the periphery are spirally organized, and they appear not to be disrupted by the phragmoplast-like junction. The microtubules in the spherical telophase cells (unlike those seen in the elongated telophase cells) are arranged differently, and no phragmoplast-junction-like structure forms in the spherical cells. The structural and functional significances of some of these new features of the organization of the microtubule cytoskeleton as revealed by the confocal microscope are discussed.
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  • 51
    ISSN: 1432-072X
    Keywords: Protein ; RNA synthesis ; Microtubules ; Microfilament-disrupting drugs ; Heat ; Cold shock ; Recovery
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effects of the microtubule-disrupting drugs, colchicine, vinblastine, podophyllotoxin, griseofulvin, and lumicolchicine (10-5 M), on protein and RNA synthesis were studied in Physarum polycephalum amoebae in culture. All, except lumicolchicine, were found to simultaneously reduce the rate of protein synthesis and stimulate RNA synthesis. These results parallel the effects seen in cells exposed to heat shock. Treatment of the cells with a microfilament-disrupting drug, cytochalasin B (10 μg/ml in ethanol), resulted in a reduced rate of protein synthesis after 2 h compared to a similar effect by vinblastine in 5–15 min. A morphological abnormality, microtubule paracystals, were seen associated with centrioles in vinblastine-treated cells in which protein synthesis had been reduced by 50%. Vinblastine and podophyllotoxin were shown to interfere with the recovery of protein synthesis after inhibition by low or elevated temperatures. The possible role of microtubules in regulating the translational response of a cell to an external environmental stimulus is discussed.
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  • 52
    ISSN: 1573-5028
    Keywords: genome organization ; in vitro transcription ; in vitro splicing ; Nicotiana ; tyrosine tRNA genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Tobacco tRNATyr genes are mainly organized as a dispersed multigene family as shown by hybridization with a tRNATyr-specific probe to Southern blots of Eco RI-digested DNA. A Nicotiana genomic library was prepared by Eco RI digestion of nuclear DNA, ligation of the fragments into the vector λgtWES·λB and in vitro packaging. The phage library was screened with a 5′-labelled synthetic oligonucleotide complementary to nucleotides 18 to 37 of cytoplasmic tobacco tRNATyr. Eleven hybridizing Eco RI fragments ranging in size from 1.7 to 7.5 kb were isolated from recombinant lambda phage and subcloned into pUC19 plasmid. Four of the sequenced tRNATyr genes code for the known tobacco tRNA1 Tyr (GψA) and seven code for tRNA2 Tyr (GψA). The two tRNA species differ in one nucleotide pair at the basis of the TψC stem. Only one tRNATyr gene (pNtY5) contains a point mutation (T54→A54). Comparison of the intervening sequences reveals that they differ considerably in length and sequence. Maturation of intron-containing pre-tRNAs was studied in HeLa and wheat germ extracts. All pre-tRNAsTyr-with one exception-are processed and spliced in both extracts. The tRNATyr gene encoded by pNtY5 is transcribed efficiently in HeLa extract but processing of the pre-tRNA is impaired.
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  • 53
    ISSN: 1573-5028
    Keywords: mRNA processing ; Nicotiana ; polymerase chain reaction ; sequencing ; transcription ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The xylanase gene from Cryptococcus albidus contains seven introns. Genomic and cDNA clones under the control of the CaMV 35S promoter were transferred into tobacco plants using Agrobacterium-mediated cell transformation. The genes were transcribed and the mRNAs were amplified by the polymerase chain reaction using primers on each side of the intron region. About 90% of the amplification products from plants transformed with the genomic clone corresponded to the size of the pre-mRNA (1.2 kb) and 10% represented the spliced product (0.85 kb). The 0.85 kb fragment was cloned and sequenced and the result indicated that the introns from the xylanase gene were accurately spliced by the plant cells.
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  • 54
    ISSN: 1573-5028
    Keywords: gene family ; Nicotiana ; oxygen evolution ; photosystem ; polymerase chain reaction
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  • 55
    ISSN: 1573-4943
    Keywords: Microtubules ; synapsin I ; cross-linking ; kinaseFa ; neurotransmission
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The ATP·Mg-dependent protein phosphatase activating factor (Fa) has been identified and purified to near homogeneity from brain. In this report, as evidenced on SDS-polyacrylamide gel electrophoresis followed by autoradiography, factorFa has further been identified as a cAMP and Ca2+-independent brain kinase that could phosphorylate synapsin I, a neuronal protein that coats synaptic vesicles, binds to cytoskeleton, and is believed to be involved in the modulation of neurotransmission. Kinetic study further indicated that factorFa could phosphorylate synapsin I with a lowK m value of about 2 µM and with a molar ratio of 1 mol of phosphate per mole of protein. Peptide mapping analysis revealed that factorFa specifically phosphorylated the tail region of synapsin I but on a unique site distinct from those phosphorylated by Ca2+/calmodulin-dependent protein kinase II and cAMP-dependent protein kinase, the two well-established synapsin I kinases. Functional study further revealed that factorFa could phosphorylate this unique specific site on the tail region of synapsin I and thereby inhibit cross-linking of synapsin I with microtubules. The results further suggest the possible involvement of factorFa as a synapsin I kinase in the regulation of axonal transport process of synaptic vesicles via the promotion of vesicles motility during neurotransmission.
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  • 56
    ISSN: 1432-2048
    Keywords: Inorganic pyrophosphate ; Nicotiana ; Sinksource ; Solanum ; Sucrose metabolism ; Transgenic plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Metabolite levels and carbohydrates were investigated in the leaves of tobacco (Nicotiana tabacum L.) and leaves and tubers of potato (Solanum tuberosum L.) plants which had been transformed with pyrophosphatase from Escherichia coli. In tobacco the leaves contained two- to threefold less pyrophosphate than controls and showed a large increase in UDP-glucose, relative to hexose phosphate. There was a large accumulation of sucrose, hexoses and starch, but the soluble sugars increased more than starch. Growth of the stem and roots was inhibited and starch, sucrose and hexoses accumulated. In potato, the leaves contained two- to threefold less pyrophosphate and an increased UDP-glucose/ hexose-phosphate ratio. Sucrose increased and starch decreased. The plants produced a larger number of smaller tubers which contained more sucrose and less starch. The tubers contained threefold higher UDP-glucose, threefold lower hexose-phosphates, glycerate-3-phosphate and phosphoenolpyruvate, and up to sixfold more fructose-2,6-bisphosphatase than the wild-type tubers. It is concluded that removal of pyrophosphate from the cytosol inhibits plant growth. It is discussed how these results provide evidence that sucrose mobilisation via sucrose synthase provides one key site at which pyrophosphate is needed for plant growth, but is certainly not the only site at which pyrophosphate plays a crucial role.
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  • 57
    ISSN: 1432-2048
    Keywords: Chitinase ; C-terminal processing ; β-1,3-glucanase ; Glycosylation ; Nicotiana ; Pathogenesis-related proteins ; Vacuole (targeting)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The class I β-1,3-glucanases are basic, vacuolar enzymes implicated in the defense of plants against pathogen infection. The tobacco (Nicotiana tabacum L.) enzyme is synthesized as a preproprotein with an N-terminal signal peptide for targeting to the lumen of the endoplasmic reticulum and an N-glycosylated C-terminal extension which is lost during protein maturation. The transport and processing of β-1,3-glucanase in cellsuspension cultures of the tobacco cultivar Havana 425 was investigated by pulse-chase labelling and cell fractionation. We verified that mature β-1,3-glucanase is localized in the vacuole of the suspension-cultured cells. Comparison of the time course of processing in homogenates, the soluble fraction, and membrane fractions indicates that proglucanase is transported from the endoplasmic reticulum via the Golgi compartment to the vacuole. Processing to the mature form occurs in the vacuole. Treatment of cells with tunicamycin, which inhibits N-glycosylation, and digestion of the 35S-labelled processing intermediates with endoglycosidase H indicate that β-1,3-glucanase has a single N-glycan attached to the C-terminal extension. Glycosylation is not required for proteolytic processing or correct targeting to the vacuole.
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  • 58
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    Theoretical and applied genetics 83 (1992), S. 515-527 
    ISSN: 1432-2242
    Keywords: Nicotiana ; Chloroplast DNA ; Restriction patterns ; Physical mapping ; Restriction site variations
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The restriction profiles of chloroplast DNA (cpDNA) from Nicotiana tabacum, N. sylvestris, N. plumbaginifolia, and N. otophora were obtained with respect to AvaI, BamHI, BglI, HindIII, PstI, PvuII, SalI, and XhoI. An efficient mapping method for the construction of cpDNA physical maps in Nicotiana was established via a computer-aided analysis of the complete cpDNA sequence of N. tabacum for probe selection. The efficiency of this approach is demonstrated by the determination of cpDNA maps from N. sylvestris, N. plumbaginifolia, and N. otophora with respect to all of the above restriction endonucleases. The size and basic structure of the cpDNA from the three species are almost identical, with an addition of approximately 80 bp in N. plumbaginifolia. The restriction patterns and hence the physical maps between N. tabacum and N. sylvestris cpDNA are identical and there is no difference in the Pvull digests of cpDNA from all four species. Restriction site variations in cpDNA from different species probably result from point mutations, which create or eliminate a particular cutting site, and they were observed spanning the whole chloroplast molecule but highly concentrated in both ends of the large, single-copy region. The results presented here will be used for the forthcoming characterization of chloroplast genomes in the interspecies somatic hybrids of Nicotiana, and will be of great value in completing the exploration of the phylogenetic relationships within this already extensively studied genus.
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  • 59
    ISSN: 1432-2242
    Keywords: Somatic cell fusion ; Asymmetric hybrids ; Atropa ; Nicotiana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mesophyll protoplasts of the kanamycin-resistant nightshade, Atropa belladonna, were fused with mesophyll protoplasts of the phosphinothricin resistant-tobacco, Nicotiana tabacum. A total of 447 colonies resistant to both inhibitors was selected. Most of them regenerated shoots with morphology similar to one of the earlier obtained and described symmetric somatic hybrids Nicotiana + Atropa. However, three colonies (0.2%) regenerated vigorously growing tobacco-like shoots; they readily rooted, and after transfer to soil, developed into normal, fertile plants. Unlike their tobacco parental line, BarD, the obtained plants are resistant to kanamycin [they root normally in the presence of kanamycin (200 mg/1)] and possess activity of neomycin phosphotransferase (NPT II) with the same electrophoretic mobility as the one of the nightshade line. According to Southern blot hybridization analysis carried out with the use of radioactively labeled cloned fragments of the Citrus lemon ribosomal DNA repeat, as well as with Nicotiana plumbaginifolia genus-specific, interspersed repeat Inp, the kanamycin-resistant plants under investigation have only species-specific hybridizing bands from tobacco. Cytological analysis of the chromosome sets shows that plants of all three lines possess 48 large chromosomes similar to Nicotiana tabacum ones (2n = 48), and one small extra chromosome (chromosome fragment) similar to Atropa belladonna ones (2n = 72). Available data allow the conclusion that highly asymmetric, normal fertile somatic hybrids with a whole diploid Nicotiana tabacum genome and only part (not more than 2.8%) of an Atropa belladonna genome have been obtained without any pretreatment of a donor genome, although both these species are somatically congruent.
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  • 60
    ISSN: 1432-2242
    Keywords: Nicotiana ; Plastid genetic markers ; Plastid segregation ; Plastid selection ; Protoplast fusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Vegetative segregation of a mixed plastid population in protoplast fusion-derived cell lines can be directed by a selection favouring the multiplication of one of the parental plastid types. This report defines some of the critical conditions leading to a homogeneous plastid population in cybrid plants generated by protoplast fusion between Nicotiana plumbaginifolia and an albino and streptomycin-resistant N. tabacum plastid mutant. Light (1,500 lx) conferred a strong selective advantage to chloroplasts versus albino plastids, while the lack of this effect in dim light (300 lx) indicated that a sufficient light intensity is essential to the phenomenon. Selection on streptomycin-containing medium in the dark, however, led to the preferential multiplication of resistant plastids. Streptomycin selection of resistant chloroplasts in the light, consequently, results in a plastid selection of doubled stringency. In another experiment a definite, but leaky, selection for chloroplast recombination (selection for greening on streptomycin-containing medium in dim light) was used to reveal various recombination products. Protoplast fusion in fact resulted in cybrid plants showing only simple chimeric segregation of unchanged parental plastids. These results demonstrate the essential requirement for stringent plastid selection, as defined by cell culture conditions, to precede the formation of shoots expected to possess the desired plastid genetic composition.
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  • 61
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    Theoretical and applied genetics 58 (1980), S. 121-127 
    ISSN: 1432-2242
    Keywords: Nicotiana ; Cytoplasmic-male-sterility ; Somatic hybridization ; Plastid segregation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Restoration of male fertility was achieved by fusing protoplasts from male sterile (CMS) Nicotiana sylvestris plants with X-irradiated protoplasts derived from fertile N. tabacum plants. The CMS N. sylvestris plants were derived from a previous somatic hybridization experiment and contained alien (Line 92) cytoplasm. About one quarter of the regenerated plants were found to be cybrids. i.e. they consisted of N. sylvestris nuclei combined with all or some components of N. tabacum cytoplasm. In one half of these cybrids male fertility was restored to different levels. The chloroplasts of the two parental donors differ in respect to tentoxin sensitivity: chloroplasts of CMS N. sylvestris are sensitive while those of N. tabacum are insensitive. It could therefore be demonstrated that there was an independent segregation of chloroplast type and male fertility/sterility: several somatic cybrids were male fertile but tentoxin sensitive and others were tentoxin insensitive yet they were male sterile. Only in about one half of the somatic cybrids was male fertility restored together with restoration to tentoxin insensitivity.
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  • 62
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    Theoretical and applied genetics 84 (1992), S. 33-38 
    ISSN: 1432-2242
    Keywords: Homologous recombination ; ATPase α-subunit ; Protoplast fusion ; Nicotiana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Segments of mitochondrial DNA (mtDNA) carrying the gene for the α-subunit of F1-ATPase (atpA) were detected by Southern hybridization with atpA from pea as probe. In the case of Nicotiana langsdorffii, we identified four fragments that are derived from combinations of two different 5′ and two different 3′ flanking regions of atpA. All four types share the coding region, suggesting that they result from homologous recombination in the coding region of atpA. By contrast, N. glauca generated only one analogous fragment, which indicated the existence of only a single type of atpA in N. glauca. In the case of somatic hybrids obtained by fusion between protoplasts from N. langsdorffii and N. glauca, analysis with EcoRI or HindIII detected three new fragments in addition to the parental fragments. These new fragments can be explained by homologous recombination within the coding region of atpA. Our results show that the coding region of atpA is involved not only in intragenomic homologous recombination but can also be involved in homologous recombination between two parental mitochondrial genomes of somatic hybrids.
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  • 63
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    Cell & tissue research 270 (1992), S. 553-558 
    ISSN: 1432-0878
    Keywords: Development, ontogenetic ; Cold exposure ; Microfilaments ; Microtubules ; Centrioles ; Drosophila melanogaster (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary When earlyDrosophila embryos were allowed to develop at 0°C, several abnormalities in the surface cap organization were observed. Scanning electron microscopy showed that exposure to cold mainly lead to the deformation of the cortical caps and to their partial fusion with adjacent caps. The process of cellularization was presumably affected and large uncellularized areas were observed. Rhodamine-phalloidin staining showed that cap deformation was closely related to the altered microfilament distribution, which was presumably responsible for the failure of large syncytial areas to cellularize. During the process of cellularization, F-actin localization did not depend on the microtubules forming the baskets around the elongating nuclei, but was related to the subpopulation of mictotubules radiating from the centrosomes toward the plasma membrane. Only these microtubules seemed to be affected by cold treatment.
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  • 64
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    Cell & tissue research 270 (1992), S. 353-363 
    ISSN: 1432-0878
    Keywords: Eye, compound ; Glia-like cells ; Pigment cells ; Cytoskeleton ; Actin filaments ; Spectrin ; Microtubules ; Apis mellifera (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The organization of the submembrane cytoskeleton of non-photoreceptive, accessory cells in the honeybee compound eye was examined using light-microscopic (phallotoxin labeling, immunohistochemistry) and electron-microscopic (decoration with myosin fragments) techniques. The crystalline cone cells contain numerous peripheral actin filaments oriented longitudinally with antiparallel polarity. Bundles of microtubules lie under the plasma membrane of primary pigment cells, in close apposition to the crystalline cone; they are interspersed with only a few actin filaments. Pigmented glial cells (secondary pigment cells) contain a two-dimensional filament/particle web lining their entire plasma membranes. Both filamentous actin and α-spectrin are localized within the cortex of these cells, indicating that they are web components. The results demonstrate that the three cell types contain different cortical cytoskeletons, implying different functional properties.
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  • 65
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    Cell & tissue research 269 (1992), S. 305-313 
    ISSN: 1432-0878
    Keywords: Cilia ; Basal apparatus ; Basal body ; Striated rootlet ; Microfilaments ; Microtubules ; Lytechinus pictus (Echinodermata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The basal apparatus of embryonic cells of the sea urchin Lytechinus pictus was examined by transmission electron microscopy and compared with the basal apparatus of other metazoan cells. The basal apparatus in these cells is associated with a specialized region of the apical cell surface that is encircled by a ring of microvilli. The basal apparatus includes several features that are common to all ciliated cells, including a basal body, basal foot, basal foot cap, and striated rootlet. However, a component not seen in the basal apparatus of other species has been observed in these cells. This structure is continuous with the striated rootlet, and its ultrastructure indicates that it is composed of the same components as the rootlet. This structure extends from the junction of the basal body and striated rootlet to the cortical region that surrounds the basal body. Based on its morphology and position, this structure is referred to as a striated side-arm. The striated side-arm is always aligned in the plane of the basal foot. Thus, both of these structures extend from the basal body in the plane of the effective stroke. It is suggested that the striated side-arm serves to stabilize the basal apparatus against force exerted by the cilium.
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  • 66
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    Cell & tissue research 208 (1980), S. 171-181 
    ISSN: 1432-0878
    Keywords: Microtubules ; Dendritic spine apparatus ; Synapse ; Development ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using techniques for enhanced microtubular preservation, including albumin pretreatment (Gray, 1975), occipital cortex of rats was studied electron microscopically at various ages of development. A close structural relationship was seen between microtubules, sacs of SER and the postsynaptic “thickening” in primordial spines and with the dense “plate” material of spine apparatuses. Stereoscopic preparations in addition show a more complicated substructure than previously described for the “plate”. Microtubules may contribute to the formation of the “plate” of the spine apparatus which in turn is associated with the postsynaptic “thickening” of the mature spine. Possible functional correlates are discussed.
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  • 67
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    Potato research 35 (1992), S. 355-364 
    ISSN: 1871-4528
    Keywords: cultivar ; nitrate ; nitrogen ; Solanum tuberosum L. ; tuber
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The duration of dormancy of seed potatoes varies between years and between origins. Therefore, the effects of conditions during crop growth on dormancy of progeny tubers were studied. The effect of nitrogen during tuber bulking on the duration of dormancy was investigated in three field experiments with two cultivars. In addition to an application of 125 kg N/ha at planting, top dressings of 0–150 kg N/ha were given about 2 weeks after tuber initiation. Haulm was pulled about 4 weeks later. The effect of nitrogen rate at planting was also examined in one experiment. Nitrogen top dressings shortened dormancy in all experiments by 5–8 days. An increased nitrogen rate at planting resulted in a shorter dormancy when the duration of dormancy was expressed in days after tuber initiation, but not when it was expressed in days after haulm pulling, probably because extra nitrogen also delayed tuber initiation.
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  • 68
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    Potato research 35 (1992), S. 365-375 
    ISSN: 1871-4528
    Keywords: cultivar ; heat sprouting ; nitrogen ; second growth ; Solanum tuberosum L. ; sprout ; tuber
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary In two indoor experiments under short day conditions, the effect of temperature during tuber bulking on dormancy of tubers was investigated for cvs Diamant and Désirée. Temperature treatments started after tuber initiation and lasted for 4 weeks, after which the haulm was removed. In Experiment 1, the day/night temperature regimes 18/12, 22/22, 26/18 and 32/12 °C (T18/12 etc.) were compared. In Experiment 2, three day temperatures (18, 24 and 30 °C) were combined with three night temperatures (12, 18 and 24 °C), resulting in nine treatments. The dormancy of cv. Diamant was shortest after very high day temperatures (30–32 °C), but intermediate day temperatures (22–26 °C) had no shortening effect compared to T18/12. Dormancy of cv. Désirée was not shortened, but rather tended to be prolonged by high temperatures (22–32 °C) during growth. High temperatures during growth resulted in more sprouts per tuber after dormancy had ended.
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  • 69
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    Potato research 23 (1980), S. 371-375 
    ISSN: 1871-4528
    Keywords: nitrogen ; phosphorus ; potassium ; calcium ; sodium ; nitrate ; sulphate ; micronutrients
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The specific gravity and dry matter content of 193 tuber samples (cultivars: Arran Banner, Spunta, Up-to-Date, Cara) from crops grown at Kokkinochoria, the major potato growing area of Cyprus, were related to each other as follows: Dry matter (%)=−269.59+268.24×specific gravity. This regression accounted for 82.4% of the variation; the fiducial limits (P〈0.05) for single determinations of specific gravity were±1.54 percentage units. The mineral contents of the tubers were similar to those reported in other countries.
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  • 70
    ISSN: 1573-9171
    Keywords: EPR ; radical ; fluorine ; conformation ; phosphorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract EPR spectra of radical adducts of phosphonyl radicals with 2-hydroperfluoro-4-methyl- and 4,4-dimethyl-2-pentene have been studied. The molecular mechanics method has been used to determine the preferred conformation of the (CF3)2CF-CF-HCF3P(O)(OMe)2 radical. The eclipsed conformation of the C–P bond and 2p z -orbital of an unpaired electron is stabilized due to steric factor and hyperconjugation.
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  • 71
    ISSN: 1615-6102
    Keywords: Cytoplasmic inheritance ; Male cytoplasmic diminution ; Male gamete (angiosperm) ; Male germ unit ; Nicotiana ; Tobacco
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The organization of the sperm cells and vegetative nucleus (male germ unit) ofNicotiana tabacum was examined 18 h after semivivo pollination using transmission electron microscopy, computerassisted serial section reconstruction and quantitative cytology. Based on a measurement of 11 cellular parameters in nine reconstructed sperm cell pairs, there are no statistically significant differences between the two cells. The Svn is characterized by a strapshaped cytoplasmic extension that is physically associated with the surface of the vegetative nucleus. The nucleus is located adjacent to the sperm crosswall, with sperm organelles being distributed between the nucleus and the extension. The Sua is a tapered cell with cytoplasmic areas at both poles and deep axial invaginations near the crosswall. This cell has a centrally-located nucleus and a largely polar distribution of organelles. Three mechanisms for cytoplasmic diminution were observed that appear to contribute actively to the loss of cytoplasmic volume and organelles: (1) enucleated cytoplasmic body production in the Sua; (2) vesiculation at the tip of the cytoplasmic projection of the Svn; and (3) vesicle-containing body accumulation in the periplasm of both the Svn and Sua.
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  • 72
    ISSN: 1615-6102
    Keywords: Actin ; Microtubules ; Cytoskeleton ; Polarity ; Pea root ; Wound
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary To assess the relative roles of cortical actin and microtubule re-orientation in the establishment of new cell polarity, we have examined the kinetics of cortical actin re-orientation around a wedge-shaped wound in pea roots. Cortical actin re-orients from a transverse alignment to an approximately longitudinal orientation between 5 and 24h after wounding, that is, after the re-alignment of microtubules, which is known to occur before 5h post-wounding. F-actin in root cortical cells does not appear to be necessary for the establishment of new cell polarity around wounds, since normal MT re-alignment, and new planes of cell division are still established around a wound in cytochalasin treated roots. The cytochalasin treatment appeared to totally disrupt cortical and cytoplasmic F-actin in cells of the root cortex. However, in the apparent absence of F-actin in these cells, the rate of wound-induced cell division, but not cell expansion, is slower, and we suggest that an effect on the phragmosomal actin is involved. Finally, we demonstrate that new cell polarity around a wound is not established if microtubules are disrupted by the herbicide oryzalin, but after re-establishment of these arrays following a wash-out of the drug, the typical new planes of cell expansion are observed. We conclude that microtubules play a critical role in establishing and maintaining cell polarity in this system, and that cortical F-actin has a minor and presently unclear function in these processes.
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  • 73
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    Protoplasma 168 (1992), S. 64-72 
    ISSN: 1615-6102
    Keywords: Cell wall ; Microtubules ; Protoplast ; Taxol ; Tobacco
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Microtubules (MTs) are important for plant cell morphogenesis because they influence the deposition of cell plate and wall components. It has been observed that tobacco protoplasts contain a disordered MT array in the cortex. Following several days in culture, these protoplasts become elongate cells with an orderly cortical MT array. The transformation of the MT array may occur by net depolymerization of the disordered MTs and repolymerization of MTs into an ordered array, or by movement of the array as an integral unit. To experimentally distinguish between these two possibilities, the drug taxol was used to stabilize MTs. Protoplasts derived from suspension cultured tobacco,Nicotiana tabacum, were grown in a medium containing the two plant hormones α-naphthaleneacetic acid and benzyladenine, in the presence or absence of 10μM taxol. Changes in cell size and shape were quantified using a video image analysis system. Cell elongation had begun within 48h of protoplast conversion, in both treatments, and continued for 7 days. Immunolocalization of tubulin showed that, in the majority of cells, MTs were disorganized immediately following protoplast conversion. After elongation, the MT arrays were observed to have reoriented to an ordered state. Taxol-treated protoplasts were found to elongate faster and to a greater extent than the non-treated controls. Additionally, the cortical array of taxol-treated protoplasts reorganized more quickly. These data indicate that the net depolymerization of disordered cortical MTs is not necessarily required for the differentiation of a protoplast into an elongate cell.
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  • 74
    ISSN: 1615-6102
    Keywords: Asters ; Microtubules ; Mitosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Using an antibody against tyrosinated tubulin and epifluorescence microscopy, mitosis was studied in three different microvessel endothelial cell types recently isolated from bovine corpus luteum. Dividing cells were flat and at certain stages individual microtubules could be followed for considerable lengths. The structure of the spindle apparatus and the course of mitosis were conventional. Microtubule asters were small from prophase until metaphase in all three cell types. However, whereas in two cell types telophase asters remained inconspicuous, prominent asters, of mostly straight microtubules, formed in telophase cells of a third cell type. Thus, aster size is heterogeneous between different endothelial cell types. Large microtubule asters are not regularly found in dividing cultured mammalian cells. The microendothelial cell types present themselves as appropriate systems for spindle research and especially for the study of aster microtubule dynamics and function.
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  • 75
    ISSN: 1615-6102
    Keywords: Actin filaments ; Microtubules ; Nicotiana tabacum ; Pollen tube ; Protoplast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Microtubules inNicotiana tabacum pollen tube subprotoplasts reassembled in wave-like to concentric cortical arrays. Crosslinks between microtubules were either 15 or 80 nm in length. Cortical actin filaments showed different distributions; no colocalization like that in pollen tubes was observed. Degradation of actin filaments by cytochalasin D had no influence on microtubule organization. Degradation of microtubules and/or actin filaments did not affect outgrowth of the subprotoplasts. Organization of the microtubules occurred independent of the presence of the generative cell and/or the vegetative nucleus. No relation of actin filament and microtubule organization with organelle distribution could be detected.
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  • 76
    ISSN: 1615-6102
    Keywords: Pollen tube ; Microtubules ; Cellular division ; Generative cell ; Sperm cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The microtubular cytoskeleton of the generative cell (GC) ofHyacinthus orientalis has been studied until the formation of the sperm cells (SCs). Immunofluorescence procedures in combination with confocal laser scanning microscopy (CLSM) has enabled the visualization of the organization of the microtubular cytoskeleton. Chemical fixation and freeze-fixation electron microscopy have been used to investigate the cytoskeleton and the ultrastructural organization of the GC and SCs. During pollen activation the GC is spindle-shaped. Microtubules (MTs) are organized as bundles and distributed in proximity of the GC plasmamembrane, forming a basket-like structure. Following migration through the pollen tube, the basket-like structure becomes more intertwined. During the nuclear division the MTs are involved in the segregation of the chromosomes and kinetochores are clearly discernible. Association with organelles is also observed. The chromosomes of the GC remain condensed until they separate in two sperm nuclei. The pre-prophase band was never observed. At the end of the GC division the microtubular network reorganizes in the two SCs.
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  • 77
    ISSN: 1615-6102
    Keywords: Dictyota dichotoma ; Microtubules ; Centrioles ; Mitotic spindle ; Cell cycle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Interphase cells ofDictyota dichotoma (Hudson) Lamour. lack cortical microtubules (Mts) but display an impressive network of cytoplasmic microtubules (c-Mts). These are focussed on two opposed perinuclear centriolar sites where centrin or a centrin-homologue is localized. Some of the Mts surround the nucleus, but the majority traverse the cytoplasm as bundles variously directed towards the plasmalemma. In apical cells, and to a lesser extent in the square or slightly elongated meristematic cells, Mts are more or less evenly arranged. In elongated cells they form thick bundles longitudinally traversing the cytoplasm; a pattern maintained in differentiated cells. In early prophase the non-perinuclear Mts disappear but by late prophase a bi-astral arrangement of short Mts is observed. They enter polar nuclear depressions and attach to differentiated regions of the nuclear envelope where polar gaps open. By metaphase the spindle Mts converge on the centrioles at the polar gaps. At anaphase, interzonal Mts are evident and the asters start to reassemble. After telophase disruption of the interzonal Mts, the daughter nuclei approach each other, but move apart again before cytokinesis. The latter movement keeps pace with the development of two interdigitating Mt systems, ensheathing both daughter nuclei. The partition membrane “bisects” this Mt “cage”. Between telophase and cytokinesis the centrosomes separate, finally occupying opposed perinuclear sites. New Mts arise at the new centrosomes, some terminating on the consolidating partition membrane. Our data show thatD. dichotoma vegetative cells display a prominent cytoplasmic Mt cytoskeleton, which undergoes continual, but definite, change in organization during the cell cycle.
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  • 78
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    Protoplasma 169 (1992), S. 148-154 
    ISSN: 1615-6102
    Keywords: Cell wall ; Micrasterias ; Microfibrils ; Microtubules ; Microtubule inhibitors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The influence of the microtubule disorganizing substances amiprophos-methyl (APM) and colchicine on secondary wall formation inMicrasterias denticulata was investigated by the freezeetch technique. The results reveal that neither microtubule inhibitor changes the pattern of microfibril deposition. The application of APM or colchicine also does not cause any structural alterations of the microfibrils or of the protoplasmic (Pf) and the exoplasmic (Ef) fracture face of the plasma membrane, thus indicating that microtubules are not involved in secondary wall formation inM. denticulata. However, since areas of the plasma membrane which collapsed upon freeze-etching are restricted to the Pf-face of cells treated with microtubule inhibitors, cortical microtubules may function as mechanical support during secondary wall formation. In the cortical cytoplasm filamentous structures are found in close spatial relationship and an almost parallel alignment to rosettes of the plasma membrane.
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  • 79
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    Protoplasma 102 (1980), S. 31-51 
    ISSN: 1615-6102
    Keywords: Microtubules ; Morphogenesis ; Cell Walls ; Roots ; Colchicine ; Cell division
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Removal and subsequent reformation of microtubules in cells of the root-tips ofAzolla pinnata R. Br. was achieved by short pulse treatments with the drug colchicine. Loss of microtubules led to the formation of multinucleate cells more frequently than to the arrest of mitosis at metaphase, and primary and secondary wall formation was also disrupted. Recovery of root development was limited. Growth of all roots ceased 5–6 days after the pulse treatment. Following the reappearance of microtubules, renewed deposition of normal wall thickenings occurred in developing xylem elements. Multinucleate cells became subdivided by walls in the apparent absence of a phragmoplast. The plane in which the new wall was formed was often located as it would have been in an untreated root, but in a number of cases abnormal or precious positioning of new walls was observed. Clusters of microtubules, matrix material, and vesicles or particles, taken to indicate microtubule initiation, were observed during the recovery from treatment.
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  • 80
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    Protoplasma 103 (1980), S. 105-114 
    ISSN: 1615-6102
    Keywords: Colchicine ; Lumicolchicine ; Microtubules ; Mitosis ; Wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Lumicolchicine was purified by preparative thin-layer chromatography. Tests for purity were ultraviolet absorption spectrophotometry, analytical thin-layer chromatography, and a bioassay using wheat roots. Wheat roots treated for 3 days with 10−3 M lumicolchicine showed no c-mitosis, but had reduced growth compared with controls.
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  • 81
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    Protoplasma 103 (1980), S. 205-229 
    ISSN: 1615-6102
    Keywords: Cell wall ; Cytochalasin B ; Microfibril orientation ; Microtubules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Cortical microtubule arrays in the radish root hair were analyzed from reconstructions of serial ultra-thin sections in order to test extant hypotheses concerning the role of microtubules in the deposition of oriented microfibrils of cellulose. Passing away from the tip, root hairs exhibit a transition from random to oriented deposition of microfibrils at approximately 25 μm. Along the root hair, passing back from the tip, the microtubules: a) increase in number to a plateau at 25 μm; b) change their length profiles from approximately 60% less than 1 μm long in the hair tip to approximately 40% less than 1 μm long at 60 μm; c) maintain a constant pattern of angular deviation from the long axis, which is similar to the deviation pattern of the oriented wall fibrils; d) maintain a constant (approximately 70% of tubules) close (within 50 nm) proximity to the plasma membrane (PM); e) maintain a low (approximately 20%) degree of inter-microtubule proximity (i.e., within 50 nm of one another); f) show evidence for some variable long range (〉50 nm) association. Fixation with glutaraldehyde in a complete microtubule polymerization medium (MTPM), or pretreatment with cytochalasin B cause an approximate twofold increase in 1. the proportion of long microtubules in the tip region and 2. microtubules within 50 nm of one another. Fixation in incomplete MTPM (without GTP) produces results similar to phosphate buffer controls. Alternative explanations for these results are examined. A new hypothesis accounting for microtubule involvement in oriented microfibril deposition is described.
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  • 82
    ISSN: 1615-6102
    Keywords: Dikaryon ; Griseofulvin ; Microtubules ; Nuclear division ; Septal development ; Septal dissolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Ultrastructural study of a dikaryon of the basidiomyceteSchizophyllum commune showed that treatment with griseofulvin affected the site of the dividing nuclei and the location and structure of the septa. The microtubules were considered to be the primary target of griseofulvin, since they participate in nuclear division and movement in the hyphae, and their assembly is known to be in other organisms than fungi inhibited by griseofulvin. It is pointed out that dikaryotic hyphae with two nuclei and a clamp connection per cell are more sensitive indicators of the effect of griseofulvin than homokaryotic hyphae, whose structure is less complex.
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  • 83
    ISSN: 1615-6102
    Keywords: Cell wall ; Chytridiomycetes ; Chytridium confervae ; Cross-wall ; Microtubules ; Ribosomes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Chytridium confervae is a eucarpic, monocentric chytrid. We have used light and electron microscopy to study the relationship between the nutrient absorbing rhizoids and the asexually reproductive sporangium during growth. We have also examined the induction of zoosporogenesis by starvation, and subsequent differentiation until zoospore release. During growth the cytoplasm of the rhizoids and the developing sporangium was continuous and similar. At the start of starvation a bundle of fibers that were visible with light microscopy appeared at the junction between the rhizoids and the sporangium. Two hours after initiation of starvation a wall, that was also visible with light microscopy, formed to separate the rhizoids from the sporangium. Electron microscopy revealed a large, ordered array of microtubules in the thallus at the same time that the fibers appeared, and a sharp difference in the density of ribosomes in the cytoplasm of the sporangium and that of the rhizoids that was apparent immediately after starvation. This cytoplasmic difference was preserved by the formation of a cross-wall that was penetrated by plasmodesmata. After the wall was formed the cytoplasm of the rhizoids senesced. Comparison ofC. confervae with other organisms that use arrays of microtubules to move organelles is made and speculation on the role of the microtubules in organelle movement and wall formation inC. confervae is offered.
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  • 84
    ISSN: 1615-6102
    Keywords: Cell shape ; Chlorosarcinopsis ; Low temperature ; Microtubules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The effect of low temperature (2 °C) on cell shape and microtubules in zoospores of the green algaChlorosarcinopsis gelatinosa has been investigated. The zoospores are 4–6 times longer than wide with a mean length of 12,5 μm and can be kept in the dark for several hours without changes in cell shape. Cell shape changes have been evaluated quantitatively by measuring changes in cell length. Low temperature induces a decrease in cell length which exhibits a two-step kinetic: during the first 30 minutes a rapid rate of decrease in cell length was measured, while during the next 4 hours a slow rate of decrease in cell length was observed. Complete regeneration of zoospore length occurs when cold-treated cells are subjected to the original zoospore induction temperature (30 °C) for two hours. Observation of numbers, disposition and types of microtubules in the zoospore during decrease in cell length has shown that within 30 minutes after cold application the secondary cytoskeletal microtubules (scmt) disappear, while flagellar root microtubules are unaffected. During this period most cells develop a prominent posterior appendage (tail). Sections demonstrate the presence of several microtubules in these tails. Flagellar root microtubules probably extend into the tails and disappearance of scmt starts at the posterior pole of the cell. Regeneration of zoospores to original cell length is coupled with reappearance of scmt starting at the anterior pole of the cell. It is concluded that secondary cytoskeletal microtubules constitute the main cytoskeleton inChlorosarcinopsis zoospores and that flagellar root microtubules contribute to only a minor extent to the cytoskeleton, because they cannot retain the cell shape. The results are discussed with respect to the functional significance of flagellar root microtubules in green algae.
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  • 85
    ISSN: 1615-6102
    Keywords: Immunofluorescence ; Microtubules ; Plant protoplasts ; Tobacco
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The arrangement of cortical microtubules in tobacco protoplasts is described using the following techniques: 1. Transmission electron microscopy (TEM) of thin sections of whole protoplasts, 2. TEM of negatively stained protoplast ghosts, and 3. Indirect immunofluorescence microscopy of protoplast ghosts. Ghosts were prepared by attaching freshly isolated protoplasts to glass coverslips or formvar/carbon-coated grids with poly-L-lysine and then bursting them either osmotically or by detergent treatment in the presence of a microtubule stabilizing buffer. Osmotic bursting of protoplasts yielded large pieces of plasma membrane with attached microtubules. These preparations proved very useful for measuring the density and length of cortical microtubules. Detergent treatment dissolved the plasma membrane and altered the distribution of cortical microtubules.
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  • 86
    ISSN: 1615-6102
    Keywords: Mebendazole ; Ascaridia ; Secretion ; Microtubules ; Anthelmintic
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    Topics: Biology
    Notes: Summary The anthelmintic compound mebendazole caused the disappearance of microtubules in the intestinal cells ofAscaridia galli. Electron microscopy revealed that soon after the microtubules disappeared there was an accumulation of secretory vesicles near the golgi areas. subsequently many of these vesicles aggregated forming dense large vesicles near the terminal web of the intestinal cells. This provides further evidence for the involvement of microtubules in the secretion of products from eukaryotic cells. It seems likely that inhibition of microtubule directed secretory functions in various cell types is an important function in the anthelmintic activity of the benzimidazole carbamates.
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  • 87
    ISSN: 1615-6102
    Keywords: Embryogenic pollen ; Nicotiana ; Pollen culture ; Selection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary By using density gradient centrifugation, employing 55% percoll and 4% sucrose as suspension medium, it is possible to select embryogenic pollen from buds after cold treatment at 10 °C for 8 or more days. These buds at the uninucleate stage of pollen were collected from plants grown in 8 hours photocycles at 18 °C and supplied with mineral salts. The embryogenic pollen are small, starch-free with a clear cytoplasm whereas large starch-filled ones are nonembryogenic. The embryogenic pollen regularly form embryos at a frequency of 2% on a mineral medium supplemented with glutamine, asparagine and sucrose at pH 6.5. These results demonstrate, for the first time, that it is possible to have embryos in appreciable frequencies in “ab initio” pollen cultures raised from cold treated anthers.
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  • 88
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    Protoplasma 167 (1992), S. 183-192 
    ISSN: 1615-6102
    Keywords: Endoplasmic reticulum ; F-actin ; Microtubules ; Orchids ; Pollen ; Mitosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The unequal first mitosis in pollen ofPhalaenopsis results in a small generative cell cut off at the distal surface of the microspore and a large vegetative cell. No preprophase band of microtubules is present, but polarization of the microspore prior to this critical division is well marked. A generative pole microtubule system (GPMS) marks the path of nuclear migration to the distal surface, and the organelles become unequally distributed. Mitochondria, plastids and dictyosomes are concentrated around the vegetative pole in the center of the microspore and are almost totally excluded from the generative pole. The prophase spindle is multipolar with a dominant convergence center at the GPMS site. The metaphase spindle is disc-shaped with numerous “minipoles” terminating in broad polar regions. In anaphase, the spindle becomes cone-shaped as the spindle elongates and the vegetative pole narrows. These changes in spindle architecture are reflected in the initial shaping of the telophase chromosome groups. F-actin is coaligned with microtubules in the spindle and is also seen as a network in the cytoplasm. An outstanding feature of orchid pollen mitosis is the abundance of endoplasmic reticulum (ER) associated with the spindle. ER extends along the kinetochore fibers, and the numerous foci of spindle fibers at the broad poles terminate in a complex of ER.
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  • 89
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    Protoplasma 102 (1980), S. 295-306 
    ISSN: 1615-6102
    Keywords: Acetabularia ; Inhibitors ; Microcinematography ; Microfilaments ; Microtubules ; Protoplasmic streaming
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The sensitivity of the dual intracellular transport system inAcetabularia mediterranea (Koop andKiermayer 1979 a) to cytochalasin B (CB, 10−5 mol), chlorisopropyl-N-phenylcarbamate (CIPC, 2.10−4 mol), and amiprophosmethyl (APM, 3.10−5 mol) has been studied by microcinematography. Vegetative cells before cap formation, 2–3 cm in length, and cells with a maximum sized cap, containing secondary nuclei, were used for the experiments. All intracellular movements ceased under the influence of CB, while in contrast to “headed streaming bands” and to the migration of the secondary nuclei, the movement of chloroplasts at “thin filaments” was found to be insensitive to Col, CIPC, and APM. All inhibitory effects of the drugs on protoplasmic streaming were completely reversible within a time of less than 10–20 minutes recovery from the drugs. The results suggest an involvement of microfilaments in all intracellular movements while, in addition, microtubules seem to be connected with the movement on “headed streaming bands”, including the migration of secondary nuclei.
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  • 90
    ISSN: 1615-6102
    Keywords: Tetrahymena ; Microtubules ; Conjugation ; Genomic exclusion ; Taxol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Indirect immunofluorescence has revealed various intracytoplasmic microtubular structures, which are transiently polymerized in specific subcellular locations during the developmental process of conjugation in the ciliateTetrahymena thermophila. These structures include: (1) micronuclear spindles, (2) perimicronuclear microtubules, (3) microtubular baskets surrounding migrating pronuclei, and (4) microtubules interconnecting the pronuclei with the conjugants' junctional zone. Furthermore, a peripheral network of intracytoplasmic microtubules related to the cell cortex is present in both vegetative cells and in conjugants. Comparative observations made on cells undergoing normal conjugation and defective conjugation (occurring either spontaneously or induced by taxol) has revealed some rules governing the pattern of deployment of conjugation-specific microtubules. The presence of perinuclear microtubular arrays during early postmeiotic stages of development is strictly limited to more anteriorly located nuclei which includes the selected haploid nucleus that further divides to form the stationary and migratory pronuclei. These perinuclear microtubules may be involved in the positional control of nuclear fates leading to effective nuclear selection. Microtubular bundles associated with pronuclei and connecting the junctional zone are only formed in the presence of functional pronuclei, and may be involved in the guidance of pronuclei leading to their fusion. The mechanism of cytoplasmic control of nuclear differentiation of derivatives of the zygotic nucleus appear to be associated with a coordinate action of two microtubular arrays: spindle microtubules of the second postzygotic division and the peripheral intracytoplasmic network of microtubules, leading to a proper subcortical positioning of the postzygotic nuclei at opposite poles of the cell.
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  • 91
    ISSN: 1573-5176
    Keywords: amino acids ; biomass ; fatty acids ; Isochrysis ; nitrogen ; starvation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Growth of cells ofIsochrysis galbana with either nitrate or ammonium as the N-source, and the effects of subsequent N-starvation of these cells, were compared. During exponential N-sufficient growth nitrate-grown cells had double the fatty acid content of the ammonium-grown cells but lower concentrations of a few amino acids. Following resuspension in N-free medium the fatty acid content of the ammonium-grown cells increased to that of the nitrate-grown cells, but there was no further increase in fatty acid content on a C-biomass or cellular basis during the following 4 days for either culture. Fatty acid synthesis was continuous during N-starvation, while it occurred during the light-phase only in exponential growth. The proportion of 18:1n9 fatty acid increased from 10 to 25% total fatty acids during N-starvation. Intracellular free amino acid content decreased in a similar manner in both cultures on N-starvation, the ratio of intracellular free amino-N/cell-C falling more rapidly than overall cellular N/C. It was concluded that optimal amino acid and fatty acid content would be attained by growth in the presence of excess nitrate. Measurements of chlorophyll and carotenoid content and ofin vivo fluorescence indicated that these parameters had potential for monitoring the C and N biomass in cultures grown under relatively constant (not necessarily continuous) illumination.
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  • 92
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    Plant and soil 139 (1992), S. 285-294 
    ISSN: 1573-5036
    Keywords: Brassica ; B. campestris ; B. carinata ; B. juncea ; B. naptus ; B. nigra ; B. oleracea ; calcium ; chloride ; potassium ; magnesium ; nitrogen ; sodium ; nutrients ; salinity ; salt-tolerance ; seawater
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The growth of six rapid-cycling lines of Brassica species, B. napus, B. campestris, B. nigra, B. juncea, B. oleracea and B. carinata was inhibited by seawater salinity. Based on the change in dry matter reduction relative to the control at varying concentrations of salts (4, 8 and 12 dS m-1), the relative salt tolerance of these species was evaluated. B. napus and B. carinata were the most tolerant and most sensitive species, respectively, while the other four species were moderately tolerant. The influence of seawater on the concentrations of 12 elements including macronutrients and micronutrients in the shoots of these Brassica plants was characterized to determine the relationship between nutritional disturbance and relative salt tolerance. It was found that seawater salinity had a significant effect on the concentrations of Ca, Mg, K, Cl, Na and total N in the shoots of these plants but only the change in Ca concentration was significantly related to the relative salt tolerance of these six rapid-cycling Brassica species according to a rank analysis of the data. This finding indicates that Ca may play a regulatory role in the responses of Brassica species to saline conditions.
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  • 93
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    Plant and soil 142 (1992), S. 157-166 
    ISSN: 1573-5036
    Keywords: Acacia ; Eucalyptus regnans ; decomposition ; litterfall ; nitrogen ; phosphorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The N and P contents of the litter layer and the return of these nutrients in litterfall were measured in seven stands of Mountain Ash (Eucalyptus regnans) ranging in age from 5 years to about 250 years. Both annual litterfall and nutrient return were correlated with stand basal area and were high compared with other productive eucalypt forests. In contrast, the fall of dead eucalypt leaves was constant with stand age, demonstrating that sites are fully occupied at an early age. Similarly, amounts of N and P in total leaf fall (overstorey plus understorey) were constant with stand age, except for low amounts in the stand aged 40 years where Acacia spp., important fixers of atmospheric N, were not prevalent. The decomposition constant (k) of organic matter in the litter layer decreased with stand age, from 0.31 year-1 at age 5 years to 0.23 year-1 at age 250 years. These constants also applied to N and P, indicating a tight coupling between organic matter decomposition and release of these nutrients from litter. The litter layer released about 30 kg ha-1 of N at age 5 years, and about 70 kg ha-1 at age 80 years. These results are discussed in relation to growth of Mountain Ash following fire, and the subsequent retention and accumulation of N during stand development.
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  • 94
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    Plant and soil 142 (1992), S. 167-176 
    ISSN: 1573-5036
    Keywords: Eucalyptus regnans ; mineralization ; nitrification ; nitrogen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Laboratory and in situ rates of N mineralization in soils from Mountain Ash (Eucalyptus regnans) forests were measured fortnightly for two years in stands aged 9, 40, 80 and about 250 years (overmature), and for one year in stands aged 5 and 46 years. Rates of anaerobic mineralization (the laboratory test) showed little seasonal or annual variation. In contrast, rates of in situ mineralization varied markedly with season, being highest in spring and summer. Anaerobic mineralization was highly correlated with stand age to 80 years, but decreased between ages 80 and 250 years. In situ mineralization also decreased between these two ages, but otherwise was not related to stand age. Hence, the correlation between anaerobic and in situ mineralization along the age sequence was weak, suggesting that the anaerobic test is of maximum utility when this pool is in balance with inputs from decomposing litter (‘steady-state’ ecosystems). Nitrification was strong in stands aged 9, 46, 80 and 250 years and weak in stands aged 5 and 40 years. Within stands, the rate of nitrification during each period of in situ containment was highly dependent on the supply of NH4 ions. Between stands, annual rates of nitrification appear to be related to the balance between the N capital of the site, its rate of turnover, and the demand for N by heterotrophs and vegetation so that if NH4 supply is depleted, little is left for autotrophic nitrifiers.
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  • 95
    Electronic Resource
    Electronic Resource
    Springer
    Plant and soil 145 (1992), S. 45-50 
    ISSN: 1573-5036
    Keywords: acidity ; manganese ; pH ; phosphorus ; soil solution wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A pot experiment was carried out in order to test the hypothesis that manganese nutrition of wheat was dependent on the phosphorus status of soil as well as on its pH and manganese status. An arable mineral soil whose lime and phosphorus status had been massively adjusted more than 18 years previously was compared with identical soil not so adjusted. Wheat plants were grown to maturity in these soils. Analyses were carried out on both soil and plant samples at intervals. Data for soil pH, soil solution concentrations of manganese and phosphorus, plant dry weight and tissue concentrations of manganese and phosphorus are presented. Concentrations of manganese were depressed in leaf tissue of plants from limed soils and also in high phosphorus soils. The depressed values for limed treatments were explained in terms of depressed soil solution manganese concentrations resulting from elevated pH. The results for high phosphorus soils could not be related to soil solution composition. It was suggested that high soil phosphorus resulted in elevated plant phosphorus which interfered in the uptake and/or translocation of manganese.
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  • 96
    Electronic Resource
    Electronic Resource
    Springer
    Plant and soil 146 (1992), S. 163-168 
    ISSN: 1573-5036
    Keywords: genetic variability ; NIR ; ploidy ; phosphorus ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract More efficient utilization of phosphorus by wheat plants is needed to extend the useful life of the phosphate reserves in the world, to reduce the cost of producing crops, and to improve the value of the grain and the straw produced. In this paper definitions of efficient use of phosphorus by wheat are reviewed, genotypic variation in phosphorus efficiency is reported, some consequences of breeding for greater efficiency are discussed, and ways to select more efficient genotypes are suggested.
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  • 97
    ISSN: 1573-5036
    Keywords: ammonium ; citrus ; nitrate ; NO3 −/NH4 + ratio ; nitrogen ; nutrient solution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In short-term water culture experiments with different 15N labeled ammonium or nitrate concentrations, citrus seedlings absorbed NH4 + at a higher rate than NO3 −. Maximum NO3 − uptake by the whole plant occurred at 120 mg L−1 NO3 −-N, whereas NH4 + absorption was saturated at 240 mg L−1 NH4 +-N. 15NH4 + accumulated in roots and to a lesser degree in both leaves and stems. However, 15NO3 − was mostly partitioned between leaves and roots. Adding increasing amounts of unlabeled NH4 + (15–60 mg L−1 N) to nutrient solutions containing 120 mg L−1 N as 15N labeled nitrate reduced 15NO3 − uptake. Maximum inhibition of 15NO3 − uptake was about 55% at 2.14 mM NH4 + (30 mg L−1 NH4 +-N) and it did not increase any further at higher NH4 + proportions. In a long-term experiment, the effects of concentration and source of added N (NO3 − or NH4 +) on nutrient concentrations in leaves from plants grown in sand were evaluated. Leaf concentration of N, P, Mg, Fe and Cu were increased by NH4 + versus NO3 − nutrition, whereas the reverse was true for Ca, K, Zn and Mn. The effects of different NO3 −-N:NH4 +-N ratios (100:0, 75:25, 50:50, 25:75 and 0:100) at 120 mg L−1 total N on leaf nutrient concentrations, fruit yield and fruit characteristics were investigated in another long-term experiment with plants grown in sand cultures. Nitrogen concentrations in leaves were highest when plants were provided with either NO3 − or NH4 + as a sole source of N. Lowest N concentration in leaves was found with a 75:25 NO3 −-N/NH4 +-N ratio. With increasing proportions of NH4 + in the N supply, leaf nutrients such as P, Mg, Fe and Cu increased, whereas Ca, K, Mn and Zn decreased. Yield in number of fruits per tree was increased significantly by supplying all N as NH4 +, although fruit weight was reduced. The number of fruits per tree was lowest with the 75:25 NO3 −-N:NH4 +-N ratio, but in this treatment fruits reached their highest weight. Rind thickness, juice acidity, and colour index of fruits decreased with increasing NH4 + in the N supply, whereas the % pulp and maturity index increased. Percent of juice in fruits and total soluble solids were only slightly affected by NO3 −:NH4 + ratio.
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  • 98
    ISSN: 1573-5044
    Keywords: carbohydrate ; germination ; Impatiens L. ; Impatiens platypetala L. ; nitrogen ; tissue culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In vitro germination of 20-day old immature ovules of Impatiens platypetala Lindl. was inhibited at concentrations as low as 50 mM sucrose or mannitol and 100 mM glucose. Younger ovules (12, 14, and 16 days old) were similarly inhibited at 100 mM sucrose. Inorganic nitrogen concentration did not affect germination regardless of ovule age, but seedling fresh weight was significantly less and abnormal development of seedlings was significantly increased by total inorganic nitrogen concentrations higher or lower than 30 mM (at a ratio of 20: 10 mM NO3 -: NH4 +) in the culture medium.
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  • 99
    Electronic Resource
    Electronic Resource
    Springer
    Plant and soil 139 (1992), S. 247-251 
    ISSN: 1573-5036
    Keywords: ammonium ; lupins ; nitrate ; nitrogen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Two cultivars of Lupinus angustifolius L. were grown in a glasshouse in solutions containing NO3 -, NH4 + or NH4NO3 with a total nitrogen concentration of 2.8 M m-3 in each treatment. One cultivar chosen (75A-258) was relatively tolerant to alkaline soils whereas the other (Yandee) was intolerant to alkalinity. Controlled experiments were used to assess the impact of cationic vs. anionic forms of nitrogen on the relative performance of these cultivars. Relative growth rates (dry weight basis) were not significantly different between the two cultivars when grown in the presence of NO3 -, NH4 + or NH4NO3. However, when NO3 - was supplied, there was a modest decline in relative growth rates in both cultivars over time. When plants grown on the three sources of nitrogen for 9 days were subsequently supplied with 15NH4NO3 or NH4 15NO3 for 30 h, NH4 + uptake was generally twice as fast as NO3 - uptake, even for plants grown in the presence of NO3 -. Low rates of NO3 - uptake accounted for the decrease in growth rates over time when plants were grown in the presence of NO3 -. It is concluded that the more rapid growth of 75A-258 than Yandee in alkaline conditions was not due to preferential uptake of NH4 + and acidification of the external medium. In support of this view, acidification of the root medium was not significantly different between cultivars when NH4 + was the sole nitrogen source.
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  • 100
    ISSN: 1573-5036
    Keywords: critical concentration ; deficiency ; diagnosis ; growth rate ; lettuce ; luxury consumption ; nitrogen ; nitrate ; nutrient requirement ; petiole sap ; phosphorus ; phosphate ; potassium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A method is described for determining the way in which growth rate varies with plant nutrient concentration using a simple nutrient interruption technique incorporating only 2 treatments. The method involves measuring the changes in growth and nutrient composition of otherwise well-nourished plants after the supply of one particular nutrient has been withheld. Critical concentrations are estimated from the relationship between the growth rate (expressed as a fraction of that for control plants of the same size which remained well-nourished throughout) and the concentration of the growth-limiting nutrient in the plants as deficiency developed. Trials of the method using young lettuce plants showed that shoot growth rate was directly proportional to total N (nitrate plus organic N) concentration, and linearly or near-linearly related to K and P concentration over a wide range; the corresponding relationship for nitrate was strongly curvi-linear. Critical concentrations (corresponding to a 10% reduction in growth rate) determined from these results were similar to critical values calculated from models derived from field data, but were generally higher than published estimates of critical concentration (based on reductions in shoot weight) for plants of a similar size. Reasons for these discrepancies are discussed. Nitrate, phosphate or potassium concentrations in sap from individual leaf petioles were highly sensitive to changes in shoot growth rate as deficiency developed, with the slope of the relationships varying with leaf position, due to differences both in their initial concentration and in the rates at which they were utilized in individual leaves. Each nutrient was always depleted more quickly in younger leaves than in older ones, providing earlier evidence of deficiency for diagnostic purposes. Although the plants were capable of accumulating nitrate, phosphate and potassium well in excess of that needed for optimum dry matter production during periods of adequate supply, the rate of mobilization of these reserves was insufficient to prevent reductions in growth rate as the plants became deficient. This brings into question the validity of the conventional concept that luxury consumption provides a store of nutrients which are freely available for use in times of shortage. The implications of these results for the use of plant analysis for assessing plant nutrient status are discussed.
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