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  • Articles  (79)
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  • 1981  (38)
  • 1980  (41)
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  • Articles  (79)
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  • Springer  (79)
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  • 1
    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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  • 2
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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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  • 3
    ISSN: 1432-2048
    Keywords: Enzymes ; Nitrogen assimilation ; Root ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The enzymes nitrate reductase (NR), glutamate dehydrogenase (GDH), glutamate synthase (GOGAT), glutamine synthetase (GS) and asparagine synthetase (AS) have been assayed in various regions along the seedling root ofZea mays L. In the intact attached root and calculated on a protein basis NR, GOGAT, and GS are found to have slightly higher specific activities in the apical 5 mm than in more mature regions of the root. GDH and AS, on the other hand, are much more active in extracts prepared from mature regions of the root than in the apical region. In excised root tips incubated in the presence of NH4 + and NO3 − there was a marked increase in GDH and AS, and a slight decrease in GOGAT and GS. Additions of NO3 − are required for NR activity but neither NO3 − nor NH4 + additions altered the activity levels of the other four enzymes. Additions of glucose to the medium inhibited the development of AS and GDH activities and resulted in higher activity levels of NR, GS and GOGAT. Glucose additions also enhanced the incorporation of acetate-14C and leucine-14C into protein. Additions of cycloheximide inhibit the development of NR, AS and GDH activities and also the incorporation of acetate-14C and leucine into protein.
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  • 4
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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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  • 5
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    Planta 148 (1980), S. 510-512 
    ISSN: 1432-2048
    Keywords: Auxin ; Cell wall pH ; Growth (roots) ; Proton flux and growth ; Root growth ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract At concentrations inhibitory to the elongation of corn (Zea mays L.) roots, the auxins, indole-3-acetic acid (IAA) and α-naphthaleneacetic acid (α-NAA), cause an increase in the pH of the bathing medium; this increase occurs with an average latent period shorter than the latent period for the inhibitory effect of these auxins on elongation. Indole-2-carboxylic acid, an inactive structural analogue of IAA, and β-naphthaleneacetic acid, an inactive analogue of α-NAA, affect neither growth nor the pH of the medium. Since acid pH is known to promote and basic pH to inhibit root elongation, the data are consistent with the hypothesis that hormone-induced modification of cell-wall pH plays a role in the control of elongation of roots, as has been proposed for elongation of stems and coleoptiles.
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  • 6
    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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  • 7
    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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  • 8
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    Planta 149 (1980), S. 91-96 
    ISSN: 1432-2048
    Keywords: Light-grown plants ; Phytochrome ; SAN 9789 ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The phytochrome system is analyzed in light-grown maize (Zea mays L.) plants, which were prevented from greening by application of the herbicide SAN 9789. The dark kinetics of phytochrome are not different in the first, second or third leaf. It is concluded that in light-grown maize plants phytochrome levels are regulated by Pr formation and Pfr and Pr destruction, rather than by Pfr→Pr dark reversion. Pr undergoes destruction after it has been cycled through Pfr. The consequences of this Pr destruction on the phytochrome system are discussed.
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  • 9
    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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  • 10
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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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  • 11
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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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  • 12
    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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  • 13
    ISSN: 1432-2048
    Keywords: Abscisic acid ; Nicotiana ; Ribonucleic acid synthesis ; Tobacco mosaic virus
    Source: Springer Online Journal Archives 1860-2000
    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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  • 14
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    Planta 151 (1981), S. 15-25 
    ISSN: 1432-2048
    Keywords: Auxin (uptake, transport) ; Benzoic acid ; N-1-Naphthylphthalamic acid ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The validity of a chemiosmotic hypothesis for uptake of weak acids as an explanation for the accumulation of auxin by cells has been explored further by comparing the uptake of indole-3-acetic acid (IAA) by 1-mm segments of corn (Zea mays L.) coleoptiles with that of benzoic acid and two neutral indoles, indoleethanol and indoleacetonitrile, which do not ionize. These substances, while structurally related to IAA lack both auxin activity and polar transport. Uptake of IAA and benzoic acid increase with decreasing external pH, whereas the uptake of the two neutral indoles is independent of external pH. Although metabolism of IAA, during 90 min or less, is minimal and without significant effect on its uptake, metabolism of benzoic acid appears responsible for the apparent saturation of benzoic acid uptake at high concentrations. An inhibitor of auxin transport, N-1-naphthylphathalamic acid (NPA), stimulates uptake of IAA but has no effect on uptake of either benzoic acid or the two neutral indoles. Thus, NPA does not affect the driving forces for accumulation of weak acids but probably specifically decreases the flux of the auxin anions relative to undissociated auxin. Since the electrochemical potential of auxin anions is usually higher in than outside cells, blocking the anion flux with NPA would enhance auxin uptake. Azide, which abolishes accumulation of both IAA and benzoic acid, may simply collapse the pH gradient across the plasma membrane. In the absence of NPA, increasing concentrations of auxins or the analogoue β-naphthaleneacetic acid (β-NAA) exert two opposing effects on the uptake of IAA-depression and stimulation. Stimulation results from saturating the anion flux. With uptake fully stimulated by NPA, however, increasing concentrations of auxins or analogues only depress uptake of [3H]IAA. These results are consistent with more than one path for auxin transport each with a different dependence on concentration. In depressing NPA-stimulated IAA uptake, the effectiveness of β-NAA≧IAA≫α-NAA≫ benzoic acid, a specificity similar to that of an auxin binding site in vitro that has been implicated by others in auxin transport. The results support the general hypothesis that cellular auxin uptake and polar transport through tissues are chemiosmotically coupled to the electrochemical potential of auxin and protons.
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  • 15
    ISSN: 1432-2048
    Keywords: Aurea mutant ; Chlorophyll-protein complexes ; Chloroplast grana ; Light-harvesting pigment-protein complex ; Nicotiana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Phenotypic characterizations of the semidominant aurea tobacco (Nicotiana tabacum L.) mutant Su/su, the homozygous mutant Su/Su and three green revertants (R1, R2, and R3) are presented. The leaf color of Su/su plants varies from yellow to light-green when grown under high and low energy fluence rates (33.0 and 3.3 W m−2), respectively. The change in visual phenotype under high-light conditions is correlated with decreased content of chlorophyll per leaf area, agranal chloroplast ultrastructure, changes in the number of chlorophyll-protein complexes, and absence of two or more of the light harvesting chlorophyll-polypeptides of 25,000–29,000 dalton. The homozygous mutant grown under low light was shown to be completely lacking in grana stacks and to be deficient in chlorophyll-protein complexes. Revertant R1 was found to be identical to wild-type plants in all parameters examined (leaf color, chloroplast ultrastructure, chlorophyll-protein complexes, chlorophyll-protein complex polypeptides) except in chlorophyll content. It did not show an increased chlorophyll and carotenoid content as did the wild-type plants when exposed to high light. Revertants R2 and R3 were similar to the heterozygous mutant Su/su in most of the parameters examined. They yellowed because of a loss of chlorophyll and an increase in the amount of carotenoids, had agranal chloroplasts, and had variant chlorophyll-protein complexes when grown under high light intensities. However, each appeared to contain some of the light-harvesting pigment-protein complex polypeptides found to be absent in Su/su when grown under high-light conditions.
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  • 16
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    Planta 153 (1981), S. 436-442 
    ISSN: 1432-2048
    Keywords: Aminopterin-resistant variants ; Cell culture ; Folate analog ; Variant selection ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Aminopterin-resistant cell lines of maize were isolated by two different procedures of callus selection and by plating suspension cultures on drugcontaining medium after mutagen treatment. Efficiencies of different methods of variant selection were compared. Four aminopterin-resistant cell lines were shown to be 10–40 times more resistant than the parental cell line, and they were also resistant to another folate analog, methotrexate. The results suggest that alterations in at least three different cell properties could be responsible for resistance; 1) increased dihydrofolate reductase activity, 2) altered aminopterin sensitivity of dihydrofolate reductase, and 3) reduced drug uptake. One of the resistant cell lines showed more than one alteration, but its resistance proved to be unstable. The results suggest that stable changes which may or may not be of genetic origin and also unstable physiological changes or a combination of both could lead to aminopterin resistance in maize cell cultures.
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  • 17
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    Planta 153 (1981), S. 443-446 
    ISSN: 1432-2048
    Keywords: ATPase ; Mitochondria ; Mycotoxin ; Pisum ; Zea ; Zearalenone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract At 5 and 10 μg ml-1 concentration, zearalenone (F-2), a mycotoxin produced by a number of species of the genus Fusarium, causes an inhibition of the oxidative phosphorylation of isolated plant mitochondria, while at 20 and 40 μg ml-1 it causes uncoupling. However, when the mitochondria are pre-incubated for 20 min with F-2, the uncoupling appears to be the prevailing effect. F-2 is also able to inhibit the mitochondrial ATPase activity (Mg2+-dependent). Conversely, F-2 (40 μg ml-1) does not alter the ATP level of maize roots and only slightly affects the ATPase activity of pea stem and maize root microsomal fractions. In addition, F-2 (10–40 μg ml-1) inhibits ATP synthesis catalyzed by rat liver mitochondria. It is suggested that the phytotoxicity of F-2, also known for its ability to collapse the transmembrane electric potential of maize roots, may be mainly linked to its ability to increase the proton permeability of the cell, similar to the common uncouplers.
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  • 18
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    Planta 153 (1981), S. 471-475 
    ISSN: 1432-2048
    Keywords: Abscisic acid ; Geotropism (root) ; Growth inhibitor ; Light and root-cap inhibitor ; Root cap (inhibitor) ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Growth inhibitors were assayed from extracts of intact (attached) and of excised (cultured) root caps of Zea mays L., cv. Merit, the roots of which show a positive geotropic response only after exposure to light. If caps are intact at the time of illumination, at least two inhibitory substances are produced, an acid inhibitor and a neutral inhibitor, whereas if caps are detached from roots, placed in culture and then illuminated only the neutral inhibitor is formed. Cycloheximide retards inhibitor production in both intact and cultured caps. When [14C]mevalonic acid is included in the culture medium and the caps are illuminated, 15–25% of the recoverable 14C cochromatographs with the neutral inhibitor, whereas in caps cultured in the dark, this radiolabelling pattern is not observed. Cyloheximide in the light reduces the incorporation of 14C into compounds cochromatographing at the Rf of the neutral inhibitor. It is suggested that the neutral inhibitor may be important in the light-induced bending of roots.
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  • 19
    ISSN: 1432-2048
    Keywords: Chloroplast ; Cybrid ; Nicotiana ; Protoplast ; Somatic hybrid ; Sterility, male
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts from a nitrate reductase-deficient mutant of Nicotiana tabacum L. were fused with protoplasts from a stamen-less, cytoplasmically malesterile cultivar of tobacco containing the cytoplasm from N. suaveolens Lehm. Plants were regenerated from the fused protoplasts and characterized with respect to stamen development, chromosome number, and chloroplast composition. Of 29 regenerated plants, stamen production was restored in 26 plants and pollen production in 22. One plant was male sterile and two plants have never flowered. Analysis of the electrophoretic mobility of ribulose-1,5-bisphosphate carboxylase (RuBPcase) showed that 19 of the plants contained RuBPcase of the N. suaveolens type, six plants contained enzyme of the N. tabacum type, and four plants contained both types. Analysis of resistance to tentoxin in seedlings from 20 of the plants demonstrated that 14 had N. suaveolens-type chloroplasts, three had N. tabacum type, and three contained both types. Many of the plants which produced stamens and pollen still contained chloroplasts of the N. suaveolens type. Thus, the trait of cytoplasmic male sterility in tobacco is not an expression of the type of chloroplast genetic material.
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  • 20
    ISSN: 1432-2048
    Keywords: Auxin (immunoassay) ; Avena ; Coleoptiles (physiological tip) ; Enzyme immunoassay ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A specific solid-phase enzyme immunoassay for the detection of as little as 3–4 pg of indole-3-acetic acid (IAA) is described. The assay involves minimal procedural efforts and requires only standard laboratory equipment. Up to 50 samples in triplicate, processed simultaneously, can be assayed and evaluated in 2.5 h. As little as 1 mg oat coleoptile tissue is sufficient for a quantitative IAA analysis and little or no extract purification is necessary. Using this assay, levels of IAA have been determined in coleoptiles of maize and oat. The distribution of IAA within single coleoptiles was quantitated and the production of IAA during the regeneration of the physiological tip in Avena coleoptiles was investigated. The changes in levels of IAA and other major phytohormones were quantitated during the growth of oat coleoptiles.
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  • 21
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    Planta 153 (1981), S. 42-48 
    ISSN: 1432-2048
    Keywords: Crown gall ; Hyperplasia ; Hypertrophy ; Teratoma ; Nicotiana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Crown-gall teratomas are tumors of higher plants with an intrinsic capacity for organogenesis. The growth pattern of tobacco (Nicotiana tabacum L.) teratoma shoots, which is highly aberrant in primary tumors, becomes normal when the shoots are grafted to healthy stock plants. However, certain abnormalities commonly persist; tumors form at the graft junctions, leaves are small, apical dominance is incomplete, the stem and proximal region of the leaf midribs swell excessively, and localized eruptions of neoplastic growth occur on the swollen tissue. Swelling of the shoots is primarily the result of cell hypertrophy in the cortex. Neoplastic divisions do not occur as a general rule; they are restricted, with the exception of tumor formation at the graft junctions, to localized eruptions of teratoid growth on the nodes and leaf midribs where cell hypertrophy is most evident. The histology of the apical meristem and histogenesis of primary tissues is normal, even in grossly distorted shoots. Similarly, there is no evidence of unregulated division in the vascular cambium. It is concluded that cell expansion and division are tightly regulated in meristematic regions of teratoma shoots whereas post-meristematic tissue is prone to excessive hypertrophy and eventual initiation of neoplastic cell division.
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  • 22
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    Planta 153 (1981), S. 511-518 
    ISSN: 1432-2048
    Keywords: Amino-acid transport ; Arginine ; Cell culture (tobacco) ; Nicotiana ; Transport (arginine)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Arginine transport in suspension-cultured cells of Nicotiana tabacum L. cv. Wisconsin-38 was investigated. Cells that were preincubated in the presence of Ca2+ for 6 h prior to transport exhibited stimulated transport rates. After the preincubation treatment, initial rates of uptake were constant for at least 45 min. Arginine accumulated in the cells against a concentration gradient; this accumulation was not the result of exchange diffusion. Arginine uptake over a concentration range of 2.5 μM to 1 mM was characterized by simple Michaelis-Menten kinetics with a Km of 0.1 mM and a Vmax of 9,000 nmol g-1 fresh weight h-1. Transport was inhibited by several compounds including carbonylcyanide-m-chlorophenylhydrazone, 2,4-dinitrophenol, N,N′-dicyclohexylcarbodiimide, and N-ethylmaleimide. Inhibition by these compounds was not the result of increased efflux resulting from membrane damage. A variety of amino acids and analogs, with the exception of D-arginine, inhibited transport, indicating that arginine transport was mediated by a general L-aminoacid permease. Competition experiments indicated that arginine and lysine exhibited cross-competition for transport, with Ki values similar to respective Km values. Arginine transport and low-affinity lysine transport are probably mediated by the same system in these cells.
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  • 23
    ISSN: 1432-2048
    Keywords: Cell-wall autohydrolysis ; β-D-Glucan ; Glucanase ; Hemicellulose ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The proteins dissociated from isolated Zea seedling cell wall using high-ionic-strength salt solutions have been found to include a number of enzymes which appear to participate in autolytic reactions of the cell wall. These enzymes caused extensive degradation of enzymatically inactive cell wall, liberating as much as 100 μg/mg dry weight over a 48-h period. Lithium chloride (3M) was shown to be most effective in yielding protein and wall-degrading activities. Molecular-sieve chromatography of the cell-wall protein resolved endo-β-D-glucanase and exo-β-1,3-glucanase (EC 3.2.1.58) activities when Avena glucan and laminarin, respectively, were employed as substrates. The exoenzyme (molecular weight around 60,000) was strongly inhibited by inorganic mercury at a concentration which suppressed the release of monosaccharide from autolytically active cell wall. The endo-β-D-glucanase (MW around 26,000), which showed a marked preference for substrates of mixed-linkage, exhibited features indicating that it initiates the autolytic solubilization of wall glucan. Cell-wall β-D-glucan, recovered as a component of an alkali-soluble cell-wall fraction, served as a substrate for the purified glucanases. Their hydrolysis pattern, assessed using gel exclusion chromatography and product analysis, confirmed that they hydrolyze β-D-glucan. The products generated by the endoglucanase were similar in molecular-size distribution to those liberated from autolytically active-wall. Exoglucanase activity was required for extensive hydrolysis of β-D-glucan in vitro. During coleoptile development the autolytic activity of the cell wall increased dramatically. This increased activity, however, did not parallel the growth potential of the tissue, but more closely reflected an increase in cell-wall β-D-glucan, the primary substrate for autolytic reactions.
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  • 24
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    Planta 147 (1980), S. 457-466 
    ISSN: 1432-2048
    Keywords: Auxin (transport, accumulation) ; Chemiosmotic hypothesis ; Coleoptile ; Transport (auxin) ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The uptake of auxin by 1-mm slices of corn (Zea mays L.) coleoptiles, a tissue known to transport auxin polarly, depends on the pH of the medium. Short-term uptake of indole-3-acetic acid (IAA) in coleoptiles increases with decreasing pH of the buffer as would be expected if the undissociated weak acid, IAA·H, were more permeable than the auxin anion, IAA-, and IAA- accumulates in the tissues because of the higher pH of the cytoplasm. Although uptake of [3H]IAA is reduced in neutral buffers, it is greater than expected if it were limited to just the extracellular space of the tissue. The radioactivity accumulated by the tissue can be quantitatively extracted by organic solvents and identified as IAA by thin-layer chromatography. The tissue radioactivity is freely mobile and can efflux from the tissue. Thus these cells in pH 5 buffer are able to retain an average internal concentration of mobile IAA that is at least several times greater than the external concentration. A prominent feature of auxin uptake from acidic buffers is enhanced accumulation at high auxin concentration. This indicates that, in addition to fluxes of IAA·H, a saturable site is involved in auxin uptake. Whenever the auxin-anion gradient is directed outward, saturating the efflux of auxin anions increases accumulation. Furthermore, the observed slowing of short-term uptake of radioactive IAA by increasing concentrations of IAA or K+ indicates either an activation of the presumptive auxin leak or saturation of another carrier-mediated uptake system such as a symport of auxin anions with protons. By contrast in neutral buffers, effects of concentration on uptake rates disappear. This implies that at neutral pH the anion leak is decreased and influx depends on the symport.
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  • 25
    ISSN: 1432-2048
    Keywords: Hydrogenperoxide ; Lignification ; Nicotiana ; Peroxidase
    Source: Springer Online Journal Archives 1860-2000
    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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  • 26
    ISSN: 1432-2048
    Keywords: Acid growth ; Auxin ; Growth ; Root ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The role of proton excretion in the growth of apical segments of maize roots has been examined. Growth is stimulated by acidic buffers and inhibited by neutral buffers. Organic buffers such as 2[N-morpholino] ethane sulphonic acid (MES) — 2-amino-2-(hydroxymethyl)propane-1,3 diol (Tris) are more effective than phosphate buffers in inhibiting growth. Fusicoccin(FC)-induced growth is also inhibited by neutral buffers. The antiauxins 4-chlorophenoxyisobutyric acid (PCIB) and 2-(naphthylmethylthio) propionic acid (NMSP) promote growth and H+-excretion over short time periods; this growth is also inhibited by neutral buffers. We conclude that growth of maize roots requires proton extrusion and that regulation of root growth by indol-3yl-acetic acid (IAA) may be mediated by control of this proton extrusion.
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  • 27
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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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  • 28
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    Planta 150 (1980), S. 406-411 
    ISSN: 1432-2048
    Keywords: Maize tissue ; RNA metabolism ; Small RNA species ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The low molecular weight RNA components of maize have been analyzed after labeling callus and leaf tissue with [3H]uridine in vitro. Electrophoresis of the isolated RNA on acrylamide slab gels reveals, apart from 5S and transfer RNA, three major and about five minor RNA species with chain lengths between 140 and 280 nucleotides. These RNA molecules are labeled as rapidly as 5S, transfer RNA, and do not represent degradation products of large ribosomal RNA molecules. Furthermore, like 5S and transfer RNA, these small RNA species are stable and show no detectable turnover within forty-eight hours. Fractionation of the tissue into crude subcellular fractions indicates a preferential association of some of the small stable RNA species with the nucleus, while others appear to be located in the cytoplasm. The low molecular weight RNA spectrum from the leaf is similar to that observed in callus, with the major small RNA species equally present in both tissues.
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  • 29
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    Planta 151 (1981), S. 68-74 
    ISSN: 1432-2048
    Keywords: Energy metabolism ; Germination (seeds) ; Protein synthesis ; Temperature and seed germination ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Poor emergence of maize seedlings, due to high soil temperatures, is a major limitation of crop potential in the lowland tropics. Ability to germinate at high temperature (〉c. 37° C) is related to the temperature sensitivity of the embryo, and there is considerable genotypic variation for this character. Respiration and mitochondrial phosphorylation proceed normally in seeds imbibing at 41° C, and ATP levels are adequate for germination. However, the specific activities of several important enzymes are lower, and the rate of protein synthesis is severely reduced compared with seeds imbibing at 28° C. The depression of the rate of protein synthesis in the embryos of several tropical hybrids imbibing at high temperature correlated with their known temperature sensitivity. It is concluded that protein synthesis is an especially temperature sensitive process in germinating maize embryos, and that this is the principal reason for the sensitivity of germinating maize seeds to high temperature.
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  • 30
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    Planta 151 (1981), S. 75-80 
    ISSN: 1432-2048
    Keywords: Embryos (protein synthesis) ; Germination (seeds) ; Protein synthesis and temperature ; Temperature and protein synthesis ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The poor germination of maize seeds at high temperatures (〉37°C) is related to the low rate of protein synthesis by the embryo. The apparatus of translation was not heat-labile when embryos were incubated for 2 h at 41°C, and cell free extracts from seeds imbibed for 16 h at this temperature were able to translate exogenous mRNA, indicating that ribosomes and other subcellular components were present and functional. Analysis of polysome profiles from embryos imbibing at high temperature indicated that the low rate of protein synthesis was due to the non-availability of active mRNA.
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  • 31
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    Planta 151 (1981), S. 141-145 
    ISSN: 1432-2048
    Keywords: Chromosome isolation ; Petroselinum ; Polyethylene-glycol ; Protoplast (uptake of chromosomes) ; Triticum ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract For mass isolation of plant metaphase chromosomes, cultured cells of wheat (Triticum monococcum) and parsley (Petroselinum hortense) were synchronized by hydroxyurea and colchicine treatment. This synchronization procedure resulted in high mitotic synchrony, especially in suspension cultures of parsley in which 80% of the cells were found to be at the metaphase stage. Mitotic protoplasts isolated from these synchronized cell cultures served as a source for isolation of chromosomes. The described isolation and purification method yielded relatively pure chromosome suspension. The uptake of the isolated plant chromosomes into recipient wheat, parsley, and maize protoplasts was induced by polyethylene-glycol treatment. Cytological studies provided evidences for uptake of plant chromosomes into plant protoplasts.
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  • 32
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    Planta 151 (1981), S. 146-150 
    ISSN: 1432-2048
    Keywords: Georeaction (root) ; Growth inhibitors ; Growth (root) ; Root (georeaction, growth) ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The growth rate of the two sides of 10-mm apical segments prepared from primary roots and of intact primary roots of maize has been analyzed in both vertical and horizontal positions, using a filming method allowing continuous growth recording. The data showed that the georeaction began by a decrease in the overall elongation rate of the roots. This inhibition is effective on the lower side of the bending zone, where the growth is practically stopped during the period of maximum rate of geocurvature. In contrast, the growth is slightly enhanced on the upper part of the elongating zone.
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  • 33
    ISSN: 1432-2048
    Keywords: Cucurbita ; Endoplasmic reticulum ; Light (blue) receptor ; Plasmalemma ; Riboflavin binding ; Zea
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    Topics: Biology
    Notes: Abstract Saturable and reversible in vitro binding of [14C]riboflavin was found to occur on subcellular, sedimentable particles from maize coleoptiles and Cucurbita hypocotyls. The KD was ca. 6 μM, the pH optimum was near 6.0, and the number of binding sites amounted to 0.1–0.5 μM on a fresh-weight basis. When the reducing agent dithionite was present, riboflavin binding increased-the KD was 2.5 μM, and the pH optimum above 8.0. The binding was specific: flavin mononucleotide (FMN) and flavin adenosine-dinucleotide (FAD) bound less tightly to these sites than riboflavin and another major soluble flavin, the previously described riboflavin-analog “FX”, occurring in grass coleoptiles. These flavin-binding sites were localized on vesicles derived from plasmalemma and endoplasmic reticulum by analyzing sucrose and metrizamide density gradients and marker enzymes.
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  • 34
    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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  • 35
    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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  • 36
    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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  • 37
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    Planta 152 (1981), S. 319-324 
    ISSN: 1432-2048
    Keywords: Nitrate influx (efflux, metabolism) ; Pennisetum ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Maize (Zea mays L.) and pearl millet (Pennisetum americanum (L.) Leeke) seedlings were exposed to [15N]nitrate for 1-h periods at eight times during a 24-h period (16–8 h light-dark for maize; 14–10 h for millet). Influx of [15N]nitrate as well as its reduction and translocation were determined during each period. The efflux of previously absorbed [14N]nitrate to the uptake solution was also estimated. No marked diurnal changes in [14N]nitrate efflux or [15N]nitrate influx were evident in maize. In contrast, [14N]nitrate efflux from millet increased and eventually exceeded [15N]nitrate influx during the late dark and early light periods, resulting in net nitrate efflux from the roots. The dissimilarity of their diurnal patterns indicates that influx and efflux are independently regulated. In both species, [15N]nitrate reduction and 15N translocation to shoots were curtailed more by darkness than was [15N]nitrate influx. In the light, maize reduced 15% and millet 24% of the incoming [15N]nitrate. In darkness, reduction dropped to 11 and 17%, respectively. Since the accumulation of reduced-15N in shoots declined abruptly in darkness, whereas that in roots was little affected, it is suggested that in darkness [15N]nitrate reduction occurred primarily in roots. The decrease in nitrate uptake and reduction in darkness was not related to efflux, which remained constant in maize and did not respond immediately to darkness in pearl millet.
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  • 38
    ISSN: 1432-2048
    Keywords: Chloroplast transfer ; Nicotiana ; Protoplasts fusion and inactivation
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    Topics: Biology
    Notes: Abstract Protoplasts of a light sensitive plastome mutant of Nicotiana tabacum (2 n=48) were irradiated and fused with iodoacetate-treated Nicotiana plumbaginifolia (2 n=20) protoplasts. Treated parental protoplasts were unable to divide. Metabolic complementation, however, helped the recovery of interspecific fusion products which survived and formed calli. Altogether 40 clones were investigated. N. plumbaginifolia plants were obtained in 15 clones (38%), somatic hybrids in 23 clones, and both types of regenerates were found in 2 clones. Irradiation therefore significantly increased the frequency of segregant formation with the non-irradiated N. plumbaginifolia nuclei (the frequency was 1.4% in the absence of irradiation). Regenerated plants in most cases (31 out of 34) contained chloroplasts from the irradiated parent. In 6 clones plants were obtained with both types of chloroplast. Thus, irradiated N. tabacum chloroplasts had an improved chance of dominating the heterokaryonderived cells, many of which contained N. plumbaginifolia nucleus. The system described should be generally applicable for the transfer of chloroplasts without the use of selectable genetic markers.
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  • 39
    ISSN: 1432-2048
    Keywords: Auxin-binding site ; Cell suspension culture ; Growth cycle ; Nicotiana
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract We studied the modulation of the number of membrane-bound naphthaleneacetic acid (NAA)-binding sites during the growth cycle of tobacco cells in batch culture. Both cell number and specific NAA-binding increased exponentially, but at different rates and for different periods. This caused a characteristic modulation of the number of binding sites per cell during the growth cycle: During the first day of the lag phase this number decreased; in the exponential phase it rose markedly, and in the stationary phase it was constant.
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  • 40
    ISSN: 1432-2048
    Keywords: Enzymes ; Nitrogen assimilation ; Root ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The enzymes nitrate reductase (NR), glutamate dehydrogenase (GDH), glutamate synthase (GOGAT), glutamine synthetase (GS) and asparagine synthetase (AS) have been assayed in various regions along the seedling root ofZea mays L. In the intact attached root and calculated on a protein basis NR, GOGAT, and GS are found to have slightly higher specific activities in the apical 5 mm than in more mature regions of the root. GDH and AS, on the other hand, are much more active in extracts prepared from mature regions of the root than in the apical region. In excised root tips incubated in the presence of NH4 + and NO3 − there was a marked increase in GDH and AS, and a slight decrease in GOGAT and GS. Additions of NO3 − are required for NR activity but neither NO3 − nor NH4 + additions altered the activity levels of the other four enzymes. Additions of glucose to the medium inhibited the development of AS and GDH activities and resulted in higher activity levels of NR, GS and GOGAT. Glucose additions also enhanced the incorporation of acetate-14C and leucine-14C into protein. Additions of cycloheximide inhibit the development of NR, AS and GDH activities and also the incorporation of acetate-14C and leucine into protein.
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  • 41
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    Planta 148 (1980), S. 510-512 
    ISSN: 1432-2048
    Keywords: Auxin ; Cell wall pH ; Growth (roots) ; Proton flux and growth ; Root growth ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract At concentrations inhibitory to the elongation of corn (Zea mays L.) roots, the auxins, indole-3-acetic acid (IAA) and α-naphthaleneacetic acid (α-NAA), cause an increase in the pH of the bathing medium; this increase occurs with an average latent period shorter than the latent period for the inhibitory effect of these auxins on elongation. Indole-2-carboxylic acid, an inactive structural analogue of IAA, and β-naphthaleneacetic acid, an inactive analogue of α-NAA, affect neither growth nor the pH of the medium. Since acid pH is known to promote and basic pH to inhibit root elongation, the data are consistent with the hypothesis that hormone-induced modification of cell-wall pH plays a role in the control of elongation of roots, as has been proposed for elongation of stems and coleoptiles.
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  • 42
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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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  • 43
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    Planta 150 (1980), S. 1-8 
    ISSN: 1432-2048
    Keywords: Calcium uptake ; Microsomes ; Mitochondria ; Transport (Ca2+) ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mitochondria from etiolated corn possess a much greater Ca2+ uptake capacity per mg protein than microsomes from the same source. Differences in energy requirements, sensitivity to specific inhibitors, and sedimentation properties enabled us to study both Ca2+ uptake mechanisms without mutual contamination. The microsomal Ca2+ uptake does not vary much among different plants as compared to the mitochondrial Ca2+ uptake; this is also true for different organs of the same plant. Mitochondrial Ca2+ uptake is more dependent on the age of the seedlings than microsomal uptake, because of changes in active Ca2+ uptake activity rather than of changes in efflux. Intactness and the oxidative and phosphorylative properties of the mitochondria remained unchanged during this time period. Na+ and Mg2+ do not induce Ca2+ release from mitochondria.
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  • 44
    ISSN: 1432-2048
    Keywords: Abscisic Acid ; Auxin ; Geotropism ; Root tip ; Zea
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    Topics: Biology
    Notes: Abstract The effect of externally applied indoleacetic acid (IAA) and abscisic acid (ABA) on the growth of roots of Zea mays L. was measured. Donor blocks of agar with IAA or ABA were placed laterally on the roots and root curvature was measured. When IAA was applied to vertical roots, a curvature directed toward the donor block was observed. This curvature corresponded to a growth inhibition at the side of the root where the donor was applied. When IAA was applied to horizontal roots from the upper side, normal geotropic downward bending was delayed or totally inhibited. The extent of retardation and the inhibition of curvature were found to depend on the concentration of IAA in the donor block. ABA neither induced curvature in vertical roots nor inhibited geotropic curvature in horizontal roots; thus the growth of roots was not inhibited by ABA. However, when, instead of donor blocks, root tips or coleoptile tips were placed onto vertical roots, a curvature of the roots was observed.
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  • 45
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    Planta 151 (1981), S. 499-505 
    ISSN: 1432-2048
    Keywords: Auxin ; Calorimetry ; Coleoptile ; Energetics ; Growth regulator ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Indole-3-acetic acid (IAA) promotes an increase in steady-state heat production by corn (Zea mays L.) coleoptile tissue; this increase is associated with an elevation in aerobic respiration rates. A detailed time dependence of the exothermic response to IAA was obtained using flow calorimetry. The latent period and magnitude of response were evaluated as a function of IAA concentration and pH. The data indicate that more than one response may occur. The optimal change in heat production was produced by an IAA concentration of 3·10-5 M. It was initiated within 5 min after the start of the IAA treatment, and reached a magnitude in excess of 25% of the tissue's basal heat production. Concentrations of IAA greater than 1·10-4 M resulted in diminished response(s), but the effect was strongly pH dependent. Several possibilities for the increased heat production triggered by IAA are discussed.
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  • 46
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    Planta 151 (1981), S. 506-511 
    ISSN: 1432-2048
    Keywords: Cell fractionation ; Cucurbita ; Malic enzyme ; Plasma membrane ; Subcellular localization ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A definite membrane fraction from Cucurbita hypocotyls, maize coleoptiles, and other plant tissues contains a NADP-dependent malic enzyme activity, up to 10% of overall tissue activity, and probably other soluble proteins. This “malic enzyme particle” is identified as plasmalemma on the basis of sedimentation behavior, density distribution in sucrose gradients, in comparison with enzyme markers, and sluggish penetration by the sugar Metrizamide. Enzyme binding to the plasma membrane is stable and scarcely sensitive to salts and EDTA, although all activity is released to the supernatant in the presence of Triton-X-100 or under hypotonic conditions. The properties of bound enzyme are similar to those of free enzyme in cell extracts. It is proposed that osmotically sensitive plasma membrane vesicles, containing cytoplasm fragments, are formed during homogenization. Low malic enzyme activities are also associated with Cucurbita proplastids.
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  • 47
    ISSN: 1432-2048
    Keywords: Lycopersicon ; Phetorespiration ; Photosynthesis ; Triacontanol ; Zea
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    Topics: Biology
    Notes: Abstract Tomato (C3-plants) and maize (C4-plants) were grown in a nutrient solution to which triacontanol was added twice a week. After about 4 weeks the triacontanol treatment caused a significant increase in the dry weight of the tomato plants. Leaf area and dry weight measurements of tomato leaves at different stages of development showed that the largest increase in growth was obtained when triacontanol treatment was initiated before bud formation. In maize, no effect of the triacontanol treatment on dry wieght was observed. Photosynthesis was inhibited by 27% in young leaves from triacontanol-treated tomato plants and 39% in the controls, when the oxygen concentration was raised from 2% to 21%. In maize no change in photosynthesis could be observed, neither after altered oxygen concentration nor after triacontanol treatment. The difference in the response of C3- and C4-plants to triacontanol indicates that it regulates processes related to photosynthesis.
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  • 48
    ISSN: 1432-2048
    Keywords: Cytoplasmic pH ; Fusicoccin ; Hydrogen-ion extrusion ; pH-stat ; Root ; Zea
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    Topics: Biology
    Notes: Abstract 31P-Nuclear-magnetic-resonance spectra of maize (Zea mays L.) root tips, that had been induced to extrude large amounts of H+ in response to fusicoccin (FC) in the presence of potassium salts, indicate that the cytoplasmic pH does not become higher than that of controls. In fact, the cytoplasmic pH may become slightly (approx. 0.1 pH unit) lower in cells extruding H+. Estimations of the buffer capacity of the cells show that without active intracellular pH regulation, H+ extrusion caused by FC would cause the intracellular pH to rise by at least 0.6 pH unit h-1. Our results indicate that intracellular pH is tightly regulated even during extreme rates of acid extrusion, and that a rise in cytoplasmic pH is not the signal linking H+ extrusion with enhanced organic-acid synthesis or other intracellular responses to H+ pumping.
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  • 49
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    Planta 152 (1981), S. 215-224 
    ISSN: 1432-2048
    Keywords: Bromodeoxyuridine tolerance ; Bromouracil excision ; Cytokinin autotrophy ; DNA synthesis ; Fluorodeoxyuridine effect ; Nicotiana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract DNA isolated from various Nicotiana tabacum cell types, differing in their degree of hormone autotrophy and incubated in the presence of bromodeoxyuridine (BrdUrd), was analyzed by isopycnic CsCl gradient centrifugation. All cell types incorporate BrdUrd into DNA in such a way that hybrid DNA is formed with 60–80% of thymine (Thy) residues replaced by bromouracil (BrUra) in the newly synthesized strand. This DNA is not replicated further under ordinary culture conditions. Whereas in “normal” hormone-dependent cells this state is final and cells necrotize, in tumor (cytokinin-auxin autotrophic) and cytokinin-autotrophic cells a mechanism is induced leading to the reduction of BrUra content in DNA. As a result a decrease in the buoyant density (in CsCl) of BrUra DNA can be observed. In the case of cytokinin-autotrophic cells supplemented with kinetin, the buoyant density of the whole DNA decreases gradually to the value of that of unsubstituted DNA, but specific radioactivities of different DNA fractions reflect the retention of the pyrimidine ring of BrUra in DNA. This is interpreted as debromination of DNA in situ. The process can be inhibited by fluorodeoxyuridine (FdUrd) and deoxycytidine (dCyd). Moreover, FdUrd (but not dCyd) allows replication of hybrid DNA in tumor cells in such a way that HH DNA with all Thy residues replaced by BrUra is formed. For cytokinin-autotrophic cells FdUrd and kinetin are required. In hormone-dependent cells replication of hybrid DNA cannot be induced under any conditions. Most of these conclusions complement our previous findings that BrdUrd tolerance in hormone-autotrophic tobacco cells in hormone controlled. It is postulated that a modulation of thymidylate synthetase specificity is one factor affecting the level of BrUra substitution in DNA. The possibility of cytokinins being involved in the control of DNA synthesis is discussed.
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  • 50
    ISSN: 1432-2048
    Keywords: Acid growth ; Geotropism ; Helianthus ; Phototropism ; Proton secretion ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract By placing seedlings of sunflower (Helianthus annuus L.) or maize (Zea mays L.) on agar plates containing a pH indicator dye it is possible to observe surface pH patterns along the growing seedling by observing color changes of the indicator dye. Using this method we find that in geotropically stimulated sunflower hypocotyls or maize coleoptiles there is enhanced proton efflux on the lower surface of the organ prior to the initiation of curvature. As curvature develops the pattern of differential acid efflux becomes more intense. A similar phenomenon is observed when these organs are exposed to unilateral illumination, i.e. enhanced acid efflux occurs on the dark side of the organ prior to the initiation of phototropic curvature and the pattern of differential acid efflux intensifies as phototropic curvature develops. These observations indicate that differential acid efflux occurs in response to tropistic stimuli and that the acid efflux pattern may mediate the development of tropistic curvatures.
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  • 51
    ISSN: 1432-2048
    Keywords: Amino acids ; Feedback inhibition ; Mutant selection ; Tissue culture ; Variants, resistant ; Zea
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    Notes: Abstract Tissue culture selection techniques were used to isolate a maize (Zea mays L.) variant D33, in which the aspartate family pathway was less sensitive to feedback inhibition by lysine. D33 was recovered by successively subculturing cultures originally derived from immature embryos on MS medium containing growth-inhibitory levels of lysine+threonine. The ability of D33 to grow vigorously on lysine+ threonine medium was retained after growth for 12 months on nonselection medium. New cultures initiated from shoot tissues of plants regenerated from D33 also were resistant to lysine+threonine inhibition. The Ki of aspartokinase for its feedback inhibitor, lysine, was about 9-fold higher in D33 than for the enzyme from unselected cultures. The free pools of lysine, threonine, isoleucine and methionine were increased 2–9-fold in D33 cultures. This was consistent with the observed change in feedback regulation of aspartokinase, the first enzyme common to the biosynthesis of these amino acids in the aspartate pathway. The accumulated evidence including the stability of resistance in the cultures, the resistance of cultures initiated from regenerated plants, the altered feedback regulation, and the increased free amino acids, indicates a mutational origin for these traits in line D33.
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  • 52
    ISSN: 1432-2048
    Keywords: Endogenous ligands ; Fusicoccin ; Proton extrusion ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The binding of fusicoccin to the microsomal preparations of maize roots in vitro is increased several-fold when segments of the tissue are washed for 2 h in distilled water before homogenization. Addition of freeze-dried wash solution to microsomal preparations of spinach leaves or fresh roots, washed roots, or coleoptiles of maize inhibited the binding of fusicoccin to particulate fractions. The freeze-dried material also blocked fusicoccin-promoted H+ extrusion from maize root segments. Roots may contain one or more water-soluble compounds competing with fusicoccin at the receptor level; such ligands might play a physiological role as modulators of the H+/K+ exchange system in higher plants.
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  • 53
    ISSN: 1432-2048
    Keywords: mRNA ; Nicotiana ; Protein synthesis ; Ribulose-1,5-bisphosphate carboxylase ; RNA (messenger) ; Translation
    Source: Springer Online Journal Archives 1860-2000
    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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  • 54
    ISSN: 1432-2048
    Keywords: Anther culture ; Fraction I protein ; Hybrid, somatic ; Nicotiana ; Ribulose-1,5-bisphosphate carboxylase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nine plants were produced from anthers of a somatic hybrid which had been obtained by fusion of Nicotiana tabacum L. and N. rustica L. protoplants. As determined by electrofocusing, the Fraction I protein of the original somatic hybrid had largesubunit polypeptides exclusively of the N. tabacum type. Two of the plants regenerated from anthers contained Fraction-I-protein large subunits exclusively of the N. rustica type. Since each plant was regenerated from a single cell, the somatic hybrid must have had cells containing both the N. tabacum and N. rustica chloroplast genome although the latter was not expressed. Possibilities to account for this non-expression of a chloroplast genome in the somatic hybrid are discussed.
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  • 55
    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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  • 56
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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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  • 57
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    Planta 147 (1980), S. 274-276 
    ISSN: 1432-2048
    Keywords: Cell division ; Dikegulac ; Lysis ; Nicotiana ; Plasmalemma ; Protoplasts ; Tonoplast
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    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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  • 58
    ISSN: 1432-2048
    Keywords: Auxin uptake ; Coleoptile ; Fluorescein ; Morphactin ; Naphthylphthalamic acid ; Triiodobenzoic acid ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Using both 1-mm segments of corn (Zea mays L.) coleoptiles and a preparation of membranes isolated from the same source, we have compared the effectiveness of several inhibitors of geotropism and polar transport in stimulating uptake of auxin (indole-3-acetic acid, IAA) into the tissue and in competing with N-1-naphthylphthalamic acid (NPA) for a membrane-bound site. Low concentrations of 2,3,5-triiodobenzoic acid (TIBA), NPA, 2-chloro-9-hydroxyfluorene-9-carboxylic acid (morphactin), and fluorescein, eosin, and mercurochrome all stimulated net uptake of [3H]IAA by corn coleoptile tissues while higher concentrations reduced the uptake of both [3H]IAA and another lipophilic weak acid, [14C]benzoic acid. Since low concentrations of fluorescein and its derivatives competed for the same membrane-bound site in vitro as did morphactin and NPA, the basis for both the specific stimulation of auxin accumulation and the inhibition of polar auxin transport by all these compounds may be their ability to interfere with the carrier-mediated efflux of auxin anions from cells. At higher concentrations, the decrease in accumulation of weak acids was nonspecific and thus may be the result of acidification of the cytoplasm and a general decrease in the driving force for uptake of the weak acids. Triiodobenzoic acid was an exception. Low concentration of TIBA (0.1–1 μM) were much less effective than NPA in competing for the NPA receptor in vitro, but little different from NPA in ability to stimulate auxin uptake. One possibility is that TIBA, a substance which is polarly transported, may compete with auxin for the polar transport site while NPA, morphactin, and the fluorescein derivatives may render this site inactive.
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  • 59
    ISSN: 1432-2048
    Keywords: Lectin binding ; Membranes (surfaces) ; Nicotiana ; Organelles (leaf) ; Protoplasts ; Spinacia
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    Topics: Biology
    Notes: Abstract Organelles isolated from leaves of spinach (Spinacia oleracea L.) were prefixed in glutaraldehyde and then incubated with ferritin conjugates of four lectins — Concanavalin A (Con A), Ricinus communis L. agglutinin, MW 120,000 (RCA), soybean agglutinin (SBA), and wheat germ agglutinin (WGA) — in order to probe their cytoplasmic surfaces for saccharide residues. In each case the major leaf organelles, including microbodies, mitochondria and chloroplast derivatives, failed to exhibit labeling when examined with the electron microscope. Tobacco (Nicotiana tabacum L.) leaf protoplasts, incubated simultaneously with and under identical conditions to the spinach organelles, showed specific labeling of their plasma membranes with all four lectin conjugates, thus establishing the efficacy of the procedure for demonstrating the presence of binding sites when they exist. Further attempts to show binding of one of the lectins, Con A, by labeling with fluorescein-Con A and by organelle agglutination, yielded results consistent with the absence of ultrastructural labeling. It is concluded that no saccharide residues recognized by the four lectins are present on the cytoplasmic surfaces of organelles and that those residues reported to be constituents of intracellular membranes, therefore, are most likely exposed on the luminal (extracytoplasmic) surfaces.
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  • 60
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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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  • 61
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    Planta 148 (1980), S. 103-107 
    ISSN: 1432-2048
    Keywords: Abscisic acid ; Cell cultures ; Nicotiana ; Variants, resistant ; Water stress
    Source: Springer Online Journal Archives 1860-2000
    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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  • 62
    ISSN: 1432-2048
    Keywords: Antigen ; Callus ; Cell suspension cultures ; Proteins (in differentiation) ; Seedling ; Vascular cylinder (stele) ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A group of antigenically distinct proteins characteristic for the tissue complex of the vascular cylinders was found in maize (Zea mays L.) seedlings using an immunofiltration technique. Specific stelar antigens present in the fully developed stele (vascular cylinder) of the primary root were also found in steles extracted from adventitious roots and from the mesocotyl but were absent, within the limits of sensitivity of the immunodiffusion tests employed, in root cortex and epidermis. Some of the stelar antigens were also evident in the meristem of the primary root and were present in traces in the scutellum, the mesocotyl node, and the primary leaves plus coleoptile. The specific stelar antigens could be traced in 13- and 15-day-old developing embryos and were definitely expressed by the 21 st day after pollination. Several stelar-specific antigens were found in embryo-derived callus tissues and in stem-derived cells maintained in serial suspension culture. Higher resolution of the stelar antigens by a modified technique of crossed immunoelectrophoresis was used to demonstrate several minor stelar antigens that were presumably characteristic exclusively of the completely differentiated stele. This technique along with sequential immunoprecipitation of labelled proteins provided a semiquantitative estimate of the specific stelar antigens in the meristem and the stele of the primary root, and in suspension-cultured cells which were devoid of noticeable signs of vascular differentiation.
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  • 63
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    Planta 148 (1980), S. 108-115 
    ISSN: 1432-2048
    Keywords: Callus ; Cell proliferation ; Cell suspension ; Coleoptile ; Cytodifferentiation ; Dedifferentiation ; Enzyme profile ; Internode ; Root tip ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The development of enzyme patterns was followed in the course of: (a) the irreversible cell differentiation via division and expansion to maturity in the root tip and coleoptile of the intact seedlings, (b) the irreversible cell dedifferentation associated with induction and establishment of callus from the growing internodes, and (c) the growth cycle (proliferation⇋stationary phase) in callus and cell-suspension cultures of maize (Zea mays L.). By measuring the activities of glycolytic, mitochondrial, microbody and hydrolytic enzymes cells proliferating in vivo and in vitro could be compared and changes related to cessation or resumption of cell division could be studied. Proliferating cells of callus and suspension cultures maintained by serial culture did not differ from those of the root meristem and coleoptile in the specific activities of hexokinase, phosphoglycerate kinase and phosphopyruvate hydratase. Proliferation in vitro resulted in an enormous increase in the ratio g glutamate-dehydrogenase/cytochrome-oxidase activity and in the level of acid-phosphatase activity, with concomitant drop in galactosidase and xylosidase activity. A 3-5-fold increase of alcohol-dehydrogenase, lactate-dehydrogenase and catalase activities was characteristic of dividing callus cells, while a ca. 100-fold increase in the fructofuranosidase-to-glucosidase activity ratio marked cell proliferation in suspension-cultured cells. Changing enzyme activities after cessation of proliferation were quite similar in root tips and coleoptiles, except those of alcohol dehydrogenase and catalase. The enzyme rearrangement during callus establishment and in the growth cycle of callus cultures was in most cases comparable to that in the intact tissues, while the changes from the dividing to the non-dividing cells in suspension cultures, in contrast, differed widely from those in the intact tissues and callus. Galactosidase and xylosidase were the only activities that showed a similar trend of changes in all the investigated, intact and in-vitro-grown cells. Thus, judged by the pattern of enzyme development, the cell suspension appears to be a unique system, virtually unrelated to the growing cells of the intact tissues. It is also very difficult to draw a definite distinction between the metabolic consequences of cell growth and enzyme modulations in cell suspensions as the cells adapt their metabolism to the environmental changes in liquid medium.
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  • 64
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    Planta 152 (1981), S. 397-407 
    ISSN: 1432-2048
    Keywords: Cytokinesis ; High voltage electron microscopy ; Mitosis ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The structure and distribution of cytoplasmic membranes during mitosis and cytokinesis in maize root tip meristematic cells was investigated by low and high voltage electron microscopy. The electron opacity of the nuclear envelope and endoplasmic reticulum (ER) was enhanced by staining the tissue in a mixture of zinc iodide and osmium tetroxide. Thin sections show the nuclear envelope to disassemble at prophase and become indistinguishable from the surrounding ER and polar aggregations of ER. In thick sections under the high voltage electron microscope the spindle is seen to be surrounded by a mass of tubular (TER) and cisternal (CER) endoplasmic reticulum derived from both the nuclear envelope and ER, which persists through metaphase and anaphase. At anaphase strands of TER traverse the spindle between the arms of the chromosomes. The octagonal nuclear pore complexes disappear by metaphase, but irregular-shaped pores persist in the membranes during mitosis. It is suggested that these form a template for pore-complex reformation during telophase. Phragmoplast formation is preceded by an aggregation of TER across the spindle at anaphase. Evidence is presented to suggest that the formation of the desmotubule of a plasmodesma is by the squeezing of a strand of endoplasmic reticulum between the vesicles of the cell plate.
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  • 65
    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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  • 66
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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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  • 67
    ISSN: 1432-2048
    Keywords: Chloroplast ; Cybrid ; Nicotiana ; Protoplast ; Somatic hybrid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts of a cytoplasmic albino mutant of Nicotiana tabacum L. characterized by a deficient chloroplast genome were fused with protoplasts of a nitrate-reductase deficient mutant (NR-) of N. tabacum. Somatic hybrids were obtained where the genome of the NR- mutant was complemented by the cytoplasmic albino mutant which could synthesize an active nitrate reductase, and the chlorophyll deficiency in the albino mutant was restored by the chloroplasts from the NR- mutant. Cybrids were also obtained in which the deficient plastids of the cytoplasmic albino mutant were replaced by normal chloroplasts from the NR- mutant. The system used permitted a simple selection of the hybrids and the cybrids. The NR- mutant was excluded at the cellular level by transfer of the cells to medium deficient in reduced nitrogen. The cytoplasmic albino mutant grew well on the selective nitrate medium. However, during callus formation, clear differences in the morphology and pigmentation of the calli were found which permitted selection for photoautotrophy at the callus level. The hybrid or cybrid nature of the plants was confirmed by examination of their morphology and chromosome number. Although the fusion partners come from the same species, only one plant showed the white-green variegated pattern typical of that of the cytoplasmic albino parent, indicating that segregation of plastids occurred during development of the calli and regeneration of the plants.
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  • 68
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    Planta 153 (1981), S. 447-452 
    ISSN: 1432-2048
    Keywords: Auxin binding ; Root (auxin) ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Auxin binding onto membrane fractions of primary roots of maize seedlings has been demonstrated using naphth-1yl-acetic acid (NAA) and indol-3yl-acetic acid (IAA) as ligands. This binding is compared with the already well characterized interaction between auxins and coleoptile membranes. The results indicate that while kinetic parameters are of the same order for root and coleoptile binding, a number of differences occur with respect to location in cells and relative affinity. The possible significance of the existence of such binding sites in root cells is discussed in relation to auxin action.
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  • 69
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    Planta 153 (1981), S. 453-458 
    ISSN: 1432-2048
    Keywords: Abscisic acid ; Gravireaction ; Growth inhibitors ; Root ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Using gas chromatography-mass spectrometry (GC/MS) techniques of analyses, it has been found that endogenous abscisic acid (ABA) becomes asymmetrically distributed in the elongation zone of horizontal Zea mays (cv. LG 11) roots which are showing a positive gravitropic response. There is a relative increase in the ABA content of the lower half and a concomitant decrease for the upper half in such roots. Asymmetric distribution of ABA is also detected in the elongation zone of half-decapped roots.
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  • 70
    ISSN: 1432-2048
    Keywords: Anaerobiosis ; Protein synthesis, organspecific ; Stress ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The pattern of protein synthesis was compared in several organs of maize (Zea mays L.) under aerobic and anaerobic conditions. Protein synthesis was measured by [35S]methionine incorporation and analysis by two-dimensional native-SDS (sodium lauryl sulfate) polyacrylamide gel electrophoresis and fluorography. The aerobic protein-synthesis profiles were very different for root, endosperm, scutellum and anther wall. However, except for some characteristic qualitative and quantitative differences, the patterns of protein synthesis during anaerobiosis were remarkably similar for these diverse organs and also for mesocotyl and coleoptile. The proteins synthesized were the anaerobic polypeptides (ANPs) which have been previously described in anaerobic roots of seedlings. Leaves exhibited no detectable protein synthesis under anaerobic conditions, and died after a short anaerobic treatment. Evidence is presented that the ANPs are not a generalized response to stress. This indicates that the ANPs are synthesized as a specific response to anaerobic conditions such as flooding.
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  • 71
    ISSN: 1432-2048
    Keywords: 1-Aminocyclopropane-1-carboxylic acid ; Ethylene ; Light and ethylene production ; Nicotiana ; Pharbitis ; Protein synthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Endogenous ethylene production of tobacco leaves was similar in light and in darkness. However, the rate of conversion of exogenously applied l-aminocyclopropane-l-carboxylic acid (ACC) to ethylene was reversibly inhibited by light. Virus-stimulated ethylene production, during the hypersensitive reaction of tobacco leaves to tobacco mosaic virus, was likewise inhibited by light. Under such circumstances ethylene production is limited at the level of the conversion of ACC to ethylene. Inhibition of the increase in ACC-stimulated ethylene production by cycloheximide and 2-(4-methyl-2,6-dinitroanilino)-N-methyl-propionamide after shifting leaf discs from light to darkness indicated that de novo protein synthsis was involved. Regulation of ACC-dependent ethylene production by reversible oxidation/reduction of essential SH groups, as suggested by Gepstein and Thimann (1980, Planta 149, 196–199) could be excluded. Instead, regulation of the ACC-converting enzyme at the level of both synthesis/degradation and activation/inactivation is suggested. Phytochrome was not involved in light inhibition, but low intensities of either red or blue light decreased the rate of ACC conversion. Dichlorophenyldimethylurea counteracted the inhibitory effect of light, indicating that (part of) the photosynthetic system is involved in the light inhibition. The ethylene production of Pharbitis cotyledons grown in darkness or light, either in the presence of absence of the inhibitor of carotenoid synthesis, SAN 9789 (norflurazon), supported this view.
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  • 72
    ISSN: 1432-2048
    Keywords: Aerenchyma ; Ethylene ; Gas space ; Oxygen and gas-space formation ; Roots, adventitious ; Silver ions ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have studied the role of ethylene in accelerating the lytic formation of gas spaces (aerenchyma) in the cortex of adventitious roots of maize (Zea mays L.) growing in poorly aerated conditions. Such roots had previously been shown to contain increased concentrations of ethylene. Ten day-old maize plants bearing seminal roots and one whorl of emerging adventitious roots were grown in nutrient solution bubbled with air, ethylene in air (0.1 to 5.0 μl l−1), or allowed to become oxygen-deficient in nonaerated (but not completely anaerobic) solution. Additions of 0.1 μl l−1 ethylene or more promoted the formation of aerenchyma, with lysis of up to 47% of the cortical cells. The effects of non-aeration were similar to those of exogenous ethylene. When silver ions, an ethylene antagonist, were present at low, non-toxic concentrations (circa 0.6 μM), aerenchyma formation was prevented in ethylene treated roots and in those exposed to oxygen deficiency. Silver ions also blocked the inhibiting effect of exogenous ethylene on root extension. By contrast, the suppression of aerenchyma formation by silver ions under oxygendeficient conditions was associated with a retardation of root extension, indicating the importance of aerenchyma for root growth in poorly aerated media. Rates of production of ethylene by excised roots were stimulated by a previous non-aeration treatment. The effectiveness of Ag+ in inhibiting equally the action on cortical cells of exogenous ethylene and of non-aeration, supports the view that gas space (aerenchyma) formation in adventitious roots ‘adpted’ to oxygendeficient environments is mediated by increased concentrations of endogenous ethylene. The possibility that extra ethylene could arise from increased biosynthesis of a precursor in root tissues with a restricted oxygen supply is discussed.
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  • 73
    ISSN: 1432-2242
    Keywords: Nicotiana ; Somatic hybrids ; Streptomycin resistance ; Plastid segregation ; Plastid DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Protoplasts of Nicotiana tabacum SRI (streptomycin resistant) and of Nicotiana knightiana (streptomycin sensitive) were fused using polyethylene glycol treatment. From three heterokaryons 500 clones were obtained. From the 43 which were further investigated, 6 resistant, 3 sensitive, and 34 chimeric (consisting of resistant and sensitive sectors) calli were found. From eight clones, a total of 39 plants were regenerated and identified as somatic hybrids. Chloroplast type (N. tabacum = NT or N. knightiana = NK) in the plants was determined on the basis of the species specific EcoRI restriction pattern of the chloroplast DNA. Regenerates contained NT (13 plants) or NK (15 plants) plastids but only the plants with the NT chloroplasts were resistant to streptomycin. This finding and our earlier data on uniparental inheritance points to the chloroplasts as the carriers of the streptomycin resistance factor.
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  • 74
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    Theoretical and applied genetics 60 (1981), S. 365-368 
    ISSN: 1432-2242
    Keywords: Disease resistance ; Isogenic lines ; Blackroot rot ; Thielaviopsis basicola ; Nicotiana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Lines isogenic or near isogenic for traits other than resistance to black root rot from Nicotiana debneyi were developed in eight cultivar backgrounds of burley tobacco (N. tabacum L.). For each cultivar background, a resistant and susceptible selection from the seventh backcross generation plus the recurrent parental cultivar were evaluated for ten agronomic and chemical traits. Resistant selections were statistically different from the susceptible entries for days to flower, total nitrogen content, and total alkaloid content. Also, resistant selections were consistently lower in yield, but the differences were statistically nonsignificant. Resistant selections were also taller in three families and in two families the resistant selections had wider leaves. Linkage of genetic material from N. debneyi with the resistance factor was suggested as the possible reason for differences between resistant and susceptible selections.
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  • 75
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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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  • 76
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    Plant systematics and evolution 138 (1981), S. 89-113 
    ISSN: 1615-6110
    Keywords: Angiosperms ; Nicotiana ; Gossypium ; Lemnaceae ; Fraction 1 protein ; differentiation and age of taxa ; migration to Australia ; land connections ; long distance dispersal
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Fraction 1 protein (F-1-protein) (ribulose bisphosphate carboxy-lase-oxygenase) contained inLemnaceae has been evolving for at least 50 million years because fossils of these plants have been identified in strata belonging to the Upper Cretaceous. Electrofocusing F-1-protein resolves the large subunit polypeptides coded by extranuclear DNA and the small subunit polypeptides coded by nuclear DNA. Four differences affecting isoelectric points of the large subunit polypeptides and eight affecting the small subunit polypeptides are now present among eleven species representing the four genera comprising theLemnaceae. In comparison, four differences in the large and 13 in the small subunit polypeptides exist among 63 species ofNicotiana; four differences in the large and eight differences in the small subunit polypeptides exist among 19 species ofGossypium. The number of differences in F-1-protein composition being of the same order of magnitude for the generaNicotiana, Gossypium, and the familyLemnaceae, we infer that these Angiosperms are of similar antiquity. Nicotiana species indigenous to Australia and Africa contain F-1-proteins whose large subunit polypeptides are different but some of whose small subunit polypeptides are like those found in species from the Western Hemisphere. The same situation is found for the F-1-protein inGossypium. These results are in harmony with the view that species ofNicotiana andGossypium have arrived in Australia via former land connections between S. America, Antarctica, and Australia.
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  • 77
    ISSN: 1615-6102
    Keywords: Nicotiana ; Dark requirement ; Protoplast regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The protoplasts ofNicotiana plumbaginifolia required darkness for cell regeneration and colony formation. Maximal plating efficiency of the protoplasts could be achieved by keeping the cultures in dark instead of light or dark/light sequence. Only two days of darkness prior to the illumination at 400 or 3,000 lux resulted in appreciable plating efficiency, than those of light from the beginning, but these values could not match the high plating efficiency in total darkness.
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  • 78
    ISSN: 1615-6102
    Keywords: Cell cycle ; Membrane charge ; Nicotiana ; Plant protoplasts ; Tobacco mosaic virus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Attachment of virions of tobacco mosaic virus to protoplasts isolated from dividing suspension cultured cells ofNicotiana sylvestris was estimated using quantitative autoradiography of individual protoplasts. Additionally, the position of each protoplast in the cell cycle was assessed by Feulgen microspectrophotometry. At pH 5.6, after preincubation with 4 μg 1−1 poly-L-ornithine, protoplasts in the G1 and G2 phases bound more virions than protoplasts in the S-phase. The possibility that such differential binding was caused by cyclical variation in the net charge on the protoplast membrane has been investigated. It was found that S-phase protoplasts ofN. sylvestris can be separarated from protoplasts of other cycle stages by partition in aqueous, two-phase, immiscible polymer systems, presumably because they differ in charge. Also, electrophoretic studies suggest that G1 phase protoplasts bear higher surface charge than some non-G1 protoplasts.
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  • 79
    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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