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  • 1
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The effects of two inhibitors of polyamine (spermidine and spermine) biosynthesis, cyclohexylamine (CHA; 5 and 10 mM) and methylglyoxal(bis-guanylhydrazone) (MGBG; 0.1, 0.5 and 1 mM), on the organogenic response in vegetative bud-forming tobacco (Nicotiana tabacum L. cv. Samsun) thin layer explants were evaluated micro- and macroscopically at different times during culture. The final number of buds formed and the percentage of organogenic explants was significantly reduced by both inhibitors, but much more so by MGBG than CHA. This inhibitory effect was already evident in MGBG-treated explants on day 5, in terms of the number of meristemoids per explant. On the contrary, in the presence of CHA, the number of meristemoids on day 5 was higher than that in the controls. Between days 9 and 13, meristemoid formation slowed down considerably in inhibitor-treated explants compared with controls. On day 13, the number of bud primordia was similar in control and CHA-treated explants, but significantly lower in MGBG-treated explants. This inhibitor also induced peculiar cytohistological events, such as a reduced formation of oval-shaped cell aggregates on the explant surface and more frequent cases of nucleolar extrusion, while CHA led to the appearance of hypertrophic epidermal cells; callus formation at the basal end of the explant and xylogenesis were also affected by the inhibitors. Ethylene biosynthesis, measured as [〈displayedItem type="mathematics" xml:id="di-fml-1" numbered="no"〉〈mediaResource alt="image" href="urn:x-wiley:00319317:PPL105223:PPL_105223_mu1"/〉C]methionine incorporation, was stimulated 2- (day 2) to 3-fold (15 h) by 0.5 mM MGBG, whereas CHA (10 mM) had little effect and aminoethoxyvinylglycine (AVG; 0.1 μM), an ethylene synthesis inhibitor, was strongly inhibitory. In control explants, the incorporation of labelled methionine into ethylene and spermidine followed an inverse trend up to day 8. In these explants, free putrescine increased 32-fold and spermidine increased about 10-fold between days 0 and 8. Trichloroacetic acid (TCA)-soluble conjugated putrescine also accumulated dramatically during culture. While CHA provoked a decline in spermidine levels, MGBG caused an unexpected increase in free spermidine and spermine titres; however, its most conspicuous effect was on the further enhancement of putrescine conjugate accumulation, while CHA and AVG had the opposite effect. Results are discussed in view of establishing a putative link between MGBG-enhanced ethylene synthesis, increased conjugate titres and inhibition of meristemoid formation.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 95 (1995), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Using tobacco root cultures, we explored the possibility of a correlation between the expression of the rol genes from the TL-DNA (rolA, rolB, rolC) of Agrobacterium rhizogenes and changes in polyamine content and metabolism. Transgenic roots were induced by inoculation of Nicotiana tabacum cv. Xanthi leaf discs with a disarmed strain of A. tumefaciens harbouring the rolA, rolB and rolC genes either singly or in combination; the presence of these genes on the plant genome was confirmed by Southern blot analysis. Transgenic roots, especially those transformed either by a combination of the three rol genes (A+B+C) or the rolC alone, grew faster than the untransformed roots. Putrescine, spermidine and traces of spermine were present in all samples, both in free and bound forms. While rolA roots showed increased levels of free and bound polyamines as compared with controls, accumulation of polyamines in rolB and rolC roots was inhibited or mantained, with the exception of a 66 and 48% increase, respectively, in the PCA-soluble conjugated fraction. In roots transgenic for all three rol genes (A+B+C), the polyamine content remained almost unaltered compared with controls, suggesting that rolB and rolC genes could nullify or compensate the rolA effects. The higher polyamine contents found in roots transformed by rolA paralleled with higher ornithine (EC 4.1.1.17) and arginine (EC 4.1.1.19) decarboxylase activities as well as higher nicotine production. It is suggested that polyamine metabolism in root cultures is altered by expression of rol genes.
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  • 3
    ISSN: 1432-203X
    Keywords: Key words Hairy roots ; Hydroxycinnamoyl amides ; Hyoscyamus muticus ; Jasmonates ; Methylputrescine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Jasmonic acid (JA) and its methyl ester (MeJA) at concentrations ranging from 0.001 to 10 μM provoked large increases in methylputrescine levels in normal and hairy roots of Hyoscyamus muticus L.; generally, levels of free putrescine and perchloric acid-soluble conjugated putrescine, spermidine and spermine also increased dramatically. More 14C-putrescine was formed when hairy roots were incubated with labelled ornithine than with arginine; conjugated 14C-putrescine was also rapidly formed. In accord with these results, ornithine decarboxylase (EC 4.1.1.17) activity was higher than that of arginine decarboxylase (EC 4.1.1.19), and MeJA enhanced these activities about two- and fourfold, respectively. Although treatment of root cultures with jasmonates enhanced precursor (putrescine, methylputrescine) levels and accumulation of secondary metabolites such as acid-soluble conjugated di-/polyamines, it provoked only modest increases in tropane alkaloid tissue levels.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1615-6102
    Keywords: Solanum melongena ; Polyamines ; Organogenesis ; Arginine decarboxylase ; Ornithine decarboxylase ; Amine oxidases
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Eggplant (Solanum melongena L. cv. Violetta lunga 2) cotyledon expiants grown on hormone-free medium (controls) or on medium containing either naphthaleneacetic acid alone (root forming) or in combination with zeatin riboside (shoot forming) showed minor differences in free polyamine titres during culture. In contrast, conjugated polyamines (particularly those in the trichloroacetic acid-soluble fraction) accumulated only in hormonetreated explants, but not in controls. The extent and the temporal changes in soluble-conjugate levels differed between root-forming and shoot-forming expiants; in the former, accumulation began earlier (within 1 day of culture) and reached the highest levels. In both organogenic programmes, maximum conjugate accumulation occurred just before and during organ emergence. Adventitious roots and shoots were formed along the cut surfaces. The regions closest to these (“borders”) displayed a significantly higher ratio of conjugated to free spermidine and/or putrescine than the nonorganogenic regions (“centres”) of the explant. Ornithine decarboxylase activity was higher than arginine decarboxylase activity both in control and hormone-treated explants. However, both activities increased markedly on day 2 of culture in the presence of hormones. Thereafter ornithine decarboxylase activity remained high in shoot-forming explants, but not in root-forming ones. Putrescine oxidising activity was also enhanced by exogenously supplied hormones starting from day 4 of culture. This activity remained high up to day 12 in the presence of auxin plus cytokinin, whereas it peaked on day 6 in auxin-treated explants. Spermidine oxidising activity was the only enzyme activity which was consistently higher in controls than in hormone-treated tissue. Differences between the two organogenic programmes with respect to temporal changes in polyamine content, and putrescine biosynthetic and oxidative activities are discussed in relation to the timing of organ formation. The latter was monitored both histologically and macroscopically.
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  • 5
    ISSN: 1615-6102
    Keywords: Ethylene ; Ethylene inhibitors ; Meristemoids ; Nicotiana tabacum ; Thin layers ; Vegetative buds
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The role of ethylene in vegetative bud regeneration was studied in cultured tobacco (Nicotiana tabacum L. cvSamsun) thinlayer expiants. The experimental approach consisted in supplementing the bud-inducing medium with an inhibitor of ethylene biosynthesis, aminoethoxyvinylglycine (AVG), an ethylene antagonist, silver thiosulphate (STS), or an ethylene-releasing compound, 2-chloroethylphosphonic acid (CEPA), at various concentrations. The organogenic response was assessed both macroscopically (percentage of bud-forming expiants, final number of buds per expiant) and cytohistologically (number, characteristics, and localisation of meristemoids and bud primordia). The time course of ethylene production during culture was also evaluated. At the end of culture (day 27) all the expiants treated with these compounds had a lower number of buds compared to controls. STS was detrimental to meristemoid initiation at all the concentrations tested. In contrast, 0.5 μM AVG, which strongly inhibited ethylene production, provoked a large increase in the formation of meristemoids early in culture and the appearance of anomalous (“twin”) buds. CEPA reduced meristemoid formation but, at the lower concentrations (1 and 10 μM) speeded up bud emergence. On the whole it mainly favoured disorganised growth and xylogenesis. The results of this work highlight the contrasting effects of ethylene in relation to the two critical stages of the organogenic process, i.e., meristemoid formation and bud primordium development.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    European archives of oto-rhino-laryngology and head & neck 256 (1999), S. 373-377 
    ISSN: 1434-4726
    Keywords: Key words Anterior commissure carcinoma ; Frontolateral laryngectomy ; Functional laryngectomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Patients with laryngeal anterior commissure, cord-commissure, bilateral anterior cord-commissure carcinomas (T1 and T2 N0 M0) were subjected to a new method of frontolateral laryngectomy using a combined endolaryngeal and external approach. The proposed surgical procedure allowed the removal of the anterior commissure and part of one or both vocal cords in a single unit, together with the cartilaginous framework, respecting the integrity of the superior portion of the thyroid cartilage. The internal procedure permitted an accurate delimitation of the posterior part of the laryngeal neoplasm. In particular, this was performed during suspension microlaryngoscopy using the CO2 laser or traditional cutting tools for section of the laryngeal visceral structures to the internal surface of the thyroid cartilage. Following this, the external approach included incision of the external perichondrium along the superior edge of the thyroid cartilage and along the median line, from the incisura to the inferior edge of the thyroid cartilage. The superior opening of the larynx is made side-to-side and the epiglottis separated at the level of the superior edge of the thyroid cartilage. The ends of the section are joined together with the superior parts of the section created during the laryngeal approach. Then progressive craniocaudal detachment of the internal perichondrium is performed backwards until the endolaryngeal sections are reached and downwards to the insertion of the cord ligaments. The inferior opening of the larynx is made by a horizontal section of the cricothyroid membrane at the level of the superior edge of the cricoid cartilage. Section of the thyroid cartilage is therefore performed in a trapezoidal shape. This section involves the inferior part of the protruding corner of the thyroid cartilage. After joining the ends of the cricothyroid section with the inferior extremities of the endolaryngeal sections, the surgical specimen is removed as a single unit. The method must be used only after accurate clinical evaluation. It is mostly recommended in subjects with cord-commissure carcinomas previously treated with radiotherapy. The results obtained were extremely satisfactory both as regards survival and functional results. In all, 27/28 patients (96.4%) were free from disease. The quality of voice was satisfactory but hoarse in 30% and breathy in 70% of the patients.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Journal of plant growth regulation 16 (1997), S. 159-167 
    ISSN: 1435-8107
    Keywords: Key Words. Hormonal effects—Hyoscyamus muticus—Roots
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract. Treatment of normal and Agrobacterium rhizogenes-transformed root cultures of Hyoscyamus muticus with three different auxins, indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), and naphthaleneacetic acid (NAA), revealed that the response varied considerably among auxins, between transformed and normal roots, and depending on the parameter. In normal roots all three auxins provoked abundant branching, with IBA and NAA being the most effective at 2.5 and 0.5 μm, respectively, whereas IAA was most effective at low concentrations (0.05 and 0.1 μm). In transformed roots exogenously supplied auxins were generally inhibitory or, at best, without effect on growth and branching. Only 0.01 μm IAA significantly enhanced lateral root number, whereas at the higher concentrations IBA, although inhibitory, was the least effective auxin. In both root types IBA had little effect on primary root growth, but normal roots were more sensitive to IAA and NAA. These results suggest a different sensitivity to auxins of normal and transformed roots since there was no significant difference in endogenous free and conjugated IAA content nor in IAA uptake capacity. Ethylene production and biosynthesis were approximately threefold higher in hairy roots, but production could be stimulated up to tenfold that of control levels in normal roots by supplying NAA or 1-aminocyclopropane-1-carboxylic acid (ACC). Treatment with 2.5 μm NAA, but not IAA or IBA, also enhanced ethylene biosynthesis in normal roots but not in transformed ones. ACC and malonyl-1-aminocyclopropane-1-carboxylic acid accumulated to detectable levels only after treatment with an auxin (NAA).
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