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  • 1
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 201 (1964), S. 1149-1150 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] This communication describes experiments with 1-methyl-3-nitro-1-nitrosoguanidine (NG). NG has already been proved to be mutagenic in E. coli3, it has cancero-static activity in mice2 and induces chromosome aberrations in Viciafaba1, although the frequency of aberrations is relatively very low, ...
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 44 (1978), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Protoplasts were isolated from two mutant cell lines of Nicotiana tabacum L. cv. Gatersleben and fused with the aid of polyethylene glycol. Both mutants lacked nitrate reductase and were thus auxotrophic for reduced nitrogen. The fusion resulted in a high frequency of hybrid cells which were detected by their regained ability to grow in media containing nitrate as sole nitrogen source. Thus, the two mutants were found to complement each other in the hybrids. In control experiments, back mutation and cross-feeding were excluded as possible explanations for the occurrence of cell lines utilizing nitrate. A total of 1061 hybrid lines capable of sustained proliferation were isolated. Some of them were further characterized with respect to nitrate reductase activity, chlorate sensitivity, chromosome number, and shoot formation.The results demonstrate that protoplast fusion can be used for the genetic analysis of cell variants of higher plants and that nitrate reductase-deficient mutants provide efficient selective systems for hybrid cells.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 36 (1966), S. 201-220 
    ISSN: 1432-2242
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Description / Table of Contents: Zusammenfassung Samen vonArabidopsis thaliana wurden mit Äthylmethansulfonat (EMS) behandelt. Es wurde die Abhängigkeit des Effekts von folgenden Faktoren untersucht: EMS-Konzentration, Behandlungszeit, Dauer der Nachquellperiode, Temperatur während der Behandlung und während des Nachquellens, Rücktrocknung der behandelten Samen, und Hydrolyse der Behandlungslösung. Die untersuchten Kriterien waren: Wachstumsrate der Primärwurzeln und andere somatische Effekte, Fertilität der M1-Pflanzen, Frequenz der rezessiven Letalmutationen (embryonale Letalmutationen und Chlorophyllmutationen). Die wichtigsten Ergebnisse sind: 1. Sowohl die Frequenz der Chlorophyllmutationen als auch die Frequenz der embryonalen Letalmutationen erhöht sich in exponentieller Abhängigkeit von der Dosis (n≈2). Das gilt unabhängig davon, ob die EMS-Konzentration oder die Behandlungszeit variiert wird. 2. Mutationsfrequenz, Sterilität und Wurzellängenreduktion erhöhen sich mit steigender Behandlungstemperatur. Für den geprüften Bereich (18° bis 36 °C) wurde ein Temperaturquotient vonQ 10=2,6 gefunden. 3. Die Steigerung der Mutationsfrequenz wird nur durch die Verringerung der Fertilität der M1-Pflanzen und nicht durch eine Verringerung des Überlebens begrenzt. Die Relationen zwischen Mutationsfrequenz, Sterilitätsgrad und Wurzellängenreduktion werden von Änderungen der Behandlungsbedingungen nicht beeinflußt. 4. Mutationsfrequenz, Sterilität und Wurzellängenreduktion sind unabhängig von der Dauer des Quellens der behandelten Samen in Wasser unter partiell anaeroben Bedingungen. Eine Verzögerung des Keimbeginns bis zu 4 Tagen nach der Behandlung hat keinen Einfluß auf die Mutationsfrequenz. 5. Durch Erhöhung der Temperatur während einer Periode von 6 h nach Behandlungsschluß werden Mutationsfrequenz, Sterilität und Wurzellängenreduktion gesteigert. 6. Der durch Trocknung der behandelten Samen verursachte Schaden wird durch 11stündiges Quellen der Samen in Wasser vor der Trocknung vollständig aufgehoben. 7. Die Effektivität von partiell hydrolysierten EMS-Lösungen entspricht der Konzentration des EMS. Der durch die Hydrolyseprodukte verursachte physiologische Schaden ist ohne praktische Bedeutung. 8. Die Beziehungen zwischen verschiedenen Maßen der Mutationsfrequenz (Frequenz der spaltenden Pflanzennachkommenschaften, Frequenz der spaltenden Schotennachkommenschaften, Frequenz der M2-Mutanten) wurden analysiert. U. a. wird gezeigt, daß zwischen der Frequenz der M2-Mutanten (m c ) und der ursprünglichen Mutationsfrequenz keine direkte Proportionalität besteht.
    Notes: Summary Seeds ofArabidopsis thaliana were treated with ethyl methanesulfonate (EMS). The dependence of the effect on the following factors was studied: EMS concentration, length of treatment, duration of posttreatment soaking, temperature during treatment and during post-treatment soaking, drying of the treated seeds and hydrolysis of treatment solution. The criteria studied included growth rate of primary roots and other somatic effects, fertility of M1 plants, frequency of recessive lethals (embryonic lethals and chlorophyll mutations). The main results are: 1. With increasing dose the frequency of chlorophyll mutations and the frequency of embryonic lethals increase exponentially (n≈2). This applies to variation of EMS concentration as well as to variation in treatment time. 2. Mutation frequency, sterility and root length reduction increase with rising treatment temperature. For the range tested (18°–36°C) a temperature quotient ofQ 10=2.6 was found. 3. The increase in mutation frequency is limited only by the decrease in fertility of M1 plants and not by a decrease in survival. The relationships between mutation frequency, degree of sterility and root length reduction are not influenced by changes in treatment conditions. 4. Mutation frequency, sterility and root length reduction are independent of the duration of soaking the treated seeds in water under partially anaerobic conditions. Delaying the start of germination up to 4 days after treatment does not influence the mutation frequency. 5. By raising the temperature during a period of 6 hrs. after treatment mutation frequency, sterility and root length reduction are increased. 6. The damage caused by drying seeds after EMS treatment is prevented completely by soaking the treated seeds in water for 11 hrs. before drying. 7. The effectiveness of partially hydrolyzed EMS solutions corresponds to the concentration of EMS. Physiological damage caused by the hydrolysis products is without practical significance. 8. The relations between various measures of mutation frequency (frequency of segregating plant progenies, frequency of segregating pod progenies, frequency of M2 mutants) have been analyzed. Among other things it is shown that there is no direct proportionality between the frequency of M2 mutants (m c ) and the initial mutation frequency.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-2048
    Keywords: Molybdenum ; Mutant (Nicotiana) ; Nicotiana (nitrate reductase) ; Nitrate reductase mutant
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two nitrate reductase-deficient mutant cell lines (CnxA68/2, CnxA101) of Nicotiana tabacum are shown to be repairable under in-vitro conditions by (i) molybdate or (ii) by preparations of active molybdenum cofactor of homologous or heterologous origin, thereby yielding about 20% and 80%, respectively, of the corresponding wild-type NADH-nitrate reductase (EC 1.6.6.1) activity. In-vitro repair of nitrate reductase activity is dependent on sulphydryl-group protecting reagents and ethylenediaminetetraacetic acid (EDTA) in the extraction medium, the nitrogen source in the growth medium and the age of the cells. The results support the conclusion that the cnxA gene controls the insertion of molybdenum into the molybdenum cofactor. They are consistent with the idea of two interlinked pathways for the metabolic processing of molybdenum acquisition, one involving the synthesis of the structural moiety of the molybdenum cofactor and the other involving processing of the molybdate anion.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 34 (1964), S. 102-120 
    ISSN: 1432-2242
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 161 (1978), S. 67-76 
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Chlorate-resistant cell lines were established from survivors after plating allodihaploid cells of Nicotiana tabacum into solid medium containing 20 mM chlorate and amino acids as sole nitrogen source. Data characterizing 9 of the most resistant lines are presented. The mutational origin of these lines was inferred on the basis of the enhancement of the variant frequency by mutagen treatment, and of the persistance of the variant phenotype in cell progeny during growth in the absence of selection for more than 3 years and in plants regenerated from two of the lines. Seven lines completely lacked in vivo nitrate reductase (NR) activity and two lines exhibited low (less than 5% of the wild type) NR activity. The abolition of NR activity was found to be not due to an impaired induction by nitrate. Data reported elsewhere show that one of the NR-negative mutants simultaneously lacks xanthine dehydrogenase activity. This pleiotropic mutation is interpreted to affect the synthesis of a molybdenum-containing cofactor, whereas the 8 other lines carry mutations specifically affecting the synthesis of the NR. Both types of NR-negative mutants were unable to grow on minimal medium containing nitrate as sole nitrogen source, but grew well on amino acids. They proved extremely sensitive to the standard medium containing nitrate and ammonium. Differences between the NR-negative mutants with respect to chlorate resistance suggest that chlorate inhibits cultured N tabacum cells not only via its NR-catalysed conversion to chlorite, but also by NR-independent mechanisms.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 161 (1978), S. 77-80 
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two types of nitrate reductase-deficient mutant cell lines (nia and cnx) of Nicotiana tabacum have been used for in vitro reconstitution of NADH-nitrate reductase. The cnx mutants simultaneously lack NADH-,FADH2-, red benzyl viologen-nitrate reductase, and xanthine dehydrogenase activities, but retain the nitrate reductase-associated NADH-cytochrome c reductase activity. These mutants are interpreted to be defective in the molybdenum-containing cofactor necessary for nitrate reductase activity. In the nia lines xanthine dehydrogenase activity is unaffected, and the loss of NADH-nitrate reductase is accompanied by a loss of all partial activities of nitrate reductase, including NADH-cytochrome c reductase. When cnx cells (induced by nitrate) were homogenized together with nia cells (induced by nitrate or uninduced), NADH-nitrate reductase activity was detectable in the cell extract. No nitrate reductase was observed when the cnx mutants were homogenized together, or after cohomogenization of the nia mutants. Thus, the inactive nitrate reductase molecule formed in the cnx mutants has been complemented in vitro with the molybdenum-containing cofactor supplied by nia extracts, thus giving rise to NADH-nitrate reductase activity. This result gives additional support to the interpretation that the active nitrate reductase of Nicotiana tabacum is composed of at least the NADH-cytochrome c reductase moiety and a molybdenum-containing cofactor which is formed by the action of the cnx gene product(s).
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  • 8
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Nicotiana tabacum mutant cell cultures lacking nitrate reductase activity were assayed for the presence of the molybdenum-cofactor using its ability to restore NADPH-nitrate reductase activity in extracts of Neurospora crassa nit-1 mycelia. The molybdenum-cofactor of the tobacco wild-type line was shown to complement efficiently the N. crassa nit-1 mutant in vitro. The molybdenum-cofactor seems to exist in a bound form, as acid-treatment was required for release of cofactor activity. Molybdate (5–10 mM), ascorbic acid, and anaerobic conditions greatly increased the activity of the cofactor, demonstrating its high lability and sensitivity to oxygen. Similar results were obtained with two tobacco nia mutants, which are defective in the apoprotein of nitrate reductase. The four cnx mutants studied were shown to contain exclusively an inactive form of the molybdenum-cofactor. This inactive cofactor could be reactivated in vitro and in vivo by unphysiologically high concentrations of molybdate (1–10 mM), thereby converting the cnx cells into highly active cofactor sources in vitro, and restoring nitrate reductase and xanthine dehydrogenase in vivo to partial acitivity. Thus the defect of the cnx mutants resides in a lack of molybdenum as a catalytically active ligand metal for the cofactor, while the structural moiety of the cofactor seems not to be impaired by the mutation. The subunit assembly of the nitrate reductase was found to be independent of the molybdenum content of the cofactor.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In vitro complementation of the nitrate reductase-deficient barley mutant nar2a extracts with molybdenum cofactor from commercial xanthine oxidase resulted in reactivation of NADH: nitrate reductase activity. Maximum reactivation was achieved with 7.5 μg/ml xanthine oxidase (final concentration), 10 mM glutathione (final concentration) and incubation for 30 min at room temperature (ca. 25°C). This in vitro complementation assay was used to determine the presence of functional apoprotein and molybdenum cofactor in 12 nitrate reductase-deficient barley mutants. Extracts of all nar1 alleles contained functional molybdenum cofactor (complemented with nar2a) but they lacked functional apoprotein (did not complement with molybdenum cofactor from xanthine oxidase). The nar2a, nar3a and nar3b extracts were able to donate functional apoprotein, but were poor sources of functional molybdenum cofactor. These data are in agreement with our previous assignment of nar1 to the barley NADH: nitrate reductase structural locus and nar2 and nar3 to molybdenum cofactor functions. Wild type cv. Steptoe barley seedlings grown in the absence of nitrate and lacking nitrate reductase activity contained low levels of molybdenum cofactor. Nitrate induction resulted in a several-fold increase in the measurable molybdenum cofactor levels that was correlated with the increase in nitrate reductase activity.
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  • 10
    ISSN: 1617-4623
    Keywords: Nicotiana plumbaginifolia ; Nitrate reductase ; Genetics ; Molybdenum cofactor biosynthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A total of 70 cnx mutants have been characterized from a collection of 211 nitrate reductase deficient (NR-) mutants isolated from mutagenized Nicotiana plumbaginifolia protoplast cultures after chlorate selection and regeneration into plants. They are presumed to be affected in the biosynthesis of the molybdenum cofactor since they are also deficient for xanthine dehydrogenase activity but contain NR apoenzyme. The remaining clones were classified as nia mutants. Sexual crosses performed between cnx mutants allowed them to be classified into six independent complementation groups. Mutants representative of these complementation groups were used for somatic hybridization experiments with the already characterized N. plumbaginifolia mutants NX1, NX24, NX23 and CNX103 belonging to the complementation groups cnxA, B, C and D respectively. On the basis of genetic analysis and somatic hybridization experiments, two new complementation groups, cnxE and F, not previously described in higher plants, were characterized. Unphysiologically high levels of molybdate can restore the NR activity of cnxA mutant seedlings in vivo, but cannot restore NR activity to any mutant from the other cnx complementation groups.
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