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  • 1
    Electronic Resource
    Electronic Resource
    Genes encoding the small subunit of RUBISCO belong to two highly conserved subfamilies in Nicotianeae (1991)
    Springer
    Journal of molecular evolution 33 (1991), S. 226-236 
    Details
    ISSN: 1432-1432
    Keywords: Concerted evolution ; Nicotianeae ; Nicotiana sylvestris ; Nicotiana tabacum ; Nicotiana tomentosiformis ; Small subunit of RUBISCO
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The sequences of seven complementary DNAs or genes encoding the small subunit (SSU) of ribulose-1,5-bisphosphate carboxylase oxygenase (RUBISCO) in several Nicotianeae were examined. Two new SSU genes isolated fromNicotiana sylvestris were included. Both sequence comparisons and Southern analyses with specific probes reveal that SSU genes fall into two homogeneous subfamilies that are highly conserved in Nicotianeae and are also present in other Solanaceae. Additional criteria such as number of introns and level of expression fitted to this classification. Homogeneity must have been maintained by gene conversion and/or an unusually high fidelity of DNA replication, whereas traces of slippage-stranded DNA mispairing and/or transposition probably explain local changes. Taken as a whole, these results show that the divergence between the two subfamilies predated the divergence between genera inside the Solanaceae, but that Nicotianeae retained the most simple SSU gene family structure.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02100673
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  • 2
    Electronic Resource
    Electronic Resource
    Ubiquitin genes are differentially regulated in protoplast-derived cultures of Nicotiana sylvestris and in response to various stresses (1992)
    Springer
    Plant molecular biology 20 (1992), S. 897-910 
    Details
    ISSN: 1573-5028
    Keywords: Cell division ; gene expression ; Nicotiana sylvestris ; protoplast ; stress ; ubiquitin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Four ubiquitin mRNA size classes were found to be differentially regulated in mesophyll protoplast-derived cultures of Nicotiana sylvestris. Three mRNA families of 1.9, 1.6 and 1.35 kb were expressed as soon as protoplasts were isolated. The 1.9 and 1.6 kb size classes were transiently expressed during the first hours of culture, whereas the level of expression of the 1.35 kb size class was maintained as long as cells kept dividing. A 0.7 kb mRNA size class started to be expressed just before the first divisions were observed. cDNAs corresponding to each of these families were isolated from a 6-h-old protoplast cDNA library and characterized. The 1.9, 1.6 and 1.35 kb mRNAs thus encode 7- or more, 6- and 5- mers, respectively, of ubiquitin whereas the 0.7 kb mRNAs encode a monomer of ubiquitin fused to a carboxyl extension protein of 52 amino acids. The expression of ubiquitin genes was studied, using probes specific for each of these transcript families, during protoplast culture and, for comparison, after various stresses including heat shock, HgCl2 treatment, a viral infection giving rise to a hypersensitive reaction, and an Agrobacterium tumefaciens infection which resulted in tumour formation. The 1.9 and 1.6 kb mRNA size classes were found to be stress-regulated, the 0.7 kb mRNA size class developmentally regulated and the 1.35 kb size class both stress- and developmentally regulated.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00027161
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  • 3
    Electronic Resource
    Electronic Resource
    A novel wound-inducible extensin gene is expressed early in newly isolated protoplasts of Nicotiana sylvestris (1995)
    Springer
    Plant molecular biology 29 (1995), S. 279-292 
    Details
    ISSN: 1573-5028
    Keywords: Agrobacterium tumefaciens ; extensin ; hydroxyproline-rich cell wall protein ; Nicotiana sylvestris ; protoplasts ; wounding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cDNA clone (6PExt 1.2) encoding a novel extensin was isolated from a cDNA library made from 6 h old mesophyll protoplasts of Nicotiana sylvestris. The screening was performed with a heterologous probe from carrot. The encoded polypeptide showed features characteristic of hydroxyproline-rich glycoproteins such as Ser-(Pro)4 repeats and a high content in Tyr and Lys residues. The presence of four Tyr-X-Tyr-Lys motifs suggests the possibility for intramolecular isodityrosine cross-links whereas three Val-Tyr-Lys motifs may participate in intermolecular cross-links. The analysis of genomic DNA gel blots using both the N. sylvestris and the carrot clones as probes showed that the 6PExt 1.2 gene belongs to a complex multigene family encoding extensin and extensin-related polypeptides in N. sylvestris as well as in related Nicotianeae including a laboratory hybrid. This was confirmed by the analysis of RNA gel blots: a set of mRNAs ranging in size from 0.3 kb to 3.5 kb was found by the carrot extensin probe. The 6PExt 1.2 probe found a 1.2 kb mRNA in protoplasts and in wounded tissues as well as a 0.9 kb mRNA which seemed to be stem-specific. The gene encoding 6PExt 1.2 was induced by wounding in protoplasts, in leaf strips and after Agrobacterium tumefaciens infection of stems.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00043652
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  • 4
    Electronic Resource
    Electronic Resource
    Isolation and characterization of a cDNA encoding a 3-hydroxy-3-methylglutaryl coenzyme A reductase from Nicotiana sylvestris (1992)
    Springer
    Plant molecular biology 20 (1992), S. 337-341 
    Details
    ISSN: 1573-5028
    Keywords: HMG-CoA reductase ; isoprenoid biosynthesis ; Nicotiana sylvestris ; protoplast ; stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cDNA library from freshly isolated mesophyll protoplasts of Nicotiana sylvestris was differentially screened using cDNAs from leaves, leaf strips submitted to the same stress as protoplasts during the isolation procedure, and cell suspension cultures. One of the selected clones (6P2) was found to encode a putative polypeptide highly homologous to previously characterized 3-hydroxy-3-methylglutaryl coenzyme A reductases. The C-terminal region of the polypeptide was highly conserved whereas its N-terminal region including the trans-membrane domains and the linker was more variable. Apart from protoplasts, the 6P2 gene was found to be expressed in apexes, anthers, roots, and in stressed leaf strips after 24h of culture, during the hypersensitive reaction to viral infection and after HgCl2 treatment. This pattern of expression is consistent with a role in plant defence mechanisms.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00014504
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  • 5
    Electronic Resource
    Electronic Resource
    Characterization of a tobacco extensin gene and regulation of its gene family in healthy plants and under various stress conditions (1997)
    Springer
    Plant molecular biology 33 (1997), S. 279-289 
    Details
    ISSN: 1573-5028
    Keywords: cell suspension culture ; cell wall ; elicitor ; extensin ; hydroxyproline-rich glycoprotein ; Nicotiana tabacum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A genomic clone (Ext 1.4) encoding an extensin was isolated from a Nicotiana tabacum genomic library. The encoded polypeptide showed features characteristic of extensins such as Ser-(Pro)4 repeats and a high content in Tyr and Lys residues. The presence of one Tyr-Leu-Tyr-Lys motif suggests the possibility for one intramolecular isodityrosine cross-link whereas numerous Val-Tyr-Lys motifs may participate in intermolecular cross-links. This extensin appears to be close to an extensin already characterized in N. tabacum but very different from the Ext 1.2 extensin of N. sylvestris. The analysis of genomic DNA gel blots using probes spanning different parts of the gene showed that the Ext 1.4 gene belongs to a complex multigene family having one member originating from N. sylvestris and three members from N. tomentosiformis. The Ext 1.4 specific probe found a 1.4 kb mRNA in stems, roots, ovaries and germinating seeds of healthy plants. The Ext 1.4 gene family is strongly induced in actively dividing cell suspension cultures and after wounding of leaves or stems in conditions where root formation occurs. On the contrary, it is not induced in leaves in response to a hyperensitive reaction to a viral infection or after elicitor treatment.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1005738815383
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  • 6
    Electronic Resource
    Electronic Resource
    Purification and characterization of 8 of the pathogenesis-related proteins in tobacco leaves reacting hypersensitively to tobacco mosaic virus (1986)
    Springer
    Plant molecular biology 6 (1986), S. 69-80 
    Details
    ISSN: 1573-5028
    Keywords: biochemical characterization ; high-performance liquid chromatography ; hypersensitive reaction ; Nicotiana tabacum ; pathogenesis-related proteins ; purification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Leaves of tobacco plants (Nicotiana tabacum cv. Samsun NN) which are reacting hypersensitively to infection with tobacco mosaic virus contain 10 major pathogenesis-related (PR) proteins which are absent, or present in small amounts in uninfected leaves. We describe here a preparative procedure of purification of the tobacco PR-proteins which involves a combination of conventional and high-performance liquid chromatography. The separation and isolation of the proteins were based on differences in net charge at different pH values, in isoelectric point and in apparent molecular weight. This procedure led to the purification to homogeneity of 8 PR-proteins, as shown by polyacrylamide slab gel electrophoresis (PAGE) of the purified proteins under denaturing and non-denaturing conditions. These were the 3 well-known proteins PR-1a,-1b and-1c, and 5 other major PR-proteins, called PR-2,-N,-O,-P and-Q, according to the nomenclature of Van Loon (39). None of the purified PR-proteins gave a positive Schiff reaction for carbohydrate content. Molecular weight determinations from gel permeation chromatography and from sodium dodecyl sulphate (SDS)-PAGE indicated that all 8 PR-proteins were monomers and that three groups could be distinguished among them. The first group is the PR-1 group containing PR-1a,-1b and-1c (12000 MW), the second consists of PR-P and PR-Q (14000 MW) and the third of PR-2, PR-N and PR-O (25000 MW). In the PR-1 group, PR-1a can be distinguished clearly from the two other members on denaturing slab gels containing both SDS and urea.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00027300
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