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  • 1
    Electronic Resource
    Electronic Resource
    Plastid gene expression in a yellow-green leaf mutant of Petunia hybrida (1983)
    Springer
    Planta 157 (1983), S. 209-217 
    Details
    ISSN: 1432-2048
    Keywords: Mutant (Petunia) ; Northern blotting and hybridization ; Petunia ; Plastid gene expression ; Protein synthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have analyzed the morphology and gene expression in plastids of a yellow-green leaf mutant of Petunia hybrida (E 5059). Under normal light intensities (20,000 lx), yellow-green leaves develop with a typical proplastid morphology (few membranes, incomplete stacking). When such plants are grown under low light intensities (3,000 lx), the newly formed leaves are green. The plastids in these green leaves have a wild-type like chloroplast morphology (thylakoids and grana structure). Pre-existing green leaves remain green in 20,000 lx, indicating that chlorophyll is not degraded. An analysis of polypeptides synthesized in isolated plastids from the yellow-green and green leaves of this mutant plant shows several differences. In the yellow-green leaf plastids only a very small amount of the large subunit of ribulose-1,5-bisphosphate carboxylase (RuBPCase) is present, while in green plastids this polypeptide is present in much higher amounts. Hybridization experiments indicated that in plastids from the yellow-green leaves the mRNA coding for the large subunit polypeptide is present in much lower amounts than in plastids from the 3,000 lx green leaves of this mutant or in chloroplasts from wild type plants. These results indicate regulation at the mRNA level. Furthermore, in yellow-green leaf plastids eleven polypeptides are present with high molecular wieght (higher than 67,000 d). Five of them are synthesized by the yellow-green leaf plastid itself. Such high molecular weight polypeptides are also synthesized by proplastids isolated from white petunia cell suspension cultures, but are not synthesized by 3,000 lx green leaf plastids, or by isolated normal leaf chloroplasts. These results indicate that the synthesis of these polypeptides is specific for the proplastid stage of chloroplast biogenesis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00405184
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  • 2
    Electronic Resource
    Electronic Resource
    Protein synthesis in Petunia hybrida chloroplasts isolated from leaves and cell cultures (1982)
    Springer
    Planta 155 (1982), S. 37-44 
    Details
    ISSN: 1432-2048
    Keywords: Cell suspension culture ; Chloroplast ; Petunia ; Protein synthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Isolation and incubation conditions were established for Petunia hybrida chloroplasts capable of performing in vitro protein and RNA synthesis. Under these conditions, chloroplasts from leaves as well as from the non-photoautotrophic mutant green cell culture AK-2401 are able to incorporate labeled amino acids into polypeptides. Intact chloroplasts can use light as an energy source; photosynthetically-inactive chloroplasts require the addition for ATP for this protein synthesis. Sodium dodecylsulphate polyacrylamide slab gel electrophoresis shows that in isolated leaf chloroplasts at least twenty-five radioactive polypeptide species are synthesized. The three major products synthesized have molecular weights of 52,000, 32,000 and 17,000. Coomassie brilliant-bluestained polypeptide patterns from plastids isolated from the mutant green cell culture AK-2401 differ considerably from those obtained from leaf chloroplasts. The pattern of radioactive polypeptides synthesized in these isolated cell culture plastids also shows differences. These results indicate that the difference in developmental stage observed between plastids from the cell culture AK-2401 and leaves is reflected in an altered expression of the chloroplast DNA.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00402929
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  • 3
    Electronic Resource
    Electronic Resource
    Physical mapping of genes for chloroplast DNA encoded subunit polypeptides of the ATPsynthase complex from Petunia hybrida (1984)
    Springer
    Current genetics 8 (1984), S. 283-290 
    Details
    ISSN: 1432-0983
    Keywords: Petunia hybrida chloroplast DNA ; E. coli minicells ; ATPsynthase ; Gene mapping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Escherichia coli minicells harbouring the cloned restriction fragment Sall S9 from P. hybrida chloroplast DNA synthesize the beta and epsilon polypeptide subunits of the CF1 component of the chloroplast ATPsynthase complex. The polypeptides were identified by molecular weight determination and immunoprecipitation. The position of the atpB and the atpE gene, encoding respectively the beta and epsilon subunit, on the Sall S9 fragment was determined in more detail by studying polypeptide synthesis directed by subclones of the S9 fragment in E. coli minicells. The atpB and atpE genes are located close to the rbcL gene, the distance between the rbcL gene and atpB gene being approximately 770 bp. Analysis of the expression of subclones of the S9 fragment in E. coli minicells also revealed that the atpE gene can be transcribed and translated independently of the expression of the atpB gene. The location of the genes coding for the alpha subunit (atpA gene) and the proteolipid subunit III of CF0 (atpH) of the ATPsynthase complex on the physical map of P. hybrida cpDNA was determined by hybridization of restriction enzyme digests of petunia cpDNA with cloned cpDNA fragments from Spirodela and wheat, containing internal parts of respectively the atpA and the atpH gene. The two genes map close together within a region of 5.2 kbp on the physical map of P. hybrida cpDNA. The distance between the atpA gene and the atpB and atpE genes is approximately 42 kbp.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00419726
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  • 4
    Electronic Resource
    Electronic Resource
    Physical mapping, nucleotide sequencing and expression in E. coli minicells of the gene for the large subunit of ribulose bisphosphate carboxylase from Petunia hybrida (1984)
    Springer
    Current genetics 8 (1984), S. 231-241 
    Details
    ISSN: 1432-0983
    Keywords: Petunia hybrida chloroplast DNA ; E. coli minicells ; rbcL gene ; Nucleotide sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The Petunia hybrida rbcL gene was identified and located on the physical map within the Sall S9 fragment of the Petunia hybrida cpDNA by heterologous hybridization with the cloned rbcL gene of spinach (pSoc3BE148). In E. coli minicells harbouring the S9 fragment inserted into pBR322, the rbcL polypeptide is synthesized as was shown by molecular weight determination, immunoprecipitation and proteolytic digestion. However, the size of the rbcL polypeptide synthesized in minicells appeared to be dependent on the orientation of the S9 fragment in pBR322. In minicells harbouring the S9 fragment inserted into pBR322 in the clockwise orientation the molecular weight of the rbcL polypeptide is approximately 53 kD, whereas in minicells harbouring the S9 fragment in the opposite orientation, the rbcL polypeptide synthesized has a molecular weight of 52 kD. The difference in molecular weight of the two rbcL polypeptides is the result of transcription and translation into the flanking pBR322 sequences. This is due to the absence of the terminal part (6 codons), including the translation stop codon, of the rbcL gene on the cloned S9 fragment as was determined by nucleotide sequencing. The observed expression of the cloned part of the rbcL gene of Petunia hybrida indicates that the E. coli minicell system can be used as a suitable and convenient system for the identification and physical mapping of chloroplast genes. Comparison of the sequence of the untranslated 3′-end of the rbcL gene of Petunia hybrida with that of Nicotiana tabacum revealed a striking similarity of the region in which stem and loop structures can be formed that are most likely involved in termination of transcription of the rbcL gene. This region appears to be highly conserved in the rbcL genes of P. hybrida, N. tabacum, S. oleracea and Z. mays.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00417821
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  • 5
    Electronic Resource
    Electronic Resource
    Light and benzylaminopurine induce changes in ultrastructure and gene expression in plastids of Petunia hybrida cell cultures (1982)
    Springer
    Current genetics 6 (1982), S. 129-135 
    Details
    ISSN: 1432-0983
    Keywords: Petunia hybrida ; Cell cultures ; Plastid gene expression ; Protein synthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The regulatory effect of light and the cytokinin 6-benzylaminopurine (BA) on the plastid ultrastructure and plastid DNA gene expression is studied in white and mutant green cell suspension cultures of Petunia hybrida. By electron microscopy we show that both light and 6-benzylaminopurine induce the formation of thylakoid membranes and grana structures in plastids of the green cultures. For membrane formation in plastids of white cultures, light in combination with BA is required. Light and benzylaminopurine also influence the plastid DNA gene expression. By in-organello protein synthesis with isolated plastids we show that light as well as benzylaminopurine affects the synthesis of plastid DNA encoded proteins. A characteristic effect of benzylaminopurine on plastids from white and green cultures is the reduction in the synthesis of the CFI subunits of 55,000 and 57,000 D, and the reduction in the synthesis of large polypeptides with a molecular weight higher than 67,000 D. In contrast to benzylaminopurine, light only affects the DNA gene expression of plastids from white cell cultures, that are in a very early stage of plastid development. Light stimulates the synthesis of polypeptides with a molecular weight of 84,000, 70,000 and 46,000 D which are encoded by cpDNA in these white culture plastids. In green cell cultures both plastids with a etioplast-like phenotype and with a chloroplast like morphology synthesize similar polypeptides, resulting in the same polypeptide pattern. Our results indicate that qualitative differences in plastid DNA gene expression as an effect of light do occur but only in plastids at very early stages of chloroplast development. We observe a gradual reduction in the number of high molecular weight polypeptides at later stages of chloroplast development. This suggests that these large polypeptides are characteristic for plastids at an early developmental stage.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00435212
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  • 6
    Electronic Resource
    Electronic Resource
    Isolation and physicochemical characterization of mitochondrial DNA from cultured cells ofPetunia hybrida (1985)
    Springer
    Theoretical and applied genetics 69 (1985), S. 223-233 
    Details
    ISSN: 1432-2242
    Keywords: P. hybrida ; mtDNA ; Cell cultures
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Mitochondrial DNA ofPetunia hybrida was purified from cell suspension cultures. Up to 50% of the DNA could be isolated as supercoiled DNA molecules by CsCl-ethidium bromide density gradient centrifugation. The DNA purified from DNase-treated mitochondria bands at a single buoyant density of 1.760 gcm−3 in neutral density gradients and runs on agarose gels as a single band with an apparent molecular weight exceeding 30 megadaltons (Md). Summing of the restriction endonuclease fragment lengths indicates a mitochondrial genome size of at least 190 Md. Electron microscopic analysis reveals the presence of a heterogeneous population of circular DNA molecules, up to 60 Md in size. Small circular DNA molecules, ranging in size from 2–30 Md are present, but unlike in cultured cells of other plant species they do not form discrete size classes and furthermore, they constitute less than 5% of the total DNA content of the mitochondria. The restriction endonuclease patterns of mitochondrial DNA do not qualitatively alter upon prolonged culture periods (up to at least two years).
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00662429
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  • 7
    Electronic Resource
    Electronic Resource
    Transcriptional and post-transcriptional regulation of chloroplast gene expression in Petunia hybrida (1986)
    Springer
    Theoretical and applied genetics 73 (1986), S. 94-101 
    Details
    ISSN: 1432-2242
    Keywords: Petunia hybrida ; rbc L ; psa A ; Chloroplast biogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary To study the control of differential gene expression during plastid biogenesis in Petunia hybrida, we have investigated the in vivo translation and transcription of the rbc L gene, coding for the large subunit of ribulose bisphosphate carboxylase (LSU), and the psa A gene, coding for P700 chlorophyll-a apoprotein (AP700). Differential expression of these plastid-encoded genes was studied in two developmentally different plastid systems, proplastid-like organelles from the green cell suspension AK2401 and mature chloroplasts from green leaves. In vivo translation of rbc L and psa A transcripts was analysed using specific antibodies. Specific transcript levels were analysed using internal fragments of the rbc L and psa A genes. A standardization procedure was used so that a direct correlation could be made between the amount of products and gene copy number. In Petunia hybrida the amount of LSU polypeptides present in both plastid types does not correspond to the amount of specific mRNA for the gene. Although the rbc L transcripts are present in both plastid types, the LSU protein is only present in green leaf plastids and not in cell culture plastids. In vitro translation of isolated rbc L transcripts give similar results, thereby suggesting that differences in the primary structure of the transcripts are responsible for the observed discrepancy. In contrast to this, the amount of AP700 polypeptides does correspond to the amount of the psa A transcripts. Therefore, our results indicate that the expression of chloroplast genes during plastid biogenesis takes place on at least two different levels: expression of the rbc L gene is regulated post-transcriptionally while expression of the psa A gene is regulated at the transcriptional level.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00273725
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  • 8
    Electronic Resource
    Electronic Resource
    A comparison of shoot regeneration from protoplasts and leaf discs of different genotypes of the cultivated tomato (1987)
    Springer
    Theoretical and applied genetics 75 (1987), S. 105-108 
    Details
    ISSN: 1432-2242
    Keywords: Tomato ; Protoplasts ; Leaf disc ; Shoot regeneration ; Inheritance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Shoot regeneration from leaf discs and leaf mesophyll protoplasts of 11 genotypes of Lycopersicon esculentum (the cultivated tomato), were compared. In both regeneration procedures genotypic differences were observed between inbred lines, and also between F1 hybrids and their parental lines. In the tested hybrid genotypes no heterosis effect with respect to shoot regeneration capacity was observed. A correlation between shoot regeneration from leaf discs and from leaf mesophyll protoplasts was apparent in the tested genotypes. This suggests that using the described procedure, shoot regeneration from leaf discs can be usef for rapid pre-screening for regeneration capacity from protoplasts of tomato genotypes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00249149
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  • 9
    Electronic Resource
    Electronic Resource
    An effective chemical mutagenesis procedure for Petunia hybrida cell suspension cultures (1979)
    Springer
    Theoretical and applied genetics 55 (1979), S. 101-106 
    Details
    ISSN: 1432-2242
    Keywords: Mutagenesis ; Petunia hybrida cell lines ; Drug resistant mutants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary By using mercury(II)-chloride (HgCl2) and Dl-6-fluorotryptophan (6FT) as positive selection conditions we were able to show that N-methyl-N′-nitro-N-nitrosoguanidine (NG) is an effective mutagen for Petunia hybrida suspension cells. A number of the 205 calli resistant to HgCl2 and 17 calli resistant to 6FT were isolated. The highest mutation frequency was 1.0 x 10−5 and 2.0 x 10−6 for HgCl2 and 6FT, respectively. A preliminary characterization of the mutants is presented. A significant increase in the number of drug-resistant calli was only obtained at NG-concentrations (5–40 μg/ml) that had no observable effect on the survival of the mutagenized cultures.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00295434
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  • 10
    Electronic Resource
    Electronic Resource
    Restriction endonuclease analysis of chloroplast DNA in interspecies somatic Hybrids of Petunia (1982)
    Springer
    Theoretical and applied genetics 62 (1982), S. 377-383 
    Details
    ISSN: 1432-2242
    Keywords: Somatic hybrid ; Petunia ; Chloroplast DNA ; Plastid segregation ; Restriction endonucleases
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Restriction endonuclease cleavage pattern analysis of chloroplast DNA (cpDNA) of three different interspecific somatic hybrid plants revealed that the cytoplasms of the hybrids contained only cpDNA of P. parodii. The somatic hybrid plants analysed were those between P. parodii (wild type) + P. hybrida (wild type); P. parodii (wild type)+P. inflata (cytoplasmic albino mutant); P. parodii (wild type) + P. parviflora (nuclear albino mutant). The presence of only P. parodii chloroplasts in the somatic hybrid of P. parodii + P. inflata is possibly due to the stringent selection used for somatic hybrid production. However, in the case of the two other somatic hybrids P. parodii + P. hybrida and P. parodii + P. parviflora it was not possible to determine whether the presence of only P. parodii chloroplasts in these somatic hybrid plants was due to the nature of the selection schemes used or simply occurred by chance. The relevance of such somatic hybrid material for the study of genomic-cytoplasmic interaction is discussed, as well as the use of restriction endonuclease fragment patterns for the analysis of taxonomic and evolutionary inter-relationships in the genus Petunia.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00275109
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