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  • 1
    Electronic Resource
    Electronic Resource
    Analysis of gene regulation in growing pollen tubes of angiosperm and gymnosperm species using microprojectile bombardment (1995)
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 93 (1995), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A microprojectile based transient expression assay was used to investigate the functional conservation of gene regulatory mechanisms in the male gametophytes of an angiosperm (Nicotiana tabacum) and two gymnospermous (Picea abies and Pinus pinaster) species. The activities of two angiosperm gene promoters, which have previously shown to be either preferentially expressed in the male gametophyte (lat52) or highly expressed in both the sporophyte and male gametophyte (ActI), were analysed. The results showed that in P. abies and P. pinaster, activity of the Act1 promoter was significantly higher than the activity of the lat52 promoter, while the converse was observed in N. tabacum. Detailed analysis of lat52 5’promoter deletions demonstrated that although the minimal -67 bp lat52 core promoter was active at low levels in all three species, upstream regulatory elements conserved among several pollen-expressed genes, including the PBI element, were not functional in P. abies and P. pinaster. These results suggest that both taxa-specific and conserved regulatory mechanisms operate to control gene expression during pollen germination and tube growth.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1399-3054.1995.tb06841.x
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  • 2
    Electronic Resource
    Electronic Resource
    Optimization of transient gene expression in pollen of Norway spruce (Picea abies) by particle acceleration (1994)
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 92 (1994), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: In order to optimize transient gene expression in Norway spruce pollen after DNA delivery with particle bombardment, effects of different conditions during homhardmenl were analysed using β-glucuroniduse (GUS) driven by the rice Act I promoter and Inciferase (LUS) driven by the tomato !at52 promoter as reporter genes. Transient gene expression was significantly increased hy using two bombardments. Also the distance from the stopping plate to the sample was critical to gam maximum gene expression. There was no significant difference between gold and tungsten particles, and the number of positively stained pollen increased with increasing DNA concentration, from 5 to 40 pg DNA added in the DNA/tungsten solution The DNA delivery to Norway spruce pollen was most efficient at a chamber pressure above 70 kPa.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1399-3054.1994.tb08829.x
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  • 3
    Electronic Resource
    Electronic Resource
    The translationally repressed pollen-specific ntp303 mRNA is stored in non-polysomal mRNPs during pollen maturation (2000)
    Springer
    Sexual plant reproduction 13 (2000), S. 135-144 
    Details
    ISSN: 1432-2145
    Keywords: Keywords Pollen development ; Pollen-specific ntp303 gene ; Stored mRNA ; Subcellular distribution ; Translational regulation ; Translational repression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  A tobacco pollen tube glycoprotein, p69 is encoded by the pollen-specific gene ntp303 that is transcribed during pollen development and pollen tube growth, but it is abundantly translated only after pollen germination. To investigate the translational repression of ntp303 mRNA during pollen development the compartmentation of ntp303 mRNA was examined and compared against another transcript (ntp52), which is efficiently translated during pollen maturation. Three subcellular fractions were isolated: a post-polysomal fraction enriched with messenger ribonucleoprotein particles, a polysomal fraction and a novel fraction of EDTA/ puromycin-resistant particles co-sedimentating with polysomes (EPP). At all developmental stages studied, ntp303 mRNA was found to be present in all fractions. Surprisingly, most of the translationally inactive ntp303 mRNA was localised in the polysomal fraction and EPPs, whereas ntp52 mRNA was distributed between the post-polysomal fraction and polysomes but was virtually undetectable in EPPs. This differential mRNA distribution pattern may help to explain the developmentally regulated translational repression of the ntp303 gene during pollen maturation, highlighting a potential role of EPPs. A model of how this differential mRNA compartmentation pattern regulates ntp303 mRNA translation is proposed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004970000047
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  • 4
    Electronic Resource
    Electronic Resource
    Activation and developmental regulation of an Arabidopsis anther-specific promoter in microspores and pollen of Nicotiana tabacum (1993)
    Springer
    Sexual plant reproduction 6 (1993), S. 217-224 
    Details
    ISSN: 1432-2145
    Keywords: Microspore development ; Pollen ; Transcriptional regulation ; Transient expression ; Microprojectile bombardment ; β-Glucuronidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Seven cytologically distinct stages during micro spore development were identified and used to define the activation of an Arabidopsis anther-specific gene (apg) in transgenic tobacco plants containing an apg promoter-gus fusion. Histochemical analysis of GUS activity showed that the apg promoter was activated in miduninucleate microspores prior to equatorial and polar nuclear migration. Quantitative analysis in isolated spores showed that GUS activity per milligram of protein decreased progressively during pollen development, but on a per spore basis showed a second peak of activity in mid-bicellular pollen. Activation of the apg promoter in isolated gametophytic cells during development was also investigated following DNA delivery by microprojectile bombardment. Levels of transient expression were detectable in uninucleate microspores, but peaked in mid-bicellular pollen, in contrast to the progressive increase in activity of the promoter of the ‘late’ pollen gene lat52. These data show that the apg promoter is activated in a biphasic pattern, and indicate that the activity of transcription factors which mediate apg promoter activity persist through microspore mitosis, but decrease during pollen maturation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00231898
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  • 5
    Electronic Resource
    Electronic Resource
    Isolation and expression of an anther-specific gene from tomato (1989)
    Springer
    Molecular genetics and genomics 217 (1989), S. 240-245 
    Details
    ISSN: 1617-4623
    Keywords: Lycopersicon esculentum ; Anther ; Microsporogenesis ; Pollen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have isolated and sequenced an anther-specific cDNA clone and a corresponding genomic clone from tomato. The gene (LAT52) encodes an 800-nucleotide-long transcript that is detectable in pollen, anthers and at 20-to 50-fold lower levels in petals. LAT52 mRNA is not detectable in pistils, sepals or non-reproductive tissues. Steady-state levels of LAT52 mRNA are detectable in immature anthers containing pollen at the tetrad stage and increase progressively throughout microsporogenesis until anthesis (pollen shed). The LAT52 gene contains 5′ and 3′ untranslated regions of 110 and approximately 150 nucleotides, respectively, and a single intron with a highly repetitive sequence. A TATA box motif is located 28 nucleotides upstream of the transcription start site. The gene encodes a putative protein of 18 kDa that is cysteine rich and has an N-terminal hydrophobic region with characteristics similar to eucaryotic secretory signal sequences. LAT52 is a single or low copy gene in tomato and shares homology with sequences in tobacco.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02464887
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  • 6
    Electronic Resource
    Electronic Resource
    Structural diversity of the patatin gene family in potato cv. Desiree (1988)
    Springer
    Molecular genetics and genomics 212 (1988), S. 325-336 
    Details
    ISSN: 1617-4623
    Keywords: Patatin ; Solanum tuberosum ; Gynogenesis ; Expression signals
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have used a combined genetical and molecular approach to study the structural diversity of the patatin gene family in tetraploid Solanum tuberosum L. cv. Desiree (2n=4x=48). Nine dihaploid derivatives (2n=2x=24) of cv. Desiree were isolated by gynogenesis through prickle pollination with S. phureja Juz et Buk. Patatin DNA sequences in Desiree and in the dihaploids were examined by probing Southern blots of restriction endonucleases Hin-dIII and XbaI digested DNA with patatin cDNA region-specific gene probes and by more detailed examination (restriction endonuclease mapping and partial DNA sequencing) of 10 patatin genomic clones in bacteriophage λ replacement vector EMBL4. This provided positive identification for most individual patatin gene family members and some estimate of their organisation and diversity. Most of the 64–72 patatin DNA copies showed little allelic variation based on HindIII and XbaI restriction fragment length polymorphism (RFLP) mapping and did not appear to be very tightly clustered. Four of the 6–8 class I patatin genes (without a characteristic 22 bp insert in their untranslated leader DNA), showed apparent allelic homogeneity, whilst the remaining class I genes comigrated with a single class II patatin gene RFLP subclass. Of the isolated clones, 4 contained apparent pseudogenes lacking 5′ control sequences and exon-1 DNA while another clone contained a patatin gene truncated at the 5′ region due to the cloning event. The remaining 5 all contained class II genes (with the 22 bp insert) and these showed varying degrees of sequence homology for 400 bp of conserved 5′ coding and non-coding DNA (from 77%–95%). In one case the extent of homology differed, with complete sequence divergence upstream of position-80 from the start of transcription. The structural diversity of the patatin gene family is discussed in relation to expression of individual patatin genes and the use of cv. Desiree as a host for potato transformation experiments.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00334703
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  • 7
    Electronic Resource
    Electronic Resource
    The 5′ flanking DNA of a patatin gene directs tuber specific expression of a chimaeric gene in potato (1987)
    Springer
    Plant molecular biology 9 (1987), S. 345-375 
    Details
    ISSN: 1573-5028
    Keywords: Patatin ; Solanum tuberosum L. ; Agrobacterium ; chimaeric gene ; chloramphenicol acetyltransferase (CAT)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A member of the patatin multigene family which encodes the major soluble tuber protein was isolated from potato cultivar Désirée. Analysis by strategic nucleotide sequencing demonstrated close homology to analogous regions of previously isolated patatin genomic clones from different cultivars. A 3.8-kb fragment containing the promoter and 5′ flanking DNA of the patatin gene was used to construct a transcriptional fusion gene with the coding DNA of the bacterial chloramphenicol acetyltransferase (CAT) gene and the polyadenylation/termination sequences of the nopaline synthase gene (nos). The chimaeric gene was reintroduced into potato cultivar Désirée by agrobacterial transformation of tissue slices. Regenerated transformed plants showed expression of the chimaeric gene (as determined by CAT activity) in tubers, but not in leaves, stems or roots of in vitro grown plants. Independent transformants did not show substantial variation in the level of induced tuber-specific CAT activity. Thus, information contained within 3.8 kb of the 5′ flanking DNA of the patatin gene analysed is sufficient to direct tuber-specific expression, largely independent of position effects.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00014911
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  • 8
    Electronic Resource
    Electronic Resource
    Developmental regulation of RI TL-DNA gene expression in roots, shoots and tubers of transformed potato (Solanum tuberosum cv. Desiree) (1986)
    Springer
    Plant molecular biology 6 (1986), S. 321-330 
    Details
    ISSN: 1573-5028
    Keywords: Agrobacterium rhizogenes ; Solanum tuberosum ; T-DNA ; gene expression ; tuberisation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Expression of TL-DNA from Agrobacterium rhizogenes plasmid pRi 1855 was examined in a transformed derivative of Solanum tuberosum cv. Desiree, D9X8a. Northern blot analysis identified at least nine TL-DNA coded transcripts in roots, shoots and tubers but their relative abundance differed within and between organs. This revealed a distinctive pattern of organ specified differential expression. Grafting experiments showed that the abnormal shape of tubers of transformed potato was probably determined by TL-DNA products synthesised within the tuber and not by diffusable products synthesised in other parts of the plant. The abundance of at least one transcript, tr5, was probably determined by culture conditions. Implications for functions and control of expression of Ri TL-DNA genes are discussed. It is suggested that Ri TL-DNA provides a convenient and extensive set of model genes to study variation and stability of expression of linked foreign genes introduced into plants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00034939
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  • 9
    Electronic Resource
    Electronic Resource
    Pollen viability and transgene expression following storage in honey (1995)
    Springer
    Transgenic research 4 (1995), S. 226-231 
    Details
    ISSN: 1573-9368
    Keywords: Transgenic plants ; pollen gene expression ; β-glucuronidase (GUS) activity ; environmental impact of transgenics ; molecular ecology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transgenic plants of tobacco andArabidopsis that produce genetically marked pollen, expressing the reporter geneuidA (gusA), were generated to determine whether pollen proteins can be expressed and stable in honey, a potential route by which foreign proteins might enter the wider environment. Hydrated tobacco pollen was found to lose viability rapidly in honey, while pollen in the natural dehydrated form remained viable for at least several days and in some cases several weeks, as determined by FDA staining activity and germinability. Dehydrated pollen was found to be capable of transient foreign gene expression, following microprojectile bombardment, after incubation in honey for at least 120 h. PCR amplification of transgene sequences in pollen of transgenic plants revealed that pollen DNA can remain relatively intact after 7 weeks in honey. GUS enzyme activity analysis and SDS-PAGE of pollen proteins revealed that foreign and native pollen proteins are stable in pollen incubated in honey for at least 6 weeks. We conclude that pollen may represent an ecologically important vector for transgenic protein products.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01969115
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  • 10
    Electronic Resource
    Electronic Resource
    Current awareness - recent symposia and seminars (1992)
    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 7 (1992), S. 215-222 
    Details
    ISSN: 0884-3996
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bio.1170070308
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