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  • 1
    Digitale Medien
    Digitale Medien
    Cloning, expression and characterization of a gene which encodes a topoisomerase I with positive supercoiling activity in pea (1998)
    Springer
    Plant molecular biology 37 (1998), S. 773-784 
    Details
    ISSN: 1573-5028
    Schlagwort(e): heterologous expression ; introduction of positive supercoils in DNA ; Pisum sativum ; RACE-PCR ; relaxation of DNA supercoils
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract We have isolated and sequenced the full length cDNA for topoisomerase I. Using degenerate primers, based on the conserved amino acid sequences of five eukaryotic topoisomerase I, a 386 bp fragment was PCR amplified using pea cDNA as template. This fragment was used as a probe to screen a pea cDNA library. Two partial cDNA clones were isolated which were truncated at the 5′ end. RACE-PCR was employed to isolate the remaining portion of the gene. The total size of the gene was 3055 bp with an open reading frame of 2676 bp. The deduced structure of pea topoisomerase I contain 892 amino acids with a calculated molecular weight of 100 kDa and an estimated pI of 9.3. A comparison of the deduced amino acid sequences of the pea topo I with the other eukaryotic topoisomerases clearly suggested that they are all related. Pea topoisomerase I has been overexpressed in E. coli system and the recombinant topoisomerase purified to homogeneity. The purified protein relaxes both positive and negative supercoiled DNA in the absence of divalent cation Mg2+. In the presence of Mg2+ ions the purified enzyme introduces positive supercoils a unique property not reported in any other organism except in archaebacterial topoisomerase I. Polyclonal antibodies were raised against recombinant topoisomerase I and western blotting with sub-cellular fractions indicated the localization of this topoisomerase in pea nuclei.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1023/A:1006086311875
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  • 2
    Digitale Medien
    Digitale Medien
    Cloning and characterization of a cDNA encoding topoisomerase II in pea and analysis of its expression in relation to cell proliferation (1999)
    Springer
    Plant molecular biology 41 (1999), S. 125-137 
    Details
    ISSN: 1573-5028
    Schlagwort(e): cell proliferation ; in vitro transcription and translation ; Pisum sativum ; plant promoter ; primer extension ; RACE-PCR ; transcript analysis
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract We have isolated and sequenced four overlapping cDNA clones to identify the full-length cDNA for topoisomerase II (PsTopII) from pea. Using degenerate primers, based on the conserved amino acid sequences of other eukaryotic type II topoisomerases, a 680 bp fragment was PCR-amplified with pea cDNA as template. This fragment was used as a probe to screen an oligo-dT-primed pea cDNA library. A partial cDNA clone was isolated that was truncated at the 3′ end. RACE-PCR was employed to isolate the remaining portion of the gene. The total size of PsTopII is 4639 bp with an open reading frame of 4392 bp. The deduced amino acid sequence shows a strong homology to other eukaryotic topoisomerase II (topo II) at the N-terminus end. The topo II transcript was abundant in proliferative tissues. We also show that the level of topo II transcripts could be stimulated by exogenous application of growth factors that induced proliferation in vitro cultures. Light irradiation to etiolated tissue strongly stimulated the expression of topo II. These results suggest that topo II gene expression is up-regulated in response to light and hormones and correlates with cell proliferation. Besides, we have also isolated and analysed the 5′-flanking region of the pea TopII gene. This is first report on the isolation of a putative promoter for topoisomerase II from plants.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1023/A:1006352820788
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  • 3
    Digitale Medien
    Digitale Medien
    Effect of mycorrhizal inoculation and soluble phosphorus fertilizer on growth and phosphorus uptake of pearl millet (1984)
    Springer
    Plant and soil 81 (1984), S. 247-256 
    Details
    ISSN: 1573-5036
    Schlagwort(e): Pearl millet ; Pennisetum americanum ; Leeke ; Phosphorus response ; VA mycorrhiza
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Summary Six mycorrhizal fungi were tested as inoculants for pearl millet (Pennisetum americanum Leeke) grown in pots maintained in a greenhouse. VAM fungi varied in their ability to stimulate plant growth and phosphorus uptake. Inoculation withGigaspora margarita, G. calospora andGlomus fasciculatum increased shoot drymatter 1.3 fold over uninoculated control. In another pot trial, inoculation withGigaspora calospora andGlomus fasciculatum resulted in dry matter and phosphorus uptake equivalent to that produced by adding phosphorus at 8 kg/ha. The influence of inoculatingGigaspora calospora on pearl millet at different levels of phosphorus fertilizer (0 to 60 kg P/ha) as triple superphosphate in sterile and unsterile alfisol soil was also studied. In sterile soil, mycorrhizal inoculation increased dry matter and phosphorus uptake at levels less than 20 kg/ha. At higher P levels the mycorrhizal effect was decreased. These studies performed in sterilized soil suggest that inoculation of pearl millet with efficient VAM fungi could be extremely useful in P deficient soils. However, its practical utility depends on screening and isolation of fungal strains which perform efficiently in natural (unsterilized) field conditions.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF02197158
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  • 4
    Digitale Medien
    Digitale Medien
    Genotype dependent variation in mycorrhizal colonization and response to inoculation of pearl millet (1985)
    Springer
    Plant and soil 86 (1985), S. 113-125 
    Details
    ISSN: 1573-5036
    Schlagwort(e): Genetic crosses ; Genotypes ; Mycorrhizal colonization ; Pearl millet ; Plant effects ; Response
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Summary Genotypes of pearl millet (Pennisetum americanum L. Leeke) were examined for differences in vesicular-arbuscular mycorrhizal (VAM) colonization and response to inoculation. For thirty genotypes tested across three field locations there was a range of mycorrhizal colonization intensity between 25 and 56%. In another experiment with two male-sterile lines, restorer lines and their derived crosses, grown in pots filled with non-sterilized soil there were significant differences between genotypes for colonization by mycorrhiza. This showed hostgenotype dependence for mycorrhizal colonization. Root growth rates, mycorrhizal root length, percentage root colonization and plant growth and P uptake were studied in ten genotypes. A set of 3 genotypes with similar root lengths varied significantly with regard to mycorrhizal root length and the percentage colonization. This supports the suggestion that VAM colonization and spread is dependent on the host genotype. The growth responses differed significantly between the genotypes and they also differed in their responses to P uptake and VAM inoculation. The utility of host-genotype dependent differences in VAM symbiosis in plant breeding is discussed.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF02185031
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