Marianne C Verberne

Leiden University, Leiden, South Holland, Netherlands

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Publications (18)95.55 Total impact

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    ABSTRACT: In Catharanthus roseus cell cultures, the monoterpenoid pathway has been shown to be a limiting factor in terpenoid indole alkaloid (TIA) production. This could be due to competition at the level of isopentenyl diphosphate::dimethylallyl diphosphate (C5) which leads to the biosynthesis of different terpenoid groups. For future engineering of the terpenoid pathway, chemical characterization of C. roseus cell cultures is a necessity. Therefore, in this study nine C. roseus cell suspension lines were characterized by analyzing the levels of the major terpenoids derived from different biosynthetic pathways which may compete for the same precursors; TIA (monoterpenoid, C10), carotenoids (tetraterpenoid, C40), and sterols (triterpenoid, C30). Among the cell lines, CRPP (S) was the most promising TIA-producing cell line which provided more TIA [24 μmol g−1 dry weight (DW)] than carotenoids (15 μmol g−1 DW) and sterols (2 μmol g−1 DW). However, when considering the distribution of the isopentenyl-precursor (C5), the carotenoids which assemble from 8× C5 represent twofold more C5-units (122 μmol g−1 DW) than the TIA in this cell line. In the CRPP (G), A12A2 (G), and A12A2 (S) cell lines, the C5 distribution was predominant toward carotenoid biosynthesis as well, resulting in a relatively high accumulation of carotenoids. The geranylgeranyl diphosphate (C20) pathway toward carotenoid production is therefore considered competitive toward TIA biosynthesis. For channeling more precursors to the TIA, the branch point for C10 and C20 seems an interesting target for metabolic engineering. Using principal component analysis of the chromatographic data, we characterized the cell lines chemically based on their metabolite levels. The information on the metabolic composition of C. roseus cell cultures is useful for developing strategies to engineer the metabolic pathways and for selection of cell lines for future studies.
    Plant Cell Tissue and Organ Culture 05/2014; 117(2):225-239. DOI:10.1007/s11240-014-0435-2 · 2.61 Impact Factor
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    Plant physiology 06/2011; 156(4):1701-5. DOI:10.1104/pp.111.178426 · 7.39 Impact Factor
  • Vincentius A. Halim, Marianne C. Verberne, Robert Verpoorte
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    ABSTRACT: The metabolite profiles of wild type tobacco plants and transgenic tobacco plants, constitutively producing salicylic acid (SA), were compared by means of liquid chromatography with online photo diode array and mass spectrometer detector (LC–PDA–MS). Statistical analysis of the peak area of 15 major peaks showed significant differences in quantities for three compounds, which are present in the extract before acid hydrolysis, containing mainly aglucones. By comparison of the retention time, UV spectra, and molecular weight with reference compounds, two of these peaks were identified as chlorogenic acid and rutin. The extracts obtained after acidic hydrolysis did not show any significant difference.
    Journal of Liquid Chromatography &amp Related Technologies 02/2007; 26(3):369-383. DOI:10.1081/JLC-120017176 · 0.64 Impact Factor
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    ABSTRACT: Phylloquinone (Vitamin K(1)) is an essential component of the photosynthetic electron transfer. As isochorismate is required for the biosynthesis of Vitamin K(1), isochorismate synthase (ICS) activity is expected to be present in all green plants. In bacteria salicylic acid (SA) is synthesized via a two step pathway involving ICS and isochorismate pyruvate lyase (IPL). The effect of the introduction in tobacco plants of the bacterial ICS and IPL genes on the endogenous isochorismate pathway was investigated. Transgenic tobacco plants in which IPL was targeted to the chloroplast suffered severe growth retardation and had low Vitamin K(1) content. Probably because isochorismate was channeled towards SA production, the plants were no longer able to produce normal levels of Vitamin K(1). Transgenic tobacco plants in which the bacterial ICS was present in the chloroplast showed higher Vitamin K(1) contents than wild type plants.
    Journal of Biotechnology 02/2007; 128(1):72-9. DOI:10.1016/j.jbiotec.2006.09.005 · 2.88 Impact Factor
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    ABSTRACT: A polyketide synthase has been suggested to play an important role in cannabinoid biosynthesis in Cannabis sativa L. This enzyme catalyzes the biosynthesis of olivetolic acid, one of the precursors for cannabinoid biosynthesis. Using a reverse transcriptase-polymerase chain reaction (RT-PCR) based on the DNA homology of chalcone synthase (EC 2.3.1.156) and valerophenone synthase (EC 2.3.1.156) of hop (Humulus lupulus), a cDNA encoding a polyketide synthase in C. sativa was identified. The coding region of the gene is 1170 bp long encoding a 389 amino acid protein of a predicted 42.7 kDa molecular mass and with a pI of 6.04. The gene shares a high homology with a chalcone synthase gene of H. lupulus, 85% and 94% homology on the level of DNA and protein, respectively. Over-expression of the construct in Escherichia coli M15 resulted in a 45 kDa protein. The protein has chalcone synthase activity as well as valerophenone synthase activity, a chalcone synthase-like activity. Using n-hexanoyl-CoA and malonyl-CoA as substrates did not give olivetol or olivetolic acid as a product.
    Plant Physiology and Biochemistry 05/2004; 42(4):291-7. DOI:10.1016/j.plaphy.2004.02.011 · 2.35 Impact Factor
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    ABSTRACT: The metabolomic analysis of wild type and constitutive salicylic acid producing tobacco plants (CSA tobacco, Nicotiana tabacum 'Samsun' NN) plants overexpressing salicylate biosynthetic genes was carried out by 1H NMR spectrometry and multivariate analysis techniques. The principle component analysis (PCA) of the 1H NMR spectra showed a clear discrimination between those samples by PC1 and PC2. The discrimination of non-inoculated, TMV-virus inoculated, and systemic leaves or veins could also be obtained by PCA analysis. Major peaks in 1H NMR spectra contributing to the discrimination were assigned as those of chlorogenic acid, malic acid, and sugars. This method allows an efficient differentiation between wild type and transgenic plants without any pre-purification steps.
    Phytochemistry 05/2004; 65(7):857-64. DOI:10.1016/j.phytochem.2004.01.019 · 3.35 Impact Factor
  • Marianne C Verberne, Jan Hoekstra, John F Bol, Huub J M Linthorst
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    ABSTRACT: The hypersensitive interaction between Tobacco mosaic virus (TMV) and tobacco results in accumulation of salicylic acid (SA), defense gene expression, and development of systemic acquired resistance (SAR) in uninfected leaves. The plant hormones SA and ethylene have been implicated in SAR. From a study with ethylene-insensitive (Tetr) tobacco, we concluded that ethylene perception is required to generate the systemic signal molecules in TMV-infected leaves that trigger SA accumulation, defense gene expression, and SAR development in uninfected leaves. Ethylene perception was not required for the responses of the plant to the systemic signal that leads to SAR development.
    The Plant Journal 08/2003; 35(1):27-32. · 6.82 Impact Factor
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    Marianne C. Verberne, Jan Hoekstra, John F. Bol, Huub J.M. Linthorst
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    ABSTRACT: The hypersensitive interaction between Tobacco mosaic virus (TMV) and tobacco results in accumulation of salicylic acid (SA), defense gene expression, and development of systemic acquired resistance (SAR) in uninfected leaves. The plant hormones SA and ethylene have been implicated in SAR. From a study with ethylene-insensitive (Tetr) tobacco, we concluded that ethylene perception is required to generate the systemic signal molecules in TMV-infected leaves that trigger SA accumulation, defense gene expression, and SAR development in uninfected leaves. Ethylene perception was not required for the responses of the plant to the systemic signal that leads to SAR development.
    The Plant Journal 06/2003; 35(1):27 - 32. DOI:10.1046/j.1365-313X.2003.01778.x · 6.82 Impact Factor
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    ABSTRACT: In higher plants, there are two pathways leading to the formation of isoprenoids: the mevalonate pathway and the recently discovered methyl-d-erythritol 4-phosphate (MEP) pathway. It was shown that the isoprenoid moiety, constituting ring C of anthraquinones (AQs) in the plant family Rubiaceae, is formed from the MEP pathway. 1-Deoxy-d-xylulose 5-phosphate synthase (DXS), catalyzing the first step of the MEP pathway, is suggested to be a regulatory enzyme. In order to evaluate the potential regulatory role of the DXS in AQ biosynthesis, we identified a full-length cDNA encoding DXS from Morinda citrifolia, a member of the Rubiaceae family, by a PCR strategy. The cDNA encodes a protein of 722 amino acids with a calculated molecular mass of 78 kDa. The deduced amino acid sequence shares high homology with amino acid sequences of DXSs from other higher plants. Southern blot indicated the existence of a small family of DXS genes in M. citrifolia. Northern blot analysis showed that the DXS transcript levels were correlated with AQ accumulation, suggesting that AQ biosynthesis is regulated at the transcriptional level of the DXS gene.
    Plant Science 06/2003; 164(6-164):911-917. DOI:10.1016/S0168-9452(02)00362-X · 4.11 Impact Factor
  • Laurentius H Nugroho, Marianne C Verberne, Robert Verpoorte
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    ABSTRACT: Chorismate mutase (CM, EC 5.4.99.5), phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) and chalcone synthase (CHS, EC 2.3.1.74) activities were studied in constitutive salicylic acid-producing (CSA) tobacco plants in relation to the accumulation of flavonoids and chlorogenic acid. The CM, PAL and CHS activities in CSA-tobacco (Nicotiana tabacum cv. Samsun NN) plants were lower than in non-transgenic tobacco plants. Flavonoid and chlorogenic acid accumulation was suppressed in CSA-tobacco plants compared to those of non-transgenic tobacco plants.
    Plant Physiology and Biochemistry 09/2002; 40(9):755-760. DOI:10.1016/S0981-9428(02)01437-7 · 2.35 Impact Factor
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    ABSTRACT: The accumulation of sesquiterpenoid phytoalexins was screened in constitutive salicylic acid producing (CSA)-tobacco leaves. Only after tobacco mosaic virus (TMV)-inoculation the sesquiterpenoid phytoalexin capsidiol was identified in CSA- and non-transgenic tobacco leaves. Capsidiol was not accumulated in CSA- and non-transgenic tobacco leaves after mechanical wounding.
    Plant Science 06/2002; 162(6):989-993. DOI:10.1016/S0168-9452(02)00049-3 · 4.11 Impact Factor
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    ABSTRACT: Large variations in nicotine and related alkaloids accumulations were found in the leaves of constitutive salicylic acid producing (CSA)-tobacco and non transgenic tobacco plants. Statistical analysis showed that there was no significant correlation between salicylic acid (SA) levels and alkaloid accumulation in CSA-tobacco plants. The accumulation of nicotine after wounding and tobacco mosaic virus (TMV)-inoculation of leaves were measured as well. The accumulation of nicotine in the leaves of CSA-tobacco was suppressed after TMV-inoculation, whereas wounding enhanced nicotine accumulation. There was no effect of TMV-inoculation on nicotine accumulation in non-transgenic tobacco but wounding also increased nicotine accumulation.
    Plant Science 04/2002; 162(4-162):575-581. DOI:10.1016/S0168-9452(01)00596-9 · 4.11 Impact Factor
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    ABSTRACT: Salicylic acid (SA) is a signalling compound in plants which is able to induce systemic acquired resistance. In the analysis of SA in plant tissues, the extraction recovery is often very low and variable. This is mainly caused by sublimation of SA, especially during evaporation of organic solvents. Techniques have been designed in order to overcome this problem. In the first part of the extraction procedure, sublimation of SA was prevented by addition of 0.2 M sodium hydroxide. At a later stage of the extraction procedure, sublimation of SA during solvent evaporation was controlled by the addition of a small amount of HPLC eluent. In this way, recoveries in the range of 71-91% for free SA and 65-79% for acid-hydrolysed SA were obtained. Recoveries could be further optimised by the use of an internal standard to correct for volume changes after the addition of the HPLC eluent.
    Phytochemical Analysis 01/2002; 13(1):45-50. DOI:10.1002/pca.615 · 2.45 Impact Factor
  • Laurentius H Nugroho, Marianne C Verberne, Robert Verpoorte
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    ABSTRACT: Salicylic acid production in roots, stems, young leaves, medium leaves and old leaves of constitutive salicylic acid producing (CSA) tobacco plants was tested. In protein extracts of CSA-tobacco plants, the formation of isochorismate and of salicylic acid (SA) were measured to determine isochorismate synthase (ICS) and isochorismate pyruvate lyase (IPL) activity. The highest production of isochorismate and SA in vitro were produced by protein extracts of the young leaves of CSA-tobacco plants. The isochorismate and SA productions in vitro by protein extracts and the accumulation of SA and SA-glucoside in CSA-tobacco plants varied among the tested lines and within one line. The accumulation of SA in vivo is determined by the activity of IPL.
    Plant Science 10/2001; 161(5):911-915. DOI:10.1016/S0168-9452(01)00486-1 · 4.11 Impact Factor
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    ABSTRACT: Cell suspensions of Catharanthus roseus produce 2,3-dihydroxybenzoic acid (2,3-DHBA) after elicitation with fungal elicitor. Together with the 2,3-DHBA production there is a strong increase in isochorismate synthase (ICS; EC 5.4.99.6) activity, an enzyme which converts chorismate into isochorismate. Bacteria also produce 2,3-DHBA which is an intermediate for the biosynthesis of siderophores. The biosynthesis pathway of 2,3-DHBA in bacteria proceeds via ICS. To investigate whether C. roseus cells use the same pathway to produce 2,3-DHBA, cells were transformed with constructs containing a C. roseus cDNA clone of ics in sense or antisense orientation. Although the transformation with ics in sense orientation resulted in constitutive ICS activity, no accumulation of 2,3-DHBA was observed. Transformation of ics in antisense orientation was not sufficient to block the ICS production. The biosynthesis of 2,3-DHBA in C. roseus probably involves a multi-enzyme pathway, which cannot simply be modulated by over-expression of ics.
    Plant Physiology and Biochemistry 07/2001; 39(7):595-602. DOI:10.1016/S0981-9428(01)01279-7 · 2.35 Impact Factor
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    ABSTRACT: After a hypersensitive response to invading pathogens, plants show elevated accumulation of salicylic acid (SA), induced expression of plant defense genes, and systemic acquired resistance (SAR) to further infection by a broad range of pathogens. There is compelling evidence that SA plays a crucial role in triggering SAR. We have transformed tobacco with two bacterial genes coding for enzymes that convert chorismate into SA by a two-step process. When the two enzymes were targeted to the chloroplasts, the transgenic (CSA, constitutive SA biosynthesis) plants showed a 500- to 1,000-fold increased accumulation of SA and SA glucoside compared to control plants. Defense genes, particularly those encoding acidic pathogenesis-related (PR) proteins, were constitutively expressed in CSA plants. This expression did not affect the plant phenotype, but the CSA plants showed a resistance to viral and fungal infection resembling SAR in nontransgenic plants.
    Nature Biotechnology 08/2000; 18(7):779-83. DOI:10.1038/77347 · 39.08 Impact Factor
  • Marianne C. Verberne, Retno A. Budi Muljono, Robert Verpoorte