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Proteomics of the effect of heavy metals: induced stress in transgenic tobacco plants, with different expression levels of trehalose-6-phosphate synthase

By 1984- Luis Filipe Pinheiro Domingues


Tese de mestrado. Biologia (Biologia Celular e Biotecnologia). Universidade de Lisboa, Faculdade de Ciências, 2012Presently, heavy metal contamination is an important pollution problem in world agriculture. The specific issues concerning heavy metal, in farming issues, are mainly due to their accumulation in the food chain with direct consequences to the consumer health. Cadmium (Cd), particularly, is one of the most challenging pollutants, due to its high water solubility, mobility, persistence and toxicity, even in low concentrations. Trehalose is a sugar that acts as an osmoprotector, and has the capacity to stabilize membranes and proteins during desiccation, heat and osmotic stress. Trehalose-6- phosphate synthase, one of the enzymes of the biosynthetic pathway has been the major target for genetic engineering, as it is considered to be the decisive factor in the trehalose biosynthesis. The aim of the present work is to study the effects of a Cd exposure of two transgenic Nicotiana tbacum lines (B5H and B1F), which had already shown enhanced tolerance to abiotic stress, in comparison to wild type. We used a proteomics approach, namely 2-DE-DIGE.and identification of differentially expressed proteins by mass spectrometry. In this work we could identify fifteen different proteins, such as carbonic anhydrase, RuBisCO and phosphoglycerate kinase, with differential expression. Results demonstrate that both B1F and B5H present an enhanced tolerance to cadmium induced stress, showing signals which denote that they may not be as affected as Wt plants by the concentration of cadmium to which they were subjected. The physiological significance of the expression profiles obtained is discussed. Genetic engineering with the trehalose-6-phosphate synthase gene seems therefore to be of use in obtaining transgenic plants of commercial use for possible growth in contaminated soils or used in phytoremediation.A contaminação dos solos por metais pesados é um importante problema ambiental. Um dos principais problemas da contaminação de solos agrícolas prende-se com a acumulação destes metais na cadeia alimentar com consequências directas, graves, na saúde do consumidor. O Cádmio (Cd), em particular, é um dos poluentes mais problemáticos, devido à sua elevada solubilidade em água, mobilidade, persistência e toxicidade, mesmo em baixas concentrações. A trealose é um açúcar que actua como osmoprotector e tem a capacidade de estabilizar membranas e proteínas durante stresses provocados por dissecação, temperatura, e stress osmótico. A Trealose-6-fosfato-sintase, uma das enzimas da via biossintética da trehalose tem sido o alvo principal para a engenharia genética, uma vez que e considerado o factor determinante na biossíntese de trealose. Com o presente trabalho pretende-se, utilizando uma abordagem proteómica, baseada em 2-DE-DIGE seguida de identificação de proteinas por espectrometria de massa, estudar os efeitos da uma exposição a Cd em duas linhas de Nicotiana tbacum (B5H e B1F), que já tinham demonstrado tolerância aumentada a stress abiótico induzido por metais pesados, em comparação com plantas Wild type. Neste trabalho foi possivel identificar 15 proteinas diferentes, tais como a anhidrase carbónica, a RuBisCO e a fosfoglicerato kinase, com expressão diferencial. Os resultados demonstram que tanto as plantas B1F como as B5H apresentam uma tolerância aumentada ao stresse induzido por cádmio., mostrando sinais que denotam que eles podem não ser tão afetados quanto plantas Wt pela concentração de cádmio a que foram submetidas. O significado fisiológico dos perfis de expressão obtidos é discutido. A engenharia genética com o gene da sintetase de trealose-6-fosfato parece, portanto, ser util na obtenção de plantas transgénicas de uso comercial para uma possivel plantação em solos contaminados ou utilizados em fitorremediação

Topics: Tabaco, Metais pesados, Transgénicos, Biotecnologia, Teses de mestrado - 2012
Year: 2012
OAI identifier: oai:repositorio.ul.pt:10451/8200
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    1. (2004). 2-D Electrophoresis.
    2. (2012). A critical role of plastidial glycolytic glyceraldehyde-3-phosphate dehydrogenase in the control of plant metabolism and development. doi
    3. (2011). Abiotic Stress In Plants – Mechanisms And Adaptations. In: doi
    4. (2012). Accumulation and translocation of heavy metals in soil and plants from fly ash contaminated area.
    5. Advancements in plant proteomics using quantitative mass spectrometry. doi
    6. (2012). An update of MALDI-TOF mass spectrometry in lipid research. doi
    7. (2012). Analysis of low molecular weight compounds by MALDI-FTICR-MS. doi
    8. (2011). Arabidopsis Thaliana Glyceraldehyde-3-Phosphate Dehydrogenase As An Oxidative Stress Sensor. doi
    9. (2012). Available at: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1913774&tool=pmcentrez&rende rtype=abstract. Accessed
    10. (2009). Avaliação da resposta ao stresse oxidativo induzido por cádmio e cobre em plantas de tabaco transformadas e não transformadas.
    11. Cadmium stress tolerance in crop plants: probing the role of sulfur. Plant signaling behavior. 2011;6(2):215–222. Available at: doi
    12. Cd-induced changes in leaf proteome of the hyperaccumulator plant Phytolacca americana. doi
    13. (2012). Comparative proteomic analysis of the short-term responses of rice roots and leaves to cadmium. doi
    14. (2012). Difference gel electrophoresis. doi
    15. (2010). Differential proteomics a study in Medicago truncatula somatic embryogenesis Differential proteomics a study in Medicago truncatula somatic embryogenesis. doi
    16. Expression of the yeast trehalose-6-phosphate synthase gene in transgenic tobacco plants: pleiotropic phenotypes include drought tolerance. doi
    17. (2012). Floridata: Nicotiana tabacum. Plant Encyclopedia.
    18. Heavy Metals Contamination Of Topsoil And Dispersion In The Vicinities Of Reclaimed Auto-Repair Workshops In doi
    19. (2012). Implications of metal accumulation mechanisms to phytoremediation. Environmental science and pollution research international. doi
    20. (2012). Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using doi
    21. In: The Arabidopsis Book.; doi
    22. Inhibition of trehalase activity enhances trehalose accumulation in transgenic plants. Plant physiology. 1997;113(1):181–90. Available at: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=158129&tool=pmcentrez&render type=abstract.
    23. (2012). Insights on the evolution of trehalose biosynthesis. BMC evolutionary biology. 2006;6:109. Available at: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1769515&tool=pmcentrez&rende rtype=abstract. Accessed
    24. (2012). Introduction to Proteomics, Tools for the New Biology. doi
    25. (2007). Introduction to proteomics: A brief historical perspective on contemporary approaches. In: doi
    26. Light regulates alternative splicing of hydroxypyruvate reductase in pumpkin. The Plant journal for cell and molecular biology. 1999;17(3):309–20. Available at: http://www.ncbi.nlm.nih.gov/pubmed/10097389.
    27. (2012). Matrix Assisted Laser Desorption/Ionization Mass Spectrometry in Peptide and Protein Analysis. In: doi
    28. mechanism and regulation of peroxiredoxins. Trends in biochemical sciences. 2003;28(1):32–40. Available at: doi
    29. (2012). Methods of quantitative proteomics and their application to plant organelle characterization. doi
    30. (2012). Multifaceted roles of glycolytic enzymes. doi
    31. (2012). Overexpression of SlGMEs leads to ascorbate accumulation with enhanced oxidative stress, cold, and salt tolerance in tomato. Plant cell reports. 2011;30(3):389–98. Available at: http://www.ncbi.nlm.nih.gov/pubmed/20981454. Accessed doi
    32. (2012). Overexpression of the trehalose-6-phosphate synthase gene OsTPS1 enhances abiotic stress tolerance in rice. doi
    33. (2003). Phosphate Dehydrogenase from Heterotrophic Cells of Wheat Interacts with 14-3-3 Proteins 1. Plant Physiology. doi
    34. (2012). phosphate synthase and trehalose-6-phosphate phosphatase in transgenic rice plants increases trehalose accumulation and abiotic stress tolerance without stunting. doi
    35. Phyto-Remediation For Rehabilitation Of Agricultural Land Contaminated By Cadmium And Copper. Phyto-remediation for rehabilitatio.
    36. Proteomic analysis as a tool for investigating arsenic stress in Pteris vittata roots colonized or not by arbuscular mycorrhizal symbiosis. doi
    37. (2012). Proteomic analysis of Arabidopsis halleri shoots in response to the heavy metals cadmium and zinc and rhizosphere microorganisms. doi
    38. (2012). Proteomic characterization of copper stress response in Cannabis sativa roots. doi
    39. (2011). Proteomics based approach to understand tissue regeneration.
    40. (2012). Quantitation in mass-spectrometry-based proteomics. Annual review of plant biology. doi
    41. (2012). Quantitative changes in protein expression of cadmium-exposed poplar plants. doi
    42. Quantitative plant proteomics. doi
    43. (1998). Regulation of Leaf Senescence by Cytokinin , Sugars , and Light Effects on NADH-Dependent Hydroxypyruvate Reductase. Plant physiology. doi
    44. (1992). Regulation of ribulose 1, 5-bisphosphate carboxylase/oxygenase activity. Annual review of plant biology. doi
    45. Remediation of heavy metals contaminated soils by ball milling. doi
    46. (2006). Responses to water withdrawal of tobacco plants genetically engineered with the AtTPS1 gene: a special reference to photosynthetic parameters. doi
    47. (2012). Rubisco activase - Rubisco’s catalytic chaperone.
    48. Rubisco activase constrains the photosynthetic potential of leaves at high temperature and CO2. doi
    49. (1999). Sample PuriÐcation and Preparation Technique Based on Nano-scale Reversed-phase Columns for the Sensitive Analysis of Complex Peptide Mixtures by Matrix-assisted Laser Desorption / Ionization Mass Spectrometry. Journal of Mass Spectrometry. doi
    50. Structure and function of GDP-mannose-3′ ,5′ -epimerase; an enzyme which performs three chemical reactions at the same active site. doi
    51. The Arabidopsis trehalose-6-P synthase AtTPS1 gene is a regulator of glucose, abscisic acid, and stress signaling. Plant physiology. doi
    52. (2012). The early responses of Arabidopsis thaliana cells to cadmium exposure explored by protein and metabolite profiling analyses. doi
    53. The effect of body protein supply on resistance to cadmium. doi
    54. (2012). The effect of silicon on the leaf proteome of rice (Oryza sativa L.) plants under cadmium-stress. doi
    55. (1994). The Role of Carbonic Anhydrase in Photosynthesis. Annual Review of Plant Physiology and Plant Molecular Biology. doi
    56. The role of trehalose biosynthesis in plants. Plant physiology. doi
    57. (2004). Tobacco ( Nicotiana tabacum L .) -A model system for tissue culture interventions and genetic engineering.
    58. Towards Understanding Plant Response to Heavy Metal Stress. In: doi
    59. (2005). Transformation of tobacco with an Arabidopsis thaliana gene involved in trehalose biosynthesis increases tolerance to several abiotic stresses. doi
    60. Trehalose accumulation in rice plants confers high tolerance levels to different abiotic stresses. doi
    61. (2008). Trehalose and Abiotic Stress in Biological Systems. doi
    62. (2012). Trehalose and its applications in plant biotechnology. doi
    63. (2012). Trehalose metabolism and signaling. Annual review of plant biology. doi
    64. (2012). Trehalose: current use and future applications. doi
    65. Two-dimensional electrophoretic analysis of rice proteins by polyethylene glycol fractionation for protein arrays. doi
    66. Unraveling the role of fungal symbionts in plant abiotic stress tolerance. Plant signaling & behavior. 2011;6(2):175–91. Available at: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=3121976&tool=pmcentrez&rende rtype=abstract.

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