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Biology

Isolamento de Viável multicelular Glândulas de tecido da planta carnívora, Nepenthes doi: 10.3791/50993 Published: December 22, 2013

Materials

Name Company Catalog Number Comments
RNAqueous-Micro Kit Life Technologies GmbH, Germany AM1931 Isolation of total RNA, 50 samples
Dynabeads mRNA DIRECT Micro Kit Life Technologies GmbH, Germany 61011 5 ml Dynabeads Oligo (dT)25
DynaMag-2 magnet Life Technologies GmbH, Germany 12321D
SuperScript CellsDirect cDNA Synthesis System Life Technologies GmbH, Germany 18080-200 25 reactions
RNaseOUT Life Technologies GmbH, Germany 10777-019 Rnase inhibitor, 5,000 units
Random Hexamers (0.4 μg/μl) QIAGEN GmbH, Germany 79236 100 μl
10 mM dNTP Mix, molecular biology grade Fermentas GmbH, Germany R0192 1 ml
Taq DNA polymerase, (recombinant) Fermentas GmbH, Germany EP0402 Supplied with 10x Taq buffer (with KCl or (NH4)2SO4) and 25 mM MgCl2
TURBO DNase Life Technologies GmbH, Germany AM2238 1,000 units
Agarose Carl Roth GmbH Co. Germany T846.3
GeneRuler Low Range DNA ladder, ready-to-use Fermentas GmbH, Germany SM1193 50 μg
Brilliant II SYBR Green QPCR Master Mix Agilent Technologies, Inc., USA 600828 Single kit
RNaseZAP Sigma-Aldrich, Taufkirchen, Germany R2020
Razor blade Carl Roth GmbH Co., Germany CK08.1
Tweezers Carl Roth GmbH Co., Germany 2801.1 / 2855.1
GE NanoVue spectrophotometer GE Healthcare Europe GmbH, Germany 28-9569-62
Mastercycler gradient Eppendorf AG, Hamburg, Germany 5331 000.010
Mx3000P Real-Time PCR System Stratagene, USA 401403
Agagel Standard Horizontal Gel Electrophoresis Chamber Biometra Discontinued
Micromanipulator aura optik gmbh aureka Equipped with a microforceps
Stereomicroscope Carl Zeiss Microscopy, Jena, Germany SteREO Lumar.V12 Equipped with an AxioCam
Lumar Filter set 01 Carl Zeiss, Microscopy, Jena, Germany 485001-0000-000 UV filter
Lumar Filter set 09 Carl Zeiss, Microscopy, Jena, Germany 485009-0000-000 non-UV filter

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References

  1. Adlassnig, W., Peroutka, M., Land, I., Lichtscheidl, I. K. Glands of carnivorous plants as a model system in cell biological research. Acta Bot. Gall. 152, 111-124 (2005).
  2. Darwin, C. Insectivorous plants. John Murray. London. (1875).
  3. Mithöfer, A. Carnivorous pitcher plants: Insights in an old topic. Phytochemistry. 72, 1678-1682 (2011).
  4. Owen Jr., T. P., Lennon, K. A., Santo, M. J., Anderson, A. N. Pathways for nutrient transport in the pitchers of the carnivorous plant. Nepenthes alata Ann. Bot. 84, 459-466 (1999).
  5. Brandt, S. P. Microgenomics: gene expression analysis at the tissue-specific and single-cell levels. J. Exp. Bot. 56, 495-505 (2005).
  6. Hölscher, D., Schneider, B. Application of laser-assisted microdissection for tissue and cell-specific analysis of RNA, proteins, and metabolites. Prog. Bot. 69, 141-167 (2008).
  7. Rottloff, S., Müller, U., Kilper, R., Mithöfer, A. Micropreparation of single secretory glands from the carnivorous plant Nepenthes. Anal. Biochem. 394, 135-137 (2009).
  8. Liu, W. H., Saint, D. H. A new quantitative method of real time reverse transcription polymerase chain reaction assay based on simulation of polymerase chain reaction kinetics. Anal. Biochem. 302, 52-59 (2002).
  9. Werker, E. Trichome diversity and development. Adv. Bot. Res. Inc. Adv. Plant Pathol. 31, 1-35 (2000).
  10. Fahn, A. Structure and function of secretory cells. Adv. Bot. Res. Inc. Adv. Plant Pathol. 31, 37-75 (2000).
  11. Juniper, B. E., Robins, R. J., Joel, D. M. The carnivorous plants. Academic Press. London. (1989).
  12. Gershenzon, J., McCaskill, D., Rajaonarivony, J., Mihaliak, C., Karp, F., Croteau, R. Isolation of secretory-cells from plant glandular trichomes and their use in biosynthetic-studies of monoterpenes and other gland products. Anal. Biochem. 200, 130-138 (1992).
  13. Rontein, D., Onillon, S., et al. CYP725A4 from yew catalyzes complex structural rearrangement of taxa-4(5),11(12)-diene into the cyclic ether 5(12)-oxa-3(11)-cyclotaxane. J. of Biol. Chem. 283, (5), 6067-6075 (2008).
  14. Aharoni, A., Jongsma, M. A., Bouwmeester, H. J. Volatile science? Engineering of terpenoid biosynthesis in plants. Trends Plant Sci. 10, 594-602 (2005).
  15. Mithöfer, A., Boland, W., Maffei, M. E. Chemical ecology of plant-insect interactions. Ann. Plant Rev. 34, 261-291 (2009).
  16. Kennedy, G. G. Tomato pests, parasitoids, and predators: Tritrophic interactions involving the genus Lycopersicon. Ann. Rev. Entomol. 48, 51-72 (2003).
  17. Mithöfer, A., Boland, W. Plant defense against herbivores: Chemical aspects. Ann. Rev. Plant Biol. 63, 431-450 (2012).
  18. Rottloff, S., Stieber, R., Maischak, H., Turini, F. G., Heubl, G., Mithöfer, A. Functional characterization of a class III acid endochitinase from the traps of the carnivorous pitcher plant genus, Nepenthes. J. Exp. Bot. 62, 4649-4647 (2011).
Isolamento de Viável multicelular Glândulas de tecido da planta carnívora,<i> Nepenthes</i
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Cite this Article

Rottloff, S., Mithöfer, A., Müller, U., Kilper, R. Isolation of Viable Multicellular Glands from Tissue of the Carnivorous Plant, Nepenthes. J. Vis. Exp. (82), e50993, doi:10.3791/50993 (2013).More

Rottloff, S., Mithöfer, A., Müller, U., Kilper, R. Isolation of Viable Multicellular Glands from Tissue of the Carnivorous Plant, Nepenthes. J. Vis. Exp. (82), e50993, doi:10.3791/50993 (2013).

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