Molecular Characterization of Transformed Plants

  • J. F. Topping
  • K. Lindsey
Part of the Springer book series (SLM)


The generation of genetically transformed plants is central to, and has indeed revolutionized, plant molecular biology. This is true for studies at both the fundamental and more applied levels of research. For researchers interested in unravelling the roles of specific genes in particular pathways of growth and development, the introduction into plants of foreign genes and gene promoters linked to reporter genes allows the detailed study of the temporal, spatial and quantitative expression of plant genes and the activities of associated regulatory sequences. In our own laboratory, we use these techniques in a programme of insertional mutagenesis to identify developmentally interesting genes (Topping et al. 1991; Lindsey et al. 1993; Topping et al. 1994). In the more applied area of genetic engineering, which is directed towards crop improvement, the introduction of novel genes encoding, for example, resistance to various pests and herbicides into economically important species, is in the long term likely to develop into a major branch of the plant breeding industry (Lindsey 1992). There are several well-characterized and very successful methods which are currently being employed to introduce specific genes and gene regulatory sequences into plants and these are described in Chapter 8 of this book.


Selectable Marker Gene Plant Molecular Biology Crude Cell Extract Microprojectile Bombardment Sterile Double Distil Water 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Chinault AC, Blakesley VA, Roessler E, Willis DG, Smith CA, Cook RG, Fenwick RG (1986) Characterization of transferable plasmids from Shigella flex-neri 2A that confers resistance to trimethoprim, streptomycin and sulfonamides. Plasmid 15:119–131PubMedCrossRefGoogle Scholar
  2. De Block M, Botterman J, Vandewiele M, Dockx J, Thoen C, Gosselé V, Rao Movva N, Thompson C, Van Montagu M, Leemans J (1987) Engineering herbicide resistance in plants by expression of a detoxifying enzyme. EMBO J 6:2513–2518PubMedGoogle Scholar
  3. D’Halluin K, Bossut M, Bonne E, Mazur B, Leemans J, Botterman J (1992) Transformation of sugarbeet {Beta vulgaris L.) and evaluation of herbicide resistance in transgenic plants. Bio/Technology 10:309–314CrossRefGoogle Scholar
  4. Hauptmann RM, Vasil V, Ozias-Akins P, Tabaeizadeh Z, Rogers SG, Fraley RT, Horsch RB, Vasil IK (1988) Evaluation of selectable markers for obtaining stable transformants in the Gramineae. Plant Physiol 86:602–606PubMedCrossRefGoogle Scholar
  5. Helmer G, Casadaban M, Bevan MW, Kayes L, Chilton MD (1984) A new chimeric gene as a marker for plant transformation: the expression of Es-cherichia coli ß-galactosidase in sunflower and tobacco cells. Bio/Technology 2:520–527CrossRefGoogle Scholar
  6. Hobbs SLA, Warkentin TD, DeLong CMO (1993) Transgene copy number can be positively or negatively associated with transgene expression. Plant Mol Biol 21:17–26PubMedCrossRefGoogle Scholar
  7. Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: ß-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3301–3307Google Scholar
  8. Jones JDG, Svab Z, Harper EC, Hurwitz CD, Maliga P (1987) A dominant nuclear streptomycin resistance marker for plant cell transformation. Mol Gen Genet 210:86–91CrossRefGoogle Scholar
  9. Jorgensen R (1990) Altered gene expression in plants due to trans-interactions between homologous genes. Trends Biotechnol 8:340–344PubMedCrossRefGoogle Scholar
  10. Kirchner G, Kinslow CJ, Bloom GC, Taylor DW (1993) Nonlethal assay system of ß-glucuronidase activity in transgenic roots of tobacco. Plant Mol Biol Rep 11:320–325CrossRefGoogle Scholar
  11. Lindsey K (1992) Genetic manipulation of crop plants. J Biotechnol 26:1–28CrossRefGoogle Scholar
  12. Lindsey K, Jones MGK (1987) Transient gene expression in electroporated protoplasts and intact cells of sugar beet. Plant Mol Biol 10:43–52CrossRefGoogle Scholar
  13. Lindsey K, Wei W, Clarke MC, McArdle HF, Rooke LM, Topping JF (1993) Tagging genomic sequences that direct transgene expression by activation of a promoter trap in plants. Transgenic Res 2:33–47PubMedCrossRefGoogle Scholar
  14. Mascarenhas JP, Hamilton DA (1992) Artifacts in the localization of GUS activity in anthers of petunia transformed with a CaMV 35S-GUS construct. Plant J 2:405–408CrossRefGoogle Scholar
  15. Matzke MA, Primig M, Trnovsky J, Matzke AJM (1989) Reversible methylation and inactivation of marker genes in sequentially transformed tobacco plants. EMBO J 8:643–649PubMedGoogle Scholar
  16. Nagy F, Odell JT, Morelli G, Chua N-H (1985) Properties of expression of the 35S promoter from CaMV in transgenic tobacco plants. In: Zaitlin M, Day P, Hollaender A (eds) Biotechnology in plant science: relevance to agriculture in the eighties. Academic Press, Orlando, pp 227–235CrossRefGoogle Scholar
  17. Ow DW, Wood KV, DeLuca M, De Wet JR, Helinski DR, Howell SH (1986) Transient and stable expression of the firefly luciferase gene in plant cells and transgenic plants. Science 234:856–859PubMedCrossRefGoogle Scholar
  18. Pridmore RD (1987) New and versatile cloning vectors with kanamycin-resistance marker. Gene 56:309–312PubMedCrossRefGoogle Scholar
  19. Reiss B, Sprengel R, Will H, Schaller H (1984) A new and sensitive method for quantitative and qualitative assay of neomycin phosphotransferase in crude cell extracts. Gene 30:211–218PubMedCrossRefGoogle Scholar
  20. Reynaerts A, De Block M, Hernalsteens J-P, van Montagu M (1988) Selectable and screenable markers. In: Gelvin SB, Schilperoort RA (eds) Plant molecular biology manual. Kluwer, Dordrecht A9.T-16Google Scholar
  21. Sambrook J, Fritsch EF, Maniatis T (eds) (1989) Molecular cloning: a laboratory manual 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring HarborGoogle Scholar
  22. Shah DM, Horsch RB, Klee HJ, Kishore GM, Winter JA, Turner NE, Hironaka CM, Sanders PR, Gasser CS, Aykent S, Siegel NR, Rogers SG, Frayley RT (1986) Engineering herbicide tolerance in transgenic plants. Science 233:478–481PubMedCrossRefGoogle Scholar
  23. Stomp A-M (1990) Use of X-Gluc for histochemical localisation of glucuronidase. In: Editorial comments. United State Biochemical, Cleveland, p 5Google Scholar
  24. Topping JF, Wei W, Lindsey K (1991) Functional tagging of regulatory elements in the plant genome. Development 112:1009–1019PubMedGoogle Scholar
  25. Topping JF, Agyeman F, Henricot B, Lindsey K (1994) Identification of molecular markers of embryogenesis in Arabidopsis thaliana by promoter trapping. Plant J 5:895–903PubMedCrossRefGoogle Scholar
  26. Twell D, Klein TM, Fromm ME, McCormick S (1989) Transient expression of chimeric genes delivered into pollen by microprojectile bombardment. Plant Physiol 91:1270–1274PubMedCrossRefGoogle Scholar
  27. Van den Elzen PJM, Townsend J, Lee KY, Bedbrook JR (1985) A chimaeric hygromycin resistance gene as a selectable marker in plant cells. Plant Mol Biol 5:299–302CrossRefGoogle Scholar
  28. Wilkinson JE, Twell D, Lindsey K (1994) Methanol does not specifically inhibit endogenous ß-glucuronidase (GUS) activity. Plant Sci 97:61–67CrossRefGoogle Scholar
  29. Wohlleben W, Arnold W, Broer I, Hillemann D, Strauch E, Pühler A (1988) Nucleotide sequence of the phosphinothricine N-acetyltransferase gene from Streptomyces viridochromogenes Tu 494 and its expression in Nicotiana ta-bacum. Gene 70:25–37PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1997

Authors and Affiliations

  • J. F. Topping
  • K. Lindsey

There are no affiliations available

Personalised recommendations