Submit Manuscript

Pakistan Journal of Biological Sciences

Year: 2009 | Volume: 12 | Issue: 2 | Page No.: 101-109
Research Article

Genetic Transformation of Nepalese Spring Wheat (Triticum aestivum L.) Cultivars with ipt Gene under the Regulation of a Senescence Enhanced Promoter from Maize

D.R. Pant
Department of Environmental Sciences, Institute of Agriculture and Animal Sciences, Rampur Campus, Rampur, Chitawan, Nepal
T. Bhattarai
Department of Botany, Amrit Campus, Kathmandu, Nepal
E. Beck
Department of Plant Physiology, University of Bayreuth, Universitaetsstrasse 30, D-95447, Bayreuth
S. Fettig
Department of Plant Physiology, University of Bayreuth, Universitaetsstrasse 30, D-95447, Bayreuth
Two Nepalese spring wheat cultivars were transformed with an ipt gene from Agrobacterium tumefaciens under the control of the senescence inducible promoter pSEE1 from maize using biolistic method. The resulting transgenic lines, one from Pasang Lahmu and seven from Annapurna-1, were studied for the expression of the transgene and the phenotype characters like chlorophyll content, chlorophylla/b ratio, PS II quantum yield and other parameters of agronomic importance. Analysis of transgene expression by RT-PCR revealed very weak or no signal at all, indicating either partial or complete silencing of the transgene in the lines tested. None of the plants exhibited a phenotype that was significantly different from the respective azygous controls.
PDF Fulltext XML References Citation

Leave a Comment

Your email address will not be published. Required fields are marked *

Article Trend

Total views 4219

References

  1. Banowetz, G.M., 1997. Cultivars of hexaploid wheat of contrasting stature and chlorophyll retention differ in cytokinin content and responsiveness. Ann. Bot., 79: 185-190.
    CrossRef
  2. Buchanan-Wollaston, V., T. Page, E. Harrison, E. Breeze and P.O. Lim et al., 2005. Comparative transcriptome analysis reveals significant differences in gene expression and signalling pathways between developmental and dark/starvation-induced senescence in Arabidopsis. Plant J., 42: 567-585.
    CrossRef
  3. Chalfie, M., Y. Tu, G. Euskirchen, W.W. Ward and D.C. Prasher, 1994. Green fluorescent protein as a marker for gene expression. Science, 263: 802-805.
    CrossRefPubMed
  4. Chen, L.F.O., J.Y. Hwang, Y.Y. Chang, C.W. Sun and S.F. Yang, 2001. Transformation of broccoli (Brassica oleracea var, italica) with isopentenyltransferase gene via Agrobacterium tumefaciens for post-harvest yellowing retardation. Mol. Breed., 7: 243-257.
    Direct Link
  5. Dangl, J.L., R.A. Dietrich and H. Thomas, 2000. Senescence and Programmed Cell Death. In: Biochemistry and Molecular Biology of Plants, Buchanan, B., W. Gruissem and R. Jones (Eds.). ASPP, Rockville, pp: 1044-1100.
  6. Daskalova, S., A. Sorokin, N.W. Scott, A. Slater and M.R. Fowler et al., 2002. Enhancement of wheat grain yields by delaying leaf senescence. Proceedings of the 10th International Association for Plant Tissue Culture and Biotechnology Congress, June 23-28, 2002, Orlando, Florida, pp: 1082-1082.
  7. Feller, U. and A. Fischer, 1994. Nitrogen metabolism in senescing leaves. Crit. Rev. Plant Sci., 13: 241-273.
    CrossRef
  8. Fettig, S. and D. Hess, 1999. Expression of a chimeric stilbene synthase gene in transgenic wheat lines. Transgenic Res., 8: 179-189.
    CrossRef
  9. Gan, S. and R.M. Amasino, 1995. Inhibition of leaf senescence by autoregulated production of cytokinin. Science, 270: 1986-1988.
    CrossRef
  10. Griffiths, C.M., S.E. Hosken, D. Oliver, J. Chojecki and H. Thomas, 1997. Sequencing, expressing pattern and RFLP mapping of a senescence-enhanced cDNA from Zea mays with high homology to oryzain Ύ and aleurain. Plant Mol. Biol., 34: 815-821.
    CrossRef
  11. Hensel, L.L., V. Grbic, D.A. Baumgarten and A.B. Bleecker, 1993. Developmental and age-related processes that influence the longevity and senescence of photosynthetic tissues in Arabidopsis. Plant Cell, 5: 553-564.
    Direct Link
  12. Huber, M., R. Hahn and D. Hess, 2002. High transformation frequencies obtained from a commercial wheat (Triticum aestivum L. cv. Combi) by microbombardment of immature embryos followed by GFP screening combined with PPT selection. Mol. Breed., 10: 19-30.
    Direct Link
  13. Jiang, C.Z., S.R. Rodermel and R.M. Shibles, 1993. Photosynthesis, rubisco activity and amount and their regulation by transcription in senescing soyabean leaves. Plant Physiol., 101: 105-112.
    Direct Link
  14. Jordi, W., A. Schaependonk, E. Davelaar, G.M. Stoopen and C.S. Pot et al., 2000. Increased cytokinin levels in transgenic PSAG12-ipt tobacco plants have large direct and indirect effects on leaf senescence, photosynthesis and N-partitioning. Plant Cell Environ., 23: 279-289.
    Direct Link
  15. Kaminek, M., B. Solcova, M. Trckova, V. Motyka, S. Daskalova and M.C. Elliott, 2003. Exploitation of Mechanisms Regulating Cytokinin Levels to Improve Cereals. In: Phytohormones in Plant Biotechnology and Agriculture, Machackova, I. and G.A. Romanov (Eds.). Kluwer Academic Publishers, Netherlands, pp: 109-128.
  16. Lim, P.O., H.J. Kim and H.G. Nam, 2007. Leaf senescence. Ann. Rev. Plant Biol., 58: 115-136.
    CrossRefDirect Link
  17. Lin, Y.J., M.L. Cao, C.G. Xu, H. Chen, J. Wei and Q.F. Zhang, 2002. Cultivating rice with delaying leaf-senescence by PSAG12-IPT gene transformation. Acta Bot. Sin., 44: 1333-1338.
    Direct Link
  18. Lohman, K.N., S. Gan, M.C. John and R.M. Amasino, 1994. Molecular analysis of natural leaf senescence in Arabidopsis thaliana. Physiol. Plant., 92: 322-328.
    CrossRef
  19. Lu, C., Q. Lu, J. Zhang and T. Kuang, 2001. Characterisation of photosynthetic pigment composition, photosystem II photochemistry and thermal energy dissipation during leaf senescence of wheat plant grown in the field. J. Exp. Bot., 52: 1805-1810.
    CrossRef
  20. McCabe, M.S., L.C. Garratt, F. Schepers, W.J.R.M. Jordi and G.M. Stoopen et al., 2001. Effects of PSAG12-IPT gene expression on development and senescence in transgenic lettuce. Plant Physiol., 127: 505-516.
    CrossRef
  21. Nooden, L.D., 1988. The Phenomena of Senescence and Aging. In: Senescence and Aging in Plants, Nooden, L.D. and A.C. Leopold (Eds.). Academic Press, San Diego, ISBN: 0125209207, pp: 1-50.
  22. Oh, S.A., J.H. Park, G.I. Lee, K.H. Paek, S.K. Park and H.G. Nam, 1999. Identification of three genetic loci controlling leaf senescence in Arabidopsis thaliana. Plant J., 12: 527-535.
    Direct Link
  23. Pellegrineschi, A., L.M. Noguera, B. Skovmand, R.M. Brito and L. Velaquez et al., 2002. Identification of highly transformable wheat genotypes for mass production of fertile transgenic plants. Genome, 45: 421-430.
    Direct Link
  24. Pellegrineschi, A., M. Reynolds, M. Pachco, R.M. Britto, R. Almeraya, K. Yamaguchi-Shinozaki and D. Hoisington, 2004. Stress induced expression in wheat of the Arabidopsis thaliana DREB1 gene delays water stress symptoms under greenhouse conditions. Genome, 47: 493-500.
    Direct Link
  25. Porra, R.J., W.A. Thompson and P.E. Kriedemann, 1989. Determination of accurate extinction coefficients and simultaneous equations for assaying chlorophylls a and b extracted with four different solvents: Verification of the concentration of chlorophyll standards by atomic absorption spectroscopy. Biochim. Biophys. Acta (BBA)-Bioenerget., 975: 384-394.
    CrossRefDirect Link
  26. Robson, P.R., I.S. Donnison, K. Wang, B. Frame, S.E. Pegg, A. Thomas and H. Thomas, 2004. Leaf senescence is delayed in maize expressing the Agrobacterium ipt gene under the control of a novel maize senescence-enhanced promoter. Plant Biotechnol. J., 2: 101-112.
    CrossRef
  27. Sambrook, J. and D.W. Russell, 2001. Molecular Cloning: A Laboratory Manual. 3rd Edn., Cold Spring Harbor Laboratory Press, New York, USA., ISBN-13: 9780879695774, Pages: 2344.
    Direct Link
  28. Sivakumar, T., Virendranath and G.C. Srivastava, 2001. Effects of benzyl adenine and abscissic acid on yield and yield componenets in triticale and wheat. J. Agron. Crop Sci., 183: 43-46.
    CrossRefDirect Link
  29. Sykorova, B., G. Kuresova, S. Daskalova, M. Trckova and K. Hoyerova et al., 2008. Senescence induced ectopic expression of the A. tumefaciens ipt gene in wheat delays leaf senescence, increases cytokinin content, nitrate influx and nitrate reductase activity, but does not affect grain yield. J. Exp. Bot.
    Direct Link
  30. Terao, T. and S. Katoh, 1996. Antenna sizes of Photosystem I and II in chlorophyll b deficient mutants of rice. Evidence for an antenna function of photosystem II centres that are inactive in electron transport. Plant Cell Physiol., 37: 307-312.
    Direct Link
  31. Thomas, H., 1992. Canopy Survival. In: Crop Photosynthesis Spatial and Temporal Determinants, Baker, N. and H. Thomas (Eds.). Elsevier, Amsterdam, pp: 11-41.
  32. Thomas, H. and C.J. Howarth, 2000. Five ways to stay green. J. Exp. Bot., 51: 329-337.
    Direct Link
  33. Thomas, H. and J. Stoddart, 1980. Leaf senescence. Ann. Rev. Plant Physiol., 31: 83-111.
    Direct Link
  34. Wilkie, S., 1997. Isolation of Total Genomic DNA. In: Plant Molecular Biology, A Laboratory Manual, Clark, M.S. (Ed.). Springer, Berlin Heidelberg, pp: 3-15.
  35. Williams, R.H. and P.M. Cartwright, 1980. The effect of applications of synthetic cytokinin on shoot dominance and grain yield in spring barley. Ann. Bot., 46: 445-452.
    Direct Link
  36. Xi, Y., O. Zhang, Y. Lin, W. Hou and M. Lu, 2004. Introduction of leaf senescence-inhibiting gene PSUB SAG12-IPT into common wheat via Agrobacterium tumefaciens soaked seeds. Scientia Agric. Sin., 37: 1235-1238.
    Direct Link
  37. Young, T.E., J. Giesler-Lee and D.R. Gallie, 2004. Senescence induced expression of cytokinin reverses pistil abortion during maize flower development. Plant J., 38: 910-922.
    Direct Link

Keywords