TRANSGENIC EGYPTIAN WHEAT (Triticum aestivum L.) WITH Arabidopsis NPR1 GENE via BIOLISTIC BOMBARDMENT

Authors

  • A. H. FAHMY Agriculture Genetic Engineering Research Institute, Agricultural Research Center, Egypt
  • K. EL MANGOURY Agriculture Genetic Engineering Research Institute, Agricultural Research Center, Egypt
  • WALAA ABOU EL-WAFA Plant Science Department, Faculty of Science, Ain Shams University, Egypt
  • HODA M. S. BARAKAT Plant Science Department, Faculty of Science, Ain Shams University, Egypt
  • SOHEIR EL-SAEED EL-KHODARY Plant Science Department, Faculty of Science, Ain Shams University, Egypt
  • J. LI Department of Plant Pathology, Kansas State University, Manhattan, KS, USA
  • S. MUTHUKRISHNAN Department of Biochemistry, Kansas State University, Manhattan, KS, USA

Abstract

AtNPR1 gene plays a crucial role in biotic resistance in plants. Immature embryo derived calli of wheat cv. Giza 164 were co-transformed with the plasmid pAHC25 containing bar and gus genes and the plasmid pJS406 containing AtNPR1 gene via Biolistic bombardment. The produced calli were subcultured on bialaphos containing medium (CIMB). Surviving calli were then regenerated on selection medium (MSRB) to produce green regenerated shoots, which were then transferred onto rooting/selection medium (FMSB). Regenerated rooted plantlets were transferred to soil pots in the control growth chamber for acclimatization. Only 48 plants succeeded in acclimatization and reached seed setting in biocontainment greenhouse. Transgene integration was confirmed by PCR and dot-blot analyses. The overall transformation frequency was 1.9%.

References

Agrios, G. N. (2005). Plant Pathology. Fifth edition, Elsevier Academic Press. New York, 5thed, ISBN: 0- 12-473276-3.

Bari, R. and J. D. Jones (2009). Role of plant hormones in plant defense responses. Plant Mol. Biology, 69: 473-488.

Bhalla, P. L. (2006). Genetic engineering of wheat-current challenges and opportunities. Trends in Biotechnology, 24: 305-311.

Chawla, H. S., L. A. Cass and J. A. Simmonds (1999). Expression of anthocyanin pigmentation in wheat tissues transformed with anthocyanin regulatory genes. Current Science, 76: 1365-1370.

Chern, M. S., H. A. Fitzgerald, R. C. Yadav, P. E. Canlas, X. Dong and P. C. Ronald (2001). Evidence for a disease-resistance pathway in rice similar to the NPR1-mediated signaling pathway in Arabidopsis. Plant Journal, 27: 101-113.

Chern, M., P. E. Canlas, H. A. Fitzgerald and P. C. Ronald (2005). Rice NRR, a negative regulator of disease resistance, interacts with Arabidopsis NPR1 and rice NH1. The Plant Journal, 43: 623-635.

Chugh, A. and P. Khurana (2003). Herbicide- resistant transgenics of bread wheat (T. aestivum) and emmer wheat (T. dicoccum( by particle bombardment and Agrobacteriummediated approaches. Current Science, 84: 78-83.

Daniell, H., M. Krishnan and B. F. McFadden (1991). Transient expression of -glucuronidase in different cellular compartments following biolistic delivery of foreign DNA into wheat leaves and calli. Plant Cell Reports, 9: 615-619.

Ding, L. P., S. C. Li, J. M. Gao, Y. S. Wang, G. X. Yang and G. Y. He (2009). Optimization of Agrobacterium- mediated transformation conditions in mature embryos of elite wheat. Molecular Biology Reports, 36: 29-36.

Fahmy, A. H., K. S. Abdalla and M. Abdallah (2007). Integration and expression of the high-molecularweight glutenin subunit Dy10 gene into Egyptian wheat. Arab J. Biotech., 10: 49-56.

Fahmy, A. H., K. El Mangoury, Walaa Abou El-Wafa, Hoda M. S. Barakat, Soheir El-Saeed El- Khodary and S. Muthukrishnan (2013). Genetic transformation of Egyptian wheat (Triticum aestivum L.) with chitinase gene via microprojectile bombardment. Egypt. J. Genet. Cytol., 42: 233- 246.

Friedrich, L., K. Lawton, R. Dietrich, M. Willits, R. Cade and J. Ryals (2001). NIM1 overexpression in Arabidopsis potentiates plant disease resistance and results in enhanced effectiveness of fungicides. Mol. Plant- Microbe Interact., 14: 1114-1124.

Jefferson, R. A., T. A. Kavanagh and M. W. Bevan (1987). GUS fusion: β- glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J., 6: 3901- 3907.

Jiang, Z. N., L. P. Xing, H. Z. Wang, L. Yu, J. L. Ni and P. D. Chen (2006). Transferring wheat pathogenesisrelated proteins 1 (TaPR -1) gene into wheat via micro-projectile bombardment. Journal of Triticeae Crops, 26: 51-57.

Koornneef, A. and C. M. J. Pieterse (2008). Cross talk in defense signaling. Plant Physiology, 146: 839- 844.

Liu, G., E. B. Holub, J. M. Alonso, J. R. Ecker and P. R. Fobert (2005). An Arabidopsis NPR1-like gene, NPR4, is required for disease resistance. Plant Journal, 41: 304- 318.

Loake, G. and M. Grant (2007). Salicylic acid in plant defense-the players and protagonists. Curr. Opin. Plant Biol., 10: 466-472.

Makandar, R., J. S. Essig, M. A. Schapaugh, H. N. Trick and J. Shah (2006). Genetically engineered resistance to Fusarium head blight in wheat by expression of Arabidopsis NPR1. Molecular Plant-Microbe Interactions, 19: 123-129.

Makandar, R., V. Nalam, R. Chaturvedi, R. Jeannotte, A. A. Sparks and J. Shah (2010). Involvement of salicylate and jasmonate signaling pathways in Arabidopsis interaction with Fusarium graminearum. Molecular Plant-Microbe Interactions, 23: 861-870.

Makandar, R., J. N. Vamsi, H. Lee, N. T. Harold, Y. Dong and J. Shah (2012). Salicylic acid regulates basal resistance to fusarium head blight in wheat. Molecular Plant- Microbe Interactions, 25: 431-439.

Murashige, T. and F. Skoog (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15: 473-497.

Punja, Z. K. (2006). Recent developments toward achieving fungal disease resistance in transgenic plants. Canadian Journal of Plant Pathology, 28: 298-308

Schroeder, H. E., A. H. Schotz, T. Wardley-Richardson, D. Spencer and T. J. V. Higgins (1993). Transformation and regeneration of two cultivars of pea (Pisum sativum L.). Plant Physiolgy, 101: 751-757.

Spoel, S. H., A. Koornneef, S. M. C. Chessens, J. P. Korzelius, J. A. VanPetl, M. J. Mueller, A. J. Buchala, J. P. Metaux, R. Brown, K. Kazoan, L. C. Van Loon, X. Dong and C. M. J. Peiterse (2003). AtNPR1 modulates cross-talk between salicylate and jasmonate dependent defense pathways through a novel function in the cytosol. Plant Cell 15: 760-770.

Spoel, S. H., J. S. Johnson and X. N. Dong (2007). Regulation of tradeoffs between plant defenses against pathogens with different lifestyles. Proc. Natl. Acad. Sci. USA, 104: 18842-18847.

Silva K. J. P., Asha Brunings, Natalia A. Peres, Z. Mou and K. M. Folta (2015). The Arabidopsis NPR1 gene confers broad-spectrum disease resistance in strawberry. Transgenic Res., 24: 693-704.

Thomma, B. P. H. J., K. Eggermont, W. F. Broekaert and P. A. Cammue (2000). Disease development of several fungi on Arabidopsis can be reduced by treatment with methyl jasmonate. Plant Physiol. Biochem., 38: 421-427.

Weeks, J. T., O. D. Anderson and A. E. Blechl (1993). Rapid production of multiple independent lines of fertile transgenic wheat (Triticum aestivum). Plant Physiology, 102: 1077-1084.

Xing, L. P., H. Z. Wang, Z. N. Jiang, J. L. Ni, A. Z. Cao, L. Yu and P. D. Chen (2008). Transformation of wheat thaumatin-like protein gene and diseases resistance analysis of the transgenic plants. Acta Agronomica Sinica, 34: 349-354.

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2017-08-06

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