Sugarcane is one of the most important worldwide cultivated agro-industrial crops that belong to the family Poaeceae and genus Saccharum. Prescribed study was conducted in Seed Biotechnology Lab, Center of Excellence in Molecular Biology, University of the Punjab, Lahore-Pakistan. The purpose of this study was to alter the genome of sugarcane line CPF246 with herbicide resistance gene GTGene driven by the sugar cane ubiquitin promoter and confirmation through PCR and Dot blot as well as protein expression analysis through ELISA. Results have shown the successful transformation and expression of transgenes against glyphosate resistance. This could prove a mile stone towards putting research efforts in the improvement of saccharum officinarum for certain traits. Outcomes of this research will be beneficial in minimizing the management cost which people do bear for weed management practices. Success of this study opens new horizons to further improve other characteristics of sugarcane using genetic modification as was done in current study.
Keywords: Saccharum officinarum, genetic modification, herbicide, GTGene, Glyphosate
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References
1. Bakker, H., Sugar Cane Cultivation and Management, Springer Science and Business Media, 2012.
2. D’Hont, A., Ison, D., Alix, K., Roux, C., and Glaszmann, J.C., Determination of basic chromosome numbers in the genus Saccharum by physical mapping of ribosomal RNA genes, Genome, 1998, vol. 41, no. 2, pp. 221–225.
3. Joyce, P., Kuwahata, M., Turner, N., and Lakshmanan, P., Selection system and co-cultivation medium are important determinants of Agrobacterium-mediated transformation of sugarcane, Plant Cell Rep., 2010, vol. 29, no. 2, pp. 173–183.
4. Enríquez-Obregó, G.A., Vázquez-Padrón, R.I., Prieto-Samsonov, D.L., Gustavo, A., and Selman-Housein, G., Herbicide-resistant sugarcane (Saccharum officinarum L.) plants by Agrobacterium-mediated transformation, Planta, 1998, vol. 206, no. 1, pp. 20–27.
5. Nasir, I.A., Tabassum, B., Qamar, Z., Javed, M.A., Tariq, M., Farooq, A.M., Butt, S.J., Qayyum, A., and Husnain, T., Herbicide-tolerant sugarcane (Saccharum officinarum L.) plants: an unconventional method of weed removal, Turkish J. Biol., 2014, vol. 38, no. 4, pp. 439–449.
6. Qureshi, M.A. and Afghan, S., Sugarcane Cultivation in Pakistan, Sugar Book Pub Pakistan Society of Sugar Technologist, 2005.
7. Srikanth, J., Subramonian, N., and Premachandran, M., Advances in transgenic research for insect resistance in sugarcane, Trop. Plant Biol., 2011, vol. 4, no. 1, pp. 52–61.
8. Gul, F., Naeem, M., and Shah, R.A., Role of gurdaspur borer (Bissetia steniellus Hampson) in sugarcane ratoon crop failure and its integrated control at Mardan, Sarhad J. Agricult., 2010, vol. 26, no. 3, pp. 387–391.
9. Khaliq, A., Ashfaq, M., Akram, W., Choi, J.K., and Lee, J.J., Effect of plant factors, sugar contents, and control methods on the top borer (Scirpophaga nivella F.) infestation in selected varieties of sugarcane, Entomol. Res., 2005, vol. 35, no. 3, pp. 153–160.
10. Nazir, A., Jariko, G.A., and Junejo, M.A., Factors affecting sugarcane production in Pakistan, Pakistan J. Com. Soc. Sci., 2013, vol. 7, no. 1, pp. 128–140.
11. Zafar, M., Tanveer, A., Cheema, Z.A., and Ashraf, M., Weed-crop competition effects on growth and yield of sugarcane planted using two methods, Pakistan J. Bot., 2010, vol. 42, no. 2, pp. 815–823.
12. McMahon, G., Lawrence, P., and O’Grady, T., Weed control in sugarcane, in Manual of Cane Growing Bureau of Sugar Experiment Stations, Indooroopilly, 2000, pp. 241–261.
13. Green, J.M., The benefits of herbicide-resistant crops, Pest Manage. Sci., 2012, vol. 68, no. 10, pp. 1323–1331.
14. Dill, G.M., Glyphosate-resistant crops: history, status and future, Pest Manage. Sci., 2005, vol. 61, no. 3, pp. 219–224.
15. Deng, L., Weng, L., and Xiao, G., Optimization of Epsps gene and development of double herbicide tolerant transgenic PGMS rice, J. Agricul. Sci. Technol., 2014, vol. 16, no. 1, pp. 217–228.
16. Keller, G., Spatola, L., Mccabe, D., Martinell, B., Swain, W., and John, M.E., Transgenic cotton resistant to herbicide bialaphos, Transgenic Res., 1997, vol. 6, no. 6, pp. 385–392.
17. Rashid, B., Saleem, Z., Husnain, T., and Riazuddin, S., Transformation and inheritance of Bt genes in Gossypium hirsutum, J. Plant Biol., 2008, vol. 51, no. 4, pp. 248–254.
18. Schmid, J. and Amrhein, N., Molecular organization of the shikimate pathway in higher plants, Phytochemistry, 1995, vol. 39, no. 4, pp. 737–749.
19. Kumar, S., Sharma, P., and Pental, D., A genetic approach to in vitro regeneration of non-regenerating cotton (Gossypium hirsutum L.) cultivars, Plant Cell Rep., 1998, vol. 18, nos. 1–2, pp. 59–63.
20. Arencibia, A.D., Carmona, E.R., Tellez, P., Chan, M.-T., Yu, S.-M., Trujillo, L.E., and Oramas, P., An efficient protocol for sugarcane (Saccharum spp. L.) transformation mediated by Agrobacterium tumefaciens, Transgenic Res., 1998, vol. 7, no. 3, pp. 213–222.
21. Birch, R., Transgenic Sugarcane: Opportunities and Limitations, 1997.
22. Taylor, P.W. and Dukic, S., Development of an in vitro culture technique for conservation of Saccharum spp. hybrid germplasm, Plant Cell, Tiss. Organ Culture, 1993, vol. 34, no. 2, pp. 217–222.
23. Gallo-Meaghe, M. and Irvine, J.E., Effects of tissue type and promoter strength on transient GUS expression in sugarcane following particle bombardment, Plant Cell Rep., 1993, vol. 12, no. 12, pp. 666–70.
24. Falco, M., Neto, A.T., and Ulian, E., Transformation and expression of a gene for herbicide resistance in a Brazilian sugarcane, Plant Cell Rep., 2000, vol. 19, no. 12, pp. 1188–1194.
25. Franks, T. and Birch, R., Gene transfer into intact sugarcane cells using microprojectile bombardment, Func. Plant Biol., 1991, vol. 18, no. 5, pp. 471–480.
26. Bower, R. and Birch, R.G., Transgenic sugarcane plants via microprojectile bombardment, Plant J., 1992, vol. 2, no. 3, pp. 409–416.
27. Arencibia, A., Vazquez, R.I., Prieto, D., Tellez, P., Carmona, E.R., Coego, A., Hernandez, L., De la Riva, G.A., and Selman-Housein, G., Transgenic sugarcane plants resistant to stem borer attack, Mol. Breed, 1997, vol. 3, no. 4, pp. 247–255.
28. Weng, L.X., Deng, H.H., Xu, J.L., Li, Q., Zhang, Y.Q., Jiang, Z.D., Li, Q.W., Chen, J.W., and Zhang, L.H., Transgenic sugarcane plants expressing high levels of modified cry1Ac provide effective control against stem borers in field trials, Trans. Res., 2011, vol. 20, no. 4, pp. 759–772.
29. Chen, L., Marmey, P., Taylor, N.J., Brizard, J.P., Espinoza, C., D’Cruz, P., Huet, H., Zhang, S., de Kochko, A., Beachy, R.N., and Fauquet, C.M., Expression and inheritance of multiple transgenes in rice plants, Nat. Biotechnol., 1998, vol. 16, no. 11, pp. 1060–1064.
30. Kaur, A., Gill, M., Gill, R., and Gosal, S., Standardization of different parameters for ‘particle gun’ mediated genetic transformation of sugarcane (Saccharum officinarum L.), Indian J. Biotech., 2007, vol. 6, no. 1, pp. 31–34.
31. Bower, R., Elliott, A.R., Potier, B.A., and Birch, R.G., High-efficiency, microprojectile-mediated cotransformation of sugarcane, using visible or selectable markers, Mol. Breed., 1996, vol. 2, no. 3, pp. 239–249.
32. Christensen, A.H. and Quail, P.H., Ubiquitin promoter-based vectors for high-level expression of selectable and/or screenable marker genes in monocotyledonous plants, Trans. Res., 1996, vol. 5, no. 3, pp. 213–218.
33. McElroy, D. and Brettell, R.I., Foreign gene expression in transgenic cereals, Trends Biotechnol., 1994, vol. 12, no. 2, pp. 62–68.
34. Joung, Y.H. and Kamo, K., Expression of a polyubiquitin promoter isolated from Gladiolus, Plant Cell Rep., 2006, vol. 25, no. 10, pp. 1081–1088.
35. Zambryski, P., Basic processes underlying Agrobacterium-mediated DNA transfer to plant cells Annu. Rev. Genet., 1988, vol. 22, no. 1, pp. 1–30.
36. Kohli, A., Leech, M., Vain, P., Laurie, D.A., and Christou, P., Transgene organization in rice engineered through direct DNA transfer supports a two-phase integration mechanism mediated by the establishment of integration hot spots. Proc. Natl. Acad. Sci. U. S. A., 1998, vol. 95, no. 12, pp. 7203–7208.
37. Guo, F.-Q., Wang, R., Chen, M., and Crawford, N.M., The Arabidopsis dual-affinity nitrate transporter gene AtNRT1. 1 (CHL1) is activated and functions in nascent organ development during vegetative and reproductive growth, Plant Cell, 2001, vol. 13, no. 8, pp. 1761–1777.
38. Kumar, R. and Sinha, R., Colloidal gold based dipstick strip for detection of genetically modified crops and produce.
39. Halder, S. and Venu, P., Bt Cry toxin expression profile in selected Pakistani cotton genotypes, Curr. Sci., 2012, vol. 102, no. 12, p. 1632.
40. Boopal, K., Hanur, V.S., Arya, V.V., and Reddy, P., Phenotypic assessment of Bt Cry2A transgenic tomato resistant to neonate larva of Helicoverpa armigera, Curr. Trends Biotechnol. Pharm., 2014, vol. 8, no. 2, pp. 124–129.
41. Bakhsh, A., Rao, A.Q., Shahid, A.A., and Husnain, T., Spatio temporal expression pattern of an insecticidal gene (cry2A) in transgenic cotton lines, Not. Sci. Biol., 2012, vol. 4, no. 4, p. 115.
42. Kiani, S., Mohamed, B.B., Shehzad, K., Jamal, A., Shahid, M.N., Shahid, A.A., and Husnain, T., Chloroplast-targeted expression of recombinant crystal-protein gene in cotton: an unconventional combat with resistant pests, J. Biotechnol., 2013, vol. 166, no. 3, pp. 88–96.
43. Bashir, K., Husnain, T., Fatima, T., Riaz, N., Makhdoom, R., and Riazuddin, S., Novel indica basmati line (B-370) expressing two unrelated genes of Bacillus thuringiensis is highly resistant to two lepidopteran insects in the field, Crop Prot., 2005, vol. 24, no. 10, pp. 870–879.