USING MANNOSE AS A POSITIVE SELECTION OF TRANSFORMED CARICA PAPAYA L. VAR ‘EKSOTIKA’
DOI:
https://doi.org/10.11113/jt.v77.6889Keywords:
‘Eksotika’ papaya, positive selection, phosphomannose isomerase (pmi), Agrobacterium-mediated transformationAbstract
The main objective of this study is to develop marker-free transgenic papaya plants via positive selection using phosphomannose isomerase (pmi) gene. Phosphomannose isomerase (PMI) is an enzyme that converts mannose-6-phosphate to fructose-6-phosphate, a glycolysis intermediate that supports the growth of plant cells. To establish a marker-free positive selection system using this PMI, the effect of mannose on the growth and development of embryogenic ‘Eksotika’ papaya callus was evaluated. One-month old embryogenic calli were cultured on Murashige and Skoog (MS) medium in which 60 g/L sucrose in the original recipe was replaced with different concentrations of mannose and sucrose. Mannose was supplied as the sole carbon source or in combination with sucrose at 0, 5, 10, 15, 20, 25 or 30 g/L. Embryogenic calli cultured on medium supplemented with a ratio of 0:60 g/L mannose: sucrose was used as a control. The results after six sub-cultures showed that most of the embryogenic calli transferred on media containing only mannose turned brown. Higher concentrations of mannose resulted in higher percentage of brown calli (dead). Mannose at 30 g/L mannose was found to be effective for screening transformed embryogenic calli. Evaluation of papaya transformation efficiency using this positive selection system was pursued using 650 one-month-old embryogenic calli Agrobacterium-transformed with pNOV2819 harboring the pmi gene. Only transformed cells are capable of utilizing mannose as a carbon source to grow. After five months on mannose selection, all 67 putative transformants obtained were PCR-positives for the pmi gene.
References
Proliferation, Maturation And Germination in Cajanus. World Journal of Agricultural Sciences. 7:86-95.
Barb, A. W., Pharr, D. M., Williamson, J. D. 2003. A Nicotiana Tabacum Cell Culture Selected For Accelerated Growth On Man Has Increased Expression Of Phosphomannose Isomerase. Journal of Plant Science. 165: 639-648.
Bojsen, K., Donaldson, I., Haldrup, A., Joersbo, M., Kreierg, J. D., Nielsen, J., Okkels, F. T., Peterson, S. G., Whenham, R. J. 1999. Positive Selection. United States Patent No. 5,994,629. November 30, 1999.
Bojsen, K., Donaldson, I., Haldrup, A., Joersboe, M., Kreierg, J. D., Nielsen, J., Okkels, F. T., Peterson, S. G., Whenham, R. J. 1998. Mannose or Xylose Based Positive Selection. United States Patent No.5,767,378. June 16, 1998.
Boscariol, R. L., Almeida, W. A., Derbyshire, M. T., Mourao, Filho, F. A., Mendes, B. M. 2003. The Use Of The PMI/Mannose Selection System To Recover Transgenic Sweet Orange Plants (Citrus sinensis L. Osbeck). Plant Cell Reports. 22: 122-8.
Chan, Y. K. 1987. Backcross Method In Improvement Of Papaya (Carica papaya L.). Malaysian Applied Biology. 16: 95-100.
Datta, K., Baisakh, N., Oliva, N., Torrizo, L., Abrigo, E., Tan, J., Rai, M., Rehana, S., Al-Babili, S., Beyer, P., Potrykus, I., Datta, S. K. 2003. Bioengineered ‘Golden’ Indica Rice Cultivars With Β-Carotene Metabolism In The Endosperm With Hygromycin And Mannose Selection Systems. Journal of Plant Biotechnology. 1: 81-90.
De Fossard, R., Myint, A. and Lee, E. 1974. A Broad Spectrum Tissue Culture Experiment With Tobacco (Nicotiana Tabacum L.) Pith Tissue Callus. Plant Physiology. 31: 125-130.
He, Z., Fu, Y., Si, H., Hu, G., Zhang, S., Yu, Y., Sun, Z. 2004. Phosphomannose-isomerase (pmi) Gene As A Selectable Marker For Rice Transformation Via Agrobacterium. Journal of Plant Science. 166: 17–22.
Hoa, T. T., Al-Babili, S., Schaub, P., Potrykus, I., Beyer, P. 2003. Golden Indica and Japonica Rice Lines Amenable To Deregulation. Journal of Plant Physiology. 133: 161-169.
Joersbo, M. 2001. Advances in the Selection Of Transgenic Plants Using Non-Antibiotic Marker Genes. Journal of Plant Physiology. 111: 269-272.
Joersbo, M., Danaldson, I., Kreiberg, J., Petersen, S.G., Brunstedt, J., Okkels, F. T. 1998. Analysis of Mannose Selection Used For Transformation Of Sugar Beet. Journal of Molecular Breeding. 4: 111-117.
Lucca, P., Ye, X. D., Potrykus, I. 2001. Effective Selection And Regeneration Of Transgenic Rice Plants With Mannose As Selective Agent. Journal of Molecular Breeding. 7: 43-49.
Murashige, T., and Skoog, F. 1962. A Revised Medium For Rapid Growth And Bioassay With Tobacco Tissue Culture. Journal of Plant Physiology. 15: 473-497.
Negrotto, D., Jolley, M., Beer S., Wenck, A., Hansen, G. 2000. The Use Of Phosphomannose-Isomerase As Selectable Marker To Recover Transgenic Maize Plants (Zea mays L.) via Agrobacterium transformation. Plant Cell Reports. 19: 798-803.
O’Kennedy, M. M., Burger, J. T., Botha, F. C. 2004. Pearl Millet Transformation System Using The Positive Selectable Marker Gene Phosphomannose Isomerase. Plant Cell Reports. 22: 684-90.
Privalle, L. S. 2002. Phosphomannose Isomerase, A Novel Plant Selection System: Potential Allergenicity Assessment. Ann New York Academy Science. 964: 129-138.
Sew, Y. S., Johari, S., Maheswary, V. and Abu Bakar, U. K. 2011. Isolation of Fruit Ripening Genes From Carica Papaya Var. Eksotika 1 Cdna Libraries. Journal of Tropical Agriculture and Food Science. 39: 203-211.
Suprasanna, P., Bapat, V. A. 2005. Selection of Transgenic Plants Using Positive Selectable Markers And Other Non-Antibiotic Marker Genes. In Ashwini Kumar, Shikha Roy eds. Plant Biotechnology & Its Applications in Tissue Culture. International, New Delhi. 54-61.
Suprasanna, P., Sagi, L., Swennen, R. 2002. Positive Selectable Markers For Routine Plant Transformation. Journal of Physiology Plant. 38: 125-128.
Todd, R., Tague, G. W. 2001. Phosphomannose Isomerase: A Versatile Selectable Marker For Arabidopsis Thaliana Germ-Line Transformation. Plant Molecular Biology. 19: 307-319.
Vilasini, P., Latipah, Z. and Salasiah, A. 2000. Induction of Somatic Embryogenesis And Plant Regeneration From Immature Embryos Of Eksotika Papaya (Carica papaya L.). Journal of Tropical Agriculture and Food Science. 28: 121-126.
Wang, A. S., Evans, R. A., Altendorf, P. R., Hanten, J. A., Doyle, M. C., and Rosichan, J. L. 2000. A Mannose Selection System For Production Of Fertile Transgenic Maize Plants From Protoplasts. Plant Cell Reports. 19: 654-660.
Wright, M., Dawson, J., Dunder, E., Suttie, J., Reed, J., Kramer, R., Chang, Y. F., Novitzky, R., Wang, H., Artim-Moore, L. 2001. Efficient Biolistic Transformation Of Maize (Zea Mays L.) And Wheat (Triticum Aestivum L.) Using The Phosphomannose Isomerase Gene, Pmi, As The Selectable Marker. Plant Cell Reports. 20: 429-436.
Zhang, P., Potrykus, I., Puonti-Kaerlas, J. 2000. Efficient Production Of Transgenic Cassava Using Negative And Positive Selection. Transgen Res. 9: 405-415.
Downloads
Published
Issue
Section
License
Copyright of articles that appear in Jurnal Teknologi belongs exclusively to Penerbit Universiti Teknologi Malaysia (Penerbit UTM Press). This copyright covers the rights to reproduce the article, including reprints, electronic reproductions, or any other reproductions of similar nature.
