Abstract
The lack of alternative selectable markers in crop transformation has been a substantial barrier for commercial application of agricultural biotechnology. We have developed an efficient selection system for wheat transformation using glyphosate-tolerant CP4 and GOX genes as a selectable marker. Immature embryos of the wheat cultivar Bobwhite were bombarded with two separate plasmids harboring the CP4/GOX and GUS genes. After a 1 week delay, the bombarded embryos were transferred to a selection medium containing 2 mM glyphosate. Embryo-derived calli were subcultured onto the same selection medium every 3 weeks consecutively for 9–12 weeks, and were then regenerated and rooted on selection media with lower glyphosate concentrations. Transgenic plants tolerant to glyphosate were recovered. ELISA assay confirmed expression of the CP4 and GOX genes in R0 plants. Southern blot analysis demonstrated that the transgenes were integrated into the wheat genomes and transmitted to the following generation. The use of CP4 and GOX genes as a selectable marker provides an efficient, effective, and alternative transformation selection system for wheat.
Similar content being viewed by others
References
Barry G, Kishore G, Padgette S, Taylor M, Kolacz K, Weldon M, Re D, Eiehholtz D, Fincher K, Hallas L (1992) In: Singh BK, Flores HE, Shannon JC (eds) Biosynthesis and molecular regulation of amino acids in plants, American Society of Plant Physiologists, pp 139–145
Becker D, Brettschneider R, Lorz H (1994) Plant J 5: 299–307
Bradford MM (1976) Anal Biochem 72: 248–256
ChenDF, Dale PJ, Mullineaux PM (1991) In: Program and Abstracts, The 3nd Intnl Congress Intnl Soc Plant Mol Biol, Tucson, USA, Abstract #418
Christou P, Ford TL, Kofron M (1991) Bio/Technology 9: 957–962
Christensen AH, Sharrock RA, Quail PH (1992) Plant Mol Biol 18: 675–689
Dale PJ, Irwin JA, Scheffler JA (1993) Plant Breed 111: 1–22
Dekeyser R, Claes B, Marichal M, Montagu MV, Caplan A (1989) Plant Physiol 90: 217–223
Dennehey BK, Petersen WL, Ford-Santino C, Pajeau M, Armstrong CL (1994) Plant Cell Tiss Org Cult 36: 1–7
Fromm ME, Morris F, Armstrong C, Williams R, Thomas J, Klein TM (1990) Bio/Technology 8: 833–839
Gordon-Kamm WJ, Spencer TM, Mangano ML, Adams TR, Daines RJ, Start WG, O'Brien JV, Chambers SA, Adams VR, Jr. Willetts NG, Rice TB, Mackey CJ, Krueger RW, Kausch AP, Lemaux PG (1990) Plant Cell 2: 603–618
Jefferson RA, Kavanagh TA, Bevan MW (1987) EMBO J 6: 3901–3907
Kay R, Chan A, Daly M, McPherson J (1987) Science 236: 1299–1302
Kishore GM, Padgette SR, Fraley RT (1992) Weed Technol 6: 626–634
McCabe DE, Swain WF, Martinell BJ, Christou P (1988) Bio/Technology 6: 923–926
Murashige T, Skoog F (1962) Physiol Plant 15: 473–497
Nehra NS, Chibbar RN, Leung N, Carwell K, Mallard C, Steinhauer L, Baga M, Kartha KK (1994) Plant J 5: 285–297
Russell JA, Roy MK, Sanford JC (1992) In Vitro Cell Devel Biol 28P: 97–105
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press
Sanford JC, Klein TM, Wolf ED, Allen N (1987) Particle Sci Technol 5: 27–37
Shure M, Wessler S, Fedoroff N (1983) Cell 35: 225–233
Simonsen CC, Levinson AD (1983) Proc Natl Acad Sci USA 80: 2495–2499
Somers DA, Rines HW, Gu W, Kaeppler H, Bushnell WR (1992) Bio/Technology 10: 1589–1594
Vasil V, Brown SM, Re D, Fromm ME, Vasil IK (1991) Bio/Technology 9: 743–747
Vasil V, Castillo AM, Fromm ME, Vasil IK (1992) Bio/Technology 10: 667–674
Vasil V, Srivastava V, Castillo AM, Fromm ME, Vasil IK (1993) Bio/Technology 11: 1153–1158
Wan Y, Lemaux PG (1994) Plant Physiol 104: 37–48
Weeks JT, Anderson OD, Blechl AE (1993) Plant Physiol 102: 1077–1084
Zhou H, Stiff CM, Konzak CF (1993) Plant Cell Rep 12: 612–616
Author information
Authors and Affiliations
Additional information
Communicated by G. Phillips
Rights and permissions
About this article
Cite this article
Zhou, H., Arrowsmith, J.W., Fromm, M.E. et al. Glyphosate-tolerant CP4 and GOX genes as a selectable marker in wheat transformation. Plant Cell Reports 15, 159–163 (1995). https://doi.org/10.1007/BF00193711
Received:
Revised:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF00193711