Abstract
Soybean (Glycine max [L.] Merrill) lectin is a seed protein that accumulates in protein bodies of cotyledons during seed development. We have constructed two expression cassettes containing the 5′ and 3′ regions of the soybean lectin gene connected by aNot I restriction site. One vector also contains the 32 amino acid signal sequence. Using polymerase chain reaction (PCR), the coding region of the β-glucuronidase (uidA) gene was inserted into theNot I site of each vector. We tested the function of the expression cassettes in transformed embryogenic cultures of soybean. Development-specific GUS expression was observed in developing somatic embryos transformed with the chimeric lectin promoter-GUS constructs as determined by histochemical assays. Our data indicate that these cassettes could be used to drive expression of foreign genes to modify embryo-specific traits of soybean as protein quality or quantity in the seed.
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Abbreviations
- Le1 :
-
gene encoding soybean lectin protein
- GUS:
-
β-glucuronidase
- uidA :
-
gene encoding β-glucuronidase (uidA is also known asgus, gusA, orGUS)
References
Chee, P.O., K.A. Fober, J.L. Slightom. 1981. Transformation of soybean (Glycine max) by infecting germinating seeds withAgrobacterium tumefaciens. Plant Physiol. 91:1212–1218.
Christou, P. 1990. Morphological description of transgenic chimeras created by the delivery, integration, and expression of foreign DNA using electric discharge particle acceleration. Ann. Bot. 66:379–386.
Crouch, M.L. 1982. Non-zygotic embryos ofBrassica napus contain embryo-specific storage proteins. Planta. 156:520–524.
Farrell, L.B. and R.N. Beachy. 1990. Manipulation of β-glucuronidase for use as a reporter in vacuolar targeting studies. Plant Mol. Biol. 15:821–825.
Finer, J.J. and M.D McMullen. 1990. Transformation of cotton (Gossypium hirsutum L.) via particle bombardment. Plant Cell Rep. 8:586–589.
Finer, J.J. and M.D. McMullen. 1991. Transformation of soybean via particle bombardment of embryogenic suspension culture tissue. In Vitro Cell. Dev. Biol. 27:175–182.
Finer, J.J. and A. Nagasawa. 1988. Development of an embryogenic suspension culture of soybean (Glycine max Merrill.). Plant Cell Tissue Organ Cult. 15:125–136.
Goldberg, R.B., S.J. Barker, L. Perez-Grau. 1989. Regulation of gene expression during plant embyogenesis. Cell 56:149–160.
Goldberg, R.B., G. Hoschek, L.O. Vodkin. 1983. An insertion sequence blocks the expression of a soybean lectin gene. Cell 33:465–475.
Hinchee, M.A.W., D.V. Conner-Ward, C.A. Newell, R.E. McDonnell, S.J. Sato, C.S. Gasser, D.A. Fischhoff, D.B. Re, R.T. Fraley, R.B. Horsch. 1988. Production of transgenic soybean plants usingAgrobacterium-mediated DNA transfer. Bio/Tech. 6:915–922.
Jefferson, R.A., T. A. Kavanagh, M.W. Bevan. 1987. GUS fusions: β-glucuronidase, as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6:3901–3907.
Lindstrom, J.T., L.O. Vodkin, R.W. Harding, R.M. Goeken. 1990. Expression of soybean lectin gene deletions in tobacco. Dev. Genet. 11, 160–167.
McCabe, D.E., W.F. Swain, B.J. Martinell, P. Christou, 1988. Stable transformation of soybean (Glycine max) by particle acceleration. Bio/Tech. 6:923–926.
Napoli, C., C. Lemieux, R. Jorgensen. 1990. Introduction of a chimeric chalcone synthase gene into petunia results in reversible co-suppression of homologous genes in trans. Plant Cell 2:279–289.
Okamura, J.K., K.D. Jofuku, R.B. Goldberg. 1986. Soybean seed lectin gene and flanking nonseed protein genes are developmentally regulated in transformed tobacco plants. Pro. Natl. Acad. Sci. USA 83:8240–8244.
Perez-Grau, L. and R.B. Goldberg. 1989. Soybean seed protein genes are regulated spatially during embryogenesis. Plant Cell 1:1095–1109.
Pull, S.P., S.G. Pueppke, T. Hymowitz, J.H. Orf. 1978. Soybean lines lacking the 120,000 dalton seed lectin. Science 200:1277–1279.
Russell, J.A., M.K. Roy, J.C. Sanford. 1992. Physical trauma and tungsten toxicity reduce the efficiency of biolistic transformation. Plant Physiol. 98:1050–1056.
Sambrook, J., E.F. Fritsch and T. Maniatis. 1989.Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, New York.
Sato, S., C. Newell, K. Kolacz, L. Tredo, J. Finer, M. Hinchee. 1993. Stable transformation via particle bombardment in two different soybean regeneration systems. 12:408–413.
Van der Krol, A.R., L.A. Mur, M. Beld, J.N.M. Mol. 1990. Flavonoid genes in petunia: Addition of a limited number of gene copies may lead to a suppression of gene expression. Plant Cell 2:291–299.
Vodkin, L.O. and N.V. Raikhel. 1986. Soybean lectin and related, proteins in seeds and roots of Le+ and Le- soybean varieties. Plant Physiol. 81:558–565.
Vodkin, L.O., P.R. Rhodes, R.B. Goldberg. 1983. A lectin gene insertion has the structural features of a transposable element. Cell 34:1023–1031.
Walling, L., G.N. Drews, R.B. Goldberg. 1986. Transcriptional and post-transcriptional regulation of soybean seed protein mRNA levels. Proc. Natl. Acad. Sci. USA 83:2123–2127.
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Cho, MJ., Widholm, J.M. & Vodkin, L.O. Cassettes for seed-specific expression tested in transformed embryogenic cultures of soybean. Plant Mol Biol Rep 13, 255–269 (1995). https://doi.org/10.1007/BF02670904
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DOI: https://doi.org/10.1007/BF02670904