Skip to main content
Log in

Molecular and genetic characterization of elite transgenic rice plants produced by electric-discharge particle acceleration

  • Published:
Theoretical and Applied Genetics Aims and scope Submit manuscript

Abstract

The recovery of transgenic rice plants expressing a number of exogenous genes was reported previously. Using immature embryo explants as the target tissue, plasmids containing both selectable and screenable marker genes were introduced into elite rice varieties via electric-discharge particle acceleration. Co-integration, copy number, expression, and inheritance of these genes were analyzed. A 100% co-integration frequency was confirmed by Southern-blot analyses of R0 plants. The majority of transgenic plants contained between one and ten copies of exogenous DNA and molecular and genetic analyses of progeny indicated that all copies in almost all R0 plants were inherited as a single dominant hemizygous locus. Co-expression of unselected genes ranged from 30–66% for gus/hmr constructs, depending on the promotor used, and up to 90% for bar/hmr constructs. The integrative structures of two unlinked transgenic loci of a rare R0 plant were analyzed in detail by Southern-blot analysis of its progeny.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Allen GC, Hall GE, Childs LC, Weissinger AK, Spiker S, Thompson WF (1993) Scaffold attachment regions increase reporter gene expression in stably transformed plant cells. The Plant Cell 5:603–613

    Google Scholar 

  • Al-Shawl R, Kinnaird J, Burke J, Bishop JO (1990) Expression of a foreign gene in a line of transgenic mice is modulated by a chromosomal position effect. Mol Cell Biol 10:1192–1198

    Google Scholar 

  • Assad FF, Tucker KL, Signer ER (1993) Epigenetic repeat-induced gene silencing (RIGS) in Arabidopsis. Plant Mol Biol 22:1067–1085

    Google Scholar 

  • Battraw M, Hall TC (1990) Histochemical analysis of CaMV 35s promoter-β-glucuronidase gene expression in transgenic rice plants. Plant Mol Biol 15:527–538

    Google Scholar 

  • Battraw M, Hall TC (1992) Expression of chimeric neomycin phosphotransferase II gene in first and second generataion transgenic rice plants. Plant Sci 86:191–202

    Google Scholar 

  • Berry-Lowe SL, McKnight TD, Shah DM, Meagher RB (1982) The nucleotide sequence, expression, and evolution of one member of a multigene family encoding the small subunit of ribulose-1, 5-biphosphate carboxylase in soybean. J Mol Appl Genet 1:483–498

    Google Scholar 

  • Callis J, Fromm M, Walbot V (1987) Introns increase gene expression in cultured maize cells. Genes Dev 1:1183–1200

    Google Scholar 

  • Christou P, Swain WF (1990) Cotransformation frequencies of foreign genes in soybean cell cultures. Theor Appl Genet 79:337–341

    Google Scholar 

  • Christou P, Swain WF, Yang N-S, McCabe D (1989) Inheritance and expression of foreign genes in transgenic soybean plants. Proc Natl Acad Sci USA 86:1991–1995

    Google Scholar 

  • Christou P, Ford TL, Kofron M (1991) Production of transgenic rice (Oryza sativa L.) plants from agronomically important indica and japonica varieties via electric-discharge particle acceleration of exogenous DNA into immature zygotic embryos. Bio/Technol. 16:957–962

    Google Scholar 

  • Church GM, Gilbert W (1984) Genomic sequencing. Proc Natl Acad Sci USA 81:1991–1995

    CAS  PubMed  Google Scholar 

  • Datta SK, Datta K, Potrykus I (1990) Embryogenesis and plant regeneration from micropores of both indica and japonica rice (Oryza sativa). Plant Sci 67:83–88

    Google Scholar 

  • De Block M, Debrouwer D (1991) Two T-DNA's co-transformed into Brassica napus by a double Agrobacterium tumefaciens infection are mainly integrated at the same locus. Theor Appl Genet 82:257–263

    Google Scholar 

  • De Block M, Botterman J, Vandewiele M, Dockx J, Thoen C, Gossele V, Rao Movva N, Van Montagu M, Leemans J (1987) Engineering herbicide resistance in plants by expression of a detoxifying enzyme. EMBO J 6:2513–2518

    Google Scholar 

  • Dellaporta SL, Wood J, Hicks JB (1984) Maize DNA miniprep. In: Malmberg R, Messing J, Sussex I (eds) Molecular biology of plants. A laboratory course manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 36–37

    Google Scholar 

  • Depicker A, Stachel S, Dhaese P, Zambryski P, Goodman HM (1982) Nonpaline synthase: transcropt mapping and DNA sequence. J Mol Appl Genet 1:499–512

    Google Scholar 

  • Gardner R, Howarth A, Hahn P, Brown-Luedi M, Shepherd R, Messing J (1981) The complete nucleotide sequence of an infectious clone of cauliflower mosiac virus by M13mp7 shotgun sequencing. Nucleic Acids Res 9:2871–2888

    Google Scholar 

  • Goto F, Toki S, Uchimiya H (1993) Inheritance of a co-transferred foreign gene in the progenies of transgenic rice plants. Transgen Res 2:300–305

    Google Scholar 

  • Hart CM, Fischer B, Neuhaus J-M, Meinss F Jr (1992) Regulated inactivation of homologous gene expression in transgenic Nicotiana sylvestris plants containing a defense-related tobacco chitinase gene. Mol Gen Genet 235:179–188

    Google Scholar 

  • Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907

    CAS  PubMed  Google Scholar 

  • Jongsma M, Koorneef M, Zabel P, Hille J (1987) Tomato protoplast transformation: physical linkage and recombination of exogenous DNA. Plant Mol Biol 8:383–394

    Google Scholar 

  • Jorgensen R (1991) Beyond antisense-How do transgenes interact with homologous plant genes? Trends Biotechnol 9:266–267

    Google Scholar 

  • Kartzke S, Saedler H, Meyer P (1990) Molecular analysis of transgenic plants derived from transformants of protoplasts at various stages of the cell cycle. Plant Sci 67:63–72

    Google Scholar 

  • Krol AR van der, Mur LA, Beld M, Mol JNM, Stuitje AR (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

    Google Scholar 

  • Leisy DJ, Hnilo J, Zhao Y, Okita T (1989) Expression of a rice glutelin Promoter in transgenic tobacco. Plant Mol Biol 14:41–50

    Google Scholar 

  • Linn F, Heidmann I, Saedler H, Meyer P (1990) Epigenetic changes in the expression of the maize A1 gene in Petunia hybrida: role of numbers of integrated gene copies and state of methylation. Mol Gen Genet 222:329–336

    Google Scholar 

  • Pater S de, Hensgens LAM, Schilperoort RA (1990) Structure and expression of a light-inducible shoot-specific rice gene. Plant Mol Biol 15:399–406

    Google Scholar 

  • Peng J, Knonowicz H, Hodges TK (1992) Transgenic indica rice plants. Theor Appl Genet 83:855–863

    Google Scholar 

  • Peng J, Lyznik LA, Lee L, Hodges TK (1990) Cotransformation of indica rice protoplasts with GUSA and Neo genes. Plant Cell Rep 9:168–172

    Google Scholar 

  • Rathore KS, Chowdhury VK, Hodges TK (1993) Use of bar as a selectable marker gene and for the production of herbicide-resistant rice plants from protoplasts. Plant Mol Biol 21:871–884

    Google Scholar 

  • Russell DR, Wallace KM, Bathe JH, Martinell BJ, McCabe DE (1993) Stable transformation of Phaseolus vulgaris via electric-discharge mediated particle acceleration. Plant Cell Rep 12:165–169

    Google Scholar 

  • Saul MW, Potrykus I (1990) Direct gene transfer to protoplasts: fate of the transferred genes. Deve Genet 11:176–181

    Google Scholar 

  • Schocher RJ, Shillito RD, Saul MW, Paszkowski, Potrykus I (1986) Co-transformation of unlinked foreign genes into plants by direct gene transfer. Biotechnology 4:1093–1096

    Google Scholar 

  • Shirstat AH, Wilford N, Croy RRD (1989) Gene copy number and levels of expression in transgenic plants of a seed-specific gene. Plant Sci 61:75–80

    Google Scholar 

  • Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517

    CAS  PubMed  Google Scholar 

  • Stockhaus J, Eckes P, Blau A, Schell J, Willimitzer L (1987) Organ-specific and dosage-dependent expression of leaf stem-specific gene from potato after tagging and transfer into potato and tobacco plants. Nucleic Acids Res 15; 3479–3491

    Google Scholar 

  • Tada Y, Sakamoto M, Fujimura T (1990) Efficient gene introduction into rice by electroporation and analysis of transgenic plants: use of electroporation buffer lacking chloride inons. Theor Appl Genet 80:475–480

    Google Scholar 

  • Timko MP, Herdies L, de Almeida E, Cashmore AR, Leemans J, Krebbers E (1988) Genetic engineering of nuclear-encoded components of the photosynthetic apparatus in Arabadopsis. In: Phillips M (ed) The impact of chemistry on biotechnology. ACS Symp Serv 362, pp 278–295

  • Van den Elzen PJM, Townsend J, Lee KY, Bedbrook JR (1985) A chimeric hygromycin resistance gene as a selectable marker in plant cells. Plant Mol Biol 5:299–302

    Google Scholar 

  • Yanisch-Perron C, Vieria J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequence of the M13 mp 18 and pUC19 vectors. Gene 33:103–119

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by I. Potrykus

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cooley, J., Ford, T. & Christou, P. Molecular and genetic characterization of elite transgenic rice plants produced by electric-discharge particle acceleration. Theoret. Appl. Genetics 90, 97–104 (1995). https://doi.org/10.1007/BF00221001

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00221001

Key words

Navigation