Skip to main content
SpringerLink
Log in

Agrobacterium-mediated transformation of Indica rice genotypes: an assessment of factors affecting the transformation efficiency

  • Published:
Plant Cell, Tissue and Organ Culture Aims and scope Submit manuscript

Abstract

An efficient Agrobacterium-mediated method for transformation of popular Bangladeshi Indica rice genotypes has been developed. Mature embryo-derived calluses as well as immature embryos were used as the target material. Transgenic plant production frequency was higher using the immature embryos than mature embryo-derived calluses. However, 3-week-old mature embryo-derived calluses served as an excellent starting material. The super-binary vector (pTOK233) was generally more effective than the binary vector (pC1301-Xa21mSS) particularly with recalcitrant Bangladeshi genotypes such as BR22. However, transformation of the Japonica cultivar Taipei-309 was equally effective with either plasmid. Inclusion of acetosyringone (200 μM) in co-cultivation media proved essential for successful transformation and the optimum co-cultivation period found was to be 3 days. A large number of morphologically normal, fertile transgenic plants were obtained which expressed gus as determined by histochemical staining. Integration of the hpt gene into the genome of transgenic plants was confirmed by molecular analysis. Mendelian inheritance of transgenes (hpt and gus gene) was observed in T<inf>1</inf> progeny.

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

Access this article

Log in via an institution

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

Abbreviations

AS:

acetosyringone

2,4-D:

2,4-dichlorophenoxyacetic acid

BA:

6-benzylaminopurine

GUS:

β-glucuronidase (gene product)

hpt :

hygromycin phosphotransferase gene

LB:

Luria–Bertani

MS:

Murashige and Skoog

NAA:

α-naphthalene acetic acid

SE:

standard errors of means

References

  • RR Aldemita TK Hodges (1996) ArticleTitleAgrobacterium tumefaciens-mediated transformation of Japonica and Indica rice varieties Planta 199 612–617

    Google Scholar 

  • M Al-Forkan B Power P Anthony KC Lowe MR Davey (2004) ArticleTitleAgrobacterium-mediated transformation of Bangladeshi Indica rices Cell. Mol. Biol. Lett. 9 287–300

    Google Scholar 

  • K Azhakanandam MS McCabe JB Power KC Lowe EC Cocking MR Davey (2000) ArticleTitleT-DNA transfer, integration, expression and inheritance in rice: effects of plant genotype and Agrobacterium super-virulence J. Plant Physiol. 157 429–439

    Google Scholar 

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

    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/Technology 9: 957–962

  • S Dai P Zheng P Marmey S Zhang W Tian S Chen RN Beachy C. Fauquet (2001) ArticleTitleComparative analysis of transgenic rice plants obtained by Agrobacterium-mediated transformation and particle bombardment Mol. Breed. 7 25–33

    Google Scholar 

  • SK Datta A Peterhans K Datta I. Potrykus (1990) ArticleTitleGenetically engineered fertile Indica rice recovered from protoplasts Bio/Technology 8 736–740

    Google Scholar 

  • MR Davey EL Rech BJ. Mulligan (1989) ArticleTitleDirect DNA transfer to plant cells Plant Mol. Biol. 13 273–285

    Google Scholar 

  • J Dong W Teng WG Buchholz TC. Hall (1996) ArticleTitleAgrobacterium-mediated transformation of Javanica rice Mol. Breed. 2 267–276

    Google Scholar 

  • K Edwards C Johnstone C. Thompson (1991) ArticleTitleA simple and rapid method for the preparation of plant genomic DNA for PCR analysis Nucleic Acids Res. 19 1349

    Google Scholar 

  • RB. Flavell (1994) ArticleTitleInactivation of gene expression in plants as a consequence of specific sequence duplication Proc. Natl. Acad. Sci. USA 91 3490–3496

    Google Scholar 

  • A Hayashimoto Z Li N. Murai (1990) ArticleTitleA polyethylene glycol-mediated protoplast transformation system for production of␣fertile transgenic rice plants Plant Physiol. 93 857–863

    Google Scholar 

  • Y Hiei T Komari T. Kubo (1997) ArticleTitleTransformation of rice mediated by Agrobacterium tumefaciens Plant Mol. Biol. 35 205–218

    Google Scholar 

  • Y Hiei S Ohta T Komari T. Kumashiro (1994) ArticleTitleEfficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA Plant J. 6 271–282

    Google Scholar 

  • A Hoekema PR Hirsch PJJ Hooykass RA Schilperoort (1983) ArticleTitleA binary plant vector strategy based on separation of vir- and T-region of the Agrobacterium tumefaciens Ti-plasmid Nature 303 179–180

    Google Scholar 

  • EE Hood GL Helmer RT Fraley MD. Chilton (1986) ArticleTitleThe hyper-virulence of Agrobacterium tumefaciens A281 is encoded in a region of pTiBo542 outside of T-DNA J.␣Bacteriol. 168 1291–1301

    Google Scholar 

  • ME Hoque (2002) Development of Agrobacterium-mediated transformation of rice Oryza sativa L genotypes grown in Bangladesh Ph.D. dissertation University of London UK

    Google Scholar 

  • S Jin T Komari MP Gordon EW. Nester (1987) ArticleTitleGenes responsible for the supervirulence phenotype of Agrobacterium tumefaciens A281 J. Bacteriol. 169 4417–4425

    Google Scholar 

  • HK Khanna SK Raina (1999) ArticleTitleAgrobacterium-mediated transformation of Indica rice cultivars using binary and superbinary vectors Aust. J. Plant Physiol. 26 311–324

    Google Scholar 

  • GS. Khush (1997) ArticleTitleOrigin, dispersal, cultivation and variation of rice Plant Mol. Biol. 35 25–34

    Google Scholar 

  • A Kohli M Leech P Vain DA Laurie P. Christou (1998) ArticleTitleTransgene organization in rice engineered through direct DNA transfer supports a two-phase integration mechanism mediated by the establishment of integration host spots Proc. Natl. Acad. Sci. USA 95 7203–7208

    Google Scholar 

  • T. Komari (1990) ArticleTitleTransformation of cultured cells of Chenopodium quinoa by binary vectors that carry a fragment of DNA from virulence region of pTiBo542 Plant Cell Rep. 9 303–306

    Google Scholar 

  • SP Kumpatla W Teng WG Buchholz TC. Hall (1997) ArticleTitleEpigenetic transcriptional silencing and 5-azacytidine-mediated reactivation of a complex transgene in rice Plant Physiol. 115 361–373

    Google Scholar 

  • T Murashige F. Skoog (1962) ArticleTitleA revised medium for rapid growth and bioassays with tobacco tissue cultures Physiol. Plant. 15 473–497

    Google Scholar 

  • WP Pawlowski DA. Somers (1996) ArticleTitleTransgene inheritance in plants genetically engineered by microprojectile bombardment Mol. Biotechnol. 6 17–30

    Google Scholar 

  • J Peng H Kononowicz TK Hodges (1992) ArticleTitleTransgenic Indica rice plants Theor. Appl. Genet. 83 855–863

    Google Scholar 

  • H Rashid S Yokoi K Toriyama K Hinata (1996) ArticleTitleTransgenic plant production mediated by Agrobacterium in Indica rice Plant Cell Rep. 15 727–730

    Google Scholar 

  • K Shimamoto R Terada T Izawa H. Fujimoto (1989) ArticleTitleFertile transgenic rice plants regenerated from transformed protoplasts Nature 338 274–276

    Google Scholar 

  • RH Smith EE. Hood (1995) ArticleTitleAgrobacterium tumefaciens transformation of monocotyledons Crop Sci. 35 301–309

    Google Scholar 

  • K Toriyama Y Arimoto H Uchimiya K. Hinata (1988) ArticleTitleTransgenic rice plants after direct gene transfer into protoplasts Bio/Technology 6 1072–1074

    Google Scholar 

  • GL Wang WY Song DL Ruan S Sideris PC. Ronald (1996) ArticleTitleThe cloned gene, Xa21, confers resistance to multiple Xanthomonas oryzae pv. oryzae isolates in transgenic plants Mol. Plant–Microbe Interact. 9 850–855

    Google Scholar 

  • Y Xu WG Buchholz RT DeRose TC. Hall (1995) ArticleTitleCharacterization of a rice gene family encoding root-specific proteins Plant Mol. Biol. 27 237–248

    Google Scholar 

  • H Yang HM Zhang MR Davey BJ Mulligan EC. Cocking (1988) ArticleTitleProduction of kanamycin resistant rice tissues following DNA uptake into protoplasts Plant Cell Rep. 7 421–425

    Google Scholar 

  • HM Zhang H Yang EL Rech TJ Golds AS Davis BJ Mulligan EC Cocking MR Davey (1988) ArticleTitleTransgenic rice plants produced by electroporation-mediated plasmid uptake into protoplasts Plant Cell Rep. 7 379–384

    Google Scholar 

  • J Zhang R Xu D Blackley MC Elliott DF. Chen (1997) ArticleTitleAgrobacterium-mediated transformation of elite Indica and Japonica rice cultivars Mol. Biotechnol. 8 223–231

    Google Scholar 

  • W Zhang R Wu (1988) ArticleTitleEfficient regeneration of transgenic plants from rice protoplasts and correctly regulated expression of the foreign gene in the plants Theor. Appl. Genet. 76 835–840

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Biotechnology Division, Bangladesh Rice Research Institute, Gazipur, 1701, Bangladesh

    Md. Enamul. Hoque

  2. Department of Agricultural Sciences, Imperial College London, Wye campus, Ashford, Kent, TN25 5AH, UK

    John W. Mansfield & Mark H. Bennett

Authors
  1. Md. Enamul. Hoque
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. John W. Mansfield
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mark H. Bennett
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Md. Enamul. Hoque.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hoque, M.E., Mansfield, J.W. & Bennett, M.H. Agrobacterium-mediated transformation of Indica rice genotypes: an assessment of factors affecting the transformation efficiency. Plant Cell Tiss Organ Cult 82, 45–55 (2005). https://doi.org/10.1007/s11240-004-6154-3

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-004-6154-3

Keywords

Search

Navigation