Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

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

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 to check access.

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

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

  2. 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

  3. 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

  4. 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

  5. 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

  6. 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

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

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

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

  10. 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

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

  12. 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

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

  14. 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

  15. 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

  16. 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

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

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

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

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

  21. 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

  22. 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

  23. 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

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

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

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

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

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

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

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

  31. 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

  32. 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

  33. 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

  34. 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

  35. 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

  36. 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

Download references

Author information

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

Keywords

  • Agrobacterium
  • Bangladeshi Indica rice
  • Oryza sativa L.
  • transgenic plants