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Molecular Breeding

, Volume 13, Issue 4, pp 301–312 | Cite as

Field evaluation and risk assessment of transgenic indica basmati rice

  • Khurram Bashir
  • Tayyab Husnain
  • Tahira Fatima
  • Zakia Latif
  • Syed Aks Mehdi
  • Sheikh Riazuddin
Article

Abstract

We report the first field trial of different transgenic lines of Indica Basmati rice (B-370) expressing cry1Ac and cry2A genes. Different transgenic lines were grown under field conditions for two consecutive years, according to RCBD and Split Plot Design respectively. All the biosafety measures were taken into consideration. Sixty neonate larvae of yellow stem borer were artificially infested into each plant in three installments. Data was recorded in terms of dead hearts and white heads at vegetative and flowering stage respectively. Transgenic lines exhibited inherent ability to protect rice plants from target insects (p<0.01). Natural infestations of rice skipper and rice leaf folder were also observed and transgenic plants were statistically superior to their untransformed counterparts. Green house whole plant bioassays were done by infesting two 2nd instar larvae of rice leaf folder per tiller. Transgenics were 96% more resistant than untransformed control plants. The presence of cry genes was observed with Dot blot, PCR and Southern blot analysis, while ELISA and Western blot analysis confirmed the expression of Cry proteins. All lines expressed higher level of Cry proteins when compared with commercially released cultivars of Bt cotton, maize and potato. It was also observed that although toxin titer substantially decreased with increasing age of the plants, it remained well within the limits to kill the target insects. Morphological studies showed significant variation for days to maturity, plant height and panicle length. Cooking qualities of seeds harvested from these lines were compared with the untransformed control. The transgenic lines had no effect on non-target insects (insects belonging to orders other than diptera and lepidoptera) and germination of three local varieties of wheat. Chances of gene spread were calculated at a level of 0.18% cross pollination in experimental lines.

cry1Ac cry2A Infestations Insect resistance Oryza sativaYSB Field testing of Bt rice 

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References

  1. Alam M.F., Abrigo E., Datta K., Datta S.K., Oliva N., Tu J. and Virmani S.S. 1999. Transgenic insect-resistant maintainer line (IR68899B) for improvement of hybrid rice. Plant Cell Rep. 18: 572–575.Google Scholar
  2. Alinia F., Ghareyazie B., Rubia L.G., Bennett J. and Cohen M.B. 2000. Effect of plant age, larval age, and fertilizer treatment on resistance of a cry1Ab transformed aromatic rice to lepidopterous stem borers and foliage feeders. J. Econ. Entomol. 93: 484–493.Google Scholar
  3. Bradford M.M. 1976. A rapid and sensitive method for the quantification of micro gram quantities of protein utilizing the principle of protein dye binding. Anzl. Biochem. 72: 248–254.Google Scholar
  4. Cheng X., Sardana R., Kaplan H. and Altosaar I. 1998. Agrobacterium-transformed rice plants expressing synthetic cryIA(b) and cryIA(c) genes are highly toxic to striped stem borer and yellow stem borer. Proc. Natl. Acad. Sci. USA 95: 2767–2772.Google Scholar
  5. Cohen M.B., Gould F. and Bentur J.S. 2000. Bt. rice: practical steps to sustainable use. IRRN 25: 4-10.Google Scholar
  6. Crowther J.R. 1995. ELISA: Theory and Practice. Methods in Molecular Biology. Volume 42. Human Press, New Jersey, USA, pp. 38-39.Google Scholar
  7. Datta K., Vasquez A., Tu J., Torrizo L., Alam M.F., Oliva N., Abrigo E., Khush G.S. and Datta S.K. 1998. Constitutive and tissue-specific differential expression of the cryIA(b) gene in transgenic rice plants conferring resistance to rice insect pests. Theor. and Appl. Genet. 97: 20–30.Google Scholar
  8. Delannay X., LaVallee B.J., Proksch R.K., Fuchs R.L., Sims S.K., Greenplate J.T., Marrone P.G., Dodson R.B., Augustine J.J., Layton J.G. and Fischhoff D.A. 1989. Field performance of transgenic tomato plants expressing Bacillus thuringiensis var kurstaki insect control protein. Bio/technology 7: 1265–1269.Google Scholar
  9. Dong Y. J., Suzuki F. T. and Terao H. 2000. Inheritance of aroma in four rice cultivars (Oryza sativa L.) IRRN 25: 15.Google Scholar
  10. Fatima T. 2001. Studies on expression of foreign genes in rice (Oryza sativa). Ph.D. Thesis, Univ. of the Punjab, Lahore, Pakistan.Google Scholar
  11. Fitt G.P., Mares C.L. and Llewellyn D.J. 1994. Field evaluation and potential ecological impact of transgenic cottons (Gossypium hirsutum) in Australia. Biocontrol Sci. & Technol. 4: 535–548.Google Scholar
  12. Fitt G.P. 1998. Efficacy of Ingard cotton – patterns and consequences, In Proceedings, 9th Australian Cotton Conference. 12–14 August 1998, Broadbeach, Queensland. Australian Cotton Growers' Research Association, Wee Waa, Australia, pp. 233–245.Google Scholar
  13. Frutos R., Rang C. and Royer M. 1999. Managing insect resistance to plants producing Bacillus thuringiensis toxins. Crit. Rev. Bio-technol. 19: 227–276.Google Scholar
  14. Fujimoto H., Itoh K., Yamamoto M., Kayozuka J. and Shimamoto K. 1993. Insect resistant rice generated by a modified delta endotoxin genes of Bacillus thuringiensis. Bio/technology 11: 1151–1155.Google Scholar
  15. Gould F. 1998. Sustainability of transgenic insecticidal cultivars: integrating pest genetics and ecology. Annu. Rev. Entomol. 43: 701–726.Google Scholar
  16. Husnain T., Jan A., Maqbool S.B., Datta S.K. and Riazuddin S. 2002. Variability in expression of insecticidal Cry1Ab gene in indica basmati rice. Euphytica: 128: 121–128.Google Scholar
  17. INGER. 1996. Standard evaluation system for Rice. IRRI, Los Banos, Philippines.Google Scholar
  18. IRRI 1972. Annual report for (1971). International Rice research Institute, Los Bonos, Philippines.Google Scholar
  19. James C. 2001. Global Status of Commercialized Transgenic Crops: ISAAA Briefs No. 24: Preview. ISAAA, Ithaca, New York, USA.Google Scholar
  20. Jenkins J.N., Parot W.L., McCarty J.C., Barton K.A. and Umbeck P.F. 1991. Field test of transgenic cotton containing a Bacillus thuringiensis gene. Technical Bulletin Mississippi Arric. Forestry Exp. Station 174: 10.Google Scholar
  21. Jiang J., Linscombe S.D., Wang J. and Oard J.H. 2000. Field evaluation of transgenic rice (Oryza sativa L.) produced by Agrobacterium and particle bombardment methods. Plant and animal genome VIII conference. Town and country Hotel, San Diego, California, USA, January 9-12, 2000.Google Scholar
  22. Juliano B.O. 1971. A simplified assay for milled rice amylose. Cereal Sci. Today 16: 334-338, 340, 360.Google Scholar
  23. Khan Z.R., Litsinger J.A., Barrion A.T., Villanueva F.F.D., Fernandez N.J. and Taylor L.D. 1991. World bibliography of rice stem borers. IRRI, Los Banos, Philippines, pp. 1794–1990.Google Scholar
  24. Koziel M.G., Beland G.L., Bowman C., Carozzi N.B., Crenshaw R., Crossland L., Dawson J., Desai N., Hill M., Kadwell S., Launis K., Lewis K., Maddox D., McPherson K., Meghji M.R., Merlin E., Rhodes R., Warren G.W., Wright M. and Evola S.V. 1993. Field performance of elite transgenic maize plants expressing an insecticidal protein derived from Bacillus thuringiensis. Bio/technology 11: 194–200.Google Scholar
  25. Little R.R., Hilder G.B. and Dawson E.H. 1958. Differencial effect of dilute alkali on 25 varieties of milled rice. Cereal Chem. 35: 111–126.Google Scholar
  26. Maqbool S.B., Husnain T., Raizuddin S. and Christou P. 1998. Effective control of yellow rice stem borer and rice leaf folder in transgenic rice indica varieties Basmati 370 and M 7 using novel 311-1-endotoxin cry2A Bacillus thuringiensis gene. Mol. Breed. 4: 501–507.Google Scholar
  27. Maqbool S.B., Raizuddin S., Loc T.N., Gatehouse A.M.R., Gatehouse J.A. and Christou P. 2001. Expression of multiple insecticidal genes confers broad resistance against a range of different rice pests. Mol. Breed. 7: 85–93.Google Scholar
  28. McGaughey W.H. 1985. Insect resistance to the biological insecticide Bacillus thuringiensis. Science 229: 193–195.Google Scholar
  29. McGaughey W.H., Gould F. and Gelernter W. 1998. Bt resistance management: a plan for reconciling the needs of the many stakeholders in Bt-based products. Nature Biotechnol. 16: 144–146.Google Scholar
  30. McGaughey W.H. and Whalon M.E. 1992. Managing insect resistance to Bacillus thuringiensis toxins. Science 258: 1451–1455.Google Scholar
  31. Mellon M. and Rissler J. (eds) 1998. Now or never: serious new plans to save a natural pest control.Union of Concerned Scientists, Cambridge, Massachusetts, USA.Google Scholar
  32. Nayak P., Basu D., Das S., Basu A., Ghosh D., Ramakrishnan N.A., Ghosh M. and Sen S.K. 1997. Transgenic elite indica rice plants expressing CryIAc delta-endotoxin of Bacillus thuringiensis are resistant against yellow stem borer (Scirpophaga incertulas). Proc. Natl. Acad. Sci. USA 94: 2111–2116.Google Scholar
  33. NBC (National Biosafety Committee Pakistan) 1999. Biosafty guidelines in genetic engineering and biotechnology. Ministry of Environ., Local Govt. Rural Develop., Govt., Pakistan.Google Scholar
  34. Niluni Z., Hizukure S., Kamagai K., Hasegawa H., Horiwaki T., Fukui T., Doi K., Nara S. and Maeda L. 1969. The effect of temperature during the maturation period on physiochemical preoperties of Potato and Rice starches.Mem. Inst. Sci Ind. Res. Osaka Univ. 36: 1-27.Google Scholar
  35. NRC (National Research Council, USA) 2000. Genetically modified pestprotected plants: science and regulation. Natl. Acad. Press, Washington D.C., USA.Google Scholar
  36. Oard J., Cohen M.A., Linscombe S., Gealy D. and Gravois K. 2000. Field evaluation of seed production, shattering, and dormancy in hybrid populations of transgenic rice (Oryza sativa) and the weed, red rice (Oryza sativa). Plant Science 157(1): 13–22.Google Scholar
  37. Orr D.B. and Landis D.L. 1997. Oviposition of European Corn Borer (Lepidoptera: Pyralidae) and impact of natural enemy populations in transgenic versus isogenic corn. J. Econ. Entomol. 90: 905–909.Google Scholar
  38. Pathak M.D. and Khan Z.R. 1994. Insect pests of rice. IRRI, Los Banos, Philippines.Google Scholar
  39. Paul C.M. 1977. Variability in amylose content of rice. MS thesis, Univ. Philippines, Los Banos, Philippines, p. 82.Google Scholar
  40. Perlak F.J., Deaton R.W., Armistrong T.A., Fuchs R.L., Sims S.R., Greenplates J.T. and Fischhoff D.A. 1990. Insect resistant cotton Plants. Bio/technology 8: 939–943.Google Scholar
  41. Perlak F.J., Stone T.B., Muskopf Y.N., Petersen L.J., Parker G.B., McPherson S.A., Wyman J., Love S., Reed G., Biever D. and Fischhoff D. A. 1993. Genetically improved potatoes: protection from damage by Colorado potato beetles. Plant Mol. Biol. 22: 313–321.Google Scholar
  42. Roger P. and Dunwell J.M. 1993. Field performance of derived generation sof transgenic tobacco. Theor. Appl. Genet. 86: 875–879.Google Scholar
  43. Salim M. and Masih R. 1987. Efficacy of insecticides against rice stem borer at NARC, Islamabad. Pak. J. Agric. Res.: 477–479.Google Scholar
  44. Shu Q.-Y., Ye G., Cui H., Cheng X., Xiang Y., Wu D., Gao M., Xia Y., Hu Cui, Sardana R. and Altossar I. 2000. Transgenic rice plants with a synthetic cry1Ab gene from Bacillus thuringiensis were highly resistant to eight lepidopteran rice pest species. Mol. Breed. 6: 433–439.Google Scholar
  45. Sims S.R. 1995. Bacillus thuringiensis var. kurstaki (CryIA)) protein expressed in transgenic cotton: effects on beneficial and other non-target insects. Southwestern Entomologist 20: 493–500.Google Scholar
  46. Stansel J.W. 1965. The influence of heredity and environment on endosperm characteristics of rice (Oryza sativa). Ph. D. thesis, Purdue Univ., Indiana, USA, p. 140.Google Scholar
  47. Tu J., Zhang G., Datta K., Xu C., He Y., Zhang Q., Khush G.S. and Datta S.K. 2000. Field performance of transgenic elite commercial hybrid rice expressing Bacillus thuringiensis δ-endotoxin. Nature Biotechnol. 18: 1101–1104.Google Scholar
  48. Vaeck M., Reynaerts A., Hoftey H., Jansens S., DeBeuckleer M., Dean C., Zabeau M., Van Montagu M. and Leemans J. 1987. Transgenic plants protected from insect attack. Nature 327: 33–37.Google Scholar
  49. Williams V.R., Wu W.T., Tsai H.Y. and Bates H.G. 1958. Varietal difference in amylose contents of rice starch. J. Agr. Food Chem. 6: 47–48.Google Scholar
  50. Ye G.-Y., Shu Q.-Y., Yao H.-W., Cui H.-R., Cheng X.-Y., Hu C., Xia Y.-W., Gao M.-W. and Altosaar I. 2001a. Field evaluation of resistance of transgenic rice containing a synthetic cryIAb gene from Bacillus thuringiensis Berliner to two stem borers.J. Econ. Entomol. 94: 271–276.Google Scholar
  51. Ye G.-Y., Tu J., Datta K., Datta S.K. 2001b. Transgenic IR72 with fused Bt Gene cry1Ab/cry1Ac from Bacillus thuringiensis is resistant against four lepidopteran species under field conditions. Plant Biotech. 18: 125–133.Google Scholar

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Khurram Bashir
    • 1
  • Tayyab Husnain
    • 1
  • Tahira Fatima
    • 1
  • Zakia Latif
    • 1
  • Syed Aks Mehdi
    • 2
  • Sheikh Riazuddin
    • 1
  1. 1.National Center of Excellence in Molecular BiologyUniversity of The PunjabLahorePakistan
  2. 2.Rice Research InstituteKala Shah Kaku, LahorePakistan

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