DOI QR코드

DOI QR Code

Overproduction of 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) confers resistance to the herbicide glyphosate in transgenic rice

  • Lee, Soo-In (Department of Agricultural Biotechnology, National Academy of Agricultural Science (NAAS), RDA) ;
  • Kim, Hyun-Uk (Department of Agricultural Biotechnology, National Academy of Agricultural Science (NAAS), RDA) ;
  • Shin, Dong-Jin (Department of Agricultural Biotechnology, National Academy of Agricultural Science (NAAS), RDA) ;
  • Kim, Jin-A (Department of Agricultural Biotechnology, National Academy of Agricultural Science (NAAS), RDA) ;
  • Hong, Joon-Ki (Department of Agricultural Biotechnology, National Academy of Agricultural Science (NAAS), RDA) ;
  • Kim, Young-Mi (Department of Agricultural Biotechnology, National Academy of Agricultural Science (NAAS), RDA) ;
  • Lee, Yeon-Hee (Department of Agricultural Biotechnology, National Academy of Agricultural Science (NAAS), RDA) ;
  • Koo, Bon-Sung (Department of Agricultural Biotechnology, National Academy of Agricultural Science (NAAS), RDA) ;
  • Kwon, Sun-Jong (Department of Agricultural Biotechnology, National Academy of Agricultural Science (NAAS), RDA) ;
  • Suh, Seok-Chul (Department of Agricultural Biotechnology, National Academy of Agricultural Science (NAAS), RDA)
  • 투고 : 2011.11.02
  • 심사 : 2011.11.15
  • 발행 : 2011.12.31

초록

Plants expressing Agrobacterium sp. strain CP4 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) are known to be resistant to glyphosate, a potent herbicide that inhibits the activity of the endogenous plant EPSPS. In order to develop herbicide-resistant rice, we prepared transgenic rice plants with CP4 EPSPS gene under the control of CaMV 35S promoter for over-expression. A recombinant plasmid was transformed into rice via Agrobacterium-mediated transformation. A large number of transgenic rice plants were obtained with glyphosate and most of the transformants showed fertile. The integration and expression of CP4 EPSPS gene from regenerated plants was analyzed by Southern and northern blot analysis. The transgenic rice plants had CP4 EPSPS enzyme activity levels more than 15-fold higher than the wild-type plants. EPSPS enzyme activity of transgenic rice plants was also identified by strip-test method. Field trial of transgenic rice plants further confirmed that they can be selectively survived at 100% by spay of glyphosate (Roundup$^{(R)}$) at a regular dose used for conventional rice weed control.

키워드

참고문헌

  1. An G, Evert PR, Mitra A, Ha SB (1988) Binary vectors. In: Gelvin SB, Schilperoort RA (eds) Plant Molecular Biology Mannual A3, pp. 1-19. Kluwer Academic Press, Dordrecht
  2. Baerson SR, Rodriguez DJ, Tran M, Feng Y, Biest NA, Dill GM (2002) Glyphosate-resistant goosegrass. Identification of a mutation in the target enzyme 5-enolpyruvylshikimate-3-phosphate synthase. Plant Physiol 129: 1265-1275 https://doi.org/10.1104/pp.001560
  3. Barry G, Kishore G, Padgette S, Taylor M, Kolacz K, Weldon M, Re D, Fincher K, Hallas L (1992) Inhibitors of amino acid biosynthesis: strategies for imparting glyphosate tolerance to crop plants. Curr Top Plant Physiol 7: 139-145
  4. Cao J, Duan X, McElroy D, Wu R (1992) Regeneration of herbicide resistant transgenic rice plants following micro-projectile-mediated transformation of suspension culture cells. Plant Cell Rep 11:586-591
  5. Chu CC, Wang CS, Sun CS, Hsu C, Yin KC, Chu CY, Bi FY (1975) Establishment of an efficient medium for anther culture of rice through comparative experiments on the nitrogen sources. Sci Sin 18:659-668
  6. Comai LD, Facciotti D, Hiatt WR, Thompson G, Rose RE, Stalker DM (1985) Expression in plants of a mutant aroA gene from Salmonella tymphimurium confers tolerance to glyphosate. Nature 317:741-744 https://doi.org/10.1038/317741a0
  7. Daniell H, Datta R, Varma S, Gray S, Lee S-B (1998) Containment of herbicide resistance through genetic engineering of the chloroplast genome. Nature Biotech 16:345-348 https://doi.org/10.1038/nbt0498-345
  8. Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation version II. Plant Mol Biol Rep 1:19-21 https://doi.org/10.1007/BF02712670
  9. Ditta G, Stanfield S, Corbin D, Helinski DR (1980) Broad host range DNA cloning system for Gram-negative bacteria: Construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci USA 77:7347-7351 https://doi.org/10.1073/pnas.77.12.7347
  10. Franz JE (1985) In The Herbicide Glyphosate (Ed. by Grossman E, Atkinson D), pp3-17. London: Butterworths
  11. Giri CC, Laxmi GV (2000) Production of transgenic rice with agronomically useful genes: an assessment. Biotechnol adv 18:653-683 https://doi.org/10.1016/S0734-9750(00)00053-7
  12. Haslam E (1993) Acid: Metabolism and Metabolites. Wiley, Chichester, UK
  13. Heckern GR, Amstrong CL, Astwood JD, Behr CF, Bookout JT, Brown SM, Cavato TA, DeBoer DL, Deng MY, George C, Hillyard JR, Hironaka CM, Howe AR, Jekse EH, Ledesma BE, Lee TC, Lirette RP, Mangano ML, Mutz JN, Qi Y, Rodriguez RE, Sihdu SR, Silvanovich A, Stoecker MA, Yingling RA, You J (2005) Development and characterization of a CP4 EPSPS-based, glyphosate-tolerant corn event. Crop Sci 45:329-339 https://doi.org/10.2135/cropsci2005.0329
  14. Hiei Y, Ohta S, Komari T, Kumashiro T (1994) Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J 6:271-282 https://doi.org/10.1046/j.1365-313X.1994.6020271.x
  15. Hinchee MA, Padgette SR, Kishore GM, Delannay X, Fraley RT (1993) Herbicide-Tolerant Crops. In: Kung S-K, Wu R (eds) Transgenic Plants vol. 1, pp 243-263. Academic Press, London
  16. Howe AR, Gasser CS, Brown SM, Padgette SR, Hart J, Parker GB, Fromm ME, Amstrong CL (2002) Glyphosate as a selectable agent for the production of fertile transgenic maize (Zea mays L.) plants. Mol Breed 10:153-164 https://doi.org/10.1023/A:1020396708088
  17. Kahrizi D, Salmanian AH, Afshari A, Moieni A, Mousavi A (2007) Simultaneous substitution of Gly96 to Ala and Ala183 to Thr in 5-enolpyruvylshikimate-3-phosphate synthase gene of E. coli (k12) and transformation of rapeseed (Brassica napus L.) on order to make tolerance to glyphosate. Plant Cell Rep 26:95-104 https://doi.org/10.1007/s00299-006-0208-4
  18. Krieb R, Zeng Q (2002) Herbicide-resistant transgenic canola PV-BNGT04 (RT73) and compositions and methods for detection thereof. Monsanto Technology L. L. C. WO 0236831A. St. Louis, MO: Monsato Technology
  19. Komari T, Hiei Y, Saito Y, Kumashiro T (1996) Vectors carrying two separate T-DNAs for co-transformation of higher plants mediated by Agrobacterium tumefaciens and segregation of transformants free from selection markers. Plant J 10:165-174 https://doi.org/10.1046/j.1365-313X.1996.10010165.x
  20. Nafziger ED, Widholm JM, Steinrucken HC, Killmer JL (1984) Selection and characterization of a carrot cell line tolerant to glyphosate. Plant Physiol 76:571-574 https://doi.org/10.1104/pp.76.3.571
  21. Lee SI, Kim HU, Lee Y-H, Suh S-C, Lim YP, Lee H-Y, Kim H-I (2001) Constitutive- and seed-specific expression of a maize lysine-feedback-insensitive dihydrodipicolinate synthase gene leads to increased free lysine levels in rice seeds. Molecular Breed 8:75-84 https://doi.org/10.1023/A:1011977219926
  22. Lin C, Feng J, Xu X, Zhao T, Ye G, Shen Z (2008) A built-in strategy for containment of transgenic plants: creation of selectively terminable transgenic rice. PLoS one 3:1-6
  23. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473-497 https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  24. Nida DL, Kolacz KH, Beuhler RE, Deaton WR, Schuler WR, Amstrong TA, Taylor ML, Evert CC, Logan GL, Padgette SR, Fuchs RL (1996) Glyphosate-tolerant cotton: genetic characterization and protein expression. J Agri Food Chem 44:1960-1966 https://doi.org/10.1021/jf9505640
  25. Padgette SR, Kolacz KH, Delannay X, Re DB, LaVallee BJ, Tinius CN, Rhodes WK, Otero YI, Barry GF, Eichholtz DA, Peschke VM, Nida DL, Taylor NB, Kishore GM (1995) Development, identification and characterization of a glyphosate-tolerant soybean line. Crop Sci 35:1451-1461 https://doi.org/10.2135/cropsci1995.0011183X003500050032x
  26. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
  27. Shah D, Horsch R, Klee HJ, Kishore GM, Winter JA, Tumer N, Hironaka CM, Sanders PR, Gasser CS, Aykent S, Siegel NR, Rogers SG, Fraley RT (1986) Engineering herbicide tolerance in transgenic plants. Science 233:478-481 https://doi.org/10.1126/science.233.4762.478
  28. Steinruecken HC, Amrhein N (1980) The herbicide glyphosate is a potent inhibitor of 5-enolpyruvyl shikimic acid 3-phosphate synthase. Biochem Biophys Res Commun 94:1207-1212 https://doi.org/10.1016/0006-291X(80)90547-1
  29. Wang HY, Li YF, Xie LX, Xu P (2003) Expression of a bacterial aroA mutant, aroA-M1, encoding 5-enolpyruvylshikimate-3-phosphate synthase for the production of glyphosate-resistant tobacco plants. J Plant Res 116:455-460 https://doi.org/10.1007/s10265-003-0120-8