biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

Biologia plantarum 51:242-248, 2007 | DOI: 10.1007/s10535-007-0048-2

Agrobacterium-mediated transformation of Cry1C, Cry2A and Cry9C genes into Gossypium hirsutum and plant regeneration

X. Guo1, C. Huang1, S. Jin1, S. Liang1, Y. Nie1, X. Zhang1,*
1 National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, Hubei, P.R. China

Three constructs harbouring novel Bacillus thuringiensis genes (Cry1C, Cry2A, Cry9C) and bar gene were transformed into four upland cotton cultivars, Ekangmian10, Emian22, Coker201 and YZ1 via Agrobacterium-mediated transformation. With the bar gene as a selectable marker, about 84.8 % of resistant calli have been confirmed positive by polymerase chain reaction (PCR) tests, and totally 50 transgenic plants were regenerated. The insertions were verified by means of Southern blotting. Bioassay showed 80 % of the transgenic plantlets generated resistance to both herbicide and insect. We optimized conditions for improving the transformation efficiency. A modified in vitro shoot-tip grafting technique was introduced to help entire transplantation. This result showed that bar gene can replace antibiotic marker genes (ex. npt II gene) used in cotton transformation.

Keywords: Agrobacterium tumefaciens; bar gene; cotton; novel Bt genes
Subjects: Agrobacterium tumefaciens; chromosome; cotton; Gossypium hirsutum; growth analysis, plant development, biomass and yield enhancement; herbicide; in vitro culture, regeneration, proliferation, differentiation; nutrient medium, Murashige and Skoog (MS); polymerase chain reaction (PCR); Southern blot analysis; transgenic plants; triterpenes, triterpenoids

Received: September 16, 2005; Accepted: May 11, 2006; Published: June 1, 2007  Show citation

ACS AIP APA ASA Harvard Chicago IEEE ISO690 MLA NLM Turabian Vancouver
Guo, X., Huang, C., Jin, S., Liang, S., Nie, Y., & Zhang, X. (2007). Agrobacterium-mediated transformation of Cry1C, Cry2A and Cry9C genes into Gossypium hirsutum and plant regeneration. Biologia plantarum, 51(2), 242-248. doi: 10.1007/s10535-007-0048-2
Share...
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)
    • Open full article

    References

    1. Agrawal, D.C., Banerjee, A.K., Kolala, R.R., Dhage, A.B., Kulkarni, A.V., Nalawade, S.M., Hazra, S., Krishnamurthy, K.V.: In vitro induction of multiple shoots and plant regeneration in cotton (Gossypium hirsutum L.).-Plant Cell Rep. 16: 647-652, 1997. Go to original source...
    2. Cao, J., Duan, X., McElroy, D., Wu, R.: Regeneration of herbicide resistant transgenic rice plants following microprojectile-mediated transformation of suspension culture cells.-Plant Cell Rep. 11: 586-591, 1992. Go to original source...
    3. Chaudhary, B.S., Kumar, K.V., Prasad, S.K., Oinam, G.S., Burma, P.K., Pental, D.: Slow desiccation leads to high-frequency shoot recovery from transformed somatic embryos of cotton (Gossypium hirsutum L. cv. Coker 310 FR).-Plant Cell Rep. 21: 955-960, 2003. Go to original source...
    4. De Block, M., Botterman, J., Vandewiele, M., Dockx, J., Thoen, C., Gossele, V., Rao, V., Movva, N., Thompson, C., Van Montagu, M.: Engineering herbicide resistance in plants by expression of a detoxifying enzyme.-EMBO J. 6: 2513-2518, 1987. Go to original source...
    5. De Block, M., De Brouwer, D., Tenning, P.: Transformation of Brassica napus and Brassica oleracea using Agrobacterium tumefaciens and the expression of the bar and neo genes in the transgenic plants.-Plant Physiol. 91: 694-701, 1989. Go to original source...
    6. Gamborg, O.L., Miller, R.A., Ojima, K.: Nutrient requirements of suspension culture of soybean roots cells.-Exp. Cell Res. 50: 150-158, 1968. Go to original source...
    7. Jin, S.X., Zhang, X.L., Liang, S.G., Nie, Y.C., Guo, X.P., Huang, C.: Factors affecting stable transformation and plant regeneration during transforming embryogenic callus of Upland cotton (Gossypium hirsutum L.) via Agrobacterium tumefaciens.-Plant Cell Tissue Organ Cult. 81: 229-237, 2005. Go to original source...
    8. Keller, G., Spatola, L., McCable, D., Martinell, B., Swain, W., John, M.E.: Transgenic cotton resistant to herbicide bialaphos.-Transgenic Res. 6: 385-392, 1997. Go to original source...
    9. Leelavathi, V.G., Sunnichan, R.K., Umria, G.P., Vijaykanth, R.K., Bhatnagar Reddy, V.: A simple and rapid Agrobacterium-mediated transformation protocol for cotton (Gossypium hirsutum L.): Embryogenic calli as a source to generate large numbers of transgenic plants.-Plant Cell Rep. 22: 465-470, 2004. Go to original source...
    10. Lin, Y.J., Zhang, Q.F.: A synthetic Bacillus thuringiensis insecticidal crystal protein Cry1C.-China Patent. 1483823.2004-03-34a.
    11. Lin, Y.J., Zhang, Q.F.: A synthetic Bacillus thuringiensis insecticidal crystal protein Cry2A.-China Patent. 1480533.2004-03-10b.
    12. Liu, H.Y., Yang, Y.H., Wu, Z.B., Wang, X.D., Yao, M.J.: Introduction of rol genes into cotton (Gossypium hirsutum L.) genome and effects of transgene expression on the plant development.-Agr. Sci. China 3: 728-737, 2004.
    13. Manickavasagam, M., Ganapathi, A., Anbazhagan, V.R., Sudhakar, B., Selvaraj, N., Vasudevan, A., Kasthurirengan, S.: Agrobacterium-mediated genetic transformation and development of herbicide-resistant sugarcane (Saccharum species hybrids) using axillary buds.-Plant Cell Rep. 23: 134-143, 2004. Go to original source...
    14. Meng, F., Shen, J., Zhou, W., Cen, H.: Long-term selection for resistance to transgenic cotton expressing Bacillus thuringiensis toxin in Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae).-Pest Manage. Sci. 60: 167-172, 2004. Go to original source...
    15. Murashige, T., Skoog, F.: A revised medium for rapid growth and bioassays with tobacco tissues cultures.-Physiol. Plant. 15: 473-479, 1962. Go to original source...
    16. Nester, E.W., Thomashow, L.S., Matthew, M., Gordon, M.: 100 Years of Bacillus thuringiensis: a Critical Scientific Assessment.-American Academy of Microbiology, Washington 2002.
    17. Paterson, A.H., Brubaker, C.L, Wendel, J.F.: A rapid method for extraction of cotton (Gossypium L.) genomic DNA suitable for RFLP or PCR analysis.-Plant mol. Biol. Rep. 11: 122-127, 1993. Go to original source...
    18. Popelka, J.C., Xu, J.P., Altpeter, F.: Generation of rye (Secale cereale L.) plants with low transgene copy number after biolistic gene transfer and production of instantly marker-free transgenic rye.-Transgenic Res. 12: 587-596, 2003. Go to original source...
    19. Sambrook, J., Fritsch, E.F., Maniatis, T.: Molecular Cloning: a Laboratory Manual. 2nd Ed.-Cold Spring Harbor Laboratory Press, Cold Spring Harbor 1989.
    20. Shu, Q.Y., Ye, G.Y., Cui, H.R., Cheng, X., Xiang, Y., Wu, D.: 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, 2000. Go to original source...
    21. Skirvin, R.M., Kenneth, D.M., Margaret, N.: Sources and frequency of somaclonal variation.-Hort. Sci. 29: 1232-1237, 1994. Go to original source...
    22. Tabashnik, B.E., Cushing, N.L., Finson, N., Johnson, M.W.: Field development of resistance to Bacillus thuringiensis in diamondback moth (Lepidoptera: Plutellidae).-J. Econ. Entomol. 83: 1671-1676, 1990. Go to original source...
    23. Thompson, C.J., Movva, N.R., Tizard, R., Crameri, R., Davies, J.E., Lauwereys, M., Botterman, J.: Characterization of the herbicide-resistance gene bar from Streptomyces hygroscopicus.-EMBO J. 6: 2519-2523, 1987. Go to original source...
    24. Wu, J.H., Zhang, X.L., Nie, Y.C., Luo, X.Y.: High-efficiency transformation of Gossypium hirsutum embryogenic calli mediated by Agrobacterium tumefaciens and regeneration of insect-resistant plants.-Plant Breed. 124: 142-146, 2005. Go to original source...
    25. Zeng, P., Vadnais, A., Zhang, J., Polacco, C.: Refined glufosinate selection in Agrobacterium-mediated transformation of soybean [Glycine max (L.) Merrill].-Plant Cell Rep. 22: 478-482, 2004. Go to original source...
    26. Zheng, S.: Chromosome variation in callus culture of Gossypium hirsutum L.-Genes 3: 211-221, 1991. Go to original source...

    Return to the content