Skip to main content
Log in

In vitro assay for 2,4-D resistance in transgenic cotton

  • Published:
In Vitro Cellular & Developmental Biology - Plant Aims and scope Submit manuscript

Summary

2,4-Dichlorophenoxyacetic acid (2,4-D) resistant plants of transgenic cotton (Gossypium hirsutum L.) were produced using Agrobacterium tumefaciens containing a plasmid carrying the neomycin phosphotransferase II (npt II) and 2,4-D monooxygenase (tfd A) genes. An in vitro assay was performed to determine the sensitivity of seed germination, and the growth of seedlings of transgenic and non-transgenic cotton to various concentrations of kanamycin and 2,4-D. The results indicated that kanamycin caused the cotyledons of non-transgenic plants to turn white, but transgenic plants grew normally. Seed germination and seedling growth of non-transgenic plants were strongly inhibited by 2,4-D, but only slightly for transgenic plants. Transgenic plants and non-transgenic plants can be clearly distinguished by the use of 2 mg l−1 2,4-D in seed germination medium. There was a high correlation between the response of seed germination and the growth of seedlings to kanamycin or 2,4-D, based on the germination ration, albino ratio, dry weight or fresh weight. On this basis, we development a rapid method for identifying transgenic plants that has been verified in the field. These findings will allow identification of cotton transformants at an early stage of plant development, saving time and improving cultivars containing the 2,4-D resistance trait.

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

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Bayley, C.; Trolinder, N. L.; Morgan, M. M.; Quisenberry, J. E.; Ow, D. W. Engineering 2,4-D resistance into cotton. Theor. Appl. Genet. 83:645–649; 1992.

    Article  CAS  Google Scholar 

  • Chen, Z. X.; Liewellyn, D. J.; Fan, Y. L.; Li, S. J.; Guo, S. D.; Jiao, G. L.; Zhao, J. X. 2,4-D resistant transgenic cotton plants produced by Agrobacterium-mediated gene transfer. Sci. Agri. Sin: 27(4):31–37; 1994.

    CAS  Google Scholar 

  • Cousins, Y. L.; Lyon, B. B.; Llewellyn, D.J. Transformation of an Australian cotton cultivar: prospects for cotton improvement through genetic engineering. Aust. J. Plant Physiol. 18:481–494; 1991.

    Article  CAS  Google Scholar 

  • Gamborg, O.; Miller, R.; Ojima, K. Nutrient requirements of suspension cultures of soybeant root cells. Exp. Cell Res. 50:151–158; 1968.

    Article  PubMed  CAS  Google Scholar 

  • International Advisory Committee. Cotton: review of world situation Washington, DC: Monogram by International Advisory Committee; 1996.

  • Jenkins, J. N.; McCarty, J. C.; Buehler, R. E.; Kiser, J.; Williams C.; Wofford, T. Resistance of cotton with δ-endotoxin genes from Bacillus thringiensis var. kurstaki on selected Lepidopteran insects. Agron. J. 89:768–780; 1997.

    Article  Google Scholar 

  • John, M. E. Cotton crop improvement through genetic engineering. Crit. Rev. Biotechnol. 17:185–209; 1997.

    Article  CAS  Google Scholar 

  • Kapaun, J. A.; Cheng, Z. M. Aminoglycoside antibiotics inhibit shoot regeneration from Siberian elm leaf explants. Hortsciences 34:727–729; 1999.

    CAS  Google Scholar 

  • Keller, G.; Spatola, L.; McCabe, D.; Martinell, B.; Swain, W.; John, M. E. Transgenic cotton resistant to herbicide bialaphos. Transgen. Res. 6:385–392; 1997.

    Article  CAS  Google Scholar 

  • Li, Y. E.; Chen, Z. X.; Wu, X.; Li, S. J.; Jiao, G. L.; Wu, J. H.; Fan, X. P.; Meng, J. H.; Zhu, Z.; Wang, W.; Zhu, Y.; Xu, H. L.; Xiao, G. F.; Li, X. H. Obtaining transgenic cotton plants with Cowpea trypsin inhibitor gene. Acta Gossypii Sinica 10:237–243; 1998.

    Google Scholar 

  • Lyon, B. R.; Cousins, Y. L.; Llewellyn, D. J.; Dennis, E. S. Cotton plants transformed with a bacterial-degradation gene are protected from accidental spray drift damage by the herbicide 2,4-dichorophen-oxyacetic acid. Transgen. Res. 2:162–169; 1993.

    Article  CAS  Google Scholar 

  • Murashige, T.; Skoog, F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15:473–479; 1962.

    Article  CAS  Google Scholar 

  • Ni, W. C.; Zhang, Z. L.; Guo, S. D. Development of transgenic insectresistant cotton plants. Sci. Agri. Sin. 31(2):8–13; 1998.

    CAS  Google Scholar 

  • Perlak, F. J.; Deaton, R. W.; Armstrong, R. L.; Fuchs, R. L.; Sims, S. R.; Greenplate, J. T.; Fischhoff, D. A. Insect resistant cotton plants. Bio/Technology 8:939–943; 1990.

    Article  PubMed  CAS  Google Scholar 

  • Rajaseharam, K.; Grulam, J. W.; Hudspeth, R. L. Herbicide-resistant Acale and Coker cottons transformation with a native gene encoding mutant forms of acetohydroxyacid synthase. Mol. Breed 2:307–319; 1996.

    Article  Google Scholar 

  • Thomas, J. C.; Adams, D. G.; Keppenne, V. D.; Wasmann, C. C.; Brown, J. K.; Kanost, M. R.; Bohnert, H. J. Proease inhibitors of Manduce sexta expressed in transgenic cotton. Plant Cell Rep. 14:758–762; 1995.

    Article  CAS  Google Scholar 

  • Umbeck, P.; Johmson, G.; Barton, K. Genetically transformed cotton (Gossypium hirsutum L.) plants. Bio/Technology 5:235–266; 1987.

    Article  Google Scholar 

  • Wilmink, A.; Dons, J. J. M. Selective agents and marker genes for use in transformation of monocotyledonous plants. Plant Mol. Biol. Rep. 11:165–185; 1993.

    CAS  Google Scholar 

  • Xie, D. X.; Fan, Y. L.; Ni, W. C.; Huang, J. Q. Transformed Bacillus thuringiensis crystal protein gene into cotton plants. China Sci. (B) 4:367–373; 1991.

    Google Scholar 

  • Zhang, B. H.; Feng, R.; Li, X. L.; Li, F. L. Anther culture and plant regeneration of cotton (Gossypium klotzschianum Anderss). Chinese Sci. Bull 41:145–148; 1996b.

    Google Scholar 

  • Zhang, B. H.; Feng, R.; Liu, F.; Yao, C. B. Direct induction of cotton somatic embryogenesis. Chinese Sei. Bull. 44:766–767; 1999.

    Article  Google Scholar 

  • Zhang, B. H.; Liu, Z. H.; Wang, H. M.; Yao, C. B. Effects of kanamycin on tissue culture and somatic embryogenesis in cotton. Plant Growth Regul. (in press); 2001.

  • Zhang, B.H.; Liu, F.; Yao, C. B. Plant regeneration via somatic embryogenesis in cotton (Gossypium hirsutum L.). Plant Cell Tiss. Organ Cult. 89–94; 2000b.

  • Zhang, B. H.; Liu, F.; Yao, C. B.; Wang, K. B. Recent progress in cotton biotechnology and genetic engineering in China. Curr. Sci. 79:37–44; 2000a.

    CAS  Google Scholar 

  • Zhang, B. H.; Wang, Q. L.; Feng, R.; Li, F. L.; Li, F. G.; Li, X. L. Somatic embryony patterns and plant regeneration in Gossypium hirsutum L. J. Agr. Biotechnol. 4(1):44–50; 1996a.

    Google Scholar 

  • Zhang, B.H.; Zhao, B. Cotton biotechnology and its application. Beijing: China Agricultural Press; 1997;1–201.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhang, BH., Wang, HM., Liu, F. et al. In vitro assay for 2,4-D resistance in transgenic cotton. In Vitro Cell.Dev.Biol.-Plant 37, 300–304 (2001). https://doi.org/10.1007/s11627-001-0053-7

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11627-001-0053-7

Key words

Navigation