Abstract
Biolistic transformation of cotton (Gossypium hirsutum L.) meristems, isolated from mature seed, is detailed in this report. A commercially available, helium-driven biolistic device (Bio-Rad PDS1000/He) was used to bombard gold particles coated with a marker gene (uidA or “β-glucuronidase”) into the shoot meristem. The penetration of gold particles was dependent on bombardment parameters and it was mostly one to two cell layers deep. Stable transformation of epidermal L1 layer was consistently observed in approximately 5% of the seedlings. Germ line transformation was observed in up to 0.71% of bombarded meristems by several laboratories. Using this method identification of germ line transformation is laborious and time-consuming. However, the protocol described here represents a simple and efficient method for generating germ line transformation events. In addition, this procedure offers a quick method to evaluate gene constructs in cotton tissues (embryos, cotyledons, leaf), especially fibers which originate as single cells from the maternal epidermis layer.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Trolinder NT, Goodin JR (1987) Somatic embryogenesis and plant regeneration in cotton (Gossypium hirsutum L.). Plant Cell Rep 6:231–234
Rajasekaran K, Grula JW, Hudspeth RL, Pofelis S, Anderson DM (1996) Herbicide-resistant Acala and Coker cottons transformed with a native gene encoding mutant forms of acetohydroxyacid synthase. Mol Breed 2:307–319
Wilkins TA, Rajasekaran K, Anderson DM (2000) Cotton biotechnology. Crit Rev Plant Sci 19:511–550
Rajasekaran K (2004) Agrobacterium-mediated genetic transformation of cotton. In: Curtis IS (ed) Transgenic crops of the world – essential protocols. Kluwer Academic Publishers, Dordrecht, The Netherlands, pp 243–254
McCabe DE, Martinell BJ (1993) Transformation of elite cotton cultivars via particle bombardment of meristems. BioTechnology 11:596–598
Keller G, Spatola L, McCabe D, Martinell B, Swain W, John ME (1997) Transgenic cotton resistant to herbicide bialaphos. Trangenic Res 6:385–392
Chlan CA, Lin J, Cary JW, Cleveland TE (1995) A procedure for biolistic transformation and regeneration of transgenic cotton from meristematic tissue. Plant Mol Biol Rep 13:31–37
Christou P (1992) Genetic transformation of crop plants using microprojectile bombardment. Plant J 2:275–281
Christou P (1996) Particle bombardment for genetic engineering of plants. R.G. Landes Company and Academic Press, Austin, TX, p 199
Aragão FJL, Vianna GR, Carvalheira SBRC, Rech EL (2005) Germ line genetic transformation in cotton (Gossypium hirsutum L.) by selection of transgenic meristematic cells with a herbicide molecule. Plant Sci 168:1227–1233
Rech EL, Vianna GR, Aragao FJL (2008) High-efficiency transformation by biolistics of soybean, common bean and cotton transgenic plants. Nat Protoc 3:410–418
Skuzeski JM, Nichols LM, Gesteland RF (1990) Analysis of leaky viral translation codons in vivo by transient expression of improved β-glucuronidase vectors. Plant Mol Biol 15:65–79
Klein TM, Gradziel T, Fromm ME, Sanford JC (1988) Factors influencing gene delivery into Zea mays cells by high-velocity microprojectiles. BioTechnology 6:559–563
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Singh M, Krikorian AD (1981) White’s standard nutrient solution. Ann Bot 47:133–139
Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5:387–405
Kosugi S, Ohashi Y, Nakajima K, Arai Y (1990) An improved assay for β-glucuronidase in transformed cells: methanol almost completely suppresses a putative endogenous β-glucuronidase activity. Plant Sci 70:133–140
Satina S, Blakeslee AF, Avery AG (1940) Demonstration of the three germ layers in the shoot apex of Datura by means of induced polyploidy in periclinal chimeras. Am J Bot 27:895–905
Tilney-Bassett RAE (1986) Plant chimeras. Edward Arnold, London, p 199
Sussex I (1989) Developmental programming of the shoot meristem. Cell 56:225–229
John ME (1997) Cotton crop improvement through genetic engineering. Crit Rev Biotech 17:185–208
Agrawal DC, Banerjee AK, Kolala RR, Dhage AB, Kulkarni AV, Nalawade SM, Hazra S, Krishnamurthy KV (1997) In vitro induction of multiple shoots and plant regeneration in cotton (Gossypium hirsutum L.). Plant Cell Rep 16:647–652
Hemphill JK, Maier CGA, Chapman KD (1998) Rapid in-vitro plant regeneration of cotton (Gossypium hirsutum L.). Plant Cell Rep 17:273–278
Bazargani MM, Tabatabaei BES, Omidi M (2011) Multiple shoot regeneration of cotton (Gossypium hirsutum L.) via shoot apex culture system. African J Biotechnol 10:2005–2011
McCabe DE, Swain WF, Martinell BJ, Christou P (1988) Stable transformation of soybean (Glycine max) by particle acceleration. BioTechnology 6:923–926
Brar GS, Cohen BA, Vick CL, Johnson GW (1994) Recovery of transgenic peanut (Arachis hypogaea L.) plants from elite cultivars utilizing Accell® technology. Plant J 5:745–753
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media New York
About this protocol
Cite this protocol
Rajasekaran, K. (2013). Biolistic Transformation of Cotton Zygotic Embryo Meristem. In: Zhang, B. (eds) Transgenic Cotton. Methods in Molecular Biology, vol 958. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-212-4_4
Download citation
DOI: https://doi.org/10.1007/978-1-62703-212-4_4
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-211-7
Online ISBN: 978-1-62703-212-4
eBook Packages: Springer Protocols