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Biolistic transformation of cotton embryogenic cell suspension cultures.
MedLine Citation:
PMID:  23143483     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Genetic transformation of cotton (Gossypium hirsutum L.) is highly dependent on the ability to regenerate fertile plants from transgenic cells through somatic embryogenesis. Induction of embryogenic cell cultures is genotype dependant. However, once embryogenic cell cultures are available, they can be effectively used for transformation by Agrobacterium or biolistic bombardment methods. Here I describe a detailed procedure to transform cotton embryogenic cell suspension cultures by biolistic bombardment. A commercially available, helium-driven biolistic device (Bio-Rad PDS1000/He) was used to bombard gold particles coated with plasmid DNA (for visual identification of transformed cells and/or selection) into embryogenic cells. Stable transformation at a high frequency (up to 4% of the transiently expressing cells) is possible. Regeneration of fertile transgenic plants from embryogenic cells takes only about 2 months. Another advantage of the embryogenic cell suspension cultures is that they are amenable for cryopreservation and long-term storage. It is highly preferable to transform commercial varieties of choice than obsolete varieties to avoid the genetic drag due to backcrossing.
Authors:
Kanniah Rajasekaran
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  958     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2013  
Date Detail:
Created Date:  2012-11-12     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  59-70     Citation Subset:  IM    
Affiliation:
Southern Regional Research Center, USDA-ARS, New Orleans, LA, USA, Rajah.Rajasekaran@ars.usda.gov.
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