Document Detail

Biolistic transformation of cotton zygotic embryo meristem.
MedLine Citation:
PMID:  23143482     Owner:  NLM     Status:  In-Data-Review    
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.
Kanniah Rajasekaran
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:  47-57     Citation Subset:  IM    
Southern Regional Research Center, USDA-ARS, New Orleans, LA, USA,
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